Hyperuricemia, caused by abnormal purine metabolism, is commonly treated with xanthine oxidase (XOD) inhibitors, uricosuric, and dietary adjustments. Recently, marine-derived bioactive peptides have gained attention as potential functional food ingredients due to their therapeutic potential. Takifugu obscurus, an economically significant offshore fish rich in crude proteins was explored in this study as a source of XOD inhibitory peptides. Enzyme hydrolysis combined with computer simulation identified TOH-A >
1 kDa and TOH-P >
1 kDa hydrolysates with high XOD inhibition rates, which were further selected for peptidomics characterization. After screening, seven peptides were synthesized, four of which (W11, DD7, WY7, and GA9) had inhibitory activity, with W11 showing the lowest IC