5-Hydroxytryptamine (5-HT) receptors and dopamine 2 (D 2 ) receptor modulate gastrointestinal motility. Gastroprokinetic agents that act on several 5-HT receptor subtypes and/or D 2 receptors are used clinically. Although the 5-HT 4 receptor is known to mediate the gastroprokinetic effects of these agents, the absence of highly selective 5-HT 4 receptor agonists has made it difficult to confirm the physiological consequences of selective 5-HT 4 receptor stimulation. In this study, we report the in vitro pharmacological profiles and the in vivo gastroprokinetic effects of 5-amino-6-chloro-N-[(1-isobutylpiperidin-4-yl)methyl]-2-methylimidazo[1,2-␣]pyridine-8-carboxamide (CJ-033,466), a novel, potent, and selective 5-HT 4 partial agonist. Compared with preceding 5-HT 4 agonists such as cisapride, mosapride, and tegaserod, CJ-033,466 had a superior in vitro profile, with nanomolar agonistic activities for the 5-HT 4 receptor and 1000-fold greater selectivity for the 5-HT 4 receptor over other 5-HT and D 2 receptors. In vivo studies in conscious dogs showed that CJ-033,466 dose-dependently stimulated gastric antral motility in both the fasted and postprandial states at the same dose range and that it was 30 times more potent than cisapride. Furthermore, CJ-033,466 accelerated the gastric emptying rate in a gastroparesis dog model at the minimally effective dose established in the gastric motility study. In conclusion, CJ-033,466 is a potent and highly selective 5-HT 4 agonist that stimulates physiologically coordinated gastric motility, and it has no activity on other 5-HT receptor subtypes and D 2 receptors. Therefore, CJ-033,466 could be used to treat gastroparesis, providing better gastroprokinetics and reduced side effects mediated by the other receptors.The neurotransmitter 5-hydroxytryptamine (5-HT) mediates a broad range of physiological and pathophysiological responses both centrally and peripherally. High amounts of 5-HT are produced in the gastrointestinal (GI) tract, and 5-HT affects GI motility by signaling through receptors that are present throughout the GI tract (Gershon, 2004;Neal and Bornstein, 2006;Costedio et al., 2007;Gershon and Tack, 2007).All classes of the extensive 5-HT receptor family, except for the ligand-gated 5-HT 3 receptor, are members of the seven transmembrane-spanning G protein-coupled receptor family. These receptors modulate signal transduction pathways via stimulating or inhibiting adenylyl cyclase, modulating cytosolic calcium concentrations, or activating multiple alternative downstream effectors.The 5-HT 4 receptor is thought to signal principally, but not T.M. and Y.O. contributed equally to the study. Article, publication date, and citation information can be found at