Abstract: Eight puerarin derivatives(PD₁-PD₈) were designed and synthesized from puerarin, and their chemical structures were determined by mass spectrometry and nuclear magnetic resonance hydrogen spectroscopy. The activation activity of each derivative on tyrosinase was measured using the method of tyrosinase dopa rate oxidation. Molecular docking simulations were utilized to explore the binding mechanism of each derivative with tyrosinase. Enzyme kinetic analysis was performed to determine the type of activation. The structures of 8 puerarin derivatives were identified as: 4'-methoxy-puerarin(PD₁), 4'-ethoxy-puerarin(PD₂), 4'-propoxyl-puerarin(PD₃), 7-O-acetyl-puerarin(PD₄), 7-O-trimethylacetyl-puerarin(PD₅), 7-methoxy-puerarin(PD₆), 7-O-ethoxycarbonylmethylene-puerarin(PD₇), 7, 4'-O-diethoxycarbonylmethylene-puerarin(PD₈). Enzyme kinetic results showed that PD₆ were mixed activation types. The results of activation activity assay showed that PD₆ was mixed activation, and the half activation concentration of PD₆(EC₅₀=0.416 mmol/L) was higher than that of puerarin(EC₅₀=0.434 mmol/L).