目的 构建透明质酸（hyaluronic acid，HA）修饰的载重楼皂苷VII（polyphyllin VII，PPVII）金属有机骨架材料IRMOF-10靶向纳米粒HA@IRMOF-10@PPVII，并系统评价其理化特性、安全性和体外抗三阴性乳腺癌（triple negative breast cancer，TNBC）活性。方法 采用三乙胺法制备金属有机骨架材料IRMOF-10，通过异位包封法包载PPVII，并利用透明质酸对载药纳米粒表面进行修饰，构建靶向纳米粒HA@IRMOF-10@PPVII；采用HPLC法测定其载药量，运用扫描电子显微镜（scanning electron microscopy，SEM）、傅里叶变换红外光谱（Fourier transform infrared spectroscopy，FTIR）分析、热重分析（thermogravimetric analysis，TG）及BET比表面积法等分析手段表征纳米粒的形貌、结构及热稳定性；通过溶血实验评估其血液安全性，采用激光共聚焦显微镜验证其靶向能力，通过CCK-8法、DAPI染色、Annexin V/PI双染、迁移实验、克隆形成实验，以及细胞摄取、活性氧（reactive oxygen species，ROS）检测、线粒体膜电位（mitochondrial membrane potential，MMP）检测等方法，综合评价纳米粒对TNBC 4T1细胞的抑制作用。结果 所制备的HA@IRMOF-10@PPVII载药量为 （39.56±1.78）%（n＝3），具备良好的生物安全性，可显著降低PPVII的溶血风险；该纳米粒能够有效增强PPVII在4T1细胞中的摄取，并显著抑制细胞迁移与克隆形成；其抗肿瘤机制与诱导细胞内ROS水平升高、MMP下降触发细胞凋亡、抑制4T1细胞的迁移与克隆密切相关。结论 HA@IRMOF-10@PPVII纳米粒制备成功，具有良好的靶向性与抗TNBC活性，能够显著增强PPVII的药效，为基于PPVII的抗肿瘤制剂研发提供了实验依据。

Objective To prepare hyaluronic acid-modified polyphyllin VII-loaded metal-organic framework material IRMOF-10 targeted nanoparticles (HA@IRMOF-10@PPVII), systematically evaluate their physicochemical properties, safety and in vitro anti-triple negative breast cancer (TNBC) activity. Methods Metal-organic framework material IRMOF-10 was synthesized by triethylamine method. PPVII was encapsulated by the heterotopic encapsulation method, and the surface of the drug-loaded nanoparticles was modified with hyaluronic acid to construct the targeted nanoparticles HA@IRMOF-10@PPVII. The drug loading was determined by HPLC. The morphology, structure and thermal stability of nanoparticles were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TG) and BET specific surface area method. Its blood safety was evaluated by hemolysis test, and its cell targeting was verified by laser confocal microscopy. CCK-8, DAPI staining, Annexin V/PI double staining, migration assay, clone formation assay, cell uptake, reactive oxygen species (ROS) detection, mitochondrial membrane potential (MMP) detection and other methods were used to verify its inhibitory effect on TNBC 4T1 cells. Results The drug loading capacity of HA@IRMOF-10@PPVII was (39.56% ± 1.78)% (n = 3). HA@IRMOF-10@PPVII had good biosafety, which could significantly reduce the hemolysis risk of PPVII. This nanoparticle can effectively improve the uptake of PPVII in 4T1 cells, and significantly inhibits cell migration and clone formation. Its anti-tumor mechanism is closely related to inducing an increase in intracellular ROS levels, a decrease in MMP triggering apoptosis, and inhibiting the migration and cloning of 4T1 cells. Conclusion The HA@IRMOF-10@PPVII nanoparticles were successfully prepared, which exhibit excellent targeting ability and anti-TNBC activity. They can significantly enhance the efficacy of PPVII, providing experimental evidence for the development of PPVII-based anti-tumor formulations.

R283.6

北京市自然科学基金项目（7262198）；国家自然科学基金项目（81703715）