Cutibacterium acnes (C. acnes, formerly classified as Propionibacterium acnes) is a Gram-positive bacterium that contributes to the development of acne vulgaris, resulting in inflammation and pustule formation on the skin. In this study, we developed and synthesized a series of antimicrobial peptides (AMPs) that are derived from the skin secretion of Rana chensinensis. Screening by MIC assay, Trp-containing peptides had potent anti-bacterial activity against C. acnes. The effect of Trp-containing peptides on the morphology was detected by scanning electron microscopy (SEM) and atomic force microscopy (AFM). The cell surface of C. acnes that was treated with Trp-containing peptides appeared rough and irregular. Further investigations into the peptides effects on cell membranes were conducted via zeta potential, Disc3(5) and Live/Dead assays. The results revealed that the AMPs bound to the cell surface, induced the disruption of the cell membranes and ultimately led to cell death. The leakage of cellular contents like nucleic acid, K and Ca also confirmed the membrane-damaging effect of Trp-containing peptides. In addition, Trp-containing peptides suppressed biofilm formation and disrupted mature biofilms of C. acnes. Among the Trp-containing peptides, I4WL5W with lowest cytotoxic effect displayed dual anti-bacterial and anti-inflammatory effects in acne mice; it decreased the bacterial load and inhibited the levels of TNF-α, IL-6 and IL-1β in the infected skin. This study demonstrated that Trp-containing peptides might be promising candidates for treating acne.