高选择性双锁型近红外靶向次氯酸荧光探针的构建及其在急性肾毒性中的生物应用

Drugs, an important method of disease prevention, treatment and diagnosis, are also one of the factors that cause acute kidney injury (AKI). Studies have shown that reactive oxygen species (such as HOCl) burst induced by drugs is a key factor leading to AKI, but the relation and the evolvement rules between HOCl and drug-induced AKI in situ are still difficult to study due to the lack of effective tools at the subcellular and in vivo levels. In view of the above problem, this project plans to construct a series of near-infrared targeted HOCl fluorescent probes with high selectivity and sensitivity to in situ tracing drug-induced AKI at the cellular, subcellular, and in vivo levels. The probes will be synthesized by covalently incorporating high-stable Si-rhodamine dyes with dithiobenzoic acid lactone recognition unit, organelle and kidney targeting groups. The near-infrared fluorescence signal changes will be triggered by the specific reactions of these probes with HOCl. Further, the AKI mouse model will be established through the common drug-gentamicin induction, and the detection performance of these probes on drug-induced AKI and remediation pathways of drugs will be systematically investigated at the in vivo level. Combined with the near-infrared excitation/ emission and high tissue permeability, the proposed probes are expected to provide new methods for the study of nephrotoxicity screening as well as the remediation mechanisms of its therapeutic drugs in the development of new drugs.

药物作为疾病预防、治疗和诊断的重要手段，也是造成急性肾损伤（AKI）的因素之一。研究表明药物诱导的活性氧（如HOCl）爆发是导致AKI的关键因素，但迄今仍缺少合适的工具在亚细胞和活体水平原位研究HOCl与药物性AKI的关联性及其演变规律。针对上述问题，本项目拟采用高稳定性的近红外硅罗丹明染料为荧光团，以二硫代苯甲酸内酯作为识别单元，通过探针与HOCl的特异开环反应引发近红外荧光信号变化。在此基础上，修饰细胞器靶向和肾靶向基团，构建高选择性和灵敏度的近红外靶向HOCl荧光探针，从而在细胞、亚细胞器和活体水平实现药物性AKI的高灵敏原位示踪。同时，以常见药物-庆大霉素为模型药物引发小鼠的肾氧化损伤，系统性考察探针在活体层次的药物性肾氧化损伤及修复途径中的检测效果。结合探针的近红外激发和发射的高组织渗透能力，拟发展的探针有望为新药开发过程中的肾毒性筛查及其修复途径机制研究提供新方法。

本项目旨在通过理性设计策略，构建一系列高保真的近红外靶向 HOCl 荧光探针，实现药物引起的 AKI 及保肾类药物修复途径过程中 HOCl 的高灵敏原位示踪。按照项目计划的指导，同时根据项目的实际进展情况做出了些许调整，申请人设计合成了高保真过氧化亚硝酰阴离子，谷胱甘肽和γ-谷氨酰转肽酶荧光探针。药物诱导肾损伤 (DIKI)和药物性肝损伤（DILI）是一种急性疾病。目前已认为过氧化亚硝酰阴离子（ONOO−，一种高活性的活性氧物种）谷胱甘肽（GSH）和γ-谷氨酰转移酶（GGT）可作为其重要的生物指标。因此，本项目拟通过合理设计策略，构建系列高选择性荧光探针，并将其用于细胞和活体水平上DIKI和DILI疾病中成像研究。我们期望该探针未来能成为临床领域诊断早期DIKI和DILI的潜在工具。