Application of Computational Simulation Integrating Inhibition Kinetics for Detecting Tyrosinase Inhibitor: Salsalate Is a New Inhibitor.

Background: Tyrosinase inhibitor developments have been widely attended by investigators for their various applications.

Objective: A combination of virtual screening of docking simulations and biochemical inhibition kinetics was performed to find a new inhibitor of tyrosinase for the clinical application of an antipigment agent.

Methods: We conducted docking simulations to detect tyrosinase key binding residues and used the detected binding residues to screen the NCBI PubChem database for probing tyrosinase binding compounds. The serial inhibition kinetics and spectrofluorimetry measurements were performed to validate the inhibitory effect on tyrosinase.

Results: We have detected 200 candidates and categorized them into four clusters. Among them, we successfully confirmed salsalate as a new inhibitor of tyrosinase measured by serial enzyme kinetics. Salsalate was detected as a reversible inhibitor of tyrosinase displaying a typical mixedtype inhibition manner (IC = 22.19 ± 1.01 mM; K = 19.98 ± 2.11 mM). Spectrofluorimetry measurement by integrating with 1-anilinonaphthalene-8-sulfonate showed that salsalate mainly induced a slight regional conformation distortion of the tyrosinase active site accompanied by a slight hydrophobic disruption.

Conclusion: Our study suggests that salsalate is a potential anti-pigment drug via inhibition of tyrosinase activity and it might be applicable for dermatologic clinical application. Also, our study enlarges an insight into the salsalate drug application.