Sacha Inchi Oil Press-Cake (SIPC), a by-product of oil processing, was used to produce a protein concentrate (SPC) which was subsequently hydrolyzed by standardized static INFOGEST 2.0 protocol. This study aimed to integrate in vitro, ex vivo and in silico methods to evaluate the release of antioxidant peptides during gastrointestinal digestion. In vitro and ex vivo methods were used to investigate the antioxidant potential of SPC digests. Bioinformatics tools (find-pep-seq, AnOxPP, AnOxPePred-1.0, HLP, PlifePred and SwissADME) were employed to characterize antioxidant peptides. The gastric and intestinal digests exhibited higher ABTS and ORAC values than those exhibited by SPC. Under basal conditions, gastric digest fractions GD1, GD2, and GD3 (<3, 3-10, and >10 kDa, respectively), separated by ultrafiltration, significantly reduced ROS levels in RAW264.7 macrophages, while under LPS stimulation, GD1 (16 µg/mL) and GD2 (500 and 1000 µg/mL) reversed the induced damage. From the de novo peptidome determined, 416 peptides were selected based on their resistance to digestion. Through in silico tools, 315 resistant peptides were identified as antioxidants. Despite low bioavailability indices predicted, the peptides SVMGPYYNSK, EWGGGGCGGGGGVSSLR, RHWLPR, LQDWYDK, and ALEETNYELEK showed potential use for extracellular targets and drug delivery systems. This research contributes to ongoing efforts to valorize agro-industrial by-products, promoting a circular economy through the integration of sustainable practices in food production and related health applications.