C5-DNA methyltransferase inhibitors: from screening to effects on zebrafish embryo development

Chembiochem. 2011 Jun 14;12(9):1337-45. doi: 10.1002/cbic.201100130. Epub 2011 Jun 1.

Abstract

DNA methylation is involved in the regulation of gene expression and plays an important role in normal developmental processes and diseases, such as cancer. DNA methyltransferases are the enzymes responsible for DNA methylation on the position 5 of cytidine in a CpG context. In order to identify and characterize novel inhibitors of these enzymes, we developed a fluorescence-based throughput screening by using a short DNA duplex immobilized on 96-well plates. We have screened 114 flavones and flavanones for the inhibition of the murine catalytic Dnmt3a/3L complex and found 36 hits with IC(50) values in the lower micromolar and high nanomolar ranges. The assay, together with inhibition tests on two other methyltransferases, structure-activity relationships and docking studies, gave insights on the mechanism of inhibition. Finally, two derivatives effected zebrafish embryo development, and induced a global demethylation of the genome, at doses lower than the control drug, 5-azacytidine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Crystallography, X-Ray
  • DNA (Cytosine-5-)-Methyltransferases / antagonists & inhibitors*
  • DNA (Cytosine-5-)-Methyltransferases / chemistry
  • Embryonic Development / drug effects*
  • Enzyme Inhibitors / pharmacology*
  • Mice
  • Models, Molecular
  • Molecular Sequence Data
  • Small Molecule Libraries / chemistry
  • Small Molecule Libraries / pharmacology*
  • Zebrafish / embryology*

Substances

  • Enzyme Inhibitors
  • Small Molecule Libraries
  • DNA (Cytosine-5-)-Methyltransferases