金屬有機(jī)籠的合成及與核酸作用研究
發(fā)布時(shí)間：2018-10-29 12:58
【摘要】：核酸是抗癌藥物的重要靶點(diǎn)之一,其中人體端粒G-四鏈體DNA二級(jí)結(jié)構(gòu)由于能夠有效抑制端粒酶的活性從而抑制腫瘤細(xì)胞的生長(zhǎng)而受到人們的廣泛關(guān)注。新型金屬有機(jī)配合物由于結(jié)構(gòu)靈活,自身攜帶正電荷易與DNA磷酸骨架相互作用等特點(diǎn)被應(yīng)用于潛在的抗癌藥物研究中。本文旨在尋找能高效穩(wěn)定G-四鏈體DNA二級(jí)結(jié)構(gòu)的新型金屬有機(jī)籠,并探究其與G-四鏈體DNA作用的各種影響因素及作用機(jī)制,主要從以下三個(gè)方面展開工作:1.柔性四面體金屬有機(jī)籠對(duì)G-四鏈體DNA的結(jié)合性能和抗癌作用。設(shè)計(jì)合成了多種新型柔性四面體Ni(II)的金屬有機(jī)籠對(duì)映體(R-1/S-1,R-2/S-2,R-3/S-3,R-4/S-4),X-射線單晶衍射確認(rèn)四面體籠狀結(jié)構(gòu)的合成且伴有高度立體化學(xué)純度(ΔΔΔΔ或ΛΛΛΛ)。通過紫外光譜滴定實(shí)驗(yàn)(UV-vis)、圓二色光譜(CD)、FRET熔點(diǎn)實(shí)驗(yàn)等方法,對(duì)金屬有機(jī)籠與G4-DNA作用強(qiáng)度和作用模式進(jìn)行系統(tǒng)地研究,并通過細(xì)胞毒性實(shí)驗(yàn)進(jìn)一步研究金屬有機(jī)籠的抗癌性質(zhì)。結(jié)果表明,金屬有機(jī)籠對(duì)G4-DNA表現(xiàn)出較強(qiáng)的結(jié)合能力(Kb=1.13×105-5.66×105 M-1),對(duì)反平行型G4-DNA有較強(qiáng)的穩(wěn)定作用(?Tm=19.2-25.1°C),并且它們的對(duì)映體ΛΛΛΛ和ΔΔΔΔ對(duì)G-四鏈體DNA有一定的對(duì)映選擇性作用;金屬有機(jī)籠與G4-DNA的作用模式為外部堆積作用,其中含萘環(huán)結(jié)構(gòu)的金屬有機(jī)籠R-3/S-3對(duì)G4-DNA的穩(wěn)定作用最強(qiáng);細(xì)胞毒性實(shí)驗(yàn)表明金屬有機(jī)籠對(duì)不同癌細(xì)胞有不同程度的抑制作用,其中R-3/S-3和R-4/S-4是最有潛力的抗癌藥物。2.剛性四面體金屬有機(jī)籠與G-四鏈體DNA的相互作用及構(gòu)效關(guān)系。合成了一系列具有不同金屬中心、配體及電荷的剛性四面體金屬有機(jī)籠1-7,通過紅外、~1H NMR、質(zhì)譜、元素分析等測(cè)試手段對(duì)其進(jìn)行詳細(xì)表征,X-射線單晶衍射證實(shí)了剛性四面體金屬有機(jī)籠1和4的確切結(jié)構(gòu)。與G4-DNA作用的一系列實(shí)驗(yàn)表明,帶8個(gè)正電荷的四面體Ni(II)金屬有機(jī)籠1-2和7對(duì)G4-DNA顯示出較強(qiáng)的結(jié)合能力(Kb=2.29×106-4.35×106 M-1)和穩(wěn)定性作用(?Tm=39.1-41.7°C),較柔性四面體金屬有機(jī)籠有很大的提高,Fe(II)的金屬有機(jī)籠3-4與G4-DNA的作用相對(duì)較弱(?Tm=18.33-24.80°C),帶4個(gè)負(fù)電荷的金屬有機(jī)籠5-6幾乎未與G4-DNA發(fā)生作用(?Tm1°C)。說明金屬有機(jī)籠的剛?cè)嵝�、金屬中心及總電荷等在與G4-DNA作用時(shí)起著非常重要的作用。3.研究剛性立方體卟啉金屬有機(jī)籠與G-四鏈體DNA的相互作用。合成了兩種具有獨(dú)特大π共軛體系并帶有16個(gè)正電荷的剛性立方體卟啉金屬有機(jī)籠8和9,對(duì)其進(jìn)行一系列譜學(xué)表征。與G4-DNA作用的實(shí)驗(yàn)表明兩種金屬有機(jī)籠不但可與G-四鏈體DNA發(fā)生很強(qiáng)程度的結(jié)合(Kb=3.89×106-5.02×106 M-1),而且對(duì)反平行構(gòu)型的四鏈體DNA表現(xiàn)出顯著提高的穩(wěn)定性作用(?Tm=32.46-41.68°C,0.50μM)。其中,基于卟啉胺(TAPP)并吡啶配體的金屬卟啉立方體籠8與G-四鏈體DNA的相互作用強(qiáng)于基于金屬卟啉胺(Zn-TAPP)并吡啶配體的立方體籠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 擄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 擄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 擄C),). The organometallic cage 5-6 with four negative charges has little interaction with G4-DNA (? Tm1 擄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 擄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.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O641.4;TQ460.1
本文編號(hào)：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 擄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 擄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 擄C),). The organometallic cage 5-6 with four negative charges has little interaction with G4-DNA (? Tm1 擄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 擄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.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O641.4;TQ460.1
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