含杂环配体的稀土有机配合物的合成、表征及相关生物活性的研究

【作者】 许东芳；

【导师】 何其庄；

【作者基本信息】 上海师范大学 ， 环境科学， 2011， 博士

【摘要】 近年来金属配合物在抗菌、抗肿瘤以及与DNA相互作用等方面的研究一直是生物无机化学研究的热点。本文以稀土金属配合物在这三方面的生物性能为着眼点,通过回流冷凝法和低温固相法以稀土,杂环化合物和第三配体合成了三大系列二十五种稀土杂环配合物。第一系列为含氮杂环配体的稀土配合物,第二系列为含硫杂环配体的稀土配合物,第三系列为含氮、硫杂环配体的稀土配合物,通过元素分析、摩尔电导、红外光谱、紫外光谱、氢谱、XRD、XPS、差热-热重以及TEM等方法对它们的组成、结构和形貌进行了表征。部分配合物的合成使用了低温固相法,由于低温固相法不使用溶剂,所以符合绿色化学的目标。确定了第一系列的化学组成为: Ln₂（ADP）（H₂O）₂ Cl₃(Ln = La³⁺,Sm³⁺,Pr³⁺,Nd³⁺, Y³⁺,Er³⁺ ; ADP =2-氨基-4,6-二甲氧基嘧啶) ;Ln₃（ADP）（MoO₄）2（CH₃OH）4Cl5•2H₂O (Ln = La³⁺, Sm³⁺, Pr³⁺,Nd³⁺, Y³⁺,Er³⁺;ADP =2-氨基-4,6-二甲氧基嘧啶, MoO4=钼酸根离子); Ln₂（ADMP）（H₂O）2Cl3 (Ln=La³⁺,ADMP=2-氨基-4,6-二甲基嘧啶); Ln₂（ADMP）（MoO4）2（CH₃OH）3Cl6•2H₂O(Ln=La³⁺,ADMP =2-氨基-4,6-二甲基嘧啶;MoO4=钼酸根离子);Ln₂（AP）（H₂O）2Cl3(Ln=La³⁺,AP=2-氨基嘧啶); Ln3（AP）（MoO₄）₂（CH₃OH）₄Cl₅(Ln = La³⁺, A P =2-氨基嘧啶;MoO4=钼酸根离子)。确定了第二系列的化学组成分别为:La（TTA）3L•H₂O (Ln = La³⁺, Ce³⁺, Nd³⁺, Eu³⁺, Er³⁺, TTA=噻吩甲酰三氟丙酮阴离子,L=对氯苯甲酸)和La（TTA）3N, （TTA=噻吩甲酰三氟丙酮阴离子,N=对羟基苯甲酸）。确定了第三系列的化学组成为:其中两个含氮、硫有机杂环配体的配合物的组成分别是La（TTA）3（Hbiim）(TTA=噻吩甲酰三氟丙酮阴离子,Hbiim =联咪唑和La（L）phen（NO₃）₃C₂H₅OH•CH₃OH,（HL=希夫碱,phen =邻菲咯啉）。另一个配合物为La（Hbiim）3N（TTA=噻吩甲酰三氟丙酮阴离子,N=对羟苯甲酸）。采用培养基扩散法和营养肉汤稀释法测定了上述稀土配合物的抗菌活性,结果表明:所合成的稀土配合物对大肠杆菌和金黄色葡萄球菌都具有抗菌性,属于广谱抗菌剂,且具有耐热性好的特点。配合物的活性都高于相应的配体。第一系列的配合物对大肠杆菌的抑菌效果要比金黄色葡萄球菌的好,抑菌圈的直径在1020mm之间,属于中等的抑菌作用;其中二元配合物的抗菌性能好于三元配合物。第二系列的配合物抗菌活性好于第一系列,抑菌圈的直径大于20mm,属于较强的抑菌作用。在第三系列中两个含氮硫杂环配体的配合物,抑菌圈的直径大于20mm,也属于较强的抑菌作用。同时对其抗菌机理做了初步探讨。采用形态学观察、MTT比色实验和流式细胞仪检测等方法研究了部分稀土配合物对K562白血病肿瘤细胞的凋亡和生长的抑制作用。倒置显微镜法和荧光显微镜法观察了经配合物诱导的K562肿瘤细胞的形态,表明配合物对K562肿瘤细胞有一定的凋亡作用;MTT比色实验结果表明:这些配合物均能够诱导K562肿瘤细胞凋亡,抑制其生长,配合物的半数抑制浓度IC50均小于10μg/mL,但是有浓度依赖性和时间依赖性,并且它们对K562肿瘤细胞的凋亡率和生长抑制率均高于其相应配体。第一系列的配合物的半数抑制浓度IC50在1¹0μg•mL-1之间,在这一系列当中,三元配合物的抗肿瘤性能好于二元配合物。第二系列配合物的半数抑制浓度IC50在1⁵μg•mL-1之间。第三系列配合物的半数抑制浓度在1¹0µg•mL-1之间。用流式细胞仪（FCM）检测了配合物La（TTA）3L•H₂O对肿瘤细胞的凋亡率,结果表明在浓度为5μg•ml-1和作用时间为48h时,La配合物La（TTA）3L•H₂O诱导K562肿瘤细胞凋亡的作用最好,并且凋亡作用发生在S期。同时对其抗肿瘤机理做了初步探讨。采用紫外吸收光谱、荧光光谱和琼脂糖凝胶电泳分析等方法研究了稀土镧配合物La（TTA）3L •H₂O与DNA的相互作用模式。结果表明:稀土配合物是以部分插入模式与DNA相互作用的,而且配合物还能够有效切割pBR322DNA,但是有浓度依赖性,其中0.1mmol•L-1为配合物对PBR322DNA的最佳切割浓度。更多还原

【Abstract】 In recent years, the study of metal complexes on antibacterial, antitumor and with DNA interaction have played importment role in the bioinorganic chemistry .On the biological properties of RE³⁺, twenty-five earth complexes with heterocyclic compounds and other ligands were synthesized by reflux condensation method and low temperature solid phase method.Their composition and structure werecharacterized by elemental analysis, molar conductance, IR, UV, 1HNMR, XRD and TEM etc. The stability of heat was proved by the data of TG-DTA. Some complexes were synthesized by low temperature solid phase method ,which was greener method without using any solvent.The compositions of the complexes containing nitrogen heterocyclic ligands were confirmed to be:Ln₂（ADP）（H₂O）2Cl3 and Ln3（ADP）（MoO4）2（CH₃OH）4Cl5•2H₂O (Ln=La³⁺,Sm³⁺,Pr³⁺,Nd³⁺,Y³⁺,Er³⁺;ADP=2-Amino-4,6-dimethoxypyrimidine); La2（ADMP）（H₂O）2Cl3 and La2（ADMP）（MoO₄）2（CH₃OH）3Cl₆•2H₂O（ADMP=2-A mino-4,6-dimethylpyrimidine）;La2（AP）（H₂O）2Cl3 and La₃（AP）（MoO₄）₂（CH₃OH）₄Cl₅ （AP =2-Aminopyrimidine）.The compositions of the complexes containing sulfur heterocyclic ligands were confirmed to be:Ln （TTA）3L] •H₂O (Ln = La³⁺, Ce³⁺, Nd³⁺, Eu³⁺, Er³⁺, TTA= Thenoyltrifluoroacetone anion , L= p-chlorbenzoicacid) ; La（TTA）3N （ TTA= Thenoyltrifluoroacetone anion,N=4-Hydroxybenzoic acid）.The compositions of the complexes containing both nitrogen and sulfur heterocyclic ligands were confirmed to be:La（TTA）₃（Hbiim）（ Hbiim=2,2-Biimdazole,TTA=Thenoyltrifluoroaceton anion）,La（L）phen（NO3）3C₂H₅OH•CH₃OH（HL= Shiff Base,phen = 1,10-Phenanthroline ）, and another complex was La（Hbiim）3N （Hbiim=2,2-biimdazole, N= 4-Hydroxybenzoic acid ）.The antibacterial activities of the complexes were evaluated by the agar diffusion method and nutrition broth dilution method. The results indicated that all the complexes exhibited antibacterial activities against Escherichia coli and Staphylococcus aureus, their antimicrobial spectrum were broad and have durable quality. In the complexes containing nitrogen heterocyclic ligands, the antibacterial activity test indicated that the complexes showed better antibacterial activity against Escherichia coli than Staphylococcus aureus, and binary complexes showed better antibacterial activity than ternary complexes.At the same time,the antibacterial activity were moderate（10-20mm）. In the complexes containing sulfur heterocyclic ligands, the antibacterial activity were good（>20mm）. In the two complexes containing both nitrogen and sulfur heterocyclic ligands, the antibacterial activity were also good（>20mm）. The antibacterial mechanism of the complexes was discussed preliminarily in this paper.The antitumor activity of some complexes against K562 tumor cell in vitro were researched by morphological observation, MTT assay and Flow Cytometry. The results showed that these complexes all could induced K562 tumor cells apoptosis and inhibited their growth,but the cell apoptosis rate and the inhibition rate both higher than their ligands and have concentration-dependent and time-dependent. The median inhibitory dose （IC50） of the complexes were below 10μg/mL.The IC50 of the complexes containing nitrogen heterocyclic ligands were between 110μg/mL, and ternary complexes showed better anti-tumor activity than binary complexes. The IC50 of the complexes containing sulfur heterocyclic ligands were between 15μg/mL. The IC50 of the complexes containing both nitrogen and sulfur heterocyclic ligands were between 15μg/mL.The complex La（TTA）3L•H₂O has induced K562 tumor cell for 48h can effect best apoptosis power, and effect S period.At the same time, the anti-tumor mechanism of the complexes was discussed preliminarily in this paper.The interaction of the complex La （TTA）3L•H₂O with DNA were studyed by UV, fluorescence spectrum and agarose gel electrophoresis etc.The results indicated that the interaction of the complex would bind DNA mainly by partially intercalation and the complex could cleave pBR322DNA effectively,but has concentration-dependent.The best concentration of cleaving pBR322DNA was 0.1mmol•L-1.更多还原