Dymek Michał, Warszycki Dawid, Podlewska Sabina, Sikora Elżbieta
Faculty of Chemical Engineering and Technology, Cracow University of Technology, Warszawska 24, 31-155 Kraków, Poland.
Department of Medicinal Chemistry, Maj Institute of Pharmacology, Polish Academy of Sciences, Smętna 12, 31-343 Kraków, Poland.
Int J Mol Sci. 2024 Dec 17;25(24):13509. doi: 10.3390/ijms252413509.
Tyrosinase is a key enzyme responsible for the formation of melanin (a natural skin pigment with ultraviolet-protection properties). However, some people experience melanin overproduction, so new, safe, and biocompatible enzyme inhibitors are sought. New tripeptide tyrosinase inhibitors were developed using molecular modeling. A combinatorial library of tripeptides was prepared and docked to the mushroom tyrosinase crystal structure and investigated with molecular dynamics. Based on the results of calculations and expert knowledge, the three potentially most active peptides (CSF, CSN, CVL) were selected. Their in vitro properties were examined, and they achieved half-maximal inhibitory concentration (IC) values of 136.04, 177.74, and 261.79 µM, respectively. These compounds attach to the binding pocket of tyrosinase mainly through hydrogen bonds and salt bridges. Molecular dynamics simulations demonstrated the stability of the peptid-tyrosinase complexes and highlighted the persistence of key interactions throughout the simulation period. The ability of these peptides to complex copper ions was also confirmed. The CSF peptide showed the highest chelating activity with copper. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay confirmed that none of the test tripeptides showed cytotoxicity toward the reconstructed human epidermis. Our results indicated that the developed tripeptides were non-toxic and effective tyrosinase inhibitors. They could be applied as raw materials in skin-brightening or anti-aging cosmetic products.
酪氨酸酶是一种负责黑色素形成的关键酶（黑色素是一种具有紫外线防护特性的天然皮肤色素）。然而，一些人会出现黑色素过度生成的情况，因此人们在寻找新型、安全且具有生物相容性的酶抑制剂。利用分子建模开发了新的三肽酪氨酸酶抑制剂。制备了一个三肽组合文库，并将其与蘑菇酪氨酸酶晶体结构对接，然后用分子动力学进行研究。根据计算结果和专业知识，选择了三种潜在活性最高的肽（CSF、CSN、CVL）。检测了它们的体外性质，它们的半数抑制浓度（IC）值分别为136.04、177.74和261.79μM。这些化合物主要通过氢键和盐桥附着在酪氨酸酶的结合口袋上。分子动力学模拟证明了肽 - 酪氨酸酶复合物的稳定性，并突出了关键相互作用在整个模拟期间的持续性。还证实了这些肽络合铜离子的能力。CSF肽对铜的螯合活性最高。3 -（4,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基四氮唑溴盐（MTT）试验证实，所测试的三肽均未对重建的人表皮显示出细胞毒性。我们的结果表明，所开发的三肽是无毒且有效的酪氨酸酶抑制剂。它们可作为亮肤或抗衰老化妆品的原料。
