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This study optimized ultrasound-assisted extraction conditions to maximize the extraction yield, total flavonoid content (TFC), total phenolic content (TPC), and DPP IV enzyme inhibitory activity from <i>Moringa oleifera</i>. The four UAE factors, solvent ratio (A), solvent–solid ratio (B), extraction temperature (C), and extraction time (D), were optimized using response surface methodology (RSM). A Box–Behnken design was used for the experimental design. The optimal conditions were found to be a 50% <i>v/v</i> solvent ratio, a 30% <i>v/w</i> solvent–solid ratio, 35 °C extraction temperature, and 45 min extraction time. The experimental value of extraction yield (<i>R</i>₁), TFC (<i>R</i>₂), TPC (<i>R</i>₃), and DPP IV enzyme inhibitory activity (<i>R</i>₄) (87.99% <i>w/w</i>, 56.63 mg QE/g extract, 97.26 mg GAE/g extract, and 93.32% inhibition, respectively) agreed with those predicted by RSM models (88.10% <i>w/w</i>, 56.61 mg QE/g extract, 97.16 mg GAE/g extract, and93.38% inhibition, respectively), thus demonstrating the appropriateness of the model used and the ability of the RSM to optimize the extraction conditions. Excellent DPP IV enzyme inhibitory activity was exhibited by <i>M. oleifera</i> compared with the standard, sitagliptin. While the modeled equation fits the data, the t-test is not significant, suggesting that the experimental values agree with those predicted by the RSM–BBD