Dry Selection and Wet Evaluation of New 1,2-Disubstituted Nitroindazolin-3-One Derivatives as Promising Agents Against Trypanosoma cruzi.

Background/Objectives: Chagas disease is endemic to 21 Latin American countries and is a great public health problem. Current chemotherapy remains unsatisfactory; consequently, the need to search for new drugs persists. The aim of this work is to develop a machine learning computational model, which allows the identification of new chemical compounds with potential trypanosomicidal activity. Methods: A large dataset of 584 compounds, obtained from the Drugs for Neglected Diseases initiative, is used to develop the computational model. AlvaDesc v3.0.14 software is used to calculate the molecular descriptors, and Scikit-learn of Python to obtain the random forest. Results: The best random forest model shows accuracy of 82.1% for the training set and near to 79% for the test set, achieving specificity values over 84.9%, and the false alarm rate values were almost under 15% for both sets. As an experiment of virtual lead generation, the present model is finally satisfactorily applied to the virtual evaluation of a series of 1,2-disubstituted nitroindazolin-3-ones obtained a good agreement between the predicted activity and the experimental assays performed. Compounds 1c and 2c stood out as the most active in the series, with half-maximal inhibitory concentration (IC50) values of 71.3 and 40.9 μM, respectively. Mechanistic analyses suggest that the presence of the nitro group may promote the generation of reactive oxygen species through enzymatic redox activation, potentially involving T. cruzi nitroreductases (TcNTRs), leading to oxidative-stress-mediated parasite death. For the most active compounds, the data are consistent with intracellular hydroxyl radical generation through enzymatic redox processes. Conclusions: Even though none of them resulted more active than nifurtimox, the current results constitute a step forward in the search for efficient ways to discover new lead antitrypanosomals.