RT Journal Article SR Electronic T1 Preclinical Evaluation of Long-Acting Muscarinic Antagonists: Comparison of Tiotropium and Investigational Drugs JF Journal of Pharmacology and Experimental Therapeutics JO J Pharmacol Exp Ther FD American Society for Pharmacology and Experimental Therapeutics SP 660 OP 668 DO 10.1124/jpet.109.152470 VO 330 IS 2 A1 Paola Casarosa A1 Thierry Bouyssou A1 Sabine Germeyer A1 Andreas Schnapp A1 Florian Gantner A1 Michael Pieper YR 2009 UL http://jpet.aspetjournals.org/content/330/2/660.abstract AB Chronic obstructive pulmonary disease (COPD) is characterized by progressive airflow limitation caused by persistent inflammatory processes in the airways. An increased cholinergic tone mediates different pathophysiological features of COPD, such as bronchoconstriction and mucus hypersecretion, mostly through activation of the human muscarinic M3 receptor (hM3) subtype. Tiotropium bromide (Spiriva) is a well established muscarinic antagonist in the pharmacological management of COPD with a once-daily posology. The rationale behind the sustained bronchodilation obtained with tiotropium consists in its slow dissociation from hM3 receptors. In this study, we performed a comprehensive preclinical comparison of tiotropium with other long-acting muscarinic antagonists (LAMAs) currently in clinical development, namely aclidinium bromide and glycopyrrolate. The different muscarinic antagonists were characterized for their 1) affinity toward the different human muscarinic receptor subtypes expressed in Chinese hamster ovary cells and kinetics of receptor dissociation, 2) potency in inhibiting the agonist-induced activation of muscarinic receptors through measurement of second messengers, and 3) efficacy and duration of bronchoprotection, as tested in a model of acetylcholine-induced bronchoconstriction in anesthetized dogs over a period of 24 h. All of the tested LAMAs showed high affinity and potency toward the hM3 receptor (tiotropium, pA2 = 10.4; aclidinium, pA2 = 9.6; and glycopyrrolate, pA2 = 9.7). However, dissociation half-lives of the LAMAs from the hM3 receptor differed significantly (tiotropium, t½ = 27 h; aclidinium, t½ = 10.7 h; and glycopyrrolate, t½ = 6.1 h). In line with their kinetic properties at the hM3, the tested LAMAs provided different levels of bronchoprotection in the in vivo setting 24 h after administration (tiotropium = 35%, aclidinium = 21%, and glycopyrrolate = 0% at 24 h) when applied at equieffective doses. The American Society for Pharmacology and Experimental Therapeutics