PDK1-IN-5 is a selective inhibitor of PDK1 that reduces phosphorylation levels to activate PDH. This compound effectively reverses the Warburg effect by increasing acetyl-CoA, reducing lactate, elevating mitochondrial ROS, and subsequently inducing apoptosis, shifting cellular energy metabolism from glycolysis to oxidative phosphorylation. PDK1-IN-5 strongly inhibits tumor growth in vivo without causing systemic toxicity and can be used in research on lung adenocarcinoma, human non-small cell lung adenocarcinoma, gastric cancer, and colorectal cancer.

PDK1-IN-5 (compound D16), over a 48-hour period, exhibits selective and potent antiproliferative activity against A549 (lung adenocarcinoma), PC9 (lung adenocarcinoma), H1975 (human non-small cell lung adenocarcinoma), HGC-27 (gastric cancer), and HCT-116 (colorectal cancer) cells, with IC50 values of 0.86, 5.99, 4.92, 2.48, and 1.15 μM, respectively. PDK1-IN-5 demonstrates strong PDK1 inhibitory activity with an inhibition rate of 53.20%. At concentrations of 0-12 μM over 0.5 hours, PDK1-IN-5 decreases PDH phosphorylation by inhibiting PDK1 activity in A549 and PC9 cells, activating PDH. In the range of 0.4-1.6 μM over 14 days, it inhibits the proliferative capacity of A549 and PC9 cells in a concentration-dependent manner. Additionally, at 0.4-1.6 μM and 3-12 μM over 24 hours, it suppresses the migration of A549 and PC9 cells in a dose-dependent manner. PDK1-IN-5, at 0.2-3.2 μM and 1.5-24 μM over 48 hours, elevates mitochondrial ROS levels and induces concentration-dependent apoptosis. By effectively inhibiting PDK1 at 0.2-3.2 μM and 1.5-24 μM over 24 and 48 hours, PDK1-IN-5 reverses the Warburg effect, shifting tumor cell metabolism from glycolysis to oxidative phosphorylation, highlighting its therapeutic potential.

PDK1-IN-5 (compound D16) (5-20 mg/kg, intraperitoneal injection, every other day for 14 days) exhibits in vivo antitumor activity by inhibiting tumor proliferation in mice xenograft models of A549 and PC9.