【作者】 李扬；

【导师】 高清志； 韩建斌；

【作者基本信息】 天津大学 ， 药学， 2021， 硕士

【摘要】 荧光成像技术是生物研究和临床诊断中最广泛,最有力的可视化技术之一,具有选择性、可见性和可调性。随着荧光探针的多样化及相关仪器的改进,荧光成像技术已广泛应用于肿瘤的成像。靶向荧光探针可特异性点亮病变细胞,提高早癌的检出率。大环主体分子以其特有的空腔结构为生物体内的一些复杂结构和过程提供重要模型,将糖类连接到各种大环主体分子中不仅可以制备出多种结构新颖的糖基缀合物,而且为大环分子提供了新的作用位点,丰富了其功能性,同时也改善了大环分子在某些应用方面的限制。本课题基于肿瘤沃伯格效应,由于肿瘤细胞与健康细胞相比特异性高表达糖转运通道蛋白GLUT,因此以GLUT1为肿瘤特异性标志物实现对肿瘤的靶向识别。以葡萄糖,甘露糖和半乳糖对杯芳烃的下缘进行衍生,同时引入荧光基团,将杯芳烃设计,合成和表征了一系列新型的硝基苯并呋喃（NBD）标记的杯[4]芳烃荧光探针（NBD-Gly-CA）。合成中所有的重要中间体和终产物都经过了¹H NMR、¹³C NMR和高分辨质谱等方法的结构确认,同时通过细胞毒性试验证明了该探针无毒,对GLUT1具有靶向性且在肿瘤细胞中可定位成像。本课题的创新之处在于:（1）利用肿瘤细胞的沃伯格效应,能够直接用在细胞系中分析和检测肿瘤细胞GLUT蛋白的表达程度,从而完成对肿瘤细胞的筛选。为肿瘤早期筛选和诊断,发现和开发新的抗肿瘤药物,预测和评估肿瘤对化疗药物的耐药性以及治疗高表达GLUT蛋白等肿瘤患者等提供了有效的手段和有用的工具。（2）以杯芳烃作为结构平台,引入不同的荧光基团,使其在肿瘤细胞中荧光成像,从而达到对肿瘤细胞的识别。（3）利用糖杯芳烃化合物具有将不同药物容纳到空腔中的能力,将铂类药物封装到杯芳烃腔内,改善铂类药物的溶解性以及抗癌活性。总之,本课题设计合成的新型杯芳烃探针实现了对肿瘤细胞的靶向成像及筛选,为靶向沃伯格效应的新型杯芳烃荧光探针的设计和开发提供了一定的科学依据。更多还原

【Abstract】 Fluorescence imaging technology is one of the most extensive and powerful visualization technologies in biological research and clinical diagnosis,with selectivity,visibility and adjustability.With the diversification of fluorescent probes and the improvement of related instruments,fluorescence imaging technology has been widely used in tumor imaging.Targeted fluorescent probes can specifically illuminate diseased cells and increase the detection rate of early cancer.With its unique cavity structure,the macrocyclic host molecule provides an important model for some complex structures and processes in the organism.Connecting sugars to various macrocyclic host molecules can not only prepare a variety of novel glycosyl conjugates.,And provide new action sites for macrocyclic molecules,enrich their functionality,and at the same time improve the limitations of macrocyclic molecules in certain applications.This topic is based on the tumor Warburg effect.Since tumor cells specifically express the sugar transport channel protein GLUT compared with healthy cells,GLUT1 is used as a tumor-specific marker to achieve targeted tumor recognition.The lower edge of the calixarene was derivatized with glucose,mannose and galactose,and a fluorescent group was introduced at the same time.The calixarene was designed,synthesized and characterized by a series of new nitrobenzofuran（NBD）labeled calixarene aromatic fluorescent probe（NBD-Gly-CA）.All the important intermediates and final products in the synthesis have been structurally confirmed by ¹H NMR,¹³C NMR and high-resolution mass spectrometry.At the same time,the cytotoxicity test proved that the probe is non-toxic,has a targeting ability to GLUT1 and can be positioned and imaged in tumor cells.The innovations of this subject are:（1）Using the Warburg effect of tumor cells,it can be directly used in cell lines to analyze and detect the expression level of GLUT protein in tumor cells,thereby completing the screening of tumor cells.It provides effective means and useful tools for the early screening and diagnosis of tumors,the discovery and development of new anti-tumor drugs,the prediction and evaluation of tumor resistance to chemotherapeutic drugs,and the treatment of tumor patients with high expression of GLUT protein.（2）Using calixarene as a structural platform,introducing different fluorophores to enable fluorescence imaging in tumor cells to achieve the role of positioning tumor cells.（3）Utilizing the ability of the sugar calixarene compound to contain different drugs in the cavity,the platinum drugs are encapsulated in the calixarene cavity to improve the solubility and anti-cancer activity of the platinum drugs.In short,the new calixarene probe designed and synthesized in this subject realizes the targeted imaging and screening of tumor cells,and provides a certain scientific basis for the design and development of a new calixarene fluorescent probe targeting the Warburg effect.更多还原