Identification of small molecules that disrupt signaling between ABL and its positive regulator RIN1

PLoS One. 2015 Mar 26;10(3):e0121833. doi: 10.1371/journal.pone.0121833. eCollection 2015.

Abstract

Constitutively active BCR-ABL kinase fusions are causative mutations in the pathogenesis of hematopoietic neoplasias including chronic myelogenous leukemia (CML). Although these fusions have been successfully targeted with kinase inhibitors, drug-resistance and relapse continue to limit long-term survival, highlighting the need for continued innovative drug discovery. We developed a time-resolved Förster resonance energy transfer (TR-FRET) -based assay to identify compounds that disrupt stimulation of the ABL kinase by blocking its ability to bind the positive regulator RIN1. This assay was used in a high throughput screen (HTS) of two small molecule libraries totaling 444,743 compounds. 708 confirmed hits were counter-screened to eliminate off-target inhibitors and reanalyzed to prioritize compounds with IC50 values below 10 μM. The CML cell line K562 was then used to identify five compounds that decrease MAPK1/3 phosphorylation, which we determined to be an indicator of RIN1-dependent ABL signaling. One of these compounds is a thiadiazole, and the other four are structurally related acyl piperidine amides. Notably, these five compounds lower cellular BCR-ABL1 kinase activity by blocking a positive regulatory interaction rather than directly inhibiting ABL catalytic function.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biflavonoids / pharmacology
  • Catechin / analogs & derivatives
  • Catechin / pharmacology
  • Fluorescence Resonance Energy Transfer
  • Fusion Proteins, bcr-abl / metabolism
  • High-Throughput Screening Assays
  • Humans
  • Intracellular Signaling Peptides and Proteins / metabolism*
  • K562 Cells
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Phosphorylation / drug effects
  • Protein Binding / drug effects
  • Protein Kinase Inhibitors / pharmacology
  • Proto-Oncogene Proteins c-abl / antagonists & inhibitors
  • Proto-Oncogene Proteins c-abl / metabolism*
  • Reproducibility of Results
  • Signal Transduction / drug effects*
  • Small Molecule Libraries / chemistry
  • Small Molecule Libraries / pharmacology*
  • Time Factors

Substances

  • Biflavonoids
  • Intracellular Signaling Peptides and Proteins
  • Protein Kinase Inhibitors
  • RIN1 protein, human
  • Small Molecule Libraries
  • theaflavin-3,3'-digallate
  • Catechin
  • Fusion Proteins, bcr-abl
  • Proto-Oncogene Proteins c-abl
  • Mitogen-Activated Protein Kinase 1