本文關(guān)鍵詞： 光動力治療 藻藍蛋白 酞菁 復(fù)合物　出處：《福州大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：光動力療法(Photodynamic Therapy,PDT)是近幾年來迅速發(fā)展起來的一種選擇性治療增生性病變的新技術(shù),已成為當前抗腫瘤研究領(lǐng)域的熱點之一。光動力療法的三要素是光、光敏劑和氧,其中光敏劑是關(guān)鍵因素。由于在近紅外區(qū)(650nm-750nm)具有良好的光物理光化學(xué)性質(zhì),酞菁(Phthalocyanine,Pc)作為第二代光敏劑的應(yīng)用前景已受到廣泛關(guān)注。但由于酞菁大環(huán)共軛體系的強疏水作用,多數(shù)酞菁化合物水溶性較低,不利于其實際應(yīng)用。另一方面,藻藍蛋白(Phycocyanin,PC)是一種具有良好水溶性的卟啉類色素蛋白,已有研究表明,藻藍蛋白具有光敏活性,作用光譜處于550nm-650nm,但光敏活性有待進一步提高。著眼于利用酞菁與藻藍蛋白的各自優(yōu)勢,進而構(gòu)建具有雙功能基的復(fù)合光敏劑,本論文圍繞藻藍蛋白與酞菁復(fù)合物的制備、光譜性質(zhì)、光動力活性開展研究,主要研究內(nèi)容和結(jié)果總結(jié)如下:1.藻藍蛋白的提取與純化本文選擇藻膽蛋白含量較高的螺旋藻作為提取原料。先采用反復(fù)凍融法從螺旋藻中得到藻藍蛋白粗提液,再將粗提液經(jīng)過(NH4)2S04鹽析進行初步的提純。鹽析后的藻藍蛋白通過DEAE Sepharose Fast Flow離子交換層析法和葡聚糖凝膠層析法進一步純化。電泳和光譜測試表明所獲得的藻藍蛋白具有較高純度,光譜純度達到4.0以上,分子量為115kDa左右。2.酞菁的合成與表征合成和表征了三種酞菁,分別是1-[4-(2-氨基乙基)苯氧基]酞菁鋅(簡稱ZnPcNH2),1,8(11),15(18),22(25)-四(3-羧基苯氧基)酞菁鋅(簡稱(4),1-[4-(2-羧基乙基)苯氧基]酞菁鋅(簡稱E1)。文獻檢索表明,ZnPcNH2酞菁配合物及其前驅(qū)體尚未見報道。3.藻藍蛋白-酞菁復(fù)合物的制備本文通過非共價結(jié)合與共價結(jié)合兩種方式來構(gòu)建藻藍蛋白-酞菁復(fù)合物。對于非共價結(jié)合途徑,首先通過光譜法研究了一系列酞菁與藻藍蛋白的相互作用。進而,選取了六種酞菁,通過溫育交換法制備了六種非共價結(jié)合復(fù)合物。對于共價結(jié)合途徑,選取了兩種氨基衍生物及兩種羧基衍生物修飾的酞菁鋅,采用戊二醛連接法以及成酰胺反應(yīng),獲得了四種共價復(fù)合物。同時,通過光譜法對復(fù)合物的組成進行了分析。4.藻藍蛋白-酞菁復(fù)合物的性質(zhì)研究研究了復(fù)合物在水溶液中電子吸收光譜、熒光光譜和存在狀態(tài)。評價了部分酞菁-藻藍蛋白復(fù)合物對人體肝癌細胞HepG2的光動力抑制活性。結(jié)果表明,無取代酞菁鋅與藻藍蛋白的復(fù)合物(ZnPc-PC,結(jié)合比為8:1)光動力抑制HepG2的IC50值為2.1μM,四羧基酞菁鋅和藻藍蛋白(C4-PC,結(jié)合比為6:1)光動力抑制HepG2的IC50值為0.93μM,上述兩種復(fù)合物不但顯示了顯著的光動力抗癌活性,且活性明顯高于藻藍蛋白和相應(yīng)的酞菁鋅,值得對其深入開展研究。
[Abstract]:Photodynamic thermotherapy (PDT) is a new technique for selective treatment of proliferative lesions developed rapidly in recent years. The three elements of photodynamic therapy are light, Guang Min and oxygen. Guang Min is the key factor. Due to its good photophysical and photochemical properties in the near infrared region of 650nm-750nm, the phthalocyanine is a phthalocyanine. The application prospect of Pc) as the second generation Guang Min has been widely concerned. However, because of the strong hydrophobic effect of phthalocyanine macrocyclic conjugation system, most phthalocyanine compounds have low water solubility, which is not conducive to their practical application. Phycocyanin (PCP) is a porphyrin pigment protein with good water solubility. It has been studied that phycocyanin has Guang Min activity. The action spectrum was at 550 nm ~ 650 nm, but Guang Min activity needed to be further improved. The aim of this study was to make use of the respective advantages of phthalocyanine and phycocyanin, and then to construct a bifunctional complex Guang Min. In this paper, the preparation, spectral properties and photodynamic activity of phycocyanin and phthalocyanine complexes were studied. The main contents and results are summarized as follows:. 1. Extraction and purification of phycocyanin. In this paper, spirulina with high phycobilis content was selected as raw material. Firstly, the crude extract of phycocyanin was obtained from Spirulina platensis by repeated freeze-thaw method. Then the crude extract was passed through NH _ 4). 2S04 was purified by salting-out. Phycocyanin was purified by DEAE Sepharose Fast. Flow ion exchange chromatography and dextran gel chromatography were further purified. Electrophoresis and spectral analysis showed that the obtained phycocyanin had high purity. The spectral purity is over 4.0, and the molecular weight of phthalocyanine is about 115kDa. The synthesis and characterization of phthalocyanine are 1-. [Zinc phthalocyanine (ZnPcNH _ 2H _ 2H _ 2H _ 2H _ 2H _ 2H _ 2H _ 2H _ 2H _ 2H _ 2H _ 2H _ 2H _ 2H _ 2O _ 2). Zinc phthalocyanine. [Zinc phthalocyanine (E1). Preparation of phycocyanin-phthalocyanine complexes. In this paper, phycocyanin-phthalocyanine complexes were constructed by non-covalent binding and covalent binding. For non-covalent combination pathways. Firstly, the interaction of a series of phthalocyanines with phycocyanin was studied by spectral method. Then, six kinds of phthalocyanines were selected and six non-covalent complexes were prepared by the method of thermo-exchange. Two amino derivatives and two carboxyl derivatives modified zinc phthalocyanine were selected and four covalent complexes were obtained by glutaraldehyde bonding method and amide-forming reaction. The composition of phycocyanin-phthalocyanine complex was analyzed by spectroscopic method. The properties of phycocyanin-phthalocyanine complex were studied. The electronic absorption spectra of the complex in aqueous solution were studied. The photodynamic inhibitory activity of partial phthalocyanine-phycocyanin complex on human hepatoma cell line HepG2 was evaluated. The IC50 value of zinc phthalocyanine and phycocyanin complex, ZnPc-PCS, was 2.1 渭 M for photodynamic inhibition of HepG2. The IC50 value of the photodynamic inhibition of HepG2 by zinc tetracarboxylphthalocyanine and phycocyanin C4-PC4 with a binding ratio of 6: 1 was 0.93 渭 M. These two complexes not only showed remarkable photodynamic anticancer activity, but also had higher activity than phycocyanin and zinc phthalocyanine.
【學(xué)位授予單位】：福州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R943;O641.4

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