Activity of Ponatinib and Vitamin K2 Against Myelodysplastic Syndrome and Acute Myeloid Leukemia Cells

Acute myeloid leukemia (AML) is a heterogeneous hematologic malignancy often characterized by poor response to conventional therapies and frequent relapse. Ponatinib, a third-generation tyrosine kinase inhibitor, has demonstrated efficacy in various leukemias but remains underexplored in AML. Vitamin K2 (VK2), a fat-soluble vitamin, has recently been implicated in inducing apoptosis in leukemia cells. This study aimed to evaluate the therapeutic potential of ponatinib and VK2, alone and in combination, across five AML cell lines.

SKM-1 (myelodysplastic syndrome cell line) and AML cell lines MOLM-14, Kasumi-1, THP-1, and MV4-11 were tested. Cells were plated in 96-well plates and exposed to ponatinib or VK2, and cell viability and cytotoxicity were assessed. After 48 or 72 h of incubation, luminescence or absorbance was recorded, and cell numbers were quantified. Cells were plated in MethoCult medium supplemented with the indicated drug concentrations and incubated for 10 days at 37°C in a humidified 5% CO₂ atmosphere. Colonies containing more than 50 cells were enumerated, and representative images were captured. Caspase-3/7 activity was measured after 48 h of drug exposure, and luminescence signals were detected. Cells were stained with the JC-1 MitoMP Detection Kit and analyzed using a fluorescence microplate reader. Mitochondrial membrane potential was determined by calculating the ratio of red to green fluorescence intensity. Intracellular adenosine triphosphate levels were quantified.

Ponatinib exhibited cell line-dependent cytotoxic effects, with MOLM-14 and MV4-11 being the most sensitive. VK2 suppressed cell viability in all tested lines, with synergistic enhancement when combined with ponatinib. The combination treatment significantly increased apoptosis, reduced colony formation, and disrupted mitochondrial membrane potential.

The combined ponatinib and VK2 treatment synergistically impairs AML cell survival by enhancing apoptosis, suppressing clonogenic growth, and disrupting mitochondrial function. This dual targeting of oncogenic kinase signaling and metabolic integrity supports the ponatinib-VK2 combination as a promising therapeutic strategy for AML.