Inhibition kinetics and molecular simulation of p-substituted cinnamic acid derivatives on tyrosinase

This study was to investigate the inhibition effects of para-substituted cinnamic acid derivatives (4-chlorocinnamic acid, 4-ethoxycinnamic acid and 4-nitrocinnamic acid) on Tyrosinase catalyzing the substrates, with the purpose of elucidating the inhibition mechanism of the tested derivatives on Tyrosinase by the UV-vis spectrum, fluorescence spectroscopy, copper interacting and molecular docking, respectively. The native-PAGE results showed that 4-chlorocinnamic acid (4-CCA), 4-ethoxycinnamic acid (4-ECA) and 4-nitrocinnamic acid (4-NCA) had inhibitory effects on Tyrosinase. Spectrophotometric analysis used to determine the inhibition capabilities of these compounds on Tyrosinase catalyzing L-tyrosine (L-Tyr) and L-3,4-Dihydroxyphenylalanine (L-DOPA) as well. The IC₅₀ values and inhibition constants were further determined. Moreover, quenching mechanisms of tested compounds to Tyrosinase belonged to static type and a red shift on fluorescence emission peak occurred when 4-NCA added. Copper interacting and molecular docking demonstrated that 4-CCA could not bind directly to the copper, but it could interact with residues in the active center of Tyrosinase. Meanwhile, 4-ECA and 4-NCA could chelate a copper ion of Tyrosinase. Anti-tyrosinase activities of para-substituted cinnamic acid derivatives would lay scientific foundation for their utilization in designing of novel Tyrosinase inhibitors.

Derivatives of cinnamic acid; Fluorescence spectra; Molecular simulation.