Evidence for a Non-β2-Adrenoceptor Binding Site in Human Lung Tissue for a Subset of β2-Adrenoceptor Agonists


The aim of this study was to compare the binding profile of a range of β2-adrenoceptor (β2-AR) agonists and antagonists in human lung tissue. Radioligand saturation and competition binding experiments were performed by filtration with a β2-AR antagonist ([3H]propranolol) or agonist ([3H]vilanterol) radioligand and membrane fragments generated from lung parenchyma in the presence of 100 μM guanosine 5’-[β,γ-imido]triphosphate (Gpp(NH)p). In membranes prepared from human lung parenchyma, carmoterol, formoterol, ICI118551, propranolol and salbutamol resulted in inhibition of [3H]vilanterol binding to levels that were significantly different from indacaterol, salmeterol and vilanterol (ANOVA, Bonferroni post-test, P < 0.001 except formoterol vs indacaterol where P < 0.01). Indacaterol and salmeterol resulted in inhibition of [3H]vilanterol binding to levels that were not significantly different from vilanterol (ANOVA, Bonferroni post-test, P > 0.05). Indacaterol, salmeterol and vilanterol resulted in full inhibition of [3H]propranolol binding to levels not significantly different from ICI118551 (ANOVA, Bonferroni post-test, P > 0.05). Indacaterol, salmeterol and vilanterol bind to an additional site in human lung parenchyma membranes that is distinct from the β2-AR.

Share and Cite:

R. Slack, "Evidence for a Non-β2-Adrenoceptor Binding Site in Human Lung Tissue for a Subset of β2-Adrenoceptor Agonists," Pharmacology & Pharmacy, Vol. 5 No. 1, 2014, pp. 30-36. doi: 10.4236/pp.2014.51006.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] E. D. Bateman, S. S. Hurd, P. J. Barnes, J. Bousquet, J. M. Drazen, et al., “Global Strategy for Asthma Management and Prevention: GINA Executive Summary,” European Respiratory Journal, Vol. 31, No. 1, 2008, pp. 143-178.
[2] M. Cazzola and M. Molimard, “The Scientific Rationale for Combining Long-Acting β2-Agonists and Muscarinic Antagonists in COPD,” Pulmonary Pharmacology & Therapeutics, Vol. 23, No. 4, 2010, pp. 257-267.
[3] V. A. Cullum, J. B. Farmer, D. Jack and G. P. Levy, “Salbutamol: A New, Selective β-Adrenoceptive Receptor Stimulant,” British Journal of Pharmacology, Vol. 35, No. 1, 1969, pp. 141-151.
[4] G. P. Anderson, “Formoterol: Pharmacology, Molecular Basis of Agonism, and Mechanism of Long Duration of a Highly Potent and Selective β2-Adrenoceptor Agonist Bronchodilator,” Life Sciences, Vol. 52, No. 26, 1993, pp. 2145-2160. http://dx.doi.org/10.1016/0024-3205(93)90729-M
[5] M. Johnson, P. R. Butchers, R. A. Coleman, A. T. Nials, P. Strong, et al., “The Pharmacology of Salmeterol,” Life Sciences, Vol. 52, No. 26, 1993, pp. 2131-2143.
[6] M. Cazzola, L. Calzetta and M. G. Matera, “β2-Adrenoceptor Agonists: Current and Future Direction,” British Journal of Pharmacology, Vol. 163, No. 1, 2011, pp. 4-17. http://dx.doi.org/10.1111/j.1476-5381.2011.01216.x
[7] G. P. Anderson, A. Lindén and K. F. Rabe, “Why Are Long-Acting Beta-Adrenoceptor Agonists Long-Acting?” European Respiratory Journal, Vol. 7, No. 3, 1994, pp. 569-578. http://dx.doi.org/10.1183/09031936.94.07030569
[8] R. A. Coleman, M. Johnson, A. T. Nials and C. J. Vardey, “Exosites: Their Current Status, and Their Relevance to the Duration of Action of Long-Acting β2-Adrenoceptor Agonists,” Trends in Pharmacological Sciences, Vol. 17, No. 9, 1996, pp. 324-330.
[9] P. Casarosa, I. Kollak, T. Kiechle, A. Ostermann, A. Schnapp, et al., “Functional and Biochemical Rationales for the 24-Hour-Long Duration of Action of Olodaterol,” Journal of Pharmacology and Experimental Therapeutics, Vol. 337, No. 3, 2011, pp. 600-609.
[10] R. M. Wallace and J. M. Young, “Temperature Dependence of the Binding of [3H]Mepyramine and Related Compounds to the Histamine H1 Receptor,” Molecular Pharmacology, Vol. 23, No. 1, 1983, pp. 60-66.
[11] A. Szczuka, M. Wennerberg, A. Packeu and G. Vauquelin, “Molecular Mechanisms for the Persistent Bronchodilatory Effect of the β2-Adrenoceptor Agonist Salmeterol,” British Journal of Pharmacology, Vol. 158, No. 1, 2009, pp. 183-194.
[12] J. C. W. Mak, B. Grandordy and P. J. Barnes, “High Affinity [3H]Formoterol Binding Sites in Lung: Characterization and Autoradiographic,” European Journal of Pharmacology: Molecular Pharmacology, Vol. 269, No. 1, 1994, pp. 35-41.
[13] R. J. Slack, V. J. Barrett, V. S. Morrison, R. G. Sturton, A. J. Emmons, et al., “In Vitro Pharmacological Characterization of Vilanterol, a Novel Long-Acting β2 Adrenoceptor Agonist with 24-Hour Duration of Action,” Journal of Pharmacology and Experimental Therapeutics, Vol. 344, No. 1, 2013, pp. 218-230.
[14] P. A. Procopiou, V. J. Barrett, N. J. Bevan, K. Biggadike, P. C. Box, et al., “Synthesis and Structure-Activity Relationships of Long-Acting β2 Adrenergic Receptor Agonists Incorporating Metabolic Inactivation: An Antedrug Approach,” Journal of Medicinal Chemistry, Vol. 53, No. 11, 2010, pp. 4522-4530.
[15] P. K. Smith, R. I. Krohn, G. T. Hermanson, A. K. Mallia, F. H. Gartner, et al., “Measurement of Protein Using Bicinchoninic Acid,” Analytical Biochemistry, Vol. 150, No. 1, 1985, pp. 76-85.
[16] J. G. Baker, “The Selectivity of β-Adrenoceptor Antagonists at the Human β1, β2 and β3 Adrenoceptors,” British Journal of Pharmacology, Vol. 144, No. 3, 2005, pp. 317-322. http://dx.doi.org/10.1038/sj.bjp.0706048
[17] S. A. Holm, “A Simple Sequentially Rejective Multiple test Procedure,” Scandinavian Journal of Statistics, Vol. 6, No. 2, 1979, pp. 65-70.

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.