目的 制备依维莫司磷脂-壳聚糖自组装纳米粒（EVR-LC/CH NPs），考察其药剂学特性、在Caco-2单层细胞模型中的跨膜转运性能，并评价其对人乳腺癌MCF-7细胞的抗肿瘤效果。方法 采用离子凝胶法制备EVR-LC/CH NPs。通过测定粒径分布、Zeta电位、透射电镜形态观察以及体外药物释放行对EVR-LC/CS NPs进行表征；利用Caco-2单层细胞模型评价EVR-LC/CH NPs的跨膜转运性能；比较EVR-LC/CH NPs与依维莫司原料药的体外抗肿瘤效果。结果EVR-LC/CH NPs的最佳处方为磷脂与药物质量比6∶1，磷脂与壳聚糖质量比20∶1，pH值为4～5。在该条件下制备的EVR-LC/CH NPs的平均粒径为（151.5±5.3）nm，PDI为0.213±0.006，Zeta电位为（39.1±0.5）mV。透射电镜下可清晰观察到其具有“核–壳”结构；体外释放研究表明，EVR-LC/CH NPs呈现双相释放模式，且具有良好的pH值响应特性；Caco-2单层细胞转运实验显示，EVR-LC/CH NPs能有效促进药物在Caco-2单层细胞中的跨膜转运；与依维莫司原料药相比，EVR-LC/CH NPs对人乳腺癌MCF-7细胞的抑制作用更显著。结论 制备的EVR-LC/CH NPs能够有效调控药物的体外释放行为，增强药物的跨膜转运能力，并显著提高药物的抗肿瘤效果。

Objective To prepare everolimus phospholipid-chitosan self-assembled nanoparticles (EVR-LC/CH NPs), investigate their pharmaceutical properties, transmembrane transport performance in the Caco-2 monolayer cell model, and to evaluate their antitumor effects on human breast cancer MCF-7 cells. Methods EVR-LC/CH NPs were prepared using the ion gelation method. The characterization of EVR-LC/CS NPs was conducted by measuring particle size distribution, Zeta potential, transmission electron microscopy morphology, and in vitro drug release behavior determination. The transmembrane transport performance of EVR-LC/CH NPs was evaluated using the Caco-2 monolayer cell model. The in vitro antitumor efficacy of EVR-LC/CH NPs was compared with that of the active pharmaceutical ingredient of everolimus. Results The optimized formulation for EVR-LC/CH NPs was as follows: the ratio of phospholipid to drug was 6:1, the ratio of phospholipid to chitosan was 20:1, and the pH value was pH 4 - 5. Under these conditions, the EVR-LC/CH NPs had an average particle size of (151.5 ± 5.3) nm, a PDI of 0.213 ± 0.006, and a Zeta potential of (39.1±0.5) mV. Transmission electron microscopy clearly observed a "core-shell" structure. The in vitro release studies showed a biphasic release pattern with excellent pH-responsive properties. Caco-2 monolayer cell transport experiments showed that EVR-LC/CH NPs effectively promoted transmembrane transport of the drug in Caco-2 monolayer cells. Compared to the active pharmaceutical ingredient of everolimus, EVR-LC/CH NPs exhibited more significant inhibitory effects on human breast cancer MCF-7 cells. Conclusion The prepared EVR LC/CH NPs can effectively regulate the in vitro release behavior of drugs, enhance the transmembrane transport ability of drugs, and significantly improve the antitumor effect of drugs.

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湖北省科技计划项目（2019CFC886）