【摘要】 首先以2,4-二甲基吡咯为原料合成氟硼二吡咯化合物(BODIPY)的核心骨架,以咔唑为原料合成4-二芳氨基甲醛衍生物,再与核心骨架发生Knoevenagel缩合反应引入到BODIPY的3、5位,得到了最大发射波长680 nm的新型BODIPY荧光染料.之后将RGD多肽与DSPE-PEG2000-MAL连接,得到具有肿瘤靶向性的两亲性聚合物嵌段(DSPE-PEG2000-MAL-RGD),随后将BODIPY荧光染料与DSPE-PEG2000-MAL-RGD进行自组装,得到水溶性良好的BODIPY-PC-RGD荧光探针复合物.通过核磁共振氢谱、荧光发射光谱、场发射扫描电镜、场发射透射电镜和荧光显微镜对荧光探针的光学性质进行测试,发现其形貌规则,大小均一,粒径大约为143 nm,具有良好的近红外荧光特性.另外,还通过细胞毒性实验、共聚焦实验及细胞流式实验对荧光探针的生物学性能进行评价.在一定浓度范围(低浓度)内,探针对U87和Hela胶质瘤细胞表现出低毒性甚至是无毒性.从共聚焦实验图像以及细胞流式实验结果可知,当细胞与BODIPY-PC-RGD孵育时,荧光探针进入细胞的量更多,这在一定程度上证明了复合物BODIPY-PC-RGD对肿瘤细胞具有一定的靶向性,其在未来肿瘤检测及诊断领域具有长远的发展潜力.更多还原

【Abstract】 First, 2,4-dimethylpyrrole and carbazole were used as the starting materials to synthesize the BODIPY framework and 4-diarylamino formaldehyde derivative, respectively. The strong electron-donating groups were introduced to the C-3,5 position of the framework by the Knoevenagel reaction, contributing to a new fluorochrome whose emission wavelength can be tuned to 680 nm. Subsequently, the long chain(DSPE-PEG2000-MAL) was connected to the target group RGD with thiol group to form an amphiphilic compound(DSPE-PEG2000-MAL-RGD). Then, BODIPY was self-assembled with DSPE-PEG2000-MAL-RGD to form nanoparticles(BODIPY-PC-RGD), which could be targeted to improve the water solubility and biocompatibility of BODIPY. The structure of BODIPY was confirmed by 1 HNMR, and its properties were tested by fluorescence microscopy, SEM, and TEM. After self-assembly, the nanoparticles were approximately 143 nm in size and had a uniform morphology. The properties of the fluorescent probe were evaluated by cytotoxicity, co-focusing, and flow cytometry experiments. The results showed that the probes exhibited weak, even negligible, cytotoxicity to the U87 and Hela cells at low concentration. When the cells were treated with BODIPY-PC-RGD, more probes could penetrate the cells, which were observed from the fluorescent areas in the graphs. The results indicated that BODIPY-PC-RGD possesses the capability of targeting cancer cells to some extent, showing its long-term development potential in the field of tumor detection and diagnosis in the future.更多还原