金属有机笼的合成及与核酸作用研究
发布时间：2018-10-29 12:58
【摘要】：核酸是抗癌药物的重要靶点之一,其中人体端粒G-四链体DNA二级结构由于能够有效抑制端粒酶的活性从而抑制肿瘤细胞的生长而受到人们的广泛关注。新型金属有机配合物由于结构灵活,自身携带正电荷易与DNA磷酸骨架相互作用等特点被应用于潜在的抗癌药物研究中。本文旨在寻找能高效稳定G-四链体DNA二级结构的新型金属有机笼,并探究其与G-四链体DNA作用的各种影响因素及作用机制,主要从以下三个方面展开工作:1.柔性四面体金属有机笼对G-四链体DNA的结合性能和抗癌作用。设计合成了多种新型柔性四面体Ni(II)的金属有机笼对映体(R-1/S-1,R-2/S-2,R-3/S-3,R-4/S-4),X-射线单晶衍射确认四面体笼状结构的合成且伴有高度立体化学纯度(ΔΔΔΔ或ΛΛΛΛ)。通过紫外光谱滴定实验(UV-vis)、圆二色光谱(CD)、FRET熔点实验等方法,对金属有机笼与G4-DNA作用强度和作用模式进行系统地研究,并通过细胞毒性实验进一步研究金属有机笼的抗癌性质。结果表明,金属有机笼对G4-DNA表现出较强的结合能力(Kb=1.13×105-5.66×105 M-1),对反平行型G4-DNA有较强的稳定作用(?Tm=19.2-25.1°C),并且它们的对映体ΛΛΛΛ和ΔΔΔΔ对G-四链体DNA有一定的对映选择性作用;金属有机笼与G4-DNA的作用模式为外部堆积作用,其中含萘环结构的金属有机笼R-3/S-3对G4-DNA的稳定作用最强;细胞毒性实验表明金属有机笼对不同癌细胞有不同程度的抑制作用,其中R-3/S-3和R-4/S-4是最有潜力的抗癌药物。2.刚性四面体金属有机笼与G-四链体DNA的相互作用及构效关系。合成了一系列具有不同金属中心、配体及电荷的刚性四面体金属有机笼1-7,通过红外、~1H NMR、质谱、元素分析等测试手段对其进行详细表征,X-射线单晶衍射证实了刚性四面体金属有机笼1和4的确切结构。与G4-DNA作用的一系列实验表明,带8个正电荷的四面体Ni(II)金属有机笼1-2和7对G4-DNA显示出较强的结合能力(Kb=2.29×106-4.35×106 M-1)和稳定性作用(?Tm=39.1-41.7°C),较柔性四面体金属有机笼有很大的提高,Fe(II)的金属有机笼3-4与G4-DNA的作用相对较弱(?Tm=18.33-24.80°C),带4个负电荷的金属有机笼5-6几乎未与G4-DNA发生作用(?Tm1°C)。说明金属有机笼的刚柔性、金属中心及总电荷等在与G4-DNA作用时起着非常重要的作用。3.研究刚性立方体卟啉金属有机笼与G-四链体DNA的相互作用。合成了两种具有独特大π共轭体系并带有16个正电荷的刚性立方体卟啉金属有机笼8和9,对其进行一系列谱学表征。与G4-DNA作用的实验表明两种金属有机笼不但可与G-四链体DNA发生很强程度的结合(Kb=3.89×106-5.02×106 M-1),而且对反平行构型的四链体DNA表现出显著提高的稳定性作用(?Tm=32.46-41.68°C,0.50μM)。其中,基于卟啉胺(TAPP)并吡啶配体的金属卟啉立方体笼8与G-四链体DNA的相互作用强于基于金属卟啉胺(Zn-TAPP)并吡啶配体的立方体笼9。
[Abstract]:Nucleic acid is one of the important targets of anticancer drugs. The secondary structure of human telomere G-quad DNA has been paid more and more attention because of its ability to inhibit the activity of telomerase and the growth of tumor cells. The novel organometallic complexes have been used in the study of potential anticancer drugs due to their flexible structure and easy interaction between their own positive charge and DNA phosphoric acid framework. The purpose of this paper is to find out a new metal-organic cage which can efficiently stabilize the secondary structure of G- quad-stranded DNA, and to explore the various influencing factors and mechanism of its interaction with G- quad-stranded DNA. The main works are as follows: 1. Binding and anticancer effects of flexible tetrahedral organometallic cages on G-tetrahedral DNA. A variety of metal-organic cage enantiomers of new flexible tetrahedral Ni (II) have been designed and synthesized (R-1 / S-1 / R-2 / S-2 / R-3 / S-3 / R-4 / S-4). X-ray single crystal diffraction confirmed the synthesis of tetrahedron cage structure with a high stereo-chemical purity (螖 or A). By means of UV titration (UV-vis) and circular dichroism spectrum (CD), FRET) melting point experiment, the interaction intensity and action mode between organometallic cages and G4-DNA were studied systematically. The anticancer properties of organometallic cages were further studied by cytotoxicity test. The results show that organometallic cages have strong binding ability to G4-DNA (Kb=1.13 脳 105-5.66 脳 105M-1) and strong stability to anti-parallel G4-DNA (? Tm=19.2-25.1 掳C),). Moreover, their enantiomers A and 螖 have a certain enantioselectivity to G-quadruplex DNA. The interaction pattern between organometallic cages and G4-DNA is external stacking, in which R-3/S-3 with naphthalene ring structure has the strongest stabilizing effect on G4-DNA. Cytotoxicity tests showed that organometallic cages had different inhibitory effects on different cancer cells, of which R-3/S-3 and R-4/S-4 were the most potential anticancer drugs. 2. Interaction and structure-activity relationship between rigid tetrahedral organometallic cages and G- quadruplex DNA. A series of rigid tetrahedral organometallic cages 1-7 with different metal centers, ligands and charges were synthesized and characterized by IR, 1H NMR, mass spectrometry and elemental analysis. X-ray single crystal diffraction confirmed the exact structure of rigid tetrahedron organometallic cages 1 and 4. A series of experiments with G4-DNA show that, 8-positively charged tetrahedral Ni (II) organometallic cages 1-2 and 7 show strong binding ability (Kb=2.29 脳 106-4.35 脳 106M-1) and stability (? Tm=39.1-41.7 掳C),) to G4-DNA. Compared with flexible tetrahedron organometallic cages, the metal-organic cages 3-4 with, Fe (II) have a relatively weak interaction with G4-DNA (? Tm=18.33-24.80 掳C),). The organometallic cage 5-6 with four negative charges has little interaction with G4-DNA (? Tm1 掳C). It shows that the rigid flexibility, metal center and total charge of organometallic cages play a very important role in the interaction with G4-DNA. The interaction between rigid cubic porphyrin metal cages and G-quadruplex DNA was studied. Two kinds of rigid cubic porphyrin metal cages 8 and 9 with 16 positive charges have been synthesized and characterized by a series of spectra. The experiments of interaction with G4-DNA show that the two organometallic cages not only bind to DNA (Kb=3.89 脳 106-5.02 脳 106M-1), but also bind to DNA. Moreover, the stability of anti-parallel four-stranded DNA was significantly improved (? Tm=32.46-41.68 掳C = 0.50 渭 M). Among them, the interaction of metalloporphyrin cube cage 8 based on (TAPP) and pyridine ligands with DNA is stronger than that based on Zn-TAPP and pyridine ligands.
【学位授予单位】：江南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O641.4;TQ460.1
本文编号：2297764
[Abstract]:Nucleic acid is one of the important targets of anticancer drugs. The secondary structure of human telomere G-quad DNA has been paid more and more attention because of its ability to inhibit the activity of telomerase and the growth of tumor cells. The novel organometallic complexes have been used in the study of potential anticancer drugs due to their flexible structure and easy interaction between their own positive charge and DNA phosphoric acid framework. The purpose of this paper is to find out a new metal-organic cage which can efficiently stabilize the secondary structure of G- quad-stranded DNA, and to explore the various influencing factors and mechanism of its interaction with G- quad-stranded DNA. The main works are as follows: 1. Binding and anticancer effects of flexible tetrahedral organometallic cages on G-tetrahedral DNA. A variety of metal-organic cage enantiomers of new flexible tetrahedral Ni (II) have been designed and synthesized (R-1 / S-1 / R-2 / S-2 / R-3 / S-3 / R-4 / S-4). X-ray single crystal diffraction confirmed the synthesis of tetrahedron cage structure with a high stereo-chemical purity (螖 or A). By means of UV titration (UV-vis) and circular dichroism spectrum (CD), FRET) melting point experiment, the interaction intensity and action mode between organometallic cages and G4-DNA were studied systematically. The anticancer properties of organometallic cages were further studied by cytotoxicity test. The results show that organometallic cages have strong binding ability to G4-DNA (Kb=1.13 脳 105-5.66 脳 105M-1) and strong stability to anti-parallel G4-DNA (? Tm=19.2-25.1 掳C),). Moreover, their enantiomers A and 螖 have a certain enantioselectivity to G-quadruplex DNA. The interaction pattern between organometallic cages and G4-DNA is external stacking, in which R-3/S-3 with naphthalene ring structure has the strongest stabilizing effect on G4-DNA. Cytotoxicity tests showed that organometallic cages had different inhibitory effects on different cancer cells, of which R-3/S-3 and R-4/S-4 were the most potential anticancer drugs. 2. Interaction and structure-activity relationship between rigid tetrahedral organometallic cages and G- quadruplex DNA. A series of rigid tetrahedral organometallic cages 1-7 with different metal centers, ligands and charges were synthesized and characterized by IR, 1H NMR, mass spectrometry and elemental analysis. X-ray single crystal diffraction confirmed the exact structure of rigid tetrahedron organometallic cages 1 and 4. A series of experiments with G4-DNA show that, 8-positively charged tetrahedral Ni (II) organometallic cages 1-2 and 7 show strong binding ability (Kb=2.29 脳 106-4.35 脳 106M-1) and stability (? Tm=39.1-41.7 掳C),) to G4-DNA. Compared with flexible tetrahedron organometallic cages, the metal-organic cages 3-4 with, Fe (II) have a relatively weak interaction with G4-DNA (? Tm=18.33-24.80 掳C),). The organometallic cage 5-6 with four negative charges has little interaction with G4-DNA (? Tm1 掳C). It shows that the rigid flexibility, metal center and total charge of organometallic cages play a very important role in the interaction with G4-DNA. The interaction between rigid cubic porphyrin metal cages and G-quadruplex DNA was studied. Two kinds of rigid cubic porphyrin metal cages 8 and 9 with 16 positive charges have been synthesized and characterized by a series of spectra. The experiments of interaction with G4-DNA show that the two organometallic cages not only bind to DNA (Kb=3.89 脳 106-5.02 脳 106M-1), but also bind to DNA. Moreover, the stability of anti-parallel four-stranded DNA was significantly improved (? Tm=32.46-41.68 掳C = 0.50 渭 M). Among them, the interaction of metalloporphyrin cube cage 8 based on (TAPP) and pyridine ligands with DNA is stronger than that based on Zn-TAPP and pyridine ligands.
【学位授予单位】：江南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O641.4;TQ460.1
【参考文献】
相关期刊论文 前2条
1 张现侠;陈灿玉;;水溶性卟啉及金属卟啉在DNA中的研究进展[J];化学学报;2012年24期
2 吕小伟,朱驹,周有骏;G四链体晶体结构及其小分子配体的研究进展[J];国外医学.药学分册;2004年06期
相关博士学位论文 前1条
1 罗旭健;靶向端粒G-四链体DNA的抗肿瘤铂化合物的合成、表征及其作用机理研究[D];中南大学;2014年
相关硕士学位论文 前1条
1 徐新新;多核金属配合物的设计合成及与DNA相互作用的研究[D];江南大学;2014年
，本文编号：2297764
本文链接：https://www.wllwen.com/kejilunwen/huaxue/2297764.html
教材专著
