Development of new anticancer thiadiazole-sulfonamides as dual EGFR/carbonic anhydrase inhibitors

Thiadiazole-sulfonamide derivatives were synthesized as dual inhibitors of epidermal growth factor receptor (EGFR) and carbonic anhydrase IX (CA-IX) to develop selective anticancer agents. Cytotoxicity was evaluated against MDA-MB-231 and MCF-7 breast cancer cells, with selectivity tested on Vero cells. Enzymatic inhibition studies were conducted against EGFR and CA-IX, using erlotinib and acetazolamide as reference drugs. Apoptosis was assessed through gene expression analysis of BAX/Bcl-2, caspase-8, and caspase-9, alongside flow cytometry for apoptosis and cell cycle analysis. Molecular docking and 200 ns molecular dynamics (MD) simulations evaluated binding interactions. Density Functional Theory (DFT) calculations and in silico ADMET predictions assessed stability, electronic properties, and safety. Compound 14 exhibited potent cytotoxicity (IC₅₀ = 5.78 μM, MDA-MB-231; 8.05 μM, MCF-7) and high selectivity (IC₅₀ = 313.08 μM, Vero). It inhibited EGFR (IC₅₀ = 5.92 nM) and CA-IX (IC₅₀ = 63 nM), surpassing reference drugs. Apoptosis induction was confirmed by a 13.97-fold increase in BAX/Bcl-2, caspase upregulation, and G1-phase arrest. Computational analyses confirmed stable binding and favorable safety. Compound 14 represents a promising dual EGFR/CA-IX inhibitor with selective anticancer activity. Further in vivo studies are warranted.