Structure-based ligand discovery targeting orthosteric and allosteric pockets of dopamine receptors. Mol Pharmacol 2013 Dec;84(6):794-807
Date
09/12/2013Pubmed ID
24021214Pubmed Central ID
PMC3834142DOI
10.1124/mol.113.088054Scopus ID
2-s2.0-84888883269 (requires institutional sign-in at Scopus site) 83 CitationsAbstract
Small molecules targeting allosteric pockets of G protein-coupled receptors (GPCRs) have a great therapeutic potential for the treatment of neurologic and other chronic disorders. Here we performed virtual screening for orthosteric and putative allosteric ligands of the human dopamine D3 receptor (D3R) using two optimized crystal-structure-based models: the receptor with an empty binding pocket (D3R(APO)), and the receptor complex with dopamine (D3R(Dopa)). Subsequent biochemical and functional characterization revealed 14 novel ligands with a binding affinity of better than 10 μM in the D3R(APO) candidate list (56% hit rate), and 8 novel ligands in the D3R(Dopa) list (32% hit rate). Most ligands in the D3R(APO) model span both orthosteric and extended pockets and behave as antagonists at D3R, with compound 7 showing the highest potency of dopamine inhibition (IC₅₀ = 7 nM). In contrast, compounds identified by the D3R(Dopa) model are predicted to occupy an allosteric site at the extracellular extension of the pocket, and they all lack the anchoring amino group. Compounds targeting the allosteric site display a variety of functional activity profiles, where behavior of at least two compounds (23 and 26) is consistent with noncompetitive allosteric modulation of dopamine signaling in the extracellular signal-regulated kinase 1 and 2 phosphorylation and β-arrestin recruitment assays. The high affinity and ligand efficiency of the chemically diverse hits identified in this study suggest utility of structure-based screening targeting allosteric sites of GPCRs.
Author List
Lane JR, Chubukov P, Liu W, Canals M, Cherezov V, Abagyan R, Stevens RC, Katritch VAuthor
Wei Liu PhD Professor in the Pharmacology and Toxicology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Allosteric SiteAnimals
Arrestins
Binding Sites
CHO Cells
Cricetinae
Cricetulus
Databases, Factual
Dopamine
High-Throughput Screening Assays
Humans
Ligands
Mitogen-Activated Protein Kinase 1
Mitogen-Activated Protein Kinase 3
Models, Molecular
Phosphorylation
Protein Binding
Protein Conformation
Radioligand Assay
Receptors, Dopamine D3
Sf9 Cells
Small Molecule Libraries
Structure-Activity Relationship
beta-Arrestins
