Akut miyeloid lösemide Syk enziminin hedeflenmesi: BI 1002494'ün ın silico ilaç yeniden konumlandırımı ve işlevsel doğrulaması

Acute myeloid leukemia (AML) is a blood cancer type characterized by the uncontrolled growth and proliferation of myeloid cells in the bone marrow. Although methods such as radiotherapy are used for treatment, limited success rates necessitate other targeted treatment studies. The spleen tyrosine kinase (Syk) enzyme, known to be associated with AML, is an enzyme important in intracellular signaling. Disorders that may occur in Syk are highly effective in the AML emergence. Therefore, inhibitors focusing on Syk are promising for AML. In the study, in silico molecular docking was performed on a candidate molecules in the opnMe database, Syk inhibitor "BI 1002494", showing high binding affinity, was selected, and a targeted "drug repurposing" was performed for Syk. The catalytic domain of Syk was produced by recombinant methods using Escherichia coli bacteria, the catalytic domain was purified by His-tag Ni-NTA affinity chromatography method and the presence of the purified protein was confirmed by Western Blot (WB) method, and Thermal Shift Assay (TSA) experiments were performed to investigate the effects of the inhibitor on the protein at the molecular level. For the cellular activity, MTT cytotoxicity assays were performed on MOLM-13 and K562 cell lines and it was observed that BI 1002494 suppressed cell proliferation, and the acquired data were similar to the FDA-approved drug R406. The results indicate that BI 1002494 is a potential therapeutic Syk inhibitor against AML. This study provides valuable contributions to targeted, in silico therapeutic approaches in the drug repurposing context.