Norcantharidin (NCTD), a demethylated derivative of cantharidin, is an anticancer active component in traditional Chinese medicine. At present, the main methods for finding its target proteins are pharmacological methods and biophysical screening, which cannot achieve the purpose of efficient and accurate screening. Here we established a new analytical method for specific fishing and assisted imaging for norcantharidin target proteins. For the AIE supramolecule probe, the benzophenone azide (BPA) fluorescent nanoparticles with strong AIE properties were encapsulated in biocompatible DSPE-PEG that covalently coupled with NCTD (named BPA@NCTD NPs). The target proteins of NCTD can be captured by BPA@NCTD NPs, and then be detected to investigate the potential signaling pathways. The screened differential proteins were analysed through the protein and signaling pathway database, and multiple signaling pathways were obtained and verified. The mechanism of norcantharidin in inhibiting the migration and invasion of A549 cells through the P53 signaling pathway was confirmed by Western blot experiments. Our research showed that AIE supramolecule probe BPA@NCTD NPs has the dual functions of specific screening of A549 cells target proteins and biological imaging, which not only offers a good anti-fluorescence quenching ability for the dynamic imaging process of NCTD, but also provides a novel and efficient specific method for efficient analysis of target proteins and signal pathways.

去甲斑蝥素（NCTD）是斑蝥素的去甲基化衍生物，是中药中的抗癌活性成分。目前寻找其靶蛋白的方法主要是药理学方法和生物物理筛选，不能达到高效、准确筛选的目的。在这里，我们建立了一种针对去甲斑蝥素目标蛋白的特异性捕捞和辅助成像的新分析方法。对于 AIE 超分子探针，具有强 AIE 特性的二苯甲酮叠氮化物 (BPA) 荧光纳米颗粒被封装在与 NCTD 共价偶联的生物相容性 DSPE-PEG 中（命名为 BPA@NCTD NPs）。BPA@NCTD NPs可以捕获NCTD的靶蛋白，然后对其进行检测以研究潜在的信号通路。通过蛋白质和信号通路数据库对筛选出的差异蛋白进行分析，得到并验证了多条信号通路。Western blot实验证实了去甲斑蝥素通过P53信号通路抑制A549细胞迁移和侵袭的机制。我们的研究表明，AIE超分子探针BPA@NCTD NPs具有特异性筛选A549细胞靶蛋白和生物成像的双重功能，不仅为NCTD的动态成像过程提供了良好的抗荧光猝灭能力，也为NCTD的动态成像过程提供了一个新的思路。高效分析目标蛋白和信号通路的新型高效特异性方法。Western blot实验证实了去甲斑蝥素通过P53信号通路抑制A549细胞迁移和侵袭的机制。我们的研究表明，AIE超分子探针BPA@NCTD NPs具有特异性筛选A549细胞靶蛋白和生物成像的双重功能，不仅为NCTD的动态成像过程提供了良好的抗荧光猝灭能力，也为NCTD的动态成像过程提供了一个新的思路。高效分析目标蛋白和信号通路的新型高效特异性方法。Western blot实验证实了去甲斑蝥素通过P53信号通路抑制A549细胞迁移和侵袭的机制。我们的研究表明，AIE超分子探针BPA@NCTD NPs具有特异性筛选A549细胞靶蛋白和生物成像的双重功能，不仅为NCTD的动态成像过程提供了良好的抗荧光猝灭能力，也为NCTD的动态成像过程提供了一个新的思路。高效分析目标蛋白和信号通路的新型高效特异性方法。