title: Annulated heterocyclic bioisosteres of norarecoline. Synthesis and molecular pharmacology at five recombinant human muscarinic acetylcholine receptors.
abstract: A series of O-alkylated analogs of 5,6,7,8-tetrahydro-4H-isoxazolo[4,5-c]azepin-3-ol (THAO) were synthesized and characterized as ligands for muscarinic acetylcholine receptors (mAChRs). O-Methyl-THAO (4a), O-ethyl-THAO (4b), O-isopropyl-THAO (4c), and O-propargyl-THAO (4d) were shown to be potent inhibitors of the binding of tritiated quinuclidinyl benzilate (QNB), pirenzepine (PZ), and oxotremorine-M (Oxo-M) to tissue membrane preparations. In the [3H]-Oxo-M binding assay, receptor affinities in the low nanomolar range were measured for 4a (IC50 = 0.010 microM), 4b (IC50 = 0.003 microM), 4c (IC50 = 0.011 microM), and 4d (IC50 = 0.0008 microM). Pharmacological effects (EC50 or Ki values) and intrinsic activities (per cent of maximal carbachol responses) were determined using five recombinant human mAChRs (m1-m5) and the functional assay, receptor selection and amplification technology (R-SAT). Compound 4c antagonized carbachol-induced responses at m1, m3, and m5. With the exception of 4b, which was an antagonist at m5, 4a,b,d showed partial agonism at m1-m5 with very similar subtype selectivity (m2 > m4 > m1 > or = m3 > m5). Agonist index values for 4a-d, which were calculated from [3H]QNB (brain) and [3H]Oxo-M (brain) binding data, were shown to be predictive of pharmacologically determined intrinsic activities at m1-m5, the same rank order of intrinsic activity being observed at all five mAChRs (4a > 4d > 4b > 4c). It is concluded that within this class of high-affinity mAChR (m1-m5) ligands, containing secondary amino groups, minor changes of the bioisosteric ester alkyl groups have marked effects on potency and, in particular, intrinsic activity.
authors.affiliation: Department of Medicinal Chemistry, Royal Danish School of Pharmacy, Copenhagen.
authors.forename: H
authors.initials: H
authors.lastname: Bräuner-Osborne
authors.forename: B
authors.initials: B
authors.lastname: Ebert
authors.forename: M R
authors.initials: MR
authors.lastname: Brann
authors.forename: E
authors.initials: E
authors.lastname: Falch
authors.forename: P
authors.initials: P
authors.lastname: Krogsgaard-Larsen
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001115:Arecoline / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism; D006571:Heterocyclic Compounds / Q000494:pharmacology*; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D018721:Muscarinic Agonists; D011485:Protein Binding; D051381:Rats; D011976:Receptors, Muscarinic / Q000378:metabolism; D011994:Recombinant Proteins / Q000819:agonists / Q000378:metabolism
chemical_list: D006571:Heterocyclic Compounds; D018721:Muscarinic Agonists; D011976:Receptors, Muscarinic; D011994:Recombinant Proteins; C094105:norarecoline; D001115:Arecoline