title: A sphingosine 1-phosphate receptor 2 selective allosteric agonist.
abstract: Molecular probe tool compounds for the Sphingosine 1-phosphate receptor 2 (S1PR2) are important for investigating the multiple biological processes in which the S1PR2 receptor has been implicated. Amongst these are NF-κB-mediated tumor cell survival and fibroblast chemotaxis to fibronectin. Here we report our efforts to identify selective chemical probes for S1PR2 and their characterization. We employed high throughput screening to identify two compounds which activate the S1PR2 receptor. SAR optimization led to compounds with high nanomolar potency. These compounds, XAX-162 and CYM-5520, are highly selective and do not activate other S1P receptors. Binding of CYM-5520 is not competitive with the antagonist JTE-013. Mutation of receptor residues responsible for binding to the zwitterionic headgroup of sphingosine 1-phosphate (S1P) abolishes S1P activation of the receptor, but not activation by CYM-5520. Competitive binding experiments with radiolabeled S1P demonstrate that CYM-5520 is an allosteric agonist and does not displace the native ligand. Computational modeling suggests that CYM-5520 binds lower in the orthosteric binding pocket, and that co-binding with S1P is energetically well tolerated. In summary, we have identified an allosteric S1PR2 selective agonist compound.
authors.affiliation: Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Hideo
authors.initials: H
authors.lastname: Satsu
authors.forename: Marie-Therese
authors.initials: MT
authors.lastname: Schaeffer
authors.forename: Miguel
authors.initials: M
authors.lastname: Guerrero
authors.forename: Adrian
authors.initials: A
authors.lastname: Saldana
authors.forename: Christina
authors.initials: C
authors.lastname: Eberhart
authors.forename: Peter
authors.initials: P
authors.lastname: Hodder
authors.forename: Charmagne
authors.initials: C
authors.lastname: Cayanan
authors.forename: Stephan
authors.initials: S
authors.lastname: Schürer
authors.forename: Barun
authors.initials: B
authors.lastname: Bhhatarai
authors.forename: Ed
authors.initials: E
authors.lastname: Roberts
authors.forename: Hugh
authors.initials: H
authors.lastname: Rosen
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Brown
mesh_terms: D000494:Allosteric Regulation; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D057166:High-Throughput Screening Assays; D006801:Humans; D007700:Kinetics; D008024:Ligands; D062105:Molecular Docking Simulation; D009154:Mutation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011758:Pyrroles / Q000737:chemistry* / Q000378:metabolism; D049349:Receptors, Lysosphingolipid / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship; D008063:Thioctic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism
chemical_list: C586763:CYM-5520; D008024:Ligands; D011758:Pyrroles; D049349:Receptors, Lysosphingolipid; C586762:XAX-162; D008063:Thioctic Acid
keywords: Allosteric; GPCR; Molecular modeling; S1PR2; Signaling; Sphingosine 1-phosphate