Novel 3-sulfonamide pyrrol-2-one derivatives containing two sulfonamide groups were synthesized via a one-pot, three-component method using trifluoroacetic acid as a catalyst. Structural confirmation was achieved using spectroscopic techniques. The compounds were tested against four selected human carbonic anhydrase isoforms (hCA I, hCA II, hCA IX, and hCA XII). Most derivatives showed significant selectivity for hCA II, with 4h, 4i, 4n, 4k, and 4j demonstrating enhanced activity due to methoxy and hydroxy group patterns. Compound 4o exhibited strong dual selectivity for hCA II and hCA IX, while 4l was the most effective inhibitor of hCA XII. Additionally, 4e showed a preference for hCA XII inhibition. Biological evaluation on MeWo, SK-BR-3, and MG-63 cancer cells showed that compound 4l was cytotoxic for MeWo cells without significantly affecting normal fibroblasts' viability. Compounds 4e, 4l, and 4o were shown to affect tumor cell viability in combination with doxorubicin in additional testing on MeWo cancer cells.