Abstract: OBJECTIVE To screen potential ligands with anti-adhesion agglutinin-like sequence protein 3 (ALS3) from a candidate drug library; clarify the antibacterial and anti-biofilm activities of epigallocatechin gallate (EGCG) and preliminarily explore its metabolomic mechanism. METHODS Using ALS3 as the target protein, molecular docking virtual screening was employed to identify ALS3 inhibitors with potential resistance to biofilm formation. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of EGCG against Candida albicans were determined using the microbroth dilution method, and the time-growth curve was plotted. The XTT assay was used to explore the effects of EGCG on the formation and dispersion of mature biofilms of C. albicans. Differential metabolites were screened by combining metabolomic sequencing to preliminarily investigate the anti-biofilm mechanism of EGCG. RESULTS A total of 69 compounds were obtained through Glide tertiary screening, and EGCG, which scored excellently and demonstrated considerable binding mode, was identified. The MIC of EGCG was 256 μg/ml, and the MBC was 1,024 μg/ml, showing time-dose dependence. The concentration of EGCG that inhibited 50% of C. albicans biofilm formation was 256 μg/ml, and when the concentration reached 1,024 μg/ml, it could inhibit 80% of biofilm formation and disperse 40% of the biofilm. Non-targeted metabolomic results indicated that, compared with the control group, C. albicans treated with EGCG had a total of 505 differential metabolites (86 upregulated and 419 downregulated), primarily enriched in metabolic pathways such as cofactor biosynthesis, ABC transporters, amino acid biosynthesis, and carbohydrate metabolism. CONCLUSIONS It is feasible to use virtual screening methods to identify lead compounds against C. albicans biofilm with ALS3 as the target; EGCG exhibits certain antibacterial and anti-biofilm activities against C. albicans and has a dispersing effect on biofilms; EGCG disrupts multiple metabolic pathways, fundamentally undermining its energy supply, structural basis, and biosynthetic capacity, while weakening its defense and exposing its vulnerability.