TR-FRET binding assay targeting unactivated form of Bruton's tyrosine kinase

Bioorg Med Chem Lett. 2015;25(10):2033-6. doi: 10.1016/j.bmcl.2015.04.001. Epub 2015 Apr 4.

Abstract

Bruton's Tyrosine Kinase (BTK) is one of the crucial kinases for the B cell maturation and mast cell activation, and specific inhibitors of BTK are considered to be attractive targets in drug discovery research. In this Letter, we have designed and synthesized a new fluorescent probe for TR-FRET-based high-throughput screening, to identify compounds that preferentially bind to an inactive conformation of BTK which has a unique structural feature. A set of kinase-focused compound library was screened using this assay method, and compound 31 was successfully identified as a potent inhibitor which preferentially bind to the inactive conformation of BTK. These results suggest that this screening method has a great potential for the discovery of novel selective BTK inhibitors.

Keywords: Bruton’s tyrosine kinase; Fluorescent probe; Kinase inhibitor; Screening; TR-FRET.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agammaglobulinaemia Tyrosine Kinase
  • Binding, Competitive
  • Biological Assay / methods*
  • Drug Delivery Systems
  • Drug Design
  • Enzyme Activation / drug effects
  • Fluorescent Dyes / chemistry*
  • Inhibitory Concentration 50
  • Models, Biological
  • Protein Binding
  • Protein Kinase Inhibitors / chemistry
  • Protein Kinase Inhibitors / isolation & purification*
  • Protein Kinase Inhibitors / pharmacology
  • Protein Structure, Tertiary
  • Protein-Tyrosine Kinases / antagonists & inhibitors*

Substances

  • Fluorescent Dyes
  • Protein Kinase Inhibitors
  • Protein-Tyrosine Kinases
  • Agammaglobulinaemia Tyrosine Kinase