Supplementary Figure Legends - PDF file 41K, Legends for supplemental figures 1-4

Supplementary Figures 1-4 - PDF file 294K, Supplementary Figure 1: PKM2 activation by screening hit XC-409; Supplementary Figure 2: Phosphorylation state of PKM2 at Tyr105 in cancer cell lines; supplementary Figure 3: Viability of a selection of lung adenocarcinoma cells in media with and without serine; Supplementary Figure 4: Sensitivity of a panel of adherent cancer cell lines to serine deprivation and treatment with PKM2 activator SGI-10067

Activators of the pyruvate kinase M2 (PKM2) are currently attracting significant interest as potential anticancer therapies. They may achieve a novel antiproliferation response in cancer cells through modulation of the classic 'Warburg effect' characteristic of aberrant metabolism. In this Letter, we describe the optimization of a weakly active screening hit to a structurally novel series of small molecule 3-(trifluoromethyl)-1H-pyrazole-5-carboxamides as potent PKM2 activators.

Abstract Inactivation of the M2 form of pyruvate kinase (PKM2) in cancer cells is associated with increased tumorigenicity. To test the hypothesis that tumor growth may be inhibited through the PKM2 pathway, we generated a series of small-molecule PKM2 activators. The compounds exhibited low nanomolar activity in both biochemical and cell-based PKM2 activity assays. These compounds did not affect the growth of cancer cell lines under normal conditions in vitro, but strongly inhibited the proliferation of multiple lung cancer cell lines when serine was absent from the cell culture media. In addition, PKM2 activators inhibited the growth of an aggressive lung adenocarcinoma xenograft. These findings show that PKM2 activation by small molecules influences the growth of cancer cells in vitro and in vivo, and suggest that such compounds may augment cancer therapies. Mol Cancer Ther; 12(8); 1453–60. ©2013 AACR.