PKM2-IN-13 is a selective inhibitor of PKM2 with an IC50 value of 55.13 μM. This compound demonstrates broad-spectrum anticancer activity while exhibiting low toxicity to normal cells. PKM2-IN-13 induces apoptosis by elevating reactive oxygen species (ROS) levels and activating Caspase-3/7. It interacts with pyruvate kinase M2 in vitro, inhibiting its glycolytic function. Additionally, PKM2-IN-13 has shown a good safety profile, with no significant adverse effects observed in in vivo experiments. It can be utilized in research studies involving oral squamous cell carcinoma, colon cancer, breast cancer, and melanoma.

PKM2:55.13 μM

PKM2-IN-13 (compound 7f), after 48 hours, induces approximately 5% hemolytic activity and effectively promotes cytotoxicity (IC 50 = 12.29 μM) in SCC-9 cells. It exhibits selective inhibitory activity against various cancer cell lines, including oral squamous carcinoma cells SCC-9 (IC 50 = 12.29 μM; SI = 1.85), SCC-4 (IC 50 = 7.93 μM), SCC-25 (IC 50 = 11.09 μM), primary gingival fibroblasts (IC 50 = 22.77 μM), and other cancer lines such as 4T1 (IC 50 = 7.63 μM), Hep-G2 (IC 50 = 15.98 μM), HCT116 (IC 50 = 6.72 μM), and B16−F10 (IC 50 = 4.93 μM). With a concentration of 24.58 μM over 0-48 hours, PKM2-IN-13 shows significant proliferation inhibition in SCC-9 cells, with disruption of cell junctions, membrane blebbing, and subsequent proliferation arrest observed as early as 6 hours after treatment, becoming more pronounced by 12 hours. At the same concentration and time frame, PKM2-IN-13 induces S-phase arrest at 24 hours in SCC-9 cells, leading to cancer cell death through an intrinsic caspase-dependent apoptotic pathway, rather than necrotic apoptosis, within 12-24 hours. The treatment also causes a significant increase in reactive oxygen species levels at 12 and 24 hours. The cytotoxic effects induced by PKM2-IN-13 (24.58 μM, 48 hours) can be mitigated by pretreatment with the antioxidant N-acetyl-L-cysteine (NAC) (10 mM) for 2 hours, suggesting a mechanism related to oxidative stress.

Administered intraperitoneally at a dosage of 100-400 mg/kg over 14 days, PKM2-IN-13 demonstrates good safety and low toxicity in C57BL/6 mice.