# In Silico Studies and Biological Evaluation of Thiosemicarbazones as Cruzain-Targeting Trypanocidal Agents for Chagas Disease

## TL;DR

Scientists tested new compounds that inhibit a key enzyme in Chagas disease parasites, showing strong effectiveness and low toxicity.

## Contribution

The study identifies thiosemicarbazone derivatives as potent cruzain inhibitors with favorable drug properties for treating Chagas disease.

## Key findings

- Compounds H7, H10, and H11 inhibited cruzain with IC50 values below 0.5 µM.
- These compounds showed low cytotoxicity in human fibroblasts with CC50 > 64 µM.
- Structure–activity analysis revealed that aromatic systems and electron-donating groups enhance cruzain binding.

## Abstract

Background/Objectives: Chagas disease remains a major unmet medical need due to the limited efficacy and safety of current therapies. Here, we investigated sixteen thiosemicarbazone (TSC) derivatives as cruzain inhibitors using an integrated in silico/in vitro workflow. Methods: Docking against cruzain (PDB 3KKU) guided hit prioritization and correlated with enzyme inhibition; validation by redocking supported the protocol’s reliability. Results: The top compounds—H7, H10 and H11—showed potent cruzain inhibition (IC50 = 0.306, 0.512 and 0.412 µM, respectively) and low-micromolar trypanocidal activity, with negligible cytotoxicity in human fibroblasts (CC50 > 64 µM) and favorable selectivity. Structure–activity insights highlighted the role of expanded aromatic systems and electron-donating groups in enhancing binding within S2/S1′ subsites, while nitro substituents were associated with higher cytotoxicity. In silico ADMET parameters supported oral drug-likeness and acceptable metabolic liabilities. Conclusions: Overall, these data position TSCs as promising anti-T. cruzi leads and underscore the value of rational design against cruzain.