PDF - 202K, (A) Representative images of wound healing assay of MDA-MB-231 cells. Right after the wound was made on the MDA-MB-231 cell monolayer, the cells were treated with TGF-beta1 (2 ng/ml) with or without ALK5 inhibitors for 53 h. Total magnification was x 100. Scale bar indicates 100 microm. Veh, SB, and EW indicate vehicle, SB-505124, and EW-7197, respectively. (B, C) Effects of EW-7197 on cell cytotoxicity (described in the Supplementary Methods). 4T1 (B) and MCF10A (C) cells were treated with indicated concentrations of EW-7197 in 0.2% HI-FBS medium for 72 h. The cell viability was determined by SRB (Sulforhodamine B) assay (described in the Supplementary Methods). Data represent the mean (plus or minus) SD (n=3).
ARTICLE ABSTRACTAdvanced tumors produce an excessive amount of transforming growth factor β (TGFβ), which promotes tumor progression at late stages of malignancy. The purpose of this study was to develop anti-TGFβ therapeutics for cancer. We synthesized a novel small-molecule TGFβ receptor I kinase (activin receptor–like kinase 5) inhibitor termed N-[[4-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-5-(6-methylpyridin-2-yl)-1H-imidazol-2-yl]methyl]-2-fluoroaniline (EW-7197), and we investigated its potential antimetastatic efficacy in mouse mammary tumor virus (MMTV)/c-Neu mice and 4T1 orthotopic–grafted mice. EW-7197 inhibited Smad/TGFβ signaling, cell migration, invasion, and lung metastasis in MMTV/c-Neu mice and 4T1 orthotopic–grafted mice. EW-7197 also inhibited the epithelial-to-mesenchymal transition (EMT) in both TGFβ-treated breast cancer cells and 4T1 orthotopic–grafted mice. Furthermore, EW-7197 enhanced cytotoxic T lymphocyte activity in 4T1 orthotopic–grafted mice and increased the survival time of 4T1-Luc and 4T1 breast tumor–bearing mice. In summary, EW-7197 showed potent in vivo antimetastatic activity, indicating its potential for use as an anticancer therapy. Mol Cancer Ther; 13(7); 1704–16. ©2014 AACR.