具有DNA和BSA键合功能的有机小分子和金属配合物的设计、合成及生物活性研究

发布时间：2018-05-28 07:00

本文选题：有机小分子 + N　；参考：《中国海洋大学》2014年博士论文

【摘要】：核酸和蛋白是构成生命体的重要基本物质,也是许多药物发挥药效的主要靶点。因此,研究有机小分子和金属配合物与核酸和蛋白之间的相互作用,对新型的高效、高选择性、低毒的抗肿瘤药物的理性设计、合成和抗肿瘤活性研究提供了重要的生物化学依据。本文以草酰胺类化合物为桥联配体,设计合成了一系列不对称N,N'-双取代草酰胺双核金属配合物,用x-射线衍射技术进行了单晶结构解析,并对有机小分子和金属配合物的DNA和BSA键合活性以及体外抗肿瘤活性进行了较为系统全面的研究,具体研究内容如下：1、有机小分子与DNA和BSA相互作用研究：选取VA二唑类化合物a-c、三氮唑类化合物d-g,以及草酰胺类化合物h和i为研究对象,通过元素分析、红外、质谱等手段对其进行了结构表征,并对化合物h和i的单晶结构进行了解析。通过紫外吸收光谱法、荧光光谱法、粘度法等方法研究了它们与鲱鱼精DNA(H5-DNA)和牛血清白蛋白(BSA)的键合活性,讨论了有机小分子化合物与DNA的结合方式和作用强弱,以及化合物猝灭BSA内源性荧光的猝灭机制和BSA结合位点。研究结果表明,与DNA插入结合的化合物的DNA键合活性比沟槽结合的化合物DNA键合活性强；结构中含有吸电子取代基,无论对以插入作用还是沟槽作用与DNA相结合的化合物,都可以增强其键合活性；所研究有机小分子化合物均可与BSA相互作用,结合位点可能为位于BSA内部疏水性口袋中的Trp-212残基。2、草酰胺配体及其双核金属配合物的设计、合成与结构：考虑到草酰胺类化合物出色的桥联功能和良好的生物活性,选取N-(5-氯-2-羟基苯基)-N'-(3-氨丙基)草酰胺(H3chapoxd)和N-(5-氯-2-羟基苯基)-N'-(3-二乙氨基丙基)草酰胺(H3chdepoxd)为桥联配体,通过改变端基配体、反应条件以及金属离子种类,采用自由配体法,合成出了9个草酰胺双核铜(Ⅱ)配合物, 即[Cu2(chapoxd)Cl(H2O)(bpy)]Cl·H2O (1)、[Cu2(chapoxd)(H2O)(bpy)](C]O4)·CH3CN (2)、[Cu2(chapoxd)(CH3CH2OH)(Me2bpy)] (C104) (3)、[Cu2(chapoxd)(H2O)(phen)]-(NO3)·H2O (4)、[Cu2(chapoxd)(H2O)(phen)](NO3) CH3OH (5)、[Cu2(chapoxd)-(H2O)(dabt)](C]O4)·2H2O (6)、[Cu2(chdepoxd)(H2O)(Me2bpy)](ClO4)·DMF (7)、 [Cu2(chdepoxd)(H2O)(dabt)](ClO4)·H2O (8). [Cu2(chdepoxd)(CH3OH)(phen)](ClO4)(9),以及1个草酰胺双核镍(Ⅱ)配合物[Ni2(chdepoxd)(bpy)2](ClO4) (10)。通过元素分析法、红外、核磁、质谱以及X-射线单晶衍射技术,对它们的结构进行了表征。3、草酰胺配体及其双核金属配合物与DNA相互作用研究：采用紫外吸收光谱法、荧光光谱法、电化学法和粘度法,对所合成的草酰胺配体H3chapoxd和H3chdepoxd及其双核金属配合物(1)～(10)的DNA键合性质进行了研究,探讨了桥联配体、端基配体、溶剂分子以及金属离子种类的改变对化合物的DNA键合活性的影响。结果表明,配体和配合物与HS-DNA的相互作用方式均为插入作用；端基配体的平面性大有利于配合物与DNA的插入作用,而端基配体空间位阻变大不利于这种插入作用;桥联配体的疏水性增加可以增强配合物与DNA的键合能力；双核镍(Ⅱ)配合物的DNA键合活性弱于双核铜(Ⅱ)配合物的。4、草酰胺配体及其双核金属配合物与BSA相互作用研究：运用紫外吸收光谱法和色氨酸荧光猝火法对所研究草酰胺配体H3chapoxd和H3chdepoxd及其双核金属配合物(1)～(10)与BSA的键合性质进行了研究。结果表明,所研究化合物均可以BSA发生相互作用;其猝灭BSA内源性荧光的机制为静态猝灭：配体及其配合物与BSA相互作用只有一个结合位点：配体的结合位点可能为BSA分子内部的疏水性Trp-212残基,而配合物则可能通过BSA表面的亲水性Trp-134残基而与BSA发生相互作用。5、有机小分子和金属配合物的体外抗肿瘤活性研究：采用磺酰罗丹明B(Sulforhodamine B, SRB)比色法,研究了有机小分子a～i、配体H3chapoxd和配体H3chdepoxd、以及双核金属配合物(1)～(10)对人肝癌细胞株(SMMC-7721)和人肺腺癌细胞株(A549)两种肿瘤细胞株的体外抗肿瘤活性。研究结果表明,所研究化合物对测试的两种细胞株均有一定程度的抑制作用,半数抑制浓度IC50值在16.9～95.1μg/mL范围内;化合物的DNA键合活性与其体外抗肿瘤活性存在明显的相关性,表明本文所研究化合物展现其体外抗肿瘤活性的作用靶点可能为DNA。
[Abstract]:Nucleic acids and proteins are important basic substances that constitute the life body, and are also the main targets of many drugs. Therefore, the study of the interaction between small organic molecules and metal complexes and nucleic acids and proteins is provided for the rational design, synthesis and antitumor activity of new highly efficient, highly selective and low toxic antitumor drugs. Important biochemical basis. In this paper, a series of asymmetric N, N'- biubstituted oxamide binuclear complexes were designed and synthesized with oxamides as bridging ligands. The structure of single crystal was analyzed by x- ray diffraction, and the DNA and BSA bonding activity of organic small molecules and metal complexes and in vitro antitumor activity were carried out. A more systematic and comprehensive study is made as follows: 1, the interaction of organic small molecules with DNA and BSA: the selection of VA two azole compound a-c, three azolol compound D-G, and oxamides h and I as the research objects, by elemental analysis, infrared and mass spectrometry. The single crystal structure of I was analyzed. The bonding activity with herring sperm DNA (H5-DNA) and bovine serum albumin (BSA) was studied by UV absorption spectrometry, fluorescence spectrometry, viscosity method and so on. The combination mode and effect of organic small molecule compounds with DNA and the quenching mechanism of quenching BSA endogenous fluorescence were discussed. The results of the BSA binding site show that the DNA bonding activity of the compound with DNA insertion is stronger than the groove bound compound DNA bond activity; the structure contains the electron absorbing substituent, and the bonding activity of the compound with the insertion or groove interaction with DNA can be enhanced; the organic small molecular chemistry is studied. The compound can interact with BSA, and the binding site may be the Trp-212 residue.2 in the hydrophobic pocket of BSA, the design, synthesis and structure of the oxamides and their binuclear complexes: considering the excellent bridging function and good biological activity of the oxamides, N- (5- chloride -2- hydroxyphenyl) -N'- (3- amyl)) is selected. H3chapoxd and N- (5- chloro -2- hydroxy phenyl) -N'- (3- two acetamyl) oxamides (H3chdepoxd) are bridged ligands. By changing the terminal ligands, reaction conditions and metal ions, 9 oxamides binuclear copper (II) complexes, [Cu2 (chapoxd) Cl (H2O) (bpy), 1, are synthesized by the free ligand method. (bpy)] (C]O4). CH3CN (2), [Cu2 (chapoxd) (Me2bpy) (C104) (C104), [Cu2 (chapoxd) (H2O), 4, 5, 6, 7, 8, 8. (CH3OH) (phen)] (ClO4) (ClO4) (9), and 1 oxamides binuclear nickel (II) nickel (II) complexes, [Ni2 (chdepoxd) (bpy) 2] (ClO4) (10). By elemental analysis, IR, NMR, MS and X- ray single crystal diffraction techniques, their structures are characterized by.3, oxamides and their binuclear metal complexes and DNA interaction: UV absorption The DNA bonding properties of the synthesized oxamide ligands H3chapoxd and H3chdepoxd and their binuclear metal complexes (1) ~ (10) were studied by spectroscopic, fluorescence, electrochemical and viscosity methods. The effects of bridging ligands, end group ligands, solvent molecules and the types of metal ions on the DNA bonding activity of the compounds were investigated. The results show that the interaction between the ligand and the ligand and HS-DNA are both inserted, and the planarity of the terminal ligand is greatly beneficial to the insertion of the complexes and DNA, while the space hindrance of the terminal ligand is not conducive to the insertion; the increase of the hydrophobicity of the bridged ligand can enhance the bonding ability of the complexes with the DNA; The DNA bond activity of the compound is weaker than the.4 of the binuclear copper (II) complex and the interaction between the oxamides and their binuclear metal complexes and BSA. The bonding properties of the studied oxamides H3chapoxd and H3chdepoxd and their binuclear metal complexes (1) ~ (10) and BSA were studied by UV absorption spectrometry and tryptophan fluorescence quenching method. The results show that all the compounds can interact with BSA; the mechanism for the quenching of BSA endogenous fluorescence is static quenching: the ligand and its complexes have only one binding site to interact with BSA: the ligand binding site may be a hydrophobic Trp-212 residue within the BSA molecule, and the complex may pass through the surface of the BSA surface. The interaction of hydrophilic Trp-134 residues and BSA with.5, the in vitro antitumor activity of organic small molecules and metal complexes: using the sulfonyl Luo Danming B (Sulforhodamine B, SRB) colorimetric method, the organic small molecules a to I, ligand H3chapoxd and ligand H3chdepoxd, and binuclear metal complexes (1) ~ (10) were used to treat human hepatocellular carcinoma cell lines (SM). MC-7721) and the in vitro antitumor activity of two human lung adenocarcinoma cell line (A549) cell lines. The results showed that the compound had a certain degree of inhibition on the two cell lines tested, and the median inhibitory concentration was within the range of 16.9 to 95.1 mu g/mL, and the DNA binding activity of the compound and its antitumor activity in vitro existed in vitro. The obvious correlation shows that the target of the compounds studied in this study is DNA..
【学位授予单位】：中国海洋大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R91;R914.5

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