Istituto di Chimica Biomolecolare, Consiglio Nazionale Ricerche, 07100 Sassari, Italy.
Dipartimento di Scienze Chimiche, Università degli Studi di Padova, 35131 Padova, Italy.
Molecules. 2020 Jun 11;25(11):2709. doi: 10.3390/molecules25112709.
The impaired activity of tyrosinase and laccase can provoke serious concerns in the life cycles of mammals, insects and microorganisms. Investigation of inhibitors of these two enzymes may lead to the discovery of whitening agents, medicinal products, anti-browning substances and compounds for controlling harmful insects and bacteria. A small collection of novel reversible tyrosinase and laccase inhibitors with a phenylpropanoid and hydroxylated biphenyl core was prepared using naturally occurring compounds and their activity was measured by spectrophotometric and electrochemical assays. Biosensors based on tyrosinase and laccase enzymes were constructed and used to detect the type of protein-ligand interaction and half maximal inhibitory concentration (IC). Most of the inhibitors showed an IC in a range of 20-423 nM for tyrosinase and 23-2619 nM for laccase. Due to the safety concerns of conventional tyrosinase and laccase inhibitors, the viability of the new compounds was assayed on PC12 cells, four of which showed a viability of roughly 80% at 40 µM. In silico studies on the crystal structure of laccase enzyme identified a hydroxylated biphenyl bearing a prenylated chain as the lead structure, which activated strong and effective interactions at the active site of the enzyme. These data were confirmed by in vivo experiments performed on the insect model .
酪氨酸酶和漆酶活性的降低会引起哺乳动物、昆虫和微生物生命周期的严重问题。对这两种酶抑制剂的研究可能会发现美白剂、药物、抗褐变物质和控制有害昆虫和细菌的化合物。使用天然存在的化合物，制备了具有苯丙烷和羟基联苯核心的新型可逆酪氨酸酶和漆酶抑制剂的小集合，并通过分光光度法和电化学测定法测量了它们的活性。构建了基于酪氨酸酶和漆酶的生物传感器，用于检测蛋白质-配体相互作用的类型和半最大抑制浓度（IC）。大多数抑制剂对酪氨酸酶的 IC 在 20-423 nM 范围内，对漆酶的 IC 在 23-2619 nM 范围内。由于传统的酪氨酸酶和漆酶抑制剂存在安全性问题，因此在 PC12 细胞上对新化合物的生存能力进行了测定，其中有 4 种在 40 µM 时的生存能力约为 80%。对漆酶酶晶体结构的计算机研究确定了带有烯丙基链的羟基联苯作为先导结构，该结构在酶的活性部位产生了强烈有效的相互作用。这些数据通过在昆虫模型上进行的体内实验得到了证实。
