Identification and characterization of a novel small-molecule inhibitor of β-catenin signaling

Am J Pathol. 2014 Jul;184(7):2111-22. doi: 10.1016/j.ajpath.2014.04.002. Epub 2014 May 10.

Abstract

Hepatocellular carcinoma (HCC), the third most common cause of cancer-related deaths worldwide, lacks effective medical therapy. Large subsets of HCC demonstrate Wnt/β-catenin activation, making this an attractive therapeutic target. We report strategy and characterization of a novel small-molecule inhibitor, ICG-001, known to affect Wnt signaling by disrupting β-catenin-CREB binding protein interactions. We queried the ZINC online database for structural similarity to ICG-001 and identified PMED-1 as the lead compound, with ≥70% similarity to ICG-001. PMED-1 significantly reduced β-catenin activity in hepatoblastoma and several HCC cells, as determined by TOPflash reporter assay, with an IC50 ranging from 4.87 to 32 μmol/L. Although no toxicity was observed in primary human hepatocytes, PMED-1 inhibited Wnt target expression in HCC cells, including those with CTNNB1 mutations, and impaired cell proliferation and viability. PMED-1 treatment decreased β-catenin-CREB binding protein interactions without affecting total β-catenin levels or activity of other common kinases. PMED-1 treatment of Tg(OTM:d2EGFP) zebrafish expressing GFP under the β-catenin/Tcf reporter led to a notable decrease in β-catenin activity. The PMED effect on β-catenin signaling lasted from 12 to 24 hours in vitro and 6 to 15 hours in vivo. Thus, using a rapid and cost-effective computational methodology, we have identified a novel and specific small-molecule inhibitor of Wnt signaling that may have implications for HCC treatment.

Publication types

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

MeSH terms

  • Animals
  • Bridged Bicyclo Compounds, Heterocyclic / pharmacology
  • CREB-Binding Protein / metabolism
  • Carcinoma, Hepatocellular / metabolism
  • Cell Line, Tumor
  • Drug Discovery
  • Humans
  • Inhibitory Concentration 50
  • Liver Neoplasms / metabolism
  • Pyrimidinones / pharmacology
  • Structure-Activity Relationship
  • Wnt Signaling Pathway / drug effects*
  • Zebrafish
  • beta Catenin / antagonists & inhibitors*
  • beta Catenin / metabolism

Substances

  • Bridged Bicyclo Compounds, Heterocyclic
  • CTNNB1 protein, human
  • ICG 001
  • Pyrimidinones
  • beta Catenin
  • CREB-Binding Protein