title: The Influence of receptor kinetics on the onset and duration of action and the therapeutic index of NVA237 and tiotropium.
abstract: Studies under nonphysiological conditions suggest that long receptor residency time is responsible for the 24-h duration of action of the long-acting muscarinic antagonist (LAMA) tiotropium. Our aim was to determine how clinically relevant dissociation rates under more physiological conditions influence the differences in onset of action between tiotropium and 3-[(cyclopentylhydroxyphenylacetyl oxy]-1,1-dimethyl-pyrrolidinium bromide (NVA237), a once-daily dry-powder formulation of the LAMA glycopyrronium bromide in development for chronic obstructive pulmonary disease. In addition, we have investigated kinetic selectivity at each of the muscarinic receptor subtypes to determine whether the improved cardiovascular therapeutic index obtained with NVA237 in animal models is attributable to differences in kinetic rate constants. The binding of radioligand [3H]N-methyl-scopolamine was measured in the presence/absence of several concentrations of unlabeled competitors, and data were analyzed using a competition kinetic model to provide on/off rates for the competitor. We found shorter dissociation half-lives for NVA237 and tiotropium under physiological (11.4 and 46.2 min, respectively) versus nonphysiological conditions (173 and 462 min, respectively). NVA237 had a more rapid onset of action (3-4.8 times) versus tiotropium, determined in an vitro calcium and rat tracheal strip assay. Simulations suggested that the more rapid onset of NVA237 action could be explained by differences in kinetic parameters. NVA237 had greater equilibrium binding and kinetic selectivity for muscarinic type 3 (M3) versus muscarinic type 2 (M2) receptors, with a faster off rate from M2 versus M3 receptors than tiotropium, potentially affording it a more favorable therapeutic index. This study suggests that the 24-h duration of action of NVA237 and tiotropium is not solely the result of their slow dissociation from the M3 receptor and highlights the importance of conducting in vitro experiments in conditions reflecting those in vivo.
authors.affiliation: Novartis Institutes for Biomedical Research, Wimblehurst Road, Horsham, West Sussex, RH12 5AB, United Kingdom.
authors.forename: David A
authors.initials: DA
authors.lastname: Sykes
authors.forename: Mark R
authors.initials: MR
authors.lastname: Dowling
authors.forename: Juliet
authors.initials: J
authors.lastname: Leighton-Davies
authors.forename: Toby C
authors.initials: TC
authors.lastname: Kent
authors.forename: Lindsay
authors.initials: L
authors.lastname: Fawcett
authors.forename: Emilie
authors.initials: E
authors.lastname: Renard
authors.forename: Alexandre
authors.initials: A
authors.lastname: Trifilieff
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Charlton
mesh_terms: D000818:Animals; D001993:Bronchodilator Agents / Q000493:pharmacokinetics / Q000494:pharmacology* / Q000627:therapeutic use*; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism; D003198:Computer Simulation; D006224:Cricetinae; D003412:Cricetulus; D003627:Data Interpretation, Statistical; D006024:Glycopyrrolate / Q000493:pharmacokinetics / Q000494:pharmacology* / Q000627:therapeutic use; D066298:In Vitro Techniques; D007700:Kinetics; D018727:Muscarinic Antagonists / Q000494:pharmacology* / Q000627:therapeutic use*; D011869:Radioligand Assay; D051381:Rats; D043585:Receptor, Muscarinic M2 / Q000037:antagonists & inhibitors; D043587:Receptor, Muscarinic M3 / Q000037:antagonists & inhibitors*; D012602:Scopolamine Derivatives / Q000493:pharmacokinetics / Q000494:pharmacology* / Q000627:therapeutic use*; D012964:Sodium / Q000494:pharmacology; D013696:Temperature; D000069447:Tiotropium Bromide; D014132:Trachea / Q000187:drug effects
chemical_list: D001993:Bronchodilator Agents; D018727:Muscarinic Antagonists; D043585:Receptor, Muscarinic M2; D043587:Receptor, Muscarinic M3; D012602:Scopolamine Derivatives; D012964:Sodium; D002118:Calcium; D006024:Glycopyrrolate; D000069447:Tiotropium Bromide