荧光传感器的合成及应用和克服MDR自组装抗癌药物的合成及表征

发布时间：2018-04-09 15:09

本文选题：荧光探针　切入点：Cu~(2+)_L-组氨酸　出处：《中国科学技术大学》2014年博士论文

【摘要】：荧光分子探针分析技术是一种重要的分子检测技术,具有灵敏度高、选择性好、检测方便、检测限低等特点。近几十年来,荧光分子探针技术在环境科学、医药以及生命科学等领域被广为应用。在论文中我们发展了一种新型的荧光可控探针Ac-SAACQ-Gly-Gly-Gly-Lys (FITC),该探针能被用于连续的选择性在体外和活细胞中检测Cu2+和L-组氨酸。分子探针结构中包含的Gly-Gly-Gly-Lys能够增加探针的水溶性和细胞渗透性；分子探针结构中SAACQ部分能够与Cu2+发生螯合作用；分子探针的荧光信号则来自于异硫氰酸荧光素(FITC),当探针分子的SAACQ部分与Cu2+结合以后由于PET现象的出现导致荧光素的荧光发生淬灭。最终我们成功实现了体外和活细胞内Cu2+以及L-组氨酸的连续性检测。定点标记蛋白在检测蛋白的表达、组合、转录,理解细胞内部随时间空间的变化具有重要的意义。在过去的十年中荧光蛋白已经广泛的应用到标记相应的fusion蛋白,并且具有很好的分辨率。但是它们具有高分子量以及对细胞环境迟钝的缺点。而化学探针则具有高标记的特点。在论文中我们通过合理改进传统的基于1,2-氨基硫醇和2-氰基苯并噻唑(CBT)的氰基的缩合反应中的底物,将该反应应用到分子成像。我们的研究体系结合了巯基与马来酰亚胺之间的亲核加成反应和CBT的-CN与半胱氨酸(Cys)的N端之间的缩合反应,最后成功的对鸡蛋膜上蛋白质的Cys残基的进行荧光标记。在临床上癌症多药耐药(MDR)的存在是癌症病人实施化学治疗失败的主要原因。多药耐药不仅对接触肿瘤细胞的药物产生耐药,而且对一些未曾接触、与之化学结构和作用机制完全不同的药物也产生交叉耐药,因此这给癌症的治疗带来了很大的障碍。而纳米载药体系的出现为克服MDR带来重大的突破。在论文中我们合理的设计了四种抗癌药物1,2,3以及对照化合物3-Scr。其中,2是化疗药物阿霉素(DOX)的衍生物。其他化合物则是紫杉醇(taxol)的衍生物。1,2,3都包含一段Arg-Val-Arg-Arg (RVRR)的肽链,该肽链能够被furin酶特异性的识别和剪切。另外,RVRR肽链片段使底物具有很好的水溶性以及细胞渗透性。当药物分子被furin酶剪切或者还原剂还原以后,缩合反应就会发生形成多聚体,进而发生自组装形成纳米颗粒,同时增加细胞内化疗药物的有效浓度。
[Abstract]:Fluorescence molecular probe analysis is an important molecular detection technology with high sensitivity, good selectivity, easy detection and low detection limit.In recent decades, fluorescent molecular probe technology has been widely used in environmental science, medicine and life sciences.In this paper, we have developed a novel fluorescent controllable probe, Ac-SAACQ-Gly-Gly-Gly-Lys FITCX, which can be used for the continuous selective detection of Cu2 and L-histidine in vitro and in living cells.The Gly-Gly-Gly-Lys contained in the molecular probe structure can increase the water solubility and cell permeability of the probe, and the SAACQ part of the molecular probe structure can chelate with Cu2.The fluorescence signal of the probe comes from the fluorescence of fluorescein isothiocyanate (FITC). When the SAACQ part of the probe molecule binds to Cu2, the fluorescence of fluorescein is quenched due to the PET phenomenon.Finally, we successfully achieved in vitro and in vivo Cu2 and L-histidine continuity detection.It is of great significance to detect the expression, combination, transcription and understanding of the changes in cell interior with time and space.In the past decade, fluorescent proteins have been widely used to label the corresponding fusion proteins and have good resolution.But they have the disadvantages of high molecular weight and being slow to the cell environment.Chemical probes are characterized by high labeling.In this paper, we apply this reaction to molecular imaging by reasonably improving the substrate in the traditional condensation reaction of cyanide based on 1- (2-aminothiol) and 2-cyanobenzothiazolium (CBT).Our system combines the nucleophilic addition reaction between mercapto and maleimide and the condensation reaction between the N-terminal of CBT and cysteine cysteine. Finally, the fluorescent labeling of the Cys residue of protein on egg membrane is carried out successfully.The presence of MDR is the main reason for the failure of chemotherapy in cancer patients.Multidrug resistance (MDR) is not only resistant to drugs in contact with tumor cells, but also cross-resistant to drugs that have not been exposed to and have completely different chemical structure and mechanism of action. Therefore, this has brought great obstacles to the treatment of cancer.The emergence of nano-drug-loaded system has brought about a great breakthrough in overcoming MDR.In this paper, we reasonably designed four anticancer drugs 1H 2C 3 and the control compound 3 Scr3.It is a derivative of doxorubicin (DOX), a chemotherapeutic drug.The other compounds are taxol taxol. the derivative of taxol.1ON2O3 contains a peptide chain of Arg-Val-Arg-Arg rvr. the peptide chain can be specifically identified and cut by furin enzyme.In addition, the RVRR peptide chain fragment makes the substrate have good water solubility and cell permeability.When the drug molecule is shredded by furin enzyme or reduced by reducing agent, the condensation reaction will take place to form a polymer, and then self-assemble to form nanoparticles, and increase the effective concentration of chemotherapeutic drugs in cells at the same time.
【学位授予单位】：中国科学技术大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R914;O657.3

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