2种咖啡酸吗啉胺的合成及其对酪氨酸酶的激活和对黑色素合成的调控
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摘要
合成了2种新型咖啡酸衍生物咖啡酸-2-氨乙基吗啉胺(C-1)和咖啡酸-N-氨丙基吗啉胺(C-2),并通过质谱、核磁共振以及红外光谱技术鉴定其结构.2种化合物对蘑菇酪氨酸酶具有良好的激活效果,其对于酪氨酸酶活性的半激活质量浓度(EC_(50))分别为0.06和0.12 mmol/L,且2种化合物的激活类型均为混合型激活.通过紫外-可见光谱法验证了C-1和C-2对酪氨酸酶的激活作用.通过荧光猝灭和分子模拟进一步分析了2种化合物与酪氨酸酶之间的相互作用,结果表明两者对酪氨酸酶的影响可能是基于化合物的碳链长度.2种化合物对人体正常肝细胞LO2无毒性,并能够增强人体黑色素瘤细胞M14中的酪氨酸酶活力.进而对M14细胞中黑色素合成相关蛋白的研究发现,2种化合物对酪氨酸酶蛋白(TYR)、酪氨酸酶家族相关蛋白(TRP-1和TRP-2)以及α-促黑色素激素(α-MSH)的表达量均起上调作用.综上所述,该化合物有望提供一种有效治疗酪氨酸酶失调的新方法.
Two new caffeic acid derivatives,caffeic acid-2-aminoethyl morpholine amine( C-1) and caffeic acid-N-propyl ammonia morpholine amine( C-2) were synthesized and their structures were characterized by LC-MS,1 H-NMR and IR. The activation effects of these compounds on tyrosinase activities were evaluated. Compounds C-1 and C-2 showed potent activation effects,and the EC_(50) values were 0. 06 mmol/L and 0. 12 mmol/L on the tyrosinase activities respectively.Moreover,the activatory mechanisms were determined to be mixed activating type,and the activation effects were then verified using ultraviolet and visible spectrometry.Interactions of the compounds with tyrosinase were further analyzed using fluorescence quenching and molecular simulation assays.The results indicated that their effects on the tyrosinase activity maybe related to the length of carbon chain of the compounds. In addition,the compounds showed no cytotoxicity on human hepatic LO2 cells,and treatment of the human M14 melanoma cells with two compounds increased intracellular tyrosinase activity.The expression levels of TYR,TRP-1,TRP-2 and α-MSH protein were up-regulated in a dose-dependent manner by compounds treatment.These results suggested that compounds C-1 and C-2 might represent a novel approach for an effective therapy for tyrosinase failure diseases.
Two new caffeic acid derivatives,caffeic acid-2-aminoethyl morpholine amine( C-1) and caffeic acid-N-propyl ammonia morpholine amine( C-2) were synthesized and their structures were characterized by LC-MS,1 H-NMR and IR. The activation effects of these compounds on tyrosinase activities were evaluated. Compounds C-1 and C-2 showed potent activation effects,and the EC_(50) values were 0. 06 mmol/L and 0. 12 mmol/L on the tyrosinase activities respectively.Moreover,the activatory mechanisms were determined to be mixed activating type,and the activation effects were then verified using ultraviolet and visible spectrometry.Interactions of the compounds with tyrosinase were further analyzed using fluorescence quenching and molecular simulation assays.The results indicated that their effects on the tyrosinase activity maybe related to the length of carbon chain of the compounds. In addition,the compounds showed no cytotoxicity on human hepatic LO2 cells,and treatment of the human M14 melanoma cells with two compounds increased intracellular tyrosinase activity.The expression levels of TYR,TRP-1,TRP-2 and α-MSH protein were up-regulated in a dose-dependent manner by compounds treatment.These results suggested that compounds C-1 and C-2 might represent a novel approach for an effective therapy for tyrosinase failure diseases.
引文
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[13]邓轶韬,丁玉梅,李华亮,等.鳄胆素联合阿霉素对人肝癌细胞SMMC-7721的抑制作用[J].厦门大学学报(自然科学版),2015,54(6):796-801.
[14]CUI T T,YI X L,ZHANG W G,et al.miR-196a-2rs11614913 polymorphism is associated with vitiligo by affecting heterodimeric molecular complexes of Tyr and Tyrp1[J].Arch Dermatol Res,2015,307(8):683-692.
[2]刘晓丹,黄磺,陈清西.苯甲酸对蘑菇酪氨酸酶抑制作用机理的研究[J].厦门大学学报(自然科学版),2003,42(1):102-107.
[3]DENG X Q,GONG S Z,ZHANG M Q.Caffeic acid activation of tyrosine enzyme catalytic reaction dynamics research[J].J Food Science,2008,29:98-101.
[4]FANG D,KUTE T,SETALURI V.Regulation of tyrosinaserelated protein-2(TYRP2)in human melanocytes:relationship to growth and morphology[J].Pigment Cell Res,2001,14:132-139.
[5]NERYA O,VAYA J,MUSA R,et al.Glabrene and isoliquiritigenin as tyrosinase inhibitors from licorice roots[J].J Agr Food Chem,2003,51(5):1201-1207.
[6]HEARING V J,TSUKAMOTO K.Enzymatic control of pigmentation in mammals[J].FASEB J,1991,5(14):2902-2909.
[7]GRIMES P E,HAMZAVI I,LEBWOHL M,et al.The efficacy of afamelanotide and narrowband UV-B phototherapy for repigmentation of vitiligo[J].JAMA Dermatol,2013,149(1):68-73.
[8]YE Y,CHOU G X,WANG H,et al.Flavonoids,apigenin and icariin exert potent melanogenic activities in murine B16melanoma cells[J].Phytomedicine,2010,18(1):32-35.
[9]MICHALUART P,MASFERRER J L,CAROTHERS A M,et al.Inhibitory effects of caffeic acid phenethyl ester on the activity and expression of cyclooxygenase-2 in human oral epithelial cells and in a rat model of inflammation[J].Cancer Res,1999,59(10):2347-2352.
[10]TOLBA M F,OMAR H A,AZAB S S,et al.Caffeic acid phenethyl ester:a review of its antioxidant activity,protective effects against ischemia-reperfusion injury and drug adverse reactions[J].Crit Rev Food Sci Nutr,2016,56(13):1549-7852.
[11]CHEN Q X,SONG K K,WANG Q,et al.Inhibitory effects on mushroom tyrosinase by some alkylbenzaldehydes[J].Journal of Enzyme Inhibition and Medicinal Chemistry,2003,18(6):491-496.
[12]KIM Y J,UYAMA H.Tyrosinase inhibitors from natural and synthetic sources:structure,inhibition mechanism and perspective for the future[J].Cell Mol Life Sci,2005,62:1707-1723.
[13]邓轶韬,丁玉梅,李华亮,等.鳄胆素联合阿霉素对人肝癌细胞SMMC-7721的抑制作用[J].厦门大学学报(自然科学版),2015,54(6):796-801.
[14]CUI T T,YI X L,ZHANG W G,et al.miR-196a-2rs11614913 polymorphism is associated with vitiligo by affecting heterodimeric molecular complexes of Tyr and Tyrp1[J].Arch Dermatol Res,2015,307(8):683-692.