Abstract
From a high content in vivo screen for modulators of developmental patterning in embryonic zebrafish, we previously identified eggmanone (EGM1, 3) as a Hedgehog (Hh) signaling inhibitor functioning downstream of Smoothened. Phenotypic optimization studies for in vitro probe development utilizing a Gli transcription-linked stable luciferase reporter cell line identified EGM1 analogs with improved potency and aqueous solubility. Mechanistic profiling of optimized analogs indicated two distinct scaffold clusters: PDE4 inhibitors able to inhibit downstream of Sufu, and PDE4-independent Hh inhibitors functioning between Smo and Sufu. Each class represents valuable in vitro probes for elucidating the complex mechanisms of Hh regulation.
Keywords:
Eggmanone; Hedgehog signaling pathway; Phenotypic screening; Probe development; Suppressor of Fused.
Published by Elsevier Ltd.
Publication types
-
Research Support, N.I.H., Extramural
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Cyclic Nucleotide Phosphodiesterases, Type 4 / metabolism*
-
Dose-Response Relationship, Drug
-
HEK293 Cells
-
Hedgehog Proteins / antagonists & inhibitors*
-
Humans
-
Molecular Structure
-
Phosphodiesterase 4 Inhibitors / chemical synthesis
-
Phosphodiesterase 4 Inhibitors / chemistry
-
Phosphodiesterase 4 Inhibitors / pharmacology*
-
Pyrimidinones / chemical synthesis
-
Pyrimidinones / chemistry
-
Pyrimidinones / pharmacology*
-
Repressor Proteins / antagonists & inhibitors
-
Repressor Proteins / deficiency
-
Signal Transduction / drug effects*
-
Smoothened Receptor / antagonists & inhibitors
-
Structure-Activity Relationship
Substances
-
Hedgehog Proteins
-
Phosphodiesterase 4 Inhibitors
-
Pyrimidinones
-
Repressor Proteins
-
SMO protein, human
-
SUFU protein, human
-
Smoothened Receptor
-
Cyclic Nucleotide Phosphodiesterases, Type 4