title: Characterization of muscarinic receptor binding by the novel radioligand, [(3)H]imidafenacin, in the bladder and other tissues of rats.
abstract: The present study aimed to directly characterize specific binding sites of tritium ([(3)H])-labeled imidafenacin, a new radioligand for labeling muscarinic receptors, in the bladder and other peripheral or central nervous tissues of rats. Muscarinic receptors in rat tissues were measured by radioligand binding assay using [(3)H]imidafenacin. Specific [(3)H]imidafenacin binding in rat tissues was saturable, reversible, and of high affinity. Estimated dissociation constants (Kd values) were significantly lower in submaxillary gland and prostate and higher in heart than in bladder, indicating lower Kd values in M1 and M3 subtype- than M2 subtype-dominating tissues. Unlabeled imidafenacin and clinically used antimuscarinic agents competed with [(3)H]imidafenacin for binding sites in bladder and other tissues in a concentration-dependent manner, which indicated pharmacological specificity of [(3)H]imidafenacin binding sites. Pretreatment with N-(2-chloroethyl)-4-piperidinyl diphenylacetate (4-DAMP mustard), an irreversible inactivating agent of M3 subtype, significantly decreased the number of [(3)H]imidafenacin binding sites in bladder, submaxillary gland, and colon, but not in heart. [(3)H]imidafenacin labeled muscarinic receptors in M1 and M3 subtype-dominating tissues with higher affinity than [N-methyl-(3)H]scopolamine methyl chloride (NMS). [(3)H]imidafenacin is a useful radioligand to label muscarinic receptors in M1- and M3-dominating tissues with high affinity.
authors.affiliation: Department of Pharmacokinetics and Pharmacodynamics, Graduate School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
authors.forename: Shiori
authors.initials: S
authors.lastname: Kuraoka
authors.affiliation: Department of Pharmacokinetics and Pharmacodynamics, Graduate School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Ito
authors.affiliation: Department of Pharmacology, School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women's University, 11-68 Koshien, Kyuban-cho, Nishinomiya 663-8179, Japan.
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Wakuda
authors.affiliation: Department of Pharmacology, School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women's University, 11-68 Koshien, Kyuban-cho, Nishinomiya 663-8179, Japan.
authors.forename: Kazumasa
authors.initials: K
authors.lastname: Shinozuka
authors.affiliation: Department of Pharmacokinetics and Pharmacodynamics, Graduate School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
authors.forename: Satomi
authors.initials: S
authors.lastname: Onoue
authors.affiliation: Department of Pharmacokinetics and Pharmacodynamics, Graduate School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan; Center for Pharma-Food Research (CPFR), Graduate School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan. Electronic address: yamada@u-shizuoka-ken.ac.jp.
authors.forename: Shizuo
authors.initials: S
authors.lastname: Yamada
mesh_terms: D000818:Animals; D007093:Imidazoles / Q000378:metabolism* / Q000493:pharmacokinetics; D008297:Male; D011869:Radioligand Assay; D051381:Rats; D017207:Rats, Sprague-Dawley; D011976:Receptors, Muscarinic / Q000378:metabolism*; D014018:Tissue Distribution; D014316:Tritium; D001743:Urinary Bladder / Q000378:metabolism*
chemical_list: D007093:Imidazoles; D011976:Receptors, Muscarinic; D014316:Tritium; C120953:imidafenacin
keywords: Bladder; Muscarinic receptor subtype; [(3)H]imidafenacin