Thiosemicarbazones are known for their biological and pharmacological activities. In this study, we have synthesized and characterized 3-Methoxybenzaldehyde thiosemicarbazone (3-MBTSc) and 4-Nitrobenzaldehyde thiosemicarbazone (4-NBTSc) using IR, ¹HNMR and ¹³C NMR. The compound’s in vitro anticancer activities against different cell lines were evaluated. Molecular docking, Insilco ADMET, and drug-likeness prediction were also done. The test compounds showed a comparative IC₅₀ and growth inhibition with the standard drug Doxorubicin. The IC₅₀ ranges from 2.82 µg/mL to 14.25 µg/mL in 3-MBTSc and 2.80 µg/mL to 7.59 µg/mL in 4-NBTSc treated cells. The MTT assay result revealed, 3-MBTSc inhibits 50.42 and 50.31 percent of cell growth in B16-F0 and EAC cell lines, respectively. The gene expression showed that tumor suppressor genes such as PTEN and BRCA1 are significantly upregulated in 7.42 and 5.33 folds, and oncogenes, PKC, and RAS are downregulated −7.96 and −7.64 folds, respectively in treated cells. The molecular docking performed on the four targeted proteins (PARP, VEGFR-1, TGF-β1, and BRAF^V600E) indicated that both 4-NBTSc and 3-MBTSc potentially bind to TGF-β1 with the best binding energy of −42.34 Kcal/mol and −32.13 Kcal/mol, respectively. In addition, the test compound possesses desirable ADMET and drug-likeness properties. Overall, both 3-MBTSc and 4-NBTSc have the potential to be multitargeting drug candidates for further study. Moreover, 3-MBTSc showed better activity than 4-NBTSc.

Background/Aim: Breast cancer is one of the world’s leading cause of deaths in women. This study evaluated the in-vitro anticancer activity of different thiosemicarbazones (HacTSc, HSTsc, 3-MBTSc, 4-NBTSc and 4-HBTSc) against MCF-7 human breast cancer cell line and MCF-10 normal cell. Materials and Methods: The ligands were prepared and characterized by UV vis, IR, ¹ H NMR. MTT assay was used to determine cell viability. Then data were analyzed using two-way ANOVA with Tukey post-hoc test. Result: The ligands have IC ₅₀ value ranging from 2.271µg/ml to 7.081µg/ml. Acetone thiosemicarbazone and 3-Methoxybenzaldehyde thiosemicarbazone, were identified as the most potent against MCF-7 breast cancer cells with IC ₅₀ value of 2.271µg/ml and 2.743µg/ml respectively. Whereas 4-Nitrobenzaldehyde thiosemicarbazone was the least potent. Also, the IC ₅₀ of the normal MCF-10 cell indicated their activity were selective. Conclusion: The activity of the ligands were dose, position and substituents dependent. Acetone thiosemicarbazone and 3-Methoxybenzaldehyde thiosemicarbazone are promising anticancer agents for further study.