Candida albicans is an endogenous opportunistic yeast that does not cause infections in healthy individuals. However, disruption of the balance between microbiome and microorganism, the commensal form becomes pathogenic with clinical manifestation. The aim ofthis study was to determine the effect of oleuropein and hydroxytyrosol and their mechanism of action in C. albicans in vitro. Using the SQM approach for modulation ofergosterol synthesiswe found that oleuropein and hydroxytyrosol in concentrations below the MIC after 18 hours of incubation at 35 °C showed their interference in ergosterol biosynthesis pathway. The testing of the effect of hydroxytrosol and oleuropein on the inhibition of blastospore germination of C. albicans showed that after 3 hours of incubation at 37 °C oleuropein and hydroxytyrosol inhibit the germination even in values below MIC affecting the cellular signal pathways (cAMP-PKA). The testing of inhibition of growth of filaments reveals that oleuropein and hydroxytyrosol after 3 hours of incubation at 37 °C inhibit the growth of filaments of C. albicans germinated cells even in sub-MIC concentrations. Oleuropein and hydroxytyrosol are extremely interesting molecules for further research into mechanisms of antifungal activity, but also for possible synergistic effect with antimycotics. Therefore, further research into mechanisms of action on virulence factors is required.