Skip to main content
SpringerLink
Log in

(±)[125Iodo]cyanopindolol, a new ligand for β-adrenoceptors: Identification and quantitation of subclasses of β-adrenoceptors in guinea pig

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

Summary

(±)[125Iodo]cyanopindolol (ICYP) is a radioligand which binds with an extraordinarily high affinity and specificity to β-adrenoceptors. In contrast to (±)[125Iodo]-hydroxybenzylpindolol (IHYP), the new ligand has neither affinity to α- nor to 5-HT-receptors. The dissociation constants of ICYP for β-adrenoceptors in various tissues range from 27 to 40 pM, thereby exceeding the affinity of IHYP by a factor of ∼ 3.

ICYP does not discriminate between β1− and β2−. Therefore, the densities of the two receptor subtypes can be determined from competition curves of ICYP by drugs previously found to show in vitro selectivity for β1−adrenoceptors.

The guinea pig left ventricle contains only β1− adrenoceptors, whereas in the lung tissue, the ratio of β1− to β2−adrenoceptors is 1 to 4. The calculated affinities of five β1− selective antagonists for β1−adrenoceptors were nearly identical in the ventricle and the lung.

Kinetic studies of ICYP binding to guinea pig lung membranes indicated that the dissociation reaction consists of two components, a fast process (t 1/2=9 min) and a slower process (t 1/2=8.8 h). A mathematical treatment revealed two possibilities of interpretation: 1. Two forms of the receptor exist which are interconvertible. 2. The (+)- and (−)-enantiomers of ICYP dissociate with different rate constants.

The low dissociation constant of ICYP in combination with its high specific radioactivity (2175 Ci mmole−1) allows binding studies to be carried out with small protein and ligand concentrations, e.g. 3 μg protein per assay in guinea pig lung membranes.

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

Abbreviations

CYP:

(±)cyanopindolol, [(±)4-(3-tert-butylamino-2-hydroxypropoxy)-1H-indole-2-carbonitrile]

ICYP:

(±)-3-[125iodo]-cyanopindolol

HYP:

(±)hydroxybenzylpindolol; IHYP, (±)[125iodo]-hydroxybenzylpindolol

3H-DHA:

(−)-[3H]dihydroalprenolol

References

  • Ablad B, Borg KD, Carlsson E, Ek L, Johnsson G, Malmfors T, Regardh CG (1975) A survey of the pharmacological properties of metoprolol in animals and man. Acta Pharmacol Toxicol (Copenh) 36, Suppl V: 7–24

    Google Scholar 

  • Aggerbeck M, Guellaen G, Hanoune J (1979) N-aralkyl substitution increases the affinity of adrenergic drugs for the α-adrenoceptor in rat liver. Br J Pharmacol 65:155–159

    Google Scholar 

  • Atlas D, Steer ML, Levitzki A (1974) Stereospecific binding of propranolol and catecholamines to beta-adrenergic receptor. Proc Natl Acad Sci USA 71:4246–4248

    Google Scholar 

  • Aurbach GD, Fedak SA, Woodard CJ, Palmer JS, Hauser D, Troxler F (1974) β-adrenergic receptor: stereospecific interaction of iodinated β-blocking agent with high affinity site. Science 186:1223–1224

    Google Scholar 

  • Barnes PJ, Dollery CT, MacDermot J (1980) Increased pulmonary α-adrenergic and reduced β-adrenergic receptors in experimental asthma. Nature 285:569–571

    Google Scholar 

  • Barovsky K, Brooker G (1980) (−)-[125I]-iodopindolol, a new highly selective radioiodinated β-adrenergic receptor antagonist: measurement of β-adrenoceptors in intact rat astrocytoma cells. J Cycl Nucleotide Res 6:297–307

    Google Scholar 

  • Barrett AM (1977) The pharmacology of atenolol. Postgrad Med J 53, Suppl 3: 58–64

    Google Scholar 

  • Bearer CF, Knapp RD, Kaumann AJ, Swartz TL, Birnbaumer L (1980) Iodohydroxybenzylpindolol: preparation, purification, localization of its iodine to the indole ring and characterization as a partial agonist. Mol Pharmacol 17:328–338

    Google Scholar 

  • Berman M (1968) The application of multicomportmental analysis to problems of clinical medicine. Ann Intern Med 68:423–448

    Google Scholar 

  • Berman M, Weiss MF (1967) SAAM Manual. NIH PHS Publication, 1703, Bethesda/Maryland, USA

  • Berthold R (1979) Br Pat Publ 2001971A and D.O.S. 2830211

  • Bieth N, Rouot B, Schwartz J, Velly J (1980) Comparison of pharmacological and binding assays for ten β-adrenoceptor blocking agents and two β-adrenoceptor agonists. Br J Pharmacol 68:563–569

    Google Scholar 

  • Boudot JP, Cavero I, Fenard S, Lefevre-Borg F, Manoury P, Roach AG (1979) Preliminary studies on SL 75212, a new potent cardioselective beta-adrenoceptor antagonist. Br J Pharmacol 66:445P

    Google Scholar 

  • Bürgisser E, Hancock AA, Lefkowitz RJ, De Lean A (1981) Anomalous equilibrium binding properties of high affinity racemic radioligands. Mol Pharmacol 19:205–216

    Google Scholar 

  • Cheng YC, Prusoff WH (1973) Relationship between the inhibition constant K1 and the concentration of inhibitor which caused 50% inhibition (I50) of an enzymatic reaction. Biochem Pharmacol 22:3099–3108

    Google Scholar 

  • Dickinson KEJ, Nahorski SR, Willcocks AL (1981) Serotonin recognition sites are labelled in cerebral cortex by the β-adrenoceptor antagonist 125I-hydroxybenzylpindolol. Br J Pharmacol 72:165P

  • Dunlop D, Shanks RG (1968) Selective blockade of adrenoceptive beta receptors in the heart. Br J Pharmacol 32:201–218

    Google Scholar 

  • Hancock AA, De Lean AL, Lefkowitz RJ (1980) Quantitative resolution of beta-adrenergic subtypes by selective ligand binding: application of a computerized model fitting technique. Mol Pharmacol 16:1–9

    Google Scholar 

  • Harden TK, Wolfe BB, Molinoff PB (1976) Binding of iodinated β-adrenergic antagonists to proteins derived from rat heart. Mol Pharmacol 12:1–15

    Google Scholar 

  • Hedberg A, Minneman KP, Molinoff PB (1980) Differential distribution of beta-1 and beta-2 adrenergic receptors in cat and guinea pig heart. J Pharmacol Exp Ther 212:503–508

    Google Scholar 

  • Hoffmann BB, Lefkowitz RJ (1980) Radioligand binding studies of adrenergic receptors: new insights into molecular and physiological regulation. Annu Rev Pharmacol Toxicol 20:581–608

    Google Scholar 

  • Hoyer D, Engel G, Berthold R. Binding characteristics of (+)-, (±)- and (−)-[125iodo]cyanopindolol to guinea pig left ventricle membranes. Naunyn-Schmiedeberg's Arch Pharmacol (in press)

  • Hunter WM, Greenwood H (1962) Preparation of 131iodine labelled human growth hormone of high specific activity. Nature 194:495–496

    Google Scholar 

  • Innis RB, Correa FMA, Snyder S (1979) Carazolol, an extremely potent β-adrenergic blocker: binding to β-adrenoceptors in brain membranes. Life Sci 24:2255–2264

    Google Scholar 

  • Kaumann AJ, Birnbaumer L, Wittmann R (1978) Heart β-adrenoceptors. In: O'Malley BW, Birnbaumer L (eds) Hormone receptors 3. Academic Press, New York, pp 133–177

    Google Scholar 

  • Kleinstein J, Glossmann H (1978) Solubilization of a mammalian β-adrenergic receptor. Naunyn-Schmiedeberg's Arch Pharmacol 305:191–200

    Google Scholar 

  • Krawietz W, Weinsteiger M, Pruchniewski M, Erdmann E (1979) Evidence for negative cooperativity among β-adrenergic receptors in cardiac and lung tissue of guinea pig. Biochem Pharmacol 28:2999–3007

    Google Scholar 

  • Laurell S, Tibbling G (1966) An enzymatic fluorometric micromethod for the determination of glycerol. Clinica Chimica Acta 13:317–322

    Google Scholar 

  • Lefkowitz RJ, Mukherjee C, Coverstone M, Caron MG (1974) Stereospecific 3H-(−)-alprenolol binding sites, β-adrenergic receptors and adenylate cyclase. Biochem Biophys Res Commun 60:703–709

    Google Scholar 

  • Lefkowitz RJ, Williams LT (1977) Catecholamine binding to the beta-adrenergic receptor. Proc Natl Acad Sci USA 74:515–519

    Google Scholar 

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

    Google Scholar 

  • Maguire ME, Ross EM, Gilman AG (1977) β-adrenergic receptor: ligand binding properties and the interaction with adenylyl cyclase. In: Greengard P, Robison GA (eds) Advances in cyclic nucleotide research 8. Raven Press, New York, pp 1–84

    Google Scholar 

  • Maguire ME, Wiklund RA, Anderson HJ, Gilman AG (1976) Binding of [125I]iodohydroxybenzylpindolol to putative β-adrenoceptors of rat glioma cells and other cell clones. J Biol Chem 251:1221–1231

    Google Scholar 

  • Malchoff CD, Marinetti GV (1976) Hormone action at the membrane level. Binding of (±)-[3H]isoproterenol to intact chicken erythrocytes and erythrocyte ghosts. Biochem Biophys Acta 436:45–52

    Google Scholar 

  • McNamara DB, Sulakhe PV, Singh JN, Dhalla NS (1974) Properties of heart sarcolemmal Na+-K+-ATPase. J Biochem 75:795–803

    Google Scholar 

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

    Google Scholar 

  • Minneman KP, Molinoff PB (1980) Classification and quantitation of β-adrenergic receptor subtypes. Biochem Pharmacol 29:1317–1323

    Google Scholar 

  • Morris TH, Kaumann AJ (1979) Characteristics of 3H-(±)-carazolol binding to myocardial β-adrenoceptors. Naunyn-Schmiedeberg's Arch Pharmacol 308:R33

    Google Scholar 

  • Pochet R, Schmitt H (1978) In vivo labeling of β-adrenergic receptors from muscle cells. In: Dumont J, Nunez J (eds) Hormone and cell regulation 2. Elsevier/North-Holland Biomedical Press, pp 133–135

  • Rodbell M (1964) Metabolism of isolated fat cells. Effects of hormones on glucose metabolism and lipolysis. J Biol Chem 239:375–380

    Google Scholar 

  • Ross EM, Maguire ME, Sturgill TW, Biltonen RL, Gilman AG (1977) Relationship between the β-adrenergic receptor and adenylate cyclase. J Biol Chem 252:5761–5775

    Google Scholar 

  • Rugg EL, Barnett DB, Nahorski SR (1978) Coexistence of β1− and β1− adrenoceptors in mammalian lung: evidence from direct binding studies. Mol Pharmacol 14:996–1005

    Google Scholar 

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

    Google Scholar 

  • Snedecor GW, Cochran WG (1973) Curvilinear regression. In: Snedecor GW, Cochran WG (eds) Statistical methods, Chapter 15. The Iowa State Univ Press, Iowa/USA, 6th edition, pp 447–471

    Google Scholar 

  • Sporn JR, Molinoff PB (1976) β-adrenoceptors in rat brain. J Cycl Nucleotide Res 2:149–161

    Google Scholar 

  • Svendsen TL, Hartling O, Trap Jensen J (1979) Immediate haemodynamic effects of propranolol, practolol, pindolol, atenolol and ICI 89-406 in healthy volunteers. Eur J Clin Pharmacol 15:223–228

    Google Scholar 

  • Schmitt H, Pochet R (1977) In vivo labelling of β-adrenergic receptors on rat glioma cells. FEBS Lett 76:302–305

    Google Scholar 

  • Terasaki WI, Brooker G (1978) 125I-Iodohydroxybenzylpindolol binding sites on intact rat glioma cells. J Biol Chem 253:5418–5425

    Google Scholar 

  • Waechter W, Muench 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 

  • Wenke M (1971) Drug receptor interactions. In: Bacq ZM (ed) Fundamentals of Biochemical Pharmacology. Pergamon Press, Oxford, pp 367–415

    Google Scholar 

  • Yamamura H, Rodbell M, Fain JN (1976) Hydroxybenzylpindolol and hydrobenzylpropranolol: Partial beta-adrenergic agonists of adenylate cyclase in the rat adipocyte. Mol Pharmacol 12:693–700

    Google Scholar 

Download references

Author information

Author notes

  1. Daniel Hoyer

    Present address: Department of Pharmacology, University of Pennsylvania, School of Medicine, 19174, Philadelphia, Pa, USA

Authors and Affiliations

  1. Pharmaceutical Division, Preclinical Research, Sandoz Ltd., CH-4002, Basle, Switzerland

    Günter Engel, Daniel Hoyer, Richard Berthold & Heribert Wagner

Authors
  1. Günter Engel
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Daniel Hoyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Richard Berthold
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Heribert Wagner
    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

Engel, G., Hoyer, D., Berthold, R. et al. (±)[125Iodo]cyanopindolol, a new ligand for β-adrenoceptors: Identification and quantitation of subclasses of β-adrenoceptors in guinea pig. Naunyn-Schmiedeberg's Arch. Pharmacol. 317, 277–285 (1981). https://doi.org/10.1007/BF00501307

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00501307

Key words

Search

Navigation