:: Lupeol (LP) and Mangiferin (MG) have beneficial effects on health, however, their pharmacokinetic properties can affect their bioavailability by oral administration, therefore, their incorporation in a hybrid matrix of ZnO and PLGA could contribute as a vehicle to improve bioavailability. Methods: Therefore, the objective of this study was to develop this matrix and evaluate its optical and bioactive properties obtained by the solvent emulsion and evaporation method, which were subjected to processes to evaluate their bioactivity as the effect of topoisomerase. Results: Functionalized treatment number 15 (TF15) showed the best results in studies of controlled release and encapsulation efficiency of lupeol (LP) and mangiferin (MG) (60.01 ± 1.24% and 57.71 ± 1.94%), the best treatment showed behaviors as a topoisomerase II inhibitor (18.60 ± 1.55). The nanoparticles developed in this study did not show a cytotoxic effect on BEAS-2B, while for HepG2 it showed a decrease in viability (IC50 1549.96 ± 174.62 µg/mL). On the other hand, although the hemolytic activity is not shown at 1 h of exposure, morphological alterations caused by TF15 are observed at concentrations of 2500 and 1250 µg/mL. Conclusion: In this context, the TF15 treatment shown by maintain its biological activity, does not present cytotoxicity for healthy cells, and decreases the growth of cancer cells.