TY - JOUR T1 - Muscarinic Receptors as Model Targets and Antitargets for Structure-Based Ligand Discovery JF - Molecular Pharmacology JO - Mol Pharmacol DO - 10.1124/mol.113.087551 SP - mol.113.087551 AU - Andrew C. Kruse AU - Dahlia R. Weiss AU - Mario Rossi AU - Jianxin Hu AU - Kelly Hu AU - Katrin Eitel AU - Peter Gmeiner AU - Jurgen Wess AU - Brian K. Kobilka AU - Brian K. Shoichet Y1 - 2013/01/01 UR - http://molpharm.aspetjournals.org/content/early/2013/07/25/mol.113.087551.abstract N2 - G protein-coupled receptors (GPCRs) regulate virtually all aspects of human physiology and represent an important class of therapeutic drug targets. Many GPCR-targeted drugs resemble endogenous agonists, often resulting in poor selectivity among receptor subtypes and restricted pharmacological profiles. The muscarinic acetylcholine receptor family exemplifies these problems; thousands of ligands are known, but few are receptor subtype-selective and almost all are cationic in nature. Using structure-based docking against the M2 and M3 muscarinic receptors, we screened 3.1 million molecules for ligands with new physical properties, chemotypes, and receptor subtype-selectivities. Of 19 docking-prioritized molecules tested against the M2 subtype, 11 had substantial activity and 8 represented new chemotypes. Intriguingly, two were uncharged ligands with low micromolar to high nanomolar Ki values, an observation with few precedents among aminergic GPCRs. To exploit a single amino-acid substitution among the binding pockets between the M2 and M3 receptors, we selected molecules predicted by docking to bind to the M3 and but not the M2 receptor. Of 16 molecules tested, eight bound to the M3 receptor. Whereas selectivity remained modest for most of these, one was a partial agonist at the M3 receptor without measurable M2 agonism. Consistent with this activity, this compound stimulated insulin release from a mouse b-cell line. These results support the ability of structure-based discovery to identify new ligands with unexplored chemotypes and physical properties, leading to new biological functions, even in an area as heavily explored as muscarinic pharmacology. ER -