Broad-Spectrum Trypanocidal Activity of the Natural Citrus Flavanone Glycosides Hesperidin and Neohesperidin Dihydrochalcone

Background and Objective: Hesperidin and neohesperidin are naturally occurring citrus flavanone glycosides that have a wide range of therapeutic uses. There is an increasing preference for natural products as chemotherapeutic agents. In this study, comprehensive research presented on the inhibitory effects of hesperidin and neohesperidin dihydrochalcone against 6 different species of Trypanosoma, as well as an analysis of their potential mechanism of action. Materials and Methods: The trypanocidal activity was evaluated using six trypanosome species cultivated in HMI-9 medium. Molecular docking procedures were conducted focusing on the Trypanosoma brucei Dihydrofolate Reductase (TbDHFR), with compound structures sourced from PubChem and optimized for physiological pH. Additionally, molecular dynamics (MD) simulations were carried out over 50 ns, assessing parameters such as RMSD, RMSF and hydrogen bonds to provide insights into the compound's stability and interactions. Data analysis utilized descriptive statistics to summarize the variations observed in the trypanocidal effects and molecular interactions. Results: The findings indicated that hesperidin lacked antitrypanosomal effects. On the other hand, neohesperidin dihydrochalcone showed significant trypanocidal activity across a wide range of Trypanosoma strains, including Trypanosoma brucei brucei GUT at 3.1, T. b. rhodesiense IL1501, T. b. gambiense IL1922, T. evansi Tansui, T. equiperdum IVM-t1 and T. congolense IL3000. The estimated IC50 values ranged from 8.88 to 22.53 mu g/mL, indicating low micromolar inhibition of Trypanosoma. Through docking and MD simulations, it is confirmed that neohesperidin dihydrochalcone binds to TbDHFR. The results of MD simulation support stable complex of neohesperidin dihydrochalcone with TbDHFR. Conclusion: The demonstrated efficacy of neohesperidin dihydrochalcone across various Trypanosoma species endorses its potential as a secure, natural agent against trypanosomiasis. It is strongly advised to further investigate the creation of analogues of neohesperidin dihydrochalcone and consider its application for the treatment of additional protozoan infections.