This study aimed to obtain tyrosinase inhibitors for treating hyperpigmentation. A series of cinnamyl ester analogues were designed and synthesized with cinnamic acid (CA) and peaonol compounds. The safety, melanin content and inhibitory effects of all target compounds were evaluated. In the enzymatic activity test, the inhibitory rate of compounds 8, 13 and 14 had stronger inhibitory activity with the IC₅₀ values ​​of 20.7 μM, 13.98 μM and 15.16 μM, respectively than the positive drug kojic acid (IC₅₀ with 30.83 μM). The cytotoxicity evaluation showed that compounds 13 and 14 have higher safety than the other compounds to the proliferation of B16F10 cells. The result of the melanocyte test supported that compound13 has stronger cellular tyrosinase inhibitory activity than kojic acid and arbutin at 100 μM and 200 μM. The enzyme kinetics mechanism revealed that compound 13 was a non-competitive inhibitor while compounds 8 and 14 were mixed inhibitors. For the experiments of melanin content and tyrosinase activity in the B16F10 melanona cells, the inhibition rates of compounds 8, 14 and 13 were with 19.62%, 20.59% and 23.83%, respectively. In addition, compound 13 revealed the highest inhibitory activity to tyrosinase in the melanocyte with inhibition rates of 23.83%, which was better than kojic acid and arbutin (19.21% and 20.45%) at the same concentration. In the anti-melanogenesis experiment, compounds 8 and 13 had better anti-melanin effects than kojic acid from 25 μM to 100 μM. In summary, the results indicated that compounds 8, 13 and 14 had better tyrosinase inhibitory activity and anti-melanogenesis activity. Especially, the compound 13 has potentiality to develop novel tyrosinase inhibitors and whitening agents. The docking studies results revealed that the functional group of compound 13 mostly depends on the phenolic hydroxyl moiety, and its hydroxyl group did not insert into the active site of tyrosinase, which was in agreement with the results of the kinetics study.

本研究旨在获得用于治疗色素沉着过度的酪氨酸酶抑制剂。以肉桂酸（CA）和牡丹醇化合物为原料设计并合成了一系列肉桂酯类似物。评估了所有目标化合物的安全性、黑色素含量和抑制作用。在酶活性试验中，化合物的抑制率8，13和14具有与IC更强的抑制活性₅₀，分别比20.7μM的值，13.98μM和15.16μM阳性药曲酸（IC ₅₀30.83 μM）。细胞毒性评价表明，化合物13和14对B16F10细胞增殖的安全性高于其他化合物。黑素细胞试验结果表明，化合物13在100 μM和200 μM时比曲酸和熊果苷具有更强的细胞酪氨酸酶抑制活性。酶动力学机制表明，化合物13是一种非竞争性抑制剂，而化合物8和14是混合抑制剂。B16F10黑色素细胞黑色素含量和酪氨酸酶活性实验中，化合物8、14和13的抑制率分别为19.62%、20.59%和23.83%。此外，化合物13对黑素细胞中酪氨酸酶的抑制活性最高，抑制率为23.83%，优于曲酸和熊果苷（19.21%和20.5%）。45%) 相同的浓度。在抗黑色素生成实验中，化合物8和13在25 μM至100 μM范围内具有比曲酸更好的抗黑色素作用。综上所述，结果表明化合物8、13和14具有较好的酪氨酸酶抑制活性和抗黑色素生成活性。特别是化合物13具有开发新型酪氨酸酶抑制剂和增白剂的潜力。对接研究结果表明，化合物13的官能团主要依赖于酚羟基部分，其羟基没有插入酪氨酸酶的活性位点，这与动力学研究结果一致。13和14具有较好的酪氨酸酶抑制活性和抗黑色素生成活性。特别是化合物13具有开发新型酪氨酸酶抑制剂和增白剂的潜力。对接研究结果表明，化合物13的官能团主要依赖于酚羟基部分，其羟基没有插入酪氨酸酶的活性位点，这与动力学研究结果一致。13和14具有较好的酪氨酸酶抑制活性和抗黑色素生成活性。特别是化合物13具有开发新型酪氨酸酶抑制剂和增白剂的潜力。对接研究结果表明，化合物13的官能团主要依赖于酚羟基部分，其羟基没有插入酪氨酸酶的活性位点，这与动力学研究结果一致。