Tyrosinase is an important rate-limiting enzyme in melanin biosynthesis. To find potential tyrosinase inhibitors with anti-melanogenic activity, a series of indole-thiazolidine-2,4-dione derivatives ~ were synthesized by incorporating indole with thiazolidine-2,4-dione into one compound and assayed for their biological activities. All compounds displayed tyrosinase inhibitory activities and had the highest anti-tyrosinase inhibitory activity with an IC value of 11.2 μM. Inhibition kinetics revealed as a mixed-type tyrosinase inhibitor. Fluorescence quenching results indicated that quenched tyrosinase fluorescence in a static process. CD spectra and 3D fluorescence spectra results suggested that the binding of with tyrosinase could change the conformation and microenvironment of tyrosinase. Molecular docking also represented the binding between and tyrosinase. Moreover, could inhibit tyrosinase activity and melanogenesis both in B16F10 cells and the zebrafish model. Therefore, compound could serve as a tyrosinase inhibitor with anti-melanogenic activity.