ZFIN ID: ZDB-PUB-170323-23
High-content screen using zebrafish (Danio rerio) embryos identifies a novel kinase activator and inhibitor
Geldenhuys, W.J., Bergeron, S.A., Mullins, J.E., Aljammal, R., Gaasch, B.L., Chen, W.C., Yun, J., Hazlehurst, L.A.
Date: 2017
Source: Bioorganic & medicinal chemistry letters   27(9): 2029-2037 (Journal)
Registered Authors: Bergeron, Sadie
Keywords: Cancer, Compound library, Kinase, Notochord, Phenotypic screen, Somites, Zebrafish
MeSH Terms:
  • Animals
  • Antineoplastic Agents/chemistry
  • Antineoplastic Agents/pharmacology
  • Benzoic Acid/chemistry
  • Benzoic Acid/pharmacology
  • Death-Associated Protein Kinases/metabolism
  • Drug Discovery/methods*
  • Drug Screening Assays, Antitumor/methods
  • Embryo, Nonmammalian/drug effects*
  • Embryo, Nonmammalian/enzymology
  • Enzyme Activation/drug effects
  • Enzyme Activators/chemistry*
  • Enzyme Activators/pharmacology*
  • Neoplasms/drug therapy
  • Neoplasms/enzymology
  • Protein Kinase Inhibitors/chemistry*
  • Protein Kinase Inhibitors/pharmacology*
  • Protein-Serine-Threonine Kinases/antagonists & inhibitors
  • Protein-Serine-Threonine Kinases/metabolism
  • Zebrafish/embryology*
  • Zebrafish Proteins/antagonists & inhibitors
  • Zebrafish Proteins/metabolism
PubMed: 28320616 Full text @ Bioorg. Med. Chem. Lett.
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 10μM 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=4.078μM) and surprisingly a DAPK1 kinase agonist/activator (EC50=39.525μM). 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.