Antimicrobial　peptides　(AMPs)　that　exert　multiple　functions　are　considered　promising　candidates　to　combat　the　bacterial　drug　resistance　crisis.　Nowadays,　targeted　peptide　modification　has　been　widely　recognised　to　improve　biological　activity　and　make　up　for　deficiencies　in　clinical　applications　such　as　toxicity.　In　this　study,　a　helix-loop　peptide　was　isolated　and　identified　from　the　skin　secretion　of　the　Wuyi　torrent　frog　Amolops　wuyiensis,　namely,　ranatuerin-2-AW　(R2AW)　(GFMDTAKNVAKNVAATLLDKLKCKITGGC).　Target　modifications　were　made　to　R2AW　to　study　the　structure–activity　relationships　and　to　optimise　its　bioactivities.　Five　analogues　were　progressively　designed　via　residue　substitution　and　truncation　and　the　antibacterial　and　anticancer　activities　were　evaluated.　We　found　that　the　serine-substitution　and　cyclic-domain-deletion　products　showed　similar　antibacterial　activity　to　the　natural　peptide　R2AW,　implying　that　the　disulphide　bridge　and　Rana　box　were　dispensable　for　the　antibacterial　activity　of　ranatuerin-2　peptides.　Notably,　the　cationicity-　and　hydrophobicity-enhanced　variant,　[Lys4,19,　Leu20]R2AW(1-22)-NH2,　exhibited　significantly　optimised　antibacterial　and　anticancer　activities.　Additionally,　it　killed　bacteria　by　membrane　disruption　at　a　highly　efficient　rate.　Moreover,　[Lys4,19,　Leu20]R2AW(1-22)-NH2　exerted　potential　in　vivo　efficacy　in　a　methicillin-resistant　Staphylococcus　aureus　(MRSA)-infected　waxworm　model.　Overall,　this　study　demonstrated　some　rational　design　ideas　for　optimising　the　dual　antibacterial　and　anticancer　activities　of　ranatuerin-2　peptides　and　it　proposes　[Lys4,19,　Leu20]R2AW(1-22)-NH2　as　an　appealing　candidate　for　therapeutic　development.