Functional Evaluation of Cooking-Mimicking Extracts From Chinese Olive (Canarium album L.) Leaves, Fruits, and Pits Using Cell-Based and In Silico Analysis

Chinese olive (CO; Canarium album L.) fruits are rich in phytochemicals with known physiological benefits. However, most studies evaluating the effects of CO have used ethyl acetate, acetone, and chloroform as the extraction solvents, which do not accurately reflect real dietary conditions. Moreover, research on CO leaves and pits remains limited. This study investigated the functional properties of extracts derived from CO leaves, fruits, and pits, prepared under simulated conventional culinary and dietary conditions (including solvents, extract combinations, and heat treatment), referred to as cooking-mimicking extracts. Our results revealed that most CO extracts reduced the production of nitric oxide in lipopolysaccharide-stimulated RAW264.7 macrophages and inhibited the proliferation of HCT116 colorectal cancer cells. Combinations of water-soluble CO extracts exerted a synergistic effect, suppressing cancer cell proliferation. To the best of our knowledge, this study is the first to identify methylellagic acid deoxyhexoside, vitexin 2-O-rhamnoside, and isovitexin 2-O-rhamnoside in CO extracts. Molecular docking predicted strong interactions with phosphofructokinase (PFK), a key glycolytic enzyme implicated in cancer cell proliferation. Treatment with CO extracts, including heat-treated forms, markedly reduced PFK activity and cell viability in HCT116 cells. These findings provide new insights into the bioactive constituents and physiological functions of CO extracts, supporting their potential application in dietotherapy.