title: In vivo structure-activity relationship studies support allosteric targeting of a dual specificity phosphatase.
abstract: Dual specificity phosphatase 6 (DUSP6) functions as a feedback attenuator of fibroblast growth factor signaling during development. In vitro high throughput chemical screening attempts to discover DUSP6 inhibitors have yielded limited success. However, in vivo whole-organism screens of zebrafish identified compound 1 (BCI) as an allosteric inhibitor of DUSP6. Here we designed and synthesized a panel of analogues to define the structure-activity relationship (SAR) of DUSP6 inhibition. In vivo high-content analysis in transgenic zebrafish, coupled with cell-based chemical complementation assays, identified structural features of the pharmacophore of 1 that were essential for biological activity. In vitro assays of DUSP hyperactivation corroborated the results from in vivo and cellular SAR. The results reinforce the notion that DUSPs are druggable through allosteric mechanisms and illustrate the utility of zebrafish as a model organism for in vivo SAR analyses.
authors.affiliation: Department of Pharmaceutical Sciences, University of Pittsburgh, 3501 Fifth Avenue, Pittsburgh, PA 15213 (USA); Present address: Walter Reed Army Institute of Research, 503 Forney Drive, Silver Spring, MD 20910 (USA).
authors.forename: Vasiliy N
authors.initials: VN
authors.lastname: Korotchenko
authors.forename: Manush
authors.initials: M
authors.lastname: Saydmohammed
authors.forename: Laura L
authors.initials: LL
authors.lastname: Vollmer
authors.forename: Ahmet
authors.initials: A
authors.lastname: Bakan
authors.forename: Kyle
authors.initials: K
authors.lastname: Sheetz
authors.forename: Karl T
authors.initials: KT
authors.lastname: Debiec
authors.forename: Kristina A
authors.initials: KA
authors.lastname: Greene
authors.forename: Christine S
authors.initials: CS
authors.lastname: Agliori
authors.forename: Ivet
authors.initials: I
authors.lastname: Bahar
authors.forename: Billy W
authors.initials: BW
authors.lastname: Day
authors.forename: Andreas
authors.initials: A
authors.lastname: Vogt
authors.forename: Michael
authors.initials: M
authors.lastname: Tsang
mesh_terms: D000494:Allosteric Regulation; D000818:Animals; D015195:Drug Design; D054642:Dual Specificity Phosphatase 6 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005346:Fibroblast Growth Factors / Q000378:metabolism; D007192:Indenes / Q000737:chemistry* / Q000494:pharmacology*; D008958:Models, Molecular; D015398:Signal Transduction / Q000187:drug effects; D013329:Structure-Activity Relationship; D015027:Zebrafish / Q000196:embryology
chemical_list: D004791:Enzyme Inhibitors; D007192:Indenes; D005346:Fibroblast Growth Factors; D048049:Extracellular Signal-Regulated MAP Kinases; D054642:Dual Specificity Phosphatase 6
keywords: BCI; FGF signaling; cognition network technology; high-content screening; zebrafish