title: Detailed Characterization of the In Vitro Pharmacological and Pharmacokinetic Properties ofN-(2-Hydroxybenzyl)-2,5-Dimethoxy-4-Cyanophenylethylamine (25CN-NBOH), a Highly Selective and Brain-Penetrant 5-HT2AReceptor Agonist.
abstract: Therapeutic interest in augmentation of 5-hydroxytryptamine2A(5-HT2A) receptor signaling has been renewed by the effectiveness of psychedelic drugs in the treatment of various psychiatric conditions. In this study, we have further characterized the pharmacological properties of the recently developed 5-HT2receptor agonistN-2-hydroxybenzyl)-2,5-dimethoxy-4-cyanophenylethylamine (25CN-NBOH) and three structural analogs at recombinant 5-HT2A, 5-HT2B, and 5-HT2Creceptors and investigated the pharmacokinetic properties of the compound. 25CN-NBOH displayed robust 5-HT2Aselectivity in [3H]ketanserin/[3H]mesulergine, [3H]lysergic acid diethylamide and [3H]Cimbi-36 binding assays (Ki2C/Ki2Aratio range of 52-81;Ki2B/Ki2Aratio of 37). Moreover, in inositol phosphate and intracellular Ca2+mobilization assays 25CN-NBOH exhibited 30- to 180-fold 5-HT2A/5-HT2Cselectivities and 54-fold 5-HT2A/5-HT2Bselectivity as measured by Δlog(Rmax/EC50) values. In an off-target screening 25CN-NBOH (10μM) displayed either substantially weaker activity or inactivity at a plethora of other receptors, transporters, and kinases. In a toxicological screening, 25CN-NBOH (100μM) displayed a benign acute cellular toxicological profile. 25CN-NBOH displayed high in vitro permeability (Papp= 29 × 10-6cm/s) and low P-glycoprotein-mediated efflux in a conventional model of cellular transport barriers. In vivo, administration of 25CN-NBOH (3 mg/kg, s.c.) in C57BL/6 mice mice produced plasma and brain concentrations of the free (unbound) compound of ∼200 nM within 15 minutes, further supporting that 25CN-NBOH rapidly penetrates the blood-brain barrier and is not subjected to significant efflux. In conclusion, 25CN-NBOH appears to be a superior selective and brain-penetrant 5-HT2Areceptor agonist compared with (±)-2,5-dimethoxy-4-iodoamphetamine (DOI), and thus we propose that the compound could be a valuable tool for future investigations of physiologic functions mediated by this receptor.
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark (A.A.J., S.L-P., G.L., H.B.-O., J.L.K.); Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, North Carolina (J.D.M., T.P.K.); and Department of Discovery Chemistry and DMPK, H. Lundbeck A/S, Valby, Denmark (C.B., J.K.).
authors.forename: Anders A
authors.initials: AA
authors.lastname: Jensen
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark (A.A.J., S.L-P., G.L., H.B.-O., J.L.K.); Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, North Carolina (J.D.M., T.P.K.); and Department of Discovery Chemistry and DMPK, H. Lundbeck A/S, Valby, Denmark (C.B., J.K.).
authors.forename: John D
authors.initials: JD
authors.lastname: McCorvy
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark (A.A.J., S.L-P., G.L., H.B.-O., J.L.K.); Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, North Carolina (J.D.M., T.P.K.); and Department of Discovery Chemistry and DMPK, H. Lundbeck A/S, Valby, Denmark (C.B., J.K.).
authors.forename: Sebastian
authors.initials: S
authors.lastname: Leth-Petersen
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark (A.A.J., S.L-P., G.L., H.B.-O., J.L.K.); Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, North Carolina (J.D.M., T.P.K.); and Department of Discovery Chemistry and DMPK, H. Lundbeck A/S, Valby, Denmark (C.B., J.K.).
authors.forename: Christoffer
authors.initials: C
authors.lastname: Bundgaard
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark (A.A.J., S.L-P., G.L., H.B.-O., J.L.K.); Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, North Carolina (J.D.M., T.P.K.); and Department of Discovery Chemistry and DMPK, H. Lundbeck A/S, Valby, Denmark (C.B., J.K.).
authors.forename: Gudrun
authors.initials: G
authors.lastname: Liebscher
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark (A.A.J., S.L-P., G.L., H.B.-O., J.L.K.); Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, North Carolina (J.D.M., T.P.K.); and Department of Discovery Chemistry and DMPK, H. Lundbeck A/S, Valby, Denmark (C.B., J.K.).
authors.forename: Terry P
authors.initials: TP
authors.lastname: Kenakin
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark (A.A.J., S.L-P., G.L., H.B.-O., J.L.K.); Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, North Carolina (J.D.M., T.P.K.); and Department of Discovery Chemistry and DMPK, H. Lundbeck A/S, Valby, Denmark (C.B., J.K.).
authors.forename: Hans
authors.initials: H
authors.lastname: Bräuner-Osborne
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark (A.A.J., S.L-P., G.L., H.B.-O., J.L.K.); Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, North Carolina (J.D.M., T.P.K.); and Department of Discovery Chemistry and DMPK, H. Lundbeck A/S, Valby, Denmark (C.B., J.K.).
authors.forename: Jan
authors.initials: J
authors.lastname: Kehler
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark (A.A.J., S.L-P., G.L., H.B.-O., J.L.K.); Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, North Carolina (J.D.M., T.P.K.); and Department of Discovery Chemistry and DMPK, H. Lundbeck A/S, Valby, Denmark (C.B., J.K.) jesper.kristensen@sund.ku.dk.
authors.forename: Jesper Langgaard
authors.initials: JL
authors.lastname: Kristensen
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000378:metabolism*; D001921:Brain / Q000187:drug effects / Q000378:metabolism*; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D057809:HEK293 Cells; D006801:Humans; D061985:Madin Darby Canine Kidney Cells; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D044402:Receptor, Serotonin, 5-HT2A / Q000378:metabolism*; D058826:Serotonin 5-HT2 Receptor Agonists / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*
chemical_list: D044402:Receptor, Serotonin, 5-HT2A; D058826:Serotonin 5-HT2 Receptor Agonists