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The red tree frog Litoria rubella from Australia has been studied for several decades showing that their dorsal skin glands secrete a number of small peptides containing a Pro-Trp sequence, known as tryptophyllin L peptides. Although peptides from many genera of Australian frogs have been reported to possess a variety of biological activities, the bioactivities of this peptide family have remained to be discovered. In this study, we investigated the antioxidant potency of a number of tryptophyllin L peptides for the first time using a joint statistical and experimental approach in which predictions based on Gaussian three-dimensional quantitative structure-activity relationship (3D-QSAR) models were employed to guide an in vitro experimental investigation. Two tryptophyllin tripeptides P-W-L (OH) and P-W-L (NH 2 ) were predicted to have the Trolox equivalent antioxidant capacity (TEAC) values of 0.80 and 0.87 μM Trolox/μM peptide, respectively. With those promising results, antioxidant capabilities of five tryptophyllin L peptides with the common core Pro-Trp-Leu were synthesized and subjected to 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing ability of plasma (FRAP) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical cation (ABTS˙ + ) radical scavenging assays. The tests indicated that all the tested tryptophyllin L peptides, noticeably S-P-W-L (OH) and F-P-W-L (NH 2 ), are strong ABTS˙ + radical scavengers and moderate scavengers in the other two assays. The results, thus, suggested that the tryptophyllin L peptides are likely to be a part of the skin antioxidant system helping the frog to cope with drastic change in oxygen exposure and humidity, as they inhabit over a large area of Australia with a wide climate variation., (© 2021 European Peptide Society and John Wiley & Sons, Ltd.)