Methionine aminopeptidase-2 (MetAP2) is over-expressed in several cancers, including the cholangiocarcinoma (CCA). We reported previously suppressive effects of fumagillin, a MetAP2 inhibitor, on growth of CCA cell lines. In the present study, we evaluated the anti-proliferative and anti-invasive activities of TNP-470, a fumagillin analogue with higher MetAP2 inhibitory activity, on CCA cell lines (KKU-M213 and KKU-M214). TNP-470 significantly inhibited growth of both lines in a dose and time dependent fashion. Moreover, a sub-toxic dose of TNP-470 significantly reduced migration and invasion of CCA cells. Exploration of the molecular mechanisms by which TNP-470 inhibited growth and metastasis of CCA cell lines demonstrated expression of c-MYC, MMP2 and MMP9 to be decreased in TNP-470 treated cells. These results suggest that TNP-470 could be a potential therapeutic agent for CCA.