High-content screen using zebrafish (Danio rerio) embryos identifies a novel kinase activator and inhibitor. Issue 9 (1st May 2017)

• Record Type: Journal Article
• Authors: Geldenhuys, Werner J.; Bergeron, Sadie A.; Mullins, Jackie E.; Aljammal, Rowaa; Gaasch, Briah L.; Chen, Wei-Chi; Yun, June; Hazlehurst, Lori A.
• Is Part Of: Bioorganic & medicinal chemistry letters. Volume 27:Issue 9(2017)
• Page Start: 2029
• Page End: 2037
• Publication Date: 2017-05-01
• Subjects: Phenotypic screen -- Cancer -- Zebrafish -- Kinase -- Compound library -- Notochord -- Somites; Bioorganic chemistry -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1016/j.bmcl.2017.02.068
• Languages: English
• ISSNs: 0960-894X

Abstract

Graphical abstract: Abstract: In this report we utilized zebrafish ( Danio rerio ) embryos in a phenotypical high-content screen (HCS) to identify novel leads in a cancer drug discovery program. We initially validated our HCS model using the flavin adenosine dinucleotide (FAD) containing endoplasmic reticulum (ER) enzyme, endoplasmic reticulum oxidoreductase (ERO1) inhibitor EN460. EN460 showed a dose response effect on the embryos with a dose of 10 μM being significantly lethal during early embryonic development. The HCS campaign which employed a small library identified a promising lead compound, a naphthyl-benzoic acid derivative coined compound 1 which had significant dosage and temporally dependent effects on notochord and muscle development in zebrafish embryos. Screening a 369 kinase member panel we show that compound 1 is a PIM3 kinase inhibitor (IC50 = 4.078 μM) and surprisingly a DAPK1 kinase agonist/activator (EC50 = 39.525 μM). To our knowledge this is the first example of a small molecule activating DAPK1 kinase. We provide a putative model for increased phosphate transfer in the ATP binding domain when compound 1 is virtually docked with DAPK1. Our data indicate that observable phenotypical changes can be used in future zebrafish screens to identify compounds acting via similar molecular signaling pathways.