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 <i>in silico</i> ADMET predictions assessed stability, electronic properties, and safety. Compound <b>14</b> 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 <b>14</b> represents a promising dual EGFR/CA-IX inhibitor with selective anticancer activity. Further <i>in vivo</i> studies are warranted.