The aim of this study was to investigate the antibacterial effect of fluoride varnish with antibacterial agents on Streptococcus mutans (S. mutans). Experimental fluoride varnish (EFV) with 5% NaF was fabricated. Five antibacterial agents were tested with 1 mM and 10 mM concentrations, respectively: Bakuchiol (BAK), Bavachalcone (BCC), Isobavachromene (IBC), and Bavachromene (BCM). MIC and MBC test was performed by antibacterial agents with and without EFV using 1.1 × 10¹⁰ CFU/ml of S. mutans. Cell viability test were performed using MTT test. Statistical analysis was done with ANOVA and Duncan's multiple range test at α=0.05. In the antibacterial agents without EFV, the MIC of BAK, BCC, and IBC was the lowest (0.015 mM), and the MBC of BAK was the lowest (0.031 mM). For the antibacterial agents with EFV, the MIC (0.031 mM) and MBC (0.062 mM) of BAK was the lowest, respectively. In MTT test, 1 mM BCM showed the highest cell viability (P < 0.05). This study suggest that BAK can be applied the fluoride varnish under the control of the concentration to develop antibacterial fluoride varnish.