Trypanosoma cruzi carbonic anhydrase inhibitors as a potential antiparasitic agent.

Chagas disease remains a significant global health challenge, underscoring the need for safer and more effective treatments. Here, we identify carbonic anhydrase inhibition as a viable chemotherapeutic strategy against Trypanosoma cruzi. Two TcCA inhibitors, 1h (IC₅₀ = 6.98 μM; SI > 71.6) and 1j (IC₅₀ = 3.69 μM; SI > 135.5), demonstrated robust and selective activity in Dm28c-luciferase-infected Vero cells and retained efficacy against the T. cruzi Y strain in cardiac muscle cells. In 3D cardiac spheroids, both TcCA inhibitors showed low cytotoxicity and significantly reduced parasite burden, achieving 87% (1h) and 74% (1j) inhibition at 2 × IC₉₀ concentration, outperforming benznidazole (Bz; 69%) at an equivalent concentration (20 μM). Remarkably, 1j exhibited a trypanocidal effect comparable to high-dose Bz (100 μM), providing sustained long-term suppression of parasite resurgence. Moreover, combining 1j with Bz resulted in additive activity, indicating promising potential for combination therapy. Overall, these findings highlight 1j as a compelling TcCA-targeting lead with efficacy approaching that of Bz, supporting further exploration of carbonic anhydrase inhibition as an effective therapeutic strategy for Chagas disease.