目的 为改善藤黄酸原料药释放的靶向性，设计制备一种由叶酸修饰的藤黄酸纳米晶-磷脂复合递药系统（folic acid-modified gambogic acid nanocrystals-phospholipid composite drug delivery system，GA-NCs@PL-FA），优化其处方工艺，并考察GA-NCs@PL-FA的理化性质、体外抗肝癌活性及体内组织分布特征。方法 通过CO₂辅助沉淀法制备藤黄酸纳米晶（gambogic acid nanocrystals，GA-NCs），再经薄膜水化法构建GA-NCs@PL-FA。结合单因素考察和星点设计-效应面法（central composite design-response surface methodology，CCD-RSM）优化处方工艺。透射电子显微镜（transmission electron microscopy，TEM）观察微观形态，测定粒径、ζ电位、包封率、载药量及稳定性；以透析袋法考察其在pH 7.4、6.5磷酸盐缓冲液（PBS，含0.5%聚山梨酯80）中的体外释放行为。利用细胞计数试剂盒-8（cell counting kit-8，CCK-8）法和细胞划痕实验，评价不同给药组对HepG2细胞活力和迁移能力的影响。采用超高效液相色谱-串联质谱法（ultra-high performance liquid chromatography-tandem mass spectrometry，UPLC-MS/MS）测定荷瘤裸鼠各组织中藤黄酸含量，分析其体内组织分布及肿瘤组织富集能力。结果 GA-NCs@PL-FA呈类球形，粒径为（183.07±0.55）nm，ζ电位为（-17.70±0.17）mV，包封率和载药量分别为（84.64±0.57）%和（4.33±0.07）%，在4、25 ℃条件下放置7 d后性质稳定；在2种释放介质中，均表现出明显的缓释效果。体外实验结果显示，与藤黄酸、GA-NCs及无叶酸修饰的藤黄酸纳米晶-磷脂复合递药系统（folic acid-free gambogic acid nanocrystals-phospholipid composite drug delivery system，GA-NCs@PL）相比，GA-NCs@PL-FA对HepG2细胞增殖和迁移的抑制作用更强，半数抑制浓度（half inhibitory concentration，IC₅₀）降至0.50 μg/mL。组织分布实验证实，GA-NCs@PL-FA可延长藤黄酸体内滞留时间，并促进其在肿瘤组织中的富集，具有良好的靶向性。结论 优化了GA-NCs@PL-FA的处方和制备工艺，体系粒径均一、载药性能较好，具有良好的缓释效果和体外抗肝癌活性，并表现出较强的肿瘤组织富集能力，可为藤黄酸的靶向制剂开发及临床应用提供参考。

Objective In order to improve the targeting release of gambogic acid, a folic acid-modified gambogic acid nanocrystals-phospholipid composite drug delivery system (GA-NCs@PL-FA) was designed and prepared, and its prescription process was optimized. The physicochemical properties, in vitro anti-hepatocarcinoma activity and in vivo tissue distribution characteristics of GA-NCs@PL-FA were investigated. Methods Gambogic acid nanocrystals (GA-NCs) were prepared by CO₂-assisted precipitation method, and then GA-NCs@PL-FA was constructed by thin film hydration method. The prescription process was optimized by single factor investigation and central composite design-response surface methodology (CCD-RSM). The microscopic morphology was observed by transmission electron microscopy (TEM), and the particle size, ζ potential, encapsulation efficiency, drug loading and stability were determined. The in vitro release behavior in pH 7.4, 6.5 phosphate buffer (PBS, containing 0.5% Tween 80) was investigated by dialysis bag method. The effects of different administration groups on the viability and migration ability of HepG2 cells were evaluated by cell counting kit-8 (CCK-8) method and cell scratch test. The content of gambogic acid in each tissue of tumor-bearing nude mice was determined by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), and its tissue distribution and tumor tissue enrichment ability were analyzed. Results The GA-NCs@PL-FA was spherical, the particle size was (183.07 ±0.55) nm, the ζ potential was (−17.70 ±0.17) mV, the encapsulation efficiency and drug loading were (84.64 ±0.57)% and (4.33 ±0.07)%, respectively. The properties were stable after 7 d at 4, 25 ℃. In both release media, it showed obvious sustained release effect. In vitro experiments showed that compared with gambogic acid, GA-NCs and folic acid-free gambogic acid nanocrystals-phospholipid composite drug delivery system (GA-NCs@PL), GA-NCs@PL-FA had a stronger inhibitory effect on the proliferation and migration of HepG2 cells, and the half inhibitory concentration (IC₅₀) was reduced to 0.50 μg/mL. The tissue distribution test confirmed that GA-NCs@PL-FA could prolong the retention time of gambogic acid in vivo and promote its enrichment in tumor tissues, with good targeting. Conclusion The formulation and preparation process of GA-NCs@PL-FA were optimized. The system had uniform particle size, good drug loading performance, good sustained release effect and in vitro anti-hepatoma activity, and showed strong tumor tissue enrichment ability. It can provide reference for the development and clinical application of gambogic acid targeted preparations.

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黑龙江省北药与功能食品特色学科项目（HLJTSXK-2022-03）；黑龙江省自然科学基金项目（LH2024H003）；国家大学生创新训练计划（202510222166）；黑龙江省教育厅创新团队项目（2024-KYYWF-0622）；佳木斯大学“东极”学术团队项目（DIXSTD202414）