Establishing the Role of Thymoquinone as Potential Inhibitor of Mutant Oncoproteins in FLT3/RAS/RAF/MEK/ERK Pathway against Acute Myeloid Leukemia

Mutations in FLT3 prompt hyperactivation of RAS/RAF/MEK/ERK growth signalling. The FLT3-D835Y mutation confers resistance to FLT3 inhibitors for acute myeloid leukemia (AML) therapy. The KRAS-G12C and BRAF-V600E are frequent mutations in several cancers. While inhibitors against RAS, RAF, MEK, and ERK are available, they are often unsuccessful against BRAF or KRAS mutations with reported toxicity rates. This study evaluates the potential of thymoquinone (TQ), a phytochemical obtained from Nigella sativa seed, to function as an inhibitor of FLT3-D835Y, KRAS-G12C, BRAF-V600E, MEK, and ERK and to modify the expressions of genes related to RAS/RAF/MEK/ERK signaling in MV4-11 AML cells. The cells were incubated with TQ and we utilized RT-qPCR to measure the target genes’ mRNA levels. Molecular docking of TQ and reference inhibitors to FLT3-D835Y, KRAS-G12C, BRAF-V600E, MEK, and ERK proteins was examined with calculations of binding energies. TQ produced significantly downregulated K-RAS, B-RAF, MEK1, and ERK2 expressions. TQ also docked to FLT3-D835Y, KRAS-G12C, BRAF-V600E, MEK1, and ERK2 with high binding affinities and low docking scores. The study identifies TQ as an inhibitor of multiple target mutations that could combat resistance to FLT3-D835Y, KRAS-G12C, and BRAF-V600E inhibitors, aiding in the improvement of AML therapy.