Skip to main content
SpringerLink
Log in

In vitro studies on the relative potency of bronchodilator agents

  • Published:
Lung Aims and scope Submit manuscript

Abstract

This study describes a rapid in vitro assay for the order of potency of bronchodilator drugs using specific binding of (−)-[3H] dihydroalprenolol ([3H]DHA) to rat lung membranes. Under linear conditions with respect to tissue, specific binding of [3H]DHA showed saturability, rapid kinetics of association and dissociation of radioligand, and sterospecificity. Nanomolar (nM) concentrations for 50% inhibition (IC50±SE) for the bronchodilator drugs examined were as follows: albuterol, 1485±170; isoproterenol, 136±53; procaterol, 162±28; terbutaline, 3310±934; and zinterol, 51±8.3. A comparison of binding studies using rat lung tissue membranes and similar preparations of rat heart and skeletal muscle demonstrated that lung tissue had 7 to 8 times more receptor sites (Bmax) for [3H]DHA than heart or skeletal muscle. Adenyl cyclase activit of the rat lung membrane preparation almost doubled in the presence of (−)-isoproterenol. Displacement of specific (3H)DHA binding in membrane preparations may provide useful data for evaluating bronchodilator compounds.

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

  1. Barnes PJ, Karliner JS, Dollery CT (1980) Human lung adrenoceptors studied by radioligand binding. Clin Sci 58:457–461

    PubMed  CAS  Google Scholar 

  2. Bennett JP Jr (1978) Methods in binding studies. In: Yamamura HI, Enna SJ, Kuhar MJ (eds) Neurotransmitter Receptor Binding. Raven Press, New York, pp 57–90

    Google Scholar 

  3. Brown BL, Albano JDM, Ekins RP, Sgherzi AM (1971) A simple and sensitive saturation assay method for the measurement of adenosine 3:5-cyclic monophosphate. Biochem J 121:561–562

    PubMed  CAS  Google Scholar 

  4. Chen FM, Yamamura HI, Roeske WR (1982) Adenylate cyclase and beta adrenergic receptor development in the mouse heart. J Pharmacol Exp Ther 221:1–13

    Google Scholar 

  5. Chen YC, Prusoff WH (1973) Relationship between the inhibition constant (Ki) and the concentration of inhibitor which causes 50 percent inhibition (I50) of an enzymatic reaction. Biochem Pharmacol 22:3099–3108

    Article  Google Scholar 

  6. Frishman WH (1981) Beta-adrenoceptor antagonists: new drugs and new indications. N Engl J Med 305:500–506

    Article  PubMed  CAS  Google Scholar 

  7. Hazeki O and Ui M (1980) Beta1 and beta2-adrenergic receptors responsible for cyclic AMP accumulation in isolated heart and lung cells. Mol Pharmacol 17:8–13

    PubMed  CAS  Google Scholar 

  8. Lands AM, Arnold AM, Luduena JP, Brown TC (1967) Differentiation of receptor systems activated by sympathomimetic amines. Nature 214:597–598

    Article  PubMed  CAS  Google Scholar 

  9. Larrson S, Svedmyr N (1977) Tremor caused by sympathomimetics is mediated by beta-2-adrenoceptors. Scand J Respir Dis 58:93–102

    Google Scholar 

  10. Lefkowitz RJ (1975) Heterogeneity of adenylate cyclase coupled beta-adrenergic receptors. Biochem Pharmacol 24:583–590

    Article  PubMed  CAS  Google Scholar 

  11. Levy B. (1966). The adrenergic blocking activity of N-tert-butyl methoxamine (butoxamine). J Pharmacol Exp Ther 151:413–422

    PubMed  CAS  Google Scholar 

  12. Lowry OH, Rosebrough NJ, Farr AI, Randall RJ (1951) Protein measurement with Folin phenol reagent. J Biol Chem. 193:265–275

    PubMed  CAS  Google Scholar 

  13. 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

    PubMed  CAS  Google Scholar 

  14. Minneman KP, Hegstrand LR, Molinoff PB (1979) Comparison of beta-adrenergic receptor subtypes in mammalian tissues. J Pharmacol Exp Ther 211:502–508

    PubMed  CAS  Google Scholar 

  15. O’Donnell SR (1972) An examination of some beta-adrenoceptor stimulants for selectivity using the isolated trachea and atria of the guinea pig. Eur J Pharmacol 19:371–379

    Article  PubMed  CAS  Google Scholar 

  16. Rugg EL Barnett DB, Nahorski SR (1978) Coexistence of B1 and B2-adrenoceptors in mammalian lung: evidence from direct binding studies. Mol Pharmacol 14:996–1005

    PubMed  CAS  Google Scholar 

  17. Williams LT, Lefkowitz RJ (1978) Receptor binding studies in adrenergic pharmacology. Raven Press, New York, pp 5–17

    Google Scholar 

  18. Williams LT, Snyderman R, Lefkowitz RJ (1976) Identification of beta-adrenergic receptors in human lymphocytes by [3H] alprenolol binding. J Clin Invest 57:149–155

    Article  PubMed  CAS  Google Scholar 

  19. Young RR, Growdon JH, Shahani BT (1975) Beta-adrenergic mechanisms in action tremor. N Engl J Med 293:950–953

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Internal Medicine, Arizona Health Sciences Center, 85724, Tucson, Arizona, USA

    Joseph H. Fleisher & Jacob L. Pinnas

Authors
  1. Joseph H. Fleisher
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jacob L. Pinnas
    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

Fleisher, J.H., Pinnas, J.L. In vitro studies on the relative potency of bronchodilator agents. Lung 163, 161–171 (1985). https://doi.org/10.1007/BF02713818

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02713818

Key words

Search

Navigation