Skip to main content
SpringerLink
Log in

Direct labelling of myocardial β 1-adrenoceptors

Comparison of binding affinity of 3H-(−)-bisoprolol with its blocking potency

  • Published:
Naunyn-Schmiedeberg's Archives of Pharmacology Aims and scope Submit manuscript

Summary

A radioligand that selectively labels β 1-adrenoceptors, 3H-(−)-bisoprolol (3H-BIS), is introduced. The binding properties of 3H-BIS to membrane particles of kitten heart are compared with the blocking properties of (−)-bisoprolol against stimulant effects of (−)-adrenaline and (−)-noradrenaline in heart preparations of kitten and guinea pig.

  1. 1.

    On kitten heart tissues (-)-bisoprolol antagonized the positive chronotropic and inotropic effects of catecholamines competitively. The effects of (-)-adrenaline were antagonized considerably less by (-)-bisoprolol than the corresponding effects of (-)-noradrenaline on sinoatrial pacemakers. The antagonism was nearly the same against both (-)-adrenaline and (-)-noradrenaline in left atria and papillary muscles. The data were analyzed with a model for 2-receptor subtypes by non-linear regression. Equilibrium dissociation constants K B (mol/l; -log K B=pK B) for a high-affinity β 1-adrenoceptor of 8.8 and for a low-affinity β 2-adrenoceptor of 7.0 were estimated in the three classes of tissues. In kitten sinoatrial pacemaker β 1-adrenoceptors contribute 76% to the stimulus induced by (-)-adrenaline and 97% to the stimulus induced by (-)-noradrenaline. In ventricle and left atrium β 1-adrenoceptors contribute 97∓99% and 100% to the stimulus caused by (-)-adrenaline and (-)-noradrenaline, respectively.

  2. 2.

    Both 3H-BIS and unlabelled (-)-bisoprolol caused competitive blockade of the positive chronotropic effects of (-)-noradrenaline in guinea-pig right atria. pK B-values of 8.7 were estimated for both unlabelled and tritiated (-)-bisoprolol. The positive chronotropic effects of (-)-adrenaline were antagonized considerably less by (-)-bisoprolol than those of (-)-noradrenaline in guinea-pig atria.

    In the presence of low concentrations of β 2-selective ICI 118,551, which did not antagonize β 1 adrenoceptor mediated effects, (−)bisoprolol antagonized positive chronotropic effects of (−)-adrenaline to the same extent as those of (−)-noradrenaline. The results are consistent with the concept of a significant role of sinoatrial β 2-adrenoceptors of guinea pig for the effects of (−)-adrenaline but not for those of (−)-noradrenaline.

  3. 3.

    3H-BIS associated and dissociated quickly with and from ventricular β 1-adrenoceptors. A k off of 1.0 min-1 was estimated. An equilibrium dissociation constant pK L *of 8.2 for 3H-BIS was estimated from saturation binding. The binding competition of the optical isomers of bupranolol with 3H-BIS revealed a degree of stereo-selectivity expected for β 1-adrenoceptors.

  4. 4.

    (-)-Bisoprolol competed with high affinity and low affinity, respectively, for β 1- and β 2-adrenoceptors of both ventricular and atrial membranes labelled with 3H-(-)-bupranolol bupranolol or 3H-(-)-propranolol. The difference in affinity of (-)-bisoprolol of 1.7±0.3 log units between β 1- and β 2-adrenoceptors agrees with the selectivity for β 1-adrenoceptors found in intact heart tissues.

  5. 5.

    (-)-Noradrenaline and (-)-adrenaline competed with low affinity with 3H-BIS for ventricular β 1-adrenoceptors. An approximately 100-fold difference between high K L-values of these catecholamines and low inotropic EC50-values is consistent with a large β 1-adrenoceptor reserve.

  6. 6.

    The binding affinity for β 1-adrenoceptors of both 3H-BIS and (-)-bisoprolol is 3∓8 times lower on membrane particles than on intact tissues, as also confirmed with the blockade of catecholamine-induced stimulation of the adenylate cyclase. By contrast, the affinities of (-)-propranolol and (-)-bupranolol are similar in intact tissues and on membrane particles. The low affinity of (-)-bisoprolol in membranes may be caused by a lesion of accessory recognition sites of the β 1-adrenoceptors due to the procedure of membrane preparation. However, the lesion of these hypothetical accessory binding sites would not lower the affinity for (-)-propranolol and (-)-bupranolol.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Arunlakshana O, Schild HO (1959) Some quantitative uses of drug antagonists. Br J Pharmacol 14:48–58

    Google Scholar 

  • Bilski A, Halliday SE, Fitzgerald JD, Wale JL (1983) The pharmacology of a β 2-selective adrenoceptor antagonist (ICI 118,551). J Cardiovasc Pharmacol 5:430–437

    Google Scholar 

  • Birnbaumer L, Duran JM, Nakahara T, Kaumann AJ (1977) Adenylyl cyclases: stimulation by hormones and regulation by nucleotides. In: Jaimeson GA, Robinson DM (eds) Mammalian cell membranes, vol 5. Responses of plasma membranes. Butterorths. London Boston pp 105–150

    Google Scholar 

  • Blinks JR (1965) convenient apparatus for recording contractions of isolated muscle. J Appl Physiol 20:755–757

    Google Scholar 

  • Carlsson E, Åblad B, Brandstrom A, Carlsson B (1972) Differentiated blockade of the chronotropic effects of various adrenergic stimuli in the cat heart. Life Sci 11:953–958

    Google Scholar 

  • Chènieux-Guicheney P, Dausse JP, Meyer P, Schmitt H (1978) Inhibition of (3H)-dihydroalprenolol binding to rat cardiac membranes by various β-blocking agents. Br J Pharmacol 63:177–182

    Google Scholar 

  • Cruickshank JM (1980) The clinical importance of cardioselectivity and lipophilicity in beta blockers. Am Heart J 100:160–178

    Google Scholar 

  • De Lean A, Hancock AA, Lefkowitz RJ (1982) Validation and statistical analysis of a computer modeling method for quantitative analysis of radioligand binding data for mixtures of pharmacological receptor subtypes. Mol Pharmacol 21:5–16

    Google Scholar 

  • Dickinson K, Richardson A, Nahorski SR (1980) Homogeneity of beta2-adrenoceptors on rat erythrocytes and reticulocytes. A comparison with heterogeneous rat lung beta-adrenoceptors. Molec Pharmacol 19:194–204

    Google Scholar 

  • Dixon WJ, Brown MB (eds) (1979) Biomedical computer programs, P series. University of California Press, Berkeley Los Angeles London

    Google Scholar 

  • Ehle B, Lemoine H, Kaumann AJ (1985) Improved evaluation of binding of ligands to membranes containing several receptor subtypes. Naunyn-Schmiedeberg's Arch Pharmacol 331:52–59

    Google Scholar 

  • Gille E, Lemoine H, Ehle B, Kaumann AJ (1985) The affinity of (−)-propranolol for β 1- and β 2-adrenoceptors of human heart. Differential antagonism of the positive inotropic effects and adenylate cyclase stimulation by (−)-noradrenaline and (−)-adrenaline. Naunyn-Schmiedeberg's Arch Pharmacol 331: 60–70

    Google Scholar 

  • Hancock AA, De Lean AL, Lefkowitz RJ (1979) Quantitative resolution of beta-adrenergic receptor subtypes by selective ligand binding: application of a computerized model fitting technique. Mol Pharmacol 16:1–9

    Google Scholar 

  • Hellenbrecht D, Lemmer B, Wiethold G, Grobecker H (1973) Measurements of hydrophobicity, surface activity, local anaesthesia, and myocardial conduction velocity as quantitative parameters of non-specific membrane affinity of nine β-adrenergic blocking agents. Naunyn-Schmiedeberg's Arch Pharmacol 277:211–226

    Google Scholar 

  • Kaumann AJ (1970) Adrenergic receptors in heart muscle: Relations among factors influencing the sensitivity of the cat papillary muscle to catecholamines. J Pharmacol Exp Ther 173:383–398

    Google Scholar 

  • Kaumann AJ (1972) Potentiation of the effects of isoprenaline and noradrenaline by hydrocortisone in cat heart muscle. Naunyn-Schmiedeberg's Arch Pharmacol 273:134–153

    Google Scholar 

  • Kaumann AJ (1978) On spare β-adrenoceptors for inotropic effects of catecholamines in kitten ventricle. Naunyn-Schmiedeberg's Arch Pharmacol 305:97–102

    Google Scholar 

  • Kaumann AJ (1982) Cultured heart cells as a model for β-adrenoceptors in a heart pacemaker. Chronotropic spare β-adrenoceptors and spare adenylyl cyclase for (−)-isoprenaline but not for (−)-dichloroisoprenaline in rat cardiocytes. Naunyn-Schmiedeberg's Arch Pharmacol 320:119–129

    Google Scholar 

  • Kaumann AJ, Birnbaumer L (1974) Characteristics of the adrenergic receptor coupled to myocardial adenylyl cyclase. Stereospecifity and determination of apparent affinity constants for beta blockers. J Biol Chem 249:7874–7885

    Google Scholar 

  • Kaumann AJ, Lemoine H (1983) Large discrepancies between membrane binding and blocking affinities in tissues for antagonists that select β-adrenoceptor subtypes but not for nonselective antagonists. Naunyn-Schmiedeberg's Arch Pharmacol 324: R 66

    Google Scholar 

  • Kaumann AJ, Marano M (1982) On equilibrium constants for complexes of drug-receptor subtypes. Selective and nonselective interactions of partial agonists with two plausible β-adrenoceptor subtypes mediating positive chronotropic effects of (−)-isoprenaline in kitten atria. Naunyn-Schmiedeberg's Arch Pharmacol 318:192–201

    Google Scholar 

  • Kaumann AJ, McInerny TK, Gilmour DP, Blinks JR (1980) Comparative assessment of β-adrenoceptor blocking agents as simple competitive antagonists in isolated heart muscle: Similarity of inotropic and chronotropic blocking potencies against isoproterenol. Naunyn-Schmiedeberg's Arch Pharmacol 311:219–236

    Google Scholar 

  • Kaumann AJ, Lemoine H, Morris TH, Schwederski U (1982) An initial characterization of human heart β-adrenoceptors and their mediation of the positive inotropic effects of catecholamines. Naunyn-Schmiedeberg's Arch Pharmacol 319: 216–221

    Google Scholar 

  • Kaumann AJ, Morris TH, Bojar H (1983) On the functional role of heterogeneous binding sites. Heart β-adrenoceptors. J Receptor Res 3:61–70

    Google Scholar 

  • Lands AM, Arnold A, McAuliff JP, Luduena FP, Brown TG (1967) Differentiation of receptor systems activated by sympathomimetic amines. Nature 214:597–598

    Google Scholar 

  • Lefkowitz RJ, Stadel JM, Caron MG (1983) Adenylate cyclase-coupled beta-adrenergic receptors: Structure and mechanisms of activation and desensitization. Ann Rev Biochem 52, 159–186

    Google Scholar 

  • Lemoine H, Kaumann AJ (1982) A novel analysis of concentration-dependence of partial agonism. Ring-demethylation of bupranolol results in a high affinity partial agonist (K 105) for myocardial and tracheal β-adrenoceptors. Naunyn-Schmiedeberg's Arch Pharmacol 320:130–144

    Google Scholar 

  • Lemoine H, Kaumann AJ (1983) A model for the interaction of competitive antagonists with two receptor-subtypes characterized by a Schild-plot with apparent slope unity. Agonist-dependent enantiomeric affinity ratios for bupranolol in tracheae but not in right atria of guinea pigs. Naunyn-Schmiedeberg's Arch Pharmacol 322:111–120

    Google Scholar 

  • Lemoine H, Ehle B, Kaumann AJ (1985) Direct labelling of β 1-adrenoceptors. Comparison of binding potency of 3H-ICI 118,551 and blocking potency of ICI 118,551. Naunyn-Schmiedeberg's Arch Pharmacol 331:40–51

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurements with the Folin phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • Manalan AS, Bosch HR, Watanabe AM (1981) Characterization of (3H)-(±)-carazolol binding to β-adrenergic receptors. Application to study of β-adrenergic receptor subtypres in canine ventricular myocardium and lung. Circulation Res 49:326–336

    Google Scholar 

  • Minneman KP, Hegstrand LR, Molinoff PB (1979) Simultancous determination of beta-1 and beta-2 adrenergic receptors in tissues containing both receptor subtypes. Mol Pharmacol 16:34–46

    Google Scholar 

  • Minneman KP, Pittman RN, Molinoff PB (1981) β-Adrenergic receptor subtypes: Properties, distribution, and regulation. Ann Rev Neurosci 4:419–461

    Google Scholar 

  • Morris TH, Kaumann AJ (1979) Characterization of 3H-(±)-carazolol binding to myocardial β-adrenoceptors. Naunyn-Schmiedeberg's Arch Pharmacol 308:R 33

    Google Scholar 

  • Morris TH, Kaumann AJ (1984) Different steric characteristics of β 1- and β 2-adrenoceptors. Naunyn Schmiedeberg's Arch Pharmacol 327:176–179

    Google Scholar 

  • Morris TH, Sandrock K, Kaumann AJ (1981) 3H-(−)-Bupranolol, a new β-adrenoceptor radioligand: Characterization of its binding to kitten heart β-adrenoceptors. Naunyn-Schmiedeberg's Arch Pharmacol 317:19–25

    Google Scholar 

  • Moustafa E, Giachetti A, Downey HF, Bashour FA (1978) Binding of (3H)-dihydroalprenolol to beta-adrenoceptors of cells isolated from adult rat heart. Naunyn-Schmiedeberg's Arch Pharmacol 303:107–109

    Google Scholar 

  • Porzig H, Becker C, Reuter H (1982) Competitive and non-competitive interactions between specific ligands and beta-adrenoceptors in living cardiac cells. Naunyn-Schmiedeberg's Arch Pharmacol 321:89–99

    Google Scholar 

  • Rugg EL, Barnett DB, Nahorski SR (1978) Coexistence of beta1 and beta2 adrenoceptors in mammalian lung: evidence from direct binding studies. Mol Pharmacol 14:996–1005

    Google Scholar 

  • Salomon Y, Londos C, Rodbell M (1974) A highly sensitive adenylate cyclase assay. Anal Biochem 58:541–548

    Google Scholar 

  • Scatchard G (1949) The attractions of proteins for small molecules and ions. Ann NY Acad Sci 51:660–672

    Google Scholar 

  • Trendelenburg U (1968) The effect of cocaine on the pacemaker of isolated guinea-pig atria. J Pharmacol Exp Ther 161:222–231

    Google Scholar 

  • Wächter W, Münch U, Lemoine H, Kaumann AJ (1980) Evidence that (+)-bupranolol interacts directly with myocardial β-adrenoceptors: Control of optical purity with differential thermal analysis. Naunyn-Schmiedeberg's Arch Pharmacol 313:1–8

    Google Scholar 

  • Walter M, Lemoine H, Kaumann AJ (1984) Stimulant and blocking effects of optical isomers of pindolol on the sinoatrial node and trachea of guinea pig. Role of β-adrenoceptor subtypes in the dissociation between blockade and stimulation. Naunyn-Schmiedeberg's Arch Pharmacol 327:159–175

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Lehrstuhl für Klinische Physiologie, Physiologisches Institut der Universität Düsseldorf, Universitätsstraße 1, D-4000, Düsseldorf, Federal Republic of Germany

    Alberto J. Kaumann & Horst Lemoine

Authors
  1. Alberto J. Kaumann
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Horst Lemoine
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kaumann, A.J., Lemoine, H. Direct labelling of myocardial β 1-adrenoceptors. Naunyn-Schmiedeberg's Arch. Pharmacol. 331, 27–39 (1985). https://doi.org/10.1007/BF00498849

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00498849

Key words

Search

Navigation