Design, synthesis and biological screening of some diaryl acetamide derivatives as potential cytotoxic agents on HL-60(TB) cell line

In this study, novel diaryl acetamide derivatives were designed and evaluated as potential cytotoxic agents against the leukemic cells HL-60 (TB) with tyrosine kinase FMS-3(FLT-3) enzyme inhibitory activity. The designed molecules were synthesized and investigated for the FLT-3 enzyme inhibition. Compounds 5a, 15b, and 16b showed significant inhibitory activity against the FLT-3 enzyme. Assessment of cytotoxicity on HL-60 cells revealed that compound 15b exhibited superior activity than Quizartinib (AC220). Additionally, compounds 5a and 15b effectively arrested the cell cycle at the G₁ phase upon testing on HL-60 cells, suggesting potential abilities in blocking HL-60 cells' proliferation. The annexin-V stain assay demonstrated that compounds 5a, 15b, and 16b induced apoptosis in HL-60 cells. Furthermore, the ELISA assay showed that the blocking of cell cycle proliferation of HL-60 cells was mediated via the induction of cell cycle regulatory proteins p53 and p21. Meanwhile, the observed apoptosis induction was mediated by increasing apoptotic mediators, Bax/Bcl-2 ratio, and up-regulating caspase-3. Molecular docking studies revealed the binding affinities and interactions of the newly synthesized compounds, along with the reference compound Quizartinib, with the FLT-3 enzyme binding site.