Objective To select potential ligands of ALS3 for drug development with anti-adhesion and/or anti-biofilm activities. Methodology ALS3 model was considered stable by DM. The main features of protein flexibility were represented by two conformers which were used in the virtual screening. Twenty-four small molecules were selected for in vitro assays. Five of them presented the best biological activity with ability to inhibit the adhesion and C. albicans biofilm formation on abiotic surface. Results To select potential ligands of ALS3 for drug development with anti-adhesion and/or anti-biofilm activities. Conclusion In silico tools application was able to select promising compounds with anti-adhesion activity, opening a new perspective of medical device treatment.