采用光谱法研究几种抗肿瘤药物与蛋白质的相互作用

发布时间：2018-06-29 10:35

本文选题：光谱分析法 + 化学计量学方法　；参考：《西华师范大学》2015年硕士论文

【摘要】：青蒿素、双氢青蒿素、斑螫素、去甲斑螫素、新橙皮苷和柚皮苷等化合物均具备生物活性和药理功效。青蒿素和双氢青蒿素具有出色的抗疟活性,成为最重要的抗疟药物,此外它们还具有抗肿瘤、抗癌等作用;斑螫素和去甲斑螫素作为抗肿瘤药物用于治疗食道癌、肝癌、胃癌等,并且在许多细胞系中去甲斑螫素的毒素比斑螫素的少,所以它能够提供更多的临床应用;新橙皮苷具备抗增值、抗过敏、抗氧化、抗雌激素、抗肿瘤、清除自由基的活性和对胃病的保护作用等药理效应,而柚皮苷具有抗氧化、抗高胆固醇、抗炎、降血糖、抗凋亡及心脏保护等多种功效,成为新型的具有应用价值的药物。本文主要探讨这些具有抗肿瘤、抗癌等作用的药物与蛋白质间的相互作用,这些研究有助于理解该类药物在体内的运输、药理学作用信息、代谢动力学及为合成高活性该类药物类似物提供前期理论基础研究。具体研究内容如下:在模拟生理条件下,通过荧光、UV-vis吸收和红外光谱分别探究BSA与双氢青蒿素(DHA)和青蒿素(ART)相互作用。通过荧光发射和瑞利散射光谱分析DHA/ART与BSA的结合特性。由分析结果可知,BSA与DHA/ART间的猝灭机理属于静态猝灭。在四个不同温度下,分别计算DHA/ART-BSA体系的结合常数和绑定位点数。依据非辐射能量转移理论,计算获得BSA与DHA/ART间的绑定距离为1.54/1.65 nm。使用UV-vis吸收、同步荧光和三维荧光光谱讨论这两种药物对BSA结构的影响。此外,在DHA/ART存在下,运用红外光谱实验分析BSA特征吸收峰的峰位和强度变化,而且还采用模拟方法计算它们对应的红外光谱数据,得到BSA二级结构含量发生变化,即它们使该蛋白质的α-螺旋含量明显降低、β-片层和无规卷曲含量增加。最后,讨论共存离子对DHA/ART-BSA体系结合常数的影响。采用上述相同的方法分别研究斑蝥素/去甲斑蝥素(CTD/NCTD)与两种蛋白质(BSA和BHb)间的相互作用。研究结果表明这两种蛋白质分子上的色氨酸残基(Trp)微环境发生改变、较少的肽链展开和它们的疏水性随这两种药物浓度增大而增加。使用多元曲线分辨-交替最小二乘(MCR-ALS)方法进一步研究这四个绑定体系不同组份的浓度分布和对应的纯光谱。结果表明,游离的CTD/NCTD和BSA/BHb的拟合光谱与它们对应的实验测定光谱能很好地相吻合。根据MCRALS模型分析这四个绑定体系对应的荧光和UV-vis扩展数据矩阵的拟合结果,发现CTD/NCTD与这两种蛋白质结合形成四个复合物(如:CTD-BSA,NCTDBSA,CTD-BHb和NCTD-BHb)。此外,荧光猝灭结果表明,这两种蛋白质与CTD/NCTD的荧光猝灭均是静态猝灭机理。分别估算BSA-CTD/NCTD和BHbCTD/NCTD复合物的结合常数Kb、绑定位点数n、绑定距离r和热力学参数。红外光谱、同步荧光和三维荧光光谱的分析结果表明这两种药物能导致BSA/BHb的构象变化。运用几种不同光谱方法研究新橙皮苷/柚皮苷(NHD/NRG)与两种蛋白质(BSA和BHb)的相互作用。结果表明,这两种蛋白质与NHD/NRG间的猝灭机理为静态猝灭。在不同温度下,计算BSA/BHb-NHD/NRG体系对应的结合常数Kb和绑定位点数n、热力学参数和绑定距离r。采用三种不同方法分别讨论在NHD/NRG溶液中BSA/BHb构象的改变。探讨共存离子对BSA-NHD/NRG和BHbNHD/NRG体系结合常数的影响。此外,建立偏最小二乘(PLS)法同时测定胃复春片中NHD和NRG的新方法。
[Abstract]:Artemisinin, dihydroartemisinin, spot stingin, DMC, neohesperidin and naringin all have bioactivity and pharmacological effects. Artemisinin and dihydroarteminin have excellent antimalarial activity and become the most important antimalarial drugs. In addition, they have antitumor and anticancer effects; spot stingin and DMC are used as anti swelling agents. Tumor drugs are used to treat cancer of the esophagus, liver cancer, gastric cancer and so on, and in many cell lines, the toxin is less than the spot stingin, so it can provide more clinical applications; neohesperidin has the pharmacological effects of anti value added, anti allergy, antioxidation, anti estrogen, antitumor, activity of removing free radical and protective effect on stomach disease. Naringin has many functions, such as antioxidation, anti cholesterol, anti-inflammatory, hypoglycemic, anti apoptosis and heart protection and so on. It has become a new type of drug with application value. This paper mainly discusses the interaction between drugs and proteins, which have antitumor and anticancer effects. These studies help to understand the transport of such drugs in the body. Pharmacological action information, metabolic kinetics and a preliminary theoretical basis for the synthesis of high activity analogues. The specific contents are as follows: under simulated physiological conditions, the interaction between BSA and dihydroartemisinin (DHA) and artemisinin (ART) is investigated by fluorescence, UV-vis absorption and infrared spectroscopy. By fluorescence emission and Rayleigh dispersion According to the analysis results, the quenching mechanism between BSA and DHA/ART belongs to static quenching. At four different temperatures, the binding constant and the number of binding sites are calculated at four different temperatures. The binding distance between BSA and DHA/ART is 1.54/1.65 nm. based on the theory of non radiation energy transfer. The effects of these two drugs on the structure of BSA were discussed using UV-vis absorption, synchronous fluorescence and three-dimensional fluorescence spectroscopy. In addition, in the presence of DHA/ART, the peaks and intensity changes of the BSA characteristic absorption peaks were analyzed by infrared spectroscopy, and the corresponding red external spectral data were calculated by the simulation method, and the structure content of the BSA two structure was changed. They decrease the alpha helix content of the protein, increase the content of the beta lamellar and random curls. Finally, the effects of coexisting ions on the binding constant of the DHA/ART-BSA system are discussed. The interaction between cantharidin / norcantharidin (CTD/NCTD) and two proteins (BSA and BHb) is studied by the same method. It is shown that the microenvironment of tryptophan residues (Trp) on these two proteins changes, less peptide chain expansion and their hydrophobicity increase with the increase of the concentration of these two drugs. The concentration distribution and corresponding pure light of different components of the four binding systems are further studied by using the multivariate curve resolution alternating least squares (MCR-ALS) method. The results show that the fitting spectra of free CTD/NCTD and BSA/BHb are in good agreement with their corresponding experimental determination spectra. According to the analysis of the fitting results of the corresponding fluorescence and UV-vis extended data matrix of the four binding systems based on the MCRALS model, it is found that CTD/NCTD is combined with these two kinds of egg white matter to form four complexes (such as CTD-BSA, NCTD). BSA, CTD-BHb and NCTD-BHb). In addition, the fluorescence quenching results show that the fluorescence quenching of these two proteins and CTD/NCTD are both static quenching mechanisms. The binding constant Kb of BSA-CTD/NCTD and BHbCTD/NCTD complexes, binding distance n, binding distance r and thermodynamic parameters, red external spectra, synchronous fluorescence and three-dimensional fluorescence spectra are analyzed. The results showed that these two drugs could lead to the conformation changes of BSA/BHb. Several different spectral methods were used to study the interaction between new hesperidin / naringin (NHD/NRG) and two proteins (BSA and BHb). The results showed that the quenching mechanism between the two proteins and NHD/NRG was static quenching. At different temperatures, the corresponding BSA/BHb-NHD/NRG system was calculated. Combining constant Kb and binding location point n, thermodynamic parameters and binding distance R. are used to discuss the changes of BSA/BHb conformation in NHD/NRG solution respectively. The influence of coexistent ions on the binding constants of BSA-NHD/NRG and BHbNHD/NRG systems is discussed. In addition, a new method for simultaneous determination of NHD and NRG in gastric spring spring tablets by partial least squares (PLS) method is established. Law.
【学位授予单位】：西华师范大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R96;O657.3

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