目的 采用等网状金属有机骨架-1（isoreticular metal-organic framework-1，IRMOF-1）包载中药抗肿瘤活性成分重楼皂苷VI（polyphyllin VI，PPVI）并包覆穿膜肽构建靶向纳米制剂（PEG-PTP/PPVI@IRMOF-1），对其进行表征和体外抗肿瘤药效初步评价。方法 通过“三乙胺沉淀法”和“反应包覆法”制备PEG-PTP/PPVI@IRMOF-1；利用粒径与ζ电位测定、扫描电子显微镜（scanning electron microscopy，SEM）分析、傅里叶变换红外光谱（Fourier-transform infrared spectroscopy，FTIR）分析、热重分析（thermogravimetric analysis，TGA）、气体吸附（Brunauer-Emmett-Teller，BET）分析、X射线光电子能谱（X-ray photoelectron spectroscopy，XPS）分析对其粒径、形貌、热稳定性、比表面积等性质进行表征；溶血实验评价纳米材料的生物安全性；CCK-8法评价材料安全性并检测纳米粒对人胰腺癌PANC-1细胞增殖的影响；荧光显微镜观察材料及纳米粒对PANC-1细胞核形态的影响；激光共聚焦实验考察纳米粒的体外靶向性；克隆形成实验、划痕实验考察纳米粒对细胞增殖、迁移的影响；流式细胞仪检测纳米粒对细胞凋亡率和细胞内活性氧（reactive oxygen species，ROS）水平的影响；激光共聚焦实验考察纳米粒对细胞线粒体膜电位（mitochondrial membrane potential，MMP）的影响。结果 成功制备了靶向纳米粒PEG-PTP/PPVI@IRMOF-1，为干燥的白色粉末状。纳米粒外观为类球形，粒径为（177.47±3.97）nm，ζ电位为（−7.94±0.14）mV；PPVI@IRMOF-1的载药量为（43.02±0.08）%；纳米材料具有良好的生物安全性；靶向肽修饰后能够明显增强肿瘤细胞对药物的摄取，显著抑制细胞增殖，促进细胞中ROS的产生和MMP的下降，诱导肿瘤细胞的凋亡。结论 制备的纳米粒形状均一，载药量较高，安全性良好，增强了PPVI的抗肿瘤疗效，能够为PPVI抗肿瘤制剂的开发提供参考。

Objective A targeted nanoformulation (PEG-PTP/PPVI@IRMOF-1) was constructed by encapsulating the traditional Chinese medicine antitumor active component polyphyllin VI (PPVI) within isoreticular metal-organic framework-1 (IRMOF-1) and coating it with a cell-penetrating peptide. The study aims to characterize the nanoformulation and conduct a preliminary evaluation of its in vitro antitumor efficacy. Methods PEG-PTP/PPVI@IRMOF-1 was fabricated via the “triethylamine precipitation method” and “reaction coating method”; The physicochemical properties of the nanoformulation, including particle size, morphology, thermal stability, and specific surface area, were characterized by particle size and ζ-potential measurement, scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), Brunauer-Emmett-Teller (BET) gas adsorption, and X-ray photoelectron spectroscopy (XPS). Hemolysis assay was used to assess the biosafety of nanomaterials. CCK-8 assay was adopted to evaluate the material safety and detect the effect of nanoparticles on the proliferation of human pancreatic cancer PANC-1 cells. The effect of materials and nanoparticles on the nuclear morphology of PANC-1 cells was observed by fluorescence microscopy; The targeting capability of the nanoparticles ex vivo was assessed by laser scanning confocal microscopy; The effects of nanoparticles on cell proliferation and migration were examined using colony formation and scratch assays, respectively; Flow cytometry was employed to determine the impact of the nanoparticles on the cellular apoptosis rate and intracellular reactive oxygen species (ROS) levels; The effect of nanoparticles on the mitochondrial membrane potential (MMP) of cells was examined by laser confocal microscopy. Results The targeted nanoformulation PEG-PTP/PPVI@IRMOF-1 was successfully fabricated as a dry white powder. The nanoparticles exhibited a quasi-spherical morphology with an average particle size of about (177.47 ± 3.97) nm and a ζ potential of (−7.94 ± 0.14) mV; The drug loading capacity (DLC) of PPVI@IRMOF-1 was (43.02 ± 0.08)%; The nanomaterial showed excellent biocompatibility; Notably, targeted peptide modification can significantly enhance the drug uptake of tumor cells, significantly inhibit cell proliferation, promote the production of ROS and the decrease of MMP in cells, and induce apoptosis of tumor cells. Conclusion The prepared PEG-PTP/PPVI@IRMOF-1 nanoparticles possess uniform morphology, high drug loading capacity, and good biocompatibility. Moreover, the nanoformulation significantly enhances the in vitro antitumor efficacy of PPVI, providing a reference for the development of PPVI-based antitumor formulations.

R283.6

北京市自然科学基金项目（7262198）；国家中医药管理局高水平建设学科—中药药剂学（zyyzdxk-2023272）