检测缺氧相关还原酶的高稳定近红外比率型荧光探针及其成像分析方法

Hypoxia-sensitive fluorescent probes can be used to monitor the hypoxic status of tumor cells via the detection of endogenous reductase. There have been a number of detection methods for hypoxia. Nevertheless, the fluorescent probes previously developed are inadequate for stability, emission wavelength, quantitative analysis. The research project seeks to develop a novel method with the highly stable, near-infrared, ratiometric properties for detection of reductase activity under hypoxic condition. The innovative research contents mainly include, to design and synthesize highly stable, near-infrared, ratiometric probes for hypoxia detection by using hemicyanine skeleton of IR 780, Changsha near-infrared fluorophores and rhodamine derivatives as the forster resonance energy transfer donor or acceptor, introducing the masking and reacting moieties, such as organic molecular probes and core-shell silica-based nanoparticals; to explore highly sensitive and selective fluorescent methods for reductase (e.g., nitroreductase, azoreductase) detection under hypoxic conditions. Additionally, their further application of hypoxic tumor cell and in vivo fluorescent imaging (e.g., zebrafish, nude mice) will be investigated. The research could provide useful methods for exploring the mechanism of hypoxic tumor. Hypoxia detection is of great importance for tumor and related diseases diagnosis.

缺氧特异性荧光探针是利用高特异性荧光探针对相关还原酶进行检测进而评价肿瘤缺氧状况的一类分析方法。本项目针对目前所报道的缺氧特异性荧光探针存在稳定性差、发射波长短和定量分析准确性差等问题，提出设计合成对肿瘤细胞缺氧相关还原酶具有高稳定性、近红外发射和比率响应的新荧光探针并建立其成像分析方法的研究课题。重点研究内容包括，IR-780降解产物半菁骨架、长沙近红外荧光染料和罗丹明染料的衍生物作为荧光供体或受体，引入相关还原酶（如硝基还原酶、偶氮还原酶等）的识别基团，设计合成有机分子荧光探针和介孔硅包覆上述荧光探针的核壳型纳米缺氧荧光探针；建立高灵敏度、高选择性的检测肿瘤细胞缺氧相关还原酶的荧光分析新方法；建立对缺氧条件下肿瘤细胞和活体（如斑马鱼、小鼠等）内相关还原酶的成像分析新方法。这些研究将为肿瘤缺氧研究提供优良的荧光探针和分析方法，对恶性肿瘤及相关疾病的诊断具有重要的科学意义和应用价值。

缺氧特异性荧光探针是利用高特异性荧光探针对相关还原酶进行检测进而评价肿瘤缺氧状况的一类分析方法。本项目针对目前所报道的缺氧特异性荧光探针存在稳定性差、发射波长短和定量分析准确性差等问题，创新性开展了一系列对肿瘤细胞缺氧相关还原酶具有高稳定性、近红外发射和比率响应的新荧光探针并建立其成像分析方法的研究，完成了预定的研究目标，取得了一系列创新性研究成果。以IR-780降解产物半菁骨架作为荧光供体或受体，引入相关检测物的识别基团，设计合成10余种近红外有机分子荧光探针；建立10余种高灵敏度、高选择性的检测肿瘤细胞相关检测物的荧光分析新方法；建立10余种肿瘤细胞和活体内相关检测物的成像分析新方法。这些研究成果将为肿瘤缺氧研究提供优良的荧光探针和分析方法，对恶性肿瘤及相关疾病的诊断具有重要的科学意义和应用价值。已在SCI源刊上发表研究论文11篇，其中SCI一区文章8篇，授权国家专利2个，培养研究生6名。