Synthesis and investigation of cytotoxic effects of some novel thiazolo [3,2-b] -1,2,4-triazole-6 (5h) -one derivatives

In this study, thirty new hybrid chalcone derivatives, namely 2-substituted-5-benzylidentiazolo[3,2-b]-1,2,4-triazol-6(5H)-one (4a-6j) which is expected to show cytotoxic activity were synthesized. Titled compounds were synthesized by the reaction of 3-substituted-1,2,4-triazole-5-thione derivatives (4-6), which were obtained from 3,4,5-trimethoxy benzoic acid, 3,4,5-trimethoxyphenyl acetic acid and 3-(3,4,5-trimethoxyphenyl)propionic acid, chloroacetic acid and various benzaldehydes (a-j) in acidic medium as a result of Claisen Schmitdh condensation reaction. The melting points of the synthesized compounds were determined and their structures were proved by IR, 1H- and 13C-NMR, mass spectroscopy and elemental analysis data. The effects of compounds on cell viability were evaluated by Sulforhodamine-B assay at 10 µM concentration in A549, P3, HCT116, MDA231 and Beas2B cell lines. Then the compound 6e (IC50: 30,4 µM; 72 hours) which has the best IC50 value among the others, has been further examined for its ATP viability test, M30 test, Annexin V test, mitochondrial membrane potential test and Caspase 3/7 test on A549 and Beas2B cell lines. As a result of these tests, it was found that compound 6e triggered apoptotic cell death and exhibited weak cytotoxic activity. Due to the weak cytotoxic activity of the compounds; the biochemical pathways that could show anticancer activity with cytostatic effect were screened by in silico methods and it was thought that the compounds might have EGFR inhibition activity. This theory was tested by Western blot test on A549 cell line with 50 µM and 100 µM concentrations of 6i and 6j compounds, and it was concluded that 6i and 6j compounds were potential EGFR inhibitors.