The present study was designed to perform structural modifications of of neobavaisoflavone(NBIF), using an in vitro enzymatic glycosylation reaction, in order to improve its water-solubility. Two novel glucosides of N...The present study was designed to perform structural modifications of of neobavaisoflavone(NBIF), using an in vitro enzymatic glycosylation reaction, in order to improve its water-solubility. Two novel glucosides of NBIF were obtained from an enzymatic glycosylation by UDP-glycosyltransferase. The glycosylated products were elucidated by LC-MS, HR-ESI-MS, and NMR analysis. The HPLC peaks were integrated and the concentrations in sample solutions were calculated. The MTT assay was used to detect the cytotoxic activity of compounds in cancer cell lines. Based on the spectroscopic analyses, the two novel glucosides were identified as neobavaisoflavone-4′-O-β-D-glucopyranoside(1) and neobavaisoflavone-4′, 7-di-O-β-D-glucopyranoside(2). Additionally, the water-solubilities of compounds 1 and 2 were approximately 175.1-and 4 031.9-fold higher than that of the substrate, respectively. Among the test compounds, only NBIF exhibited weak cytotoxicity against four human cancer cell lines, with IC50 values ranging from 63.47 to 72.81 μmol·L^(-1). These results suggest that in vitro enzymatic glycosylation is a powerful approach to structural modification, improving water-solubility.展开更多
新补骨脂异黄酮为中药补骨脂中的黄酮类成分,具有抗菌、抗炎、抗癌、抗骨质疏松等多种药理作用。该文研究了新补骨脂异黄酮在Caco-2细胞模型中的吸收机制。采用Thermo Syncronis C18柱,流动相为甲醇-0.1%甲酸水溶液(90∶10),流速0.2 m L...新补骨脂异黄酮为中药补骨脂中的黄酮类成分,具有抗菌、抗炎、抗癌、抗骨质疏松等多种药理作用。该文研究了新补骨脂异黄酮在Caco-2细胞模型中的吸收机制。采用Thermo Syncronis C18柱,流动相为甲醇-0.1%甲酸水溶液(90∶10),流速0.2 m L·min^(-1),电喷雾电离正离子模式(ESI+),以柳胺酚为内标物质建立新补骨脂异黄酮在Caco-2细胞孵育液中浓度的检测方法,考察时间、浓度、P-gp蛋白抑制剂维拉帕米、MRP-2蛋白抑制剂MK-571和BCRP蛋白抑制剂Ko143对新补骨脂异黄酮吸收的影响。研究结果表明,新补骨脂异黄酮在10~2 000μg·L^(-1)具有良好的线性,且专属性、基质效应、提取回收率、精密度、准确度和稳定性均符合测定要求。新补骨脂异黄酮在Caco-2细胞模型中的转运与时间和浓度呈正相关。15,30,50μmol·^(L-1)的新补骨脂异黄酮ER分别为1.64,1.94,0.99。与对照组相比,盐酸维拉帕米,MK-571,Ko143均可促进新补骨脂异黄酮的转运,其中盐酸维拉帕米和Ko143的作用较为显著。新补骨脂异黄酮在Caco-2细胞模型中的吸收以主动转运为主,同时存在被动转运,可能存在肠道转运蛋白的外排机制。展开更多
基金financially supported by National Natural Science Foundation of China(No.81302671)the Education Department of Anhui Natural Science Research Project China(No.KJ2015A273)Zhejiang Provincial Natural Science Foundation of China(No.LY13H10004)
文摘The present study was designed to perform structural modifications of of neobavaisoflavone(NBIF), using an in vitro enzymatic glycosylation reaction, in order to improve its water-solubility. Two novel glucosides of NBIF were obtained from an enzymatic glycosylation by UDP-glycosyltransferase. The glycosylated products were elucidated by LC-MS, HR-ESI-MS, and NMR analysis. The HPLC peaks were integrated and the concentrations in sample solutions were calculated. The MTT assay was used to detect the cytotoxic activity of compounds in cancer cell lines. Based on the spectroscopic analyses, the two novel glucosides were identified as neobavaisoflavone-4′-O-β-D-glucopyranoside(1) and neobavaisoflavone-4′, 7-di-O-β-D-glucopyranoside(2). Additionally, the water-solubilities of compounds 1 and 2 were approximately 175.1-and 4 031.9-fold higher than that of the substrate, respectively. Among the test compounds, only NBIF exhibited weak cytotoxicity against four human cancer cell lines, with IC50 values ranging from 63.47 to 72.81 μmol·L^(-1). These results suggest that in vitro enzymatic glycosylation is a powerful approach to structural modification, improving water-solubility.
