High-content screen using zebrafish (Danio rerio) embryos identifies a novel kinase activator and inhibitor

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS

卷:27

Article

Geldenhuys, Werner J.¹  Bergeron, Sadie A.²  Mullins, Jackie E.²  Aljammal, Rowaa¹  Gaasch, Briah L.¹  Chen, Wei-Chi¹  Yun, June³  Hazlehurst, Lori A.¹ 

[1] West Virginia Univ, Sch Pharm, Dept Pharmaceut Sci, Morgantown, WV 26506 USA

[2] West Virginia Univ, Eberly Coll Arts & Sci, Dept Biol, Morgantown, WV 26506 USA

[3] Northeast Ohio Med Univ, Coll Med, Dept Integrat Med Sci, Rootstown, OH 44272 USA

关键词: Phenotypic screen;    Cancer;    Zebrafish;    Kinase;    Compound library;    Notochord;    Somites;   

DOI  :  10.1016/j.bmcl.2017.02.068

来源: Elsevier

【 摘 要 】

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 101 mu M 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 mu M) and surprisingly a DAPK1 kinase agonist/activator (EC50 = 39.525 mu M). 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. (C) 2017 Elsevier Ltd. All rights reserved.

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