Development of highly potent melanogenesis inhibitor by in vitro, in vivo and computational studies.
Development of highly potent melanogenesis inhibitor by in vitro, in vivo and computational studies.
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DOI：
10.2147/dddt.s137550
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Seo SY
中科院分区：
文献类型：
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作者：
Abbas Q;Ashraf Z;Hassan M;Nadeem H;Latif M;Afzal S;Seo SY
本工作描述了作为黑素生成抑制剂的几个羟基化酰胺衍生物的合成。体外、体内和计算研究证明，与标准曲酸相比，化合物6d是高度有效的黑素生成抑制剂。标题酰胺4a-e和6a-e的合成路线简单，产率高。大多数合成的化合物显示出良好的蘑菇酪氨酸酶抑制活性，但化合物6d显示出优异的活性（IC 50 0.15 μM），与标准曲酸（IC 50 16.69 μM）相比。采用Lineweaver-Burk作图法确定了化合物4c和6d的动力学机理，发现化合物4c和6d表现为非竞争性抑制，而化合物6a和6 b表现为混合型抑制。动力学机制进一步揭示了化合物6d与酪氨酸酶形成不可逆复合物。化合物4c、6a、6 b和6d的Ki值分别为0.188、0.84、2.20和0.217 µM。在A375人黑色素瘤细胞中的人酪氨酸酶抑制活性的结果显示，化合物6d在50 μg/mL的浓度下显示91.9%的抑制活性。化合物6d在斑马鱼胚胎中的体内细胞毒性评价显示其对斑马鱼无毒性。在斑马鱼中进行的黑色素脱色试验表明，与相同浓度的曲酸相比，化合物6d具有更大的降低黑色素含量的潜力。计算研究也支持湿实验室的发现，因为化合物6d显示出与靶蛋白（PDBID：2 Y 9 X）的最高结合亲和力，结合能值为−7.90 kcal/mol。分子动力学模拟研究也证明酰胺6d与酪氨酸酶形成最稳定的复合物。基于我们的体外、体内和计算机研究，我们提出化合物6d是一个有希望的候选者，用于开发安全的美容剂。
The present work describes the synthesis of few hydroxylated amide derivatives as melanogenesis inhibitors. In vitro, in vivo and computational studies proved that compound 6d is a highly potent melanogenesis inhibitor compared to standard kojic acid. The title amides 4a–e and 6a–e were synthesized following simple reaction routes with excellent yields. Most of the synthesized compounds exhibited good mushroom tyrosinase inhibitory activity, but compound 6d showed excellent activity (IC50 0.15 µM) compared to standard kojic acid (IC50 16.69 µM). Lineweaver–Burk plots were used for the determination of kinetic mechanism, and it was found that compounds 4c and 6d showed non-competitive inhibition while 6a and 6b showed mixed-type inhibition. The kinetic mechanism further revealed that compound 6d formed irreversible complex with the target enzyme tyrosinase. The Ki values determined for compounds 4c, 6a, 6b and 6d are 0.188, 0.84, 2.20 and 0.217 µM respectively. Results of human tyrosinase inhibitory activity in A375 human melanoma cells showed that compound 6d exhibited 91.9% inhibi-tory activity at a concentration of 50 µg/mL. In vivo cytotoxicity evaluation of compound 6d in zebrafish embryos showed that it is non-toxic to zebrafish. Melanin depigmentation assay performed in zebrafish indicated that compound 6d possessed greater potential in decreasing melanin contents compared to kojic acid at the same concentration. Computational studies also supported the wet lab findings as compound 6d showed a highest binding affinity with the target protein (PDBID: 2Y9X) with a binding energy value of −7.90 kcal/mol. Molecular dynamic simulation studies also proved that amide 6d formed the most stable complex with tyrosinase. Based upon our in vitro, in vivo and computational studies, we propose that compound 6d is a promising candidate for the development of safe cosmetic agent.