基于卟啉及酞菁衍生物磷光探针的设计、合成及应用研究

发布时间：2018-03-04 12:45

本文选题：金属卟啉衍生物　切入点：金属酞菁衍生物　出处：《太原理工大学》2017年硕士论文　论文类型：学位论文

【摘要】：癌症是全球范围内威胁人类健康的一大杀手,它和心血管疾病已经成为医学上难以克服的两大难题,因此癌症的诊断和治疗成为材料学、化学和生物医学等领域的研究热点之一。癌症的诊断技术包括各种非入侵式的肿瘤成像技术,例如核磁共振成像、超声波成像、荧光成像等,通过这些方法可诊断肿瘤组织的形态及代谢情况。其中,荧光成像技术灵敏度高,副作用小,不含放射性物质且操作简单,因此安全性更好。就治疗技术而言,光动力治疗(PDT)是一种新兴的肿瘤治疗手段,它具有肿瘤靶向性好,副作用小,不损伤病人正常器官等特点,因而近些年来PDT发展迅速。光动力治疗需具备三个基本要素:光敏剂、激发光与分子氧。卟啉衍生物是一类传统的光敏剂,已经被很多国家批准应用于临床实践中。其中铂卟啉和钯卟啉的衍生物具有磷光量子产率高、三线态寿命长、斯托克斯位移大、光稳定性好等优点,因此其作为磷光探针在荧光成像上有很高的研究价值。酞菁作为光动力治疗的第二类光敏剂有其独特优势:化学性质稳定,合成方法简单,在可见光区600—700 nm的摩尔吸光系数是卟啉的20-50倍,无毒,对肿瘤组织具有选择性和较强杀伤力,且对周围正常组织影响较小,因此适用于癌症光动力学治疗。但是文献报道的大多数卟啉衍生物和酞菁配合物在水中溶解度较低,并且目前临床应用的光敏剂存在组织选择性差和聚集诱导猝灭等问题,在很大程度上限制了其在生物医学领域的应用。因此我们致力于合成多个系列基于金属卟啉和酞菁的的诊疗一体化高水溶性荧光纳米探针,并探究其在癌细胞诊疗方面的应用。本论文的主要研究内容如下:1.设计合成了金属卟啉小分子(PtTPP和PdTPP)和金属卟啉聚合物(Pt-P和Pd-P);将PtTPP,PdTPP,Pt-P,Pd-P通过再沉淀法制备成纳米颗粒(NPs);利用核磁共振(NMR)、红外分光光度计(FT-IR)、动态光散射粒度仪(DLS)、紫外可见吸收分光光度计(UV-vis)、荧光分光光度计(PL)等对化合物的结构、尺寸与性能进行测试表征;激光扫描共聚焦荧光显微镜成像实验结果表明Pt-PNPs对HeLa细胞具有良好的标记功能,而且Pt-PNPs随着氧浓度的降低荧光寿命有所延长;但是这些纳米颗粒均在细胞外发生不同程度的团聚,影响细胞成像质量,从而有必要进一步提高金属卟啉聚合物纳米颗粒的水溶性,增大其生物相容性。2.利用b-环糊精(b-CD)对金属卟啉化合物进行修饰合成了PtTCPP@b-CD和PdTCPP@b-CD,成功改善了金属卟啉类化合物的水溶性,之后利用CTAB对PtTCPP@b-CD和PdTCPP@b-CD进行表面电位改性,从Zeta电位测试结果得知表面电荷由负性转变为正性。同样利用1H NMR、FT-IR、DLS、UV-vis、PL等测试方法对化合物的结构与性能进行详细测试表征;MTT实验结果表明PtTCPP@b-CD@CTAB和PdTCPP@b-CD@CTAB纳米探针生物毒性较小;激光扫描共聚焦荧光显微镜成像实验结果表明这些纳米探针对癌细胞具有靶向性识别作用,同时还有高浓度荧光发射增强性质,这些研究结果证明了PtTCPP@b-CD@CTAB和PdTCPP@b-CD@CTAB纳米探针具有癌细胞诊断应用的潜力。3.利用b-CD修饰金属酞菁合成了PtTCPc@b-CD和PdTCPc@b-CD,大大提高了金属酞菁化合物的水溶性,同样利用CTAB对PtTCPc@b-CD和PdTCPc@b-CD进行表面电位改性(由负性转为正性)。利用1H NMR、FT-IR、DLS、UV-vis、PL等对化合物的结构与性能进行详细测试表征;MTT实验结果表明PtTCPc@b-CD@CTAB和PdTCPc@b-CD@CTAB生物毒性较小;激光扫描共聚焦荧光显微镜成像实验表明PdTCPc@b-CD@CTAB纳米探针具有良好的近红外细胞成像功能。
[Abstract]:Cancer is a major killer worldwide threat to human health and cardiovascular disease, it has become the two largest medical problem difficult to overcome, so the diagnosis and treatment of cancer has become one of the research hotspot in materials science, chemistry and biomedical fields. Diagnosis of cancer including tumor non invasive imaging techniques, such as magnetic resonance imaging, ultrasonic imaging, fluorescence imaging, diagnosis of morphology and metabolism of tumor tissue by these methods. The fluorescence imaging technique with high sensitivity, small side effect, non radioactive material and has the advantages of simple operation, better safety. Therefore treatment technology, photodynamic therapy (PDT) is a new tumor the treatment means, it has tumor targeting, small side effect, no damage to patients with normal organs and other characteristics, so in recent years the rapid development of PDT. Photodynamic therapy requires three base The elements: photosensitizer, luminescence and molecular oxygen. The porphyrin derivative is a kind of traditional photosensitizer, has been approved by many countries used in clinical practice. The platinum and palladium porphyrin porphyrin derivatives with high phosphorescence quantum yield of three line state, long service life, large Stokes shift, has the advantages of good light stability, so as to the phosphorescence probes in fluorescence imaging has high research value. Second kind of photosensitizer phthalocyanine as photodynamic therapy has its unique advantages: chemical stability, simple synthetic method, in 600 - 700 nm Moore visible light absorption coefficient is 20-50 times, porphyrin non-toxic, selectivity and strong lethality of tumor tissue, and less impact on the surrounding normal tissue, so it is suitable for photodynamic therapy of cancer. But most of the porphyrin derivatives and phthalocyanine complexes reported low solubility in water, and now The clinical application of the photosensitizer problems of poor tissue selectivity and aggregation induced quenching, which limits its application in the biomedical field to a great extent. Therefore, we are committed to the synthesis of multiple series of metal porphyrin and phthalocyanine and the integration of highly water-soluble fluorescent probes based on, and explore its application in the diagnosis of cancer cells. The research contents of this thesis are as follows: 1. the synthesis of metal porphyrin molecules (PtTPP and PdTPP) and the design of metal porphyrin polymers (Pt-P and Pd-P); PtTPP, PdTPP, Pt-P, Pd-P were prepared by reprecipitation method into nanoparticles (NPs); the use of nuclear magnetic resonance (NMR), infrared spectrophotometer (FT-IR), dynamic light scattering particle analyzer (DLS), UV VIS spectrophotometer (UV-vis), fluorescence spectrophotometer (PL) on the compound structure, size and performance were characterized; laser scanning confocal fluorescence microscope The experimental results show that Pt-PNPs has good markers of HeLa cells, and Pt-PNPs with the decrease of oxygen concentration and fluorescence increased life; but these nanoparticles are outside the cells of different degrees of aggregation, affecting cell imaging quality, so it is necessary to further improve the metal porphyrin polymer nanoparticles increases water solubility, biocompatibility.2. b- (b-CD) using cyclodextrin to metal porphyrin compounds were synthesized and modified PtTCPP@b-CD PdTCPP@b-CD, succeeded in improving the metalloporphyrin solubility in water, after the use of CTAB surface potential on PtTCPP@b-CD and PdTCPP@b-CD modified Zeta potential, from the test results that the surface charge changed from negative to positive. By utilizing the same 1H NMR, FT-IR, DLS, UV-vis, structure and properties of PL test method for the characterization of compounds with MTT; the experimental results show that PtTCPP@b-CD @CTAB and PdTCPP@b-CD@CTAB nano probe biological toxicity is small; laser scanning confocal fluorescence microscopy imaging experimental results show that the nano probe has a role to recognition of cancer cells, as well as high concentration enhanced fluorescence emission properties, these results show that PtTCPP@b-CD@CTAB and PdTCPP@b-CD@CTAB nano probe has the potential application of.3. diagnosis of cancer cells by using b-CD modified metal phthalocyanine PtTCPc@b-CD and PdTCPc@b-CD were synthesized, greatly improves the water solubility of metal phthalocyanine compounds, also use CTAB on PtTCPc@b-CD and PdTCPc@b-CD surface potential modification (from negative to positive). Using 1H NMR, FT-IR, DLS, UV-vis, PL etc. the structure and properties of these compounds were characterized with MTT; the experimental results show that PtTCPc@b-CD@CTAB and PdTCPc@b-CD@CTAB toxicity is small; laser scanning confocal fluorescence microscopy imaging experiment It is shown that the PdTCPc@b-CD@CTAB nanoprobe has a good imaging function of near infrared cells.

【学位授予单位】：太原理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R91;R96

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