Introduction: Acne Vulgaris is a multifactorial disorder affecting various parts of the pilosebaceous unit and ranks second in the occurrence of skin diseases. The primary line of therapy for acne vulgaris involves antibiotics. However, prolonged and monotherapy use of antibiotics may lead to resistance. Therefore, an alternative approach is necessary for managing acne vulgaris, such as utilizing anti-acne compounds. Aims: The aim of this study is to evaluate the differences in the effectiveness of the anti-acne compounds niacinamide and melaleuca alternifolia in inhibiting the growth of Propionibacterium acnes, both as individual agents and when combined with other anti-acne compounds. Method: This research employs a true experimental design with a posttest-only control group design. The statistical analysis utilized is the One Way ANOVA test, with the diffusion disc method employed for testing. Results: The results of antibacterial activity testing indicate that both melaleuca alternifolia and niacinamide are effective in inhibiting the growth of Propionibacterium acnes. The One Way ANOVA analysis yields a p-value of 0.000, smaller than α = 0.005, signifying a significant difference in mean values among the sample groups. Conclusion: The anti-acne compounds niacinamide and melaleuca alternifolia exhibit antibacterial activity in inhibiting the growth of Propionibacterium acnes. Nevertheless, melaleuca alternifolia demonstrates higher effectiveness compared to niacinamide in inhibiting the growth of Propionibacterium acnes.