MMP inhibitor(Paratek Pharmaceuticals)

Colorectal cancer (CRC) progression is often driven by hypoxia-induced signaling that promotes metastasis through epithelial-mesenchymal transition and invadopodia formation. PX-12, an irreversible thioredoxin-1 (Trx-1) inhibitor, has shown anticancer potential, but its anti-invasive effects under hypoxic conditions remain unclear. This study investigated the cytotoxic, anti-migratory, anti-invasive, and anti-invadopodia activities of PX-12 in hypoxia-induced CRC cells. Human HCT116 cells were exposed to dimethyloxalylglycine (DMOG) to induce hypoxia. Cytotoxicity was assessed using MTT assays, while migration and invasion were evaluated through wound-healing and Transwell assays. Invadopodia formation was quantified using Oregon Green-labeled gelatin degradation assays. Protein expression of HIF-1α and Trx-1 was analyzed via Jess Simple Western system. PX-12 significantly reduced HCT116 cell viability in a dose-dependent manner, with lower cytotoxicity under DMOG-induced hypoxia. PX-12 markedly suppressed cell migration and invasion, particularly under hypoxia-like conditions. Gelatin degradation analysis revealed a significant reduction in invadopodia formation with PX-12 treatment, paralleling effects of the MMP inhibitor GM6001. Although DMOG markedly upregulated HIF-1α expression, PX-12 did not alter Trx-1 protein levels, suggesting that its inhibitory effects may occurs via enzymatic blockade rather than changes in protein expression. PX-12 exerts potent anti-invasive effects by inhibiting migration, invasion, and invadopodia formation in hypoxia-mimicking CRC cells. These findings highlight PX-12 as therapeutic agent targeting hypoxia-driven migratory and invasive behaviors in CRC.