【作者】 高苏亚；

【导师】 李华；

【作者基本信息】 西北大学 ， 分析化学， 2012， 博士

【摘要】 药物分子与生物相关物质之间的相互作用研究具有非常重要的意义。随着研究者研究水平的不断提高,分析仪器的不断更新和新药的不断问世,药物与生物分子之间的研究方法也不断增多和更新。本文采用紫外-可见分光光度法、荧光光谱法、毛细管电泳法以及区段灌注技术、计算机分子模拟技术等方法研究了姜黄素、坦索罗辛、双醋瑞因、生物碱类及苯甲酸类药物与DNA、牛血清白蛋白、环糊精和金属离子等之间的相互作用,测定了结合常数和相关热力学参数,推测了一定条件下的作用机理。在研究方法和数据处理中有一定的改进和扩展,并将其研究方法应用于相应的药物分析之中,包括药物的质量控制和手性药物对映体的拆分。本论文分为两部分。第一部分是绪论,引用大量文献详细综述了分子间相互作用研究的进展及重要意义,研究方法及其应用,药物分子与生物相关物质间相互作用的研究现状等。第二部分是研究报告,具体研究内容如下：1.光谱法研究姜黄素与有关物质的相互作用及应用采用双等色分光光度法测定了在pH=6.5时姜黄素与Fe2+的络合稳定常数,同时对实验设计提出新的改进方法,并将Excel应用于迭代法的数据处理中,大大简化了实验操作和数据处理过程。由此建立了姜黄素-分光光度法测定复方硫酸亚铁叶酸片中Fe2+的含量,方法简便可靠。采用双倒数法获得该络合物与ssDNA的结合常数,探讨了其作用机理。采用荧光结合Excel迭代数据处理方法和紫外光谱法系统研究了姜黄素与BSA、p-CD、HP-p-CD之间的相互作用机理,制备并考察了CUR-HP-β-CD包合物的光谱性质、水溶性及稳定性。2.三种生物碱与BSA及环糊精相互作用的方法学研究及应用采用经典的荧光法研究了咖啡因、可可碱和茶碱与BSA的作用机理；同时分别采用以药物为添加剂的“DP”方式和以BSA为添加剂的“PD”方式的亲和毛细管电泳法(ACE)研究了三种生物碱与BSA的相互作用。两种方法的实验结果基本一致。采用紫外分析法测定了三种生物碱与p-CD的相互作用,利用其识别作用建立了以p-CD为添加剂的CZE法对茶叶、可乐饮料、咖啡、药品等样品中三种生物碱同时定量的分析方法。3.坦索罗辛与环糊精/BSA相互作用的方法学研究及应用采用分光光度法结合分子模拟技术研究了坦索罗辛和β-CD、HP-β-CD的相互作用,以此建立了CZE法同时分离测定坦索罗辛原药及其缓释胶囊中坦索罗辛和相关中间体的方法。采用CZE和ACE原理,探讨了坦索罗辛消旋体的手性拆分、对映体与手性选择剂β-CD、HP-β-CD和BSA之间的相互作用。利用毛细管区段灌注技术和区段前药物代替标记物的方法成功地解决了坦索罗辛及其中间体与BSA作用的背景干扰问题。4.苯甲酸类与BSA相互作用的光度法研究及其HPCE分析应用采用紫外光度法的静电模型研究了苯甲酸、水杨酸、乙酰水杨酸(阿司匹林)与BSA的相互作用。实验过程中以BSA溶液作参比,测定药物-BSA混合体系的吸光度值,可消除残余BSA对药物吸光度测量的干扰,此方法可推广到其它类似的有光谱重叠现象的相互作用体系。采用β-CD修饰的MECC法同时分离了5种苯甲酸类化合物,并应用MECC-内标法分离测定了板蓝根药材中芳香酸类化合物。5.双醋瑞因与环糊精/BSA相互作用的光谱法研究双醋瑞因是一种新型抗炎药物,可显著缓解和改善骨关节炎患者的关节功能。利用简单的紫外分光光度法测得双醋瑞因与β-CD、HP-β-CD之间的包合作用；利用荧光分析手段测得双醋瑞因对BSA的荧光有静态猝灭效应,同时用改进的Excel迭代方法计算了双醋瑞因与BSA在不同温度下的结合位点数、结合常数和相关热力学参数,并推测出其作用力类型主要为疏水和静电作用。更多还原

【Abstract】 It is very important to investigate the interaction between drug molecules and biological related material. With the improvement of researchers to study, analytical instrument unceasing renewal and the new drugs coming out constantly, the research methods of interactions between drugs and biological molecules are increasing and being updated. In this thesis the interactions between drugs (curcumin, tamsulosin, diacerein, alkaloids and benzoic acid drugs) and some substances (DNA, bovine serum albumin, cyclodextrins and metal ions) are studied by the methods such as ultraviolet-visible spectrophotometry, fluorescence spectrometry, capillary electrophoresis method and partial filling technique, computer molecular modeling, etc. The binding constant and related thermodynamics parameters are determined. And the mechanism of interactions is speculated at the certain conditions. The certain improvement and expansion are applied in research methods. In additional, these methods are applied in the corresponding analysis of drugs, including drug quality control and separation of chiral drugs enantiomer.This thesis includes two parts. Part one is an introduction, which reports in detail the development and important significance of the interactions, methods and their application, the present study about drugs and biological material by citing with lots of references. Part two is a research report. And the specific research contents are listed as follows:1. Investigation on the interaction between curcumin and related substances by spectroscopic methods and their applicationsThe stability constant of curcumin-Fe2+complex is determined by spectrophotometric dual isosbestic points when the value of pH is6.5in buffer solution. And the new improvement is put forward in experimental design. Excel is used in the iteration data processing, greatly simplifying the experiment and data processing. The curcumin-spectrophotometry is applied to determine Fe2+in compound ferrous sulfate folic acid tablets, and this method is simple and reliable. The binding constant of the curcumin-Fe2+complex with salmon sperm DNA (ssDNA) is determined by double multiplicative inverse curve, and its mechanism is discussed. By fluorescence combined with Excel iteration method in data processing and ultraviolet spectrum are applied to investigate in detail the interactions between curcumin and bovine serum albumin (BSA), beta-cyclodextrin (β-CD), hycroxypropyl-beta-cyclodextrin (HP-(3-CD). The curcumin-HP-(3-CD complex is prepared and its properties are investigated such as infrared spectrum, water-solubility and stability, etc.2. Methodology study on the interaction between three alkaloids with BSA/CDs and applicationFirstly, classical fluorescence spectroscopy is used to study the interaction mechanism between BSA and drugs (caffeine, theobromine and theophylline). Secondly, affinity capillary electrophoresis (ACE) is also used for investigation the interactions through two modes:"Dp" for drugs as additive and "PD" for BSA as additive, respectively. Their experimental results are consistent. Thirdly, the interactions between three alkaloids with β-CD are analyzed by UV spectrometer. Finally, based on their recognition function, the quantitative analysis is established for determining three alkaloids in samples (tea, cola drink, coffee and drugs) by capillary zone electrophoresis (CZE) with β-CD as additive in the buffer solution.3. Methodology study on the interaction between tamsulosin with BSA/CDs and applicationThe interactions between tamsulosin with β-CD and HP-P-CD are investigated by spectrophotometry combined with molecular modeling. Then using the recognition, CZE method is established for simultaneous separation and determination tanmulosin in its bulk drug and sustained-release capsule. Based on the principles of CZE and ACE, the enantioseparation of tamsulosin and the interactions between the enantiomers with chiral selectors are investigated, the chiral selectors including β-CD, HP-β-CD, BSA, dextran, etc. Additionally, in experimental investigation the interactions tamsulosin and intermediates with BSA, the background interference problem is solved successfully by using partial filling technique and drugs instead of neutral markers in capillary electrophoresis.4. Interactions between benzoic acid drugs with BSA by spectrophotometry and their quantitative analysis by high performance capillary electrophoresis (HPCE)The electrostatic mode is applied to study the interactions between benzoic acids and BSA by spectrophotometry. BSA solution is used as reference solution in experimental process to determine the absorbance of drug-BSA system, which can eliminate the interference about residual BSA on absorbance measure of drugs. This method can be extended to other similar spectrum overlapping interaction system. In additional, five benzoic acids are separated simultaneously by micellar electrokinetic capillary chromatography (MECC) modified with β-CD. Then the MECC-internal standard method is applied to separate and determine aromatic acid compounds in the radix isatidis, the results are feasible and satisfactory.5. Interactions between diacerein and CDs/BSA by spectroscopic methodDiacerein is a kind of new anti-inflammatory. It can ease and improve significantly the patients’arthralgia with osteoarthritis. The inclusion interactions between diacerein with β-CD and HP-β-CD are observed by simple UV spectrophtotometry. The results show the inclusion is feasible and its degradation will be improved. The interaction of diacerein-BSA system is investigated by using fluorescence analysis. The results show that diacerein can quench the fluorescence of BSA with a static quenching mechanism. At the same time Excel iterative method is used to data processing in determining the binding constant of diacerein-BSA complex at different temperature. Then the interactional type is speculated from the relevant thermodynamic parameters, which is mainly hydrophobic interaction and electrostatic effect.更多还原