目的 合成两亲性聚合物聚乙二醇-聚L-天冬氨酸［methoxy poly （ethylene glycol）-b-poly （β-benzyl L-aspartate），PEG-b-PBLA］和pH敏感性聚合物［octadecylamine-p （API-Asp）₁₀，OAPI］，并利用PEG-b-PBLA包载OAPI和抗肿瘤药物姜黄素，构建pH敏感性载药胶束，考察其理化性质，并评价其体内外抗非小细胞肺癌（non-small cell lung cancer，NSCLC）效果。方法 利用甲氧基聚乙二醇胺［methoxy poly （ethylene glycol） amine，mPEG₂₀₀₀-NH₂］引发L-天冬氨酸-4-苄酯-N-羧基环内酸酐［benzyl （S）-2，5-dioxooxazolidine-4-acetate，BLA-NCA］开环聚合制备两亲性聚合物PEG-b-PBLA。利用硬脂胺引发BLA-NCA开环聚合和氨解反应合成具有pH敏感特性的聚合物OAPI。采用透析法制备载姜黄素的具pH敏感特性胶束（C-NP），并对其粒径、稳定性和pH响应性体外药物释放等性质进行了考察。最后，利用NSCLC A549细胞、三维肿瘤球模型以及A549细胞建立的荷瘤小鼠模型考察了C-NP胶束的体内外抗肿瘤效果。结果 透射电子显微镜结果显示，所制备的C-NP呈类球形，具有明显的核壳结构，且分布均匀，其粒径为（165.32±1.53）nm，ζ电位为（-5.24±0.42）mV，载药量为（5.27±0.14）%，包封率为（88.46±3.28）%，临界胶束浓度为4.82 μg/mL。体外稳定性实验显示在生理条件下，C-NP具有较好的稳定性，敏感性实验显示在酸性条件下，C-NP不稳定，具pH敏感特性。体外药物释放结果表明，在pH 5.0的介质中孵育72 h后，C-NP的累积释放量高达（81.44±5.90）%，表明C-NP胶束具有良好的pH敏感性。细胞毒性实验、肿瘤球生长抑制实验证明C-NP能增强姜黄素体外抗肿瘤活性。细胞共定位实验显示，相比较pH不敏感组C-B-NP，C-NP可以在酸性溶酶体中快速分解，从而逃逸发挥抗肿瘤效应。体内药效学实验结果表明，C-NP组显著抑制了荷瘤裸鼠的肿瘤生长。结论 成功构建了载有姜黄素的pH敏感性胶束C-NP，其具有良好的理化性质和pH敏感性，且通过多种模型初步证实其对NSCLC非小细胞肺癌具有较强的抑制作用。

Objective To synthesize the amphiphilic polymer methoxy poly(ethylene glycol)-b-poly(β-benzyl L-aspartate) (PEG-b- PBLA) and pH-sensitive polypeptide (OAPI), then, PEG-b-PBLA was used to construct pH-sensitive drug-carrying micelles containing OAPI and curcumin, finally, the physicochemical properties and anti-non-small cell lung cancer (NSCLC) effects of the micelles were evaluated in vivo and in vitro. Methods The methoxy poly(ethylene glycol) amine (mPEG₂₀₀₀-NH₂) was utilized to initiate benzyl (S)-2,5-dioxooxazolidine-4-acetate (BLA-NCA) to synthesize the amphiphilic polymer PEG-b-PBLA by the ring-opening polymerization, and the pH-sensitive polypeptide OAPI was synthesized by ring-opening polymerization of BLA-NCA triggered by stearamine and ammonolysis reaction. Then, the pH sensitive micelle (C-NP) containing curcumin was prepared by dialysis method. The particle size, stability and pH-sensitive drug release properties of C-NP were investigated. Finally, the in vitro and in vivo anti-tumor effects of C-NP micelle were evaluated by using A549 cells, three-dimensional tumor ball models and tumor bearing mice model of NSCLC. Results The prepared C-NP showed a uniform spherical shape with obvious core-shell structure, uniform distribution under transmission electron microscopy. Its particle size was (165.32 ±1.53) nm, ζ potential was (-5.24 ±0.42) mV, drug loading was (5.27 ±0.14)%, and encapsulation effenciency was (88.46 ±3.28)%. The critical micellar concentration was 4.82 μg/mL. In vitro stability experiments showed that under physiological conditions, C-NP had good stability, sensitivity experiments showed that under acidic conditions, C-NP was unstable, with pH sensitive characteristics. In vitro release results showed that the cumulative release of C-NP reached (81.44 ±5.90)% after 72 h incubation in pH 5.0 medium, indicating that C-NP micelles had good pH sensitivity. The cytotoxicity, tumor spheroid growth inhibition experiments revealed C-NP can enhance the anti-tumor activity in vitro. The cell colocalization experiment showed that, compared with the pH-insensitive group C-B-NP, C-NP could be decomposed rapidly in acid lysosomes, thus escaping to play an anti-tumor effect. In vivo pharmacodynamic experiment results showed that C-NP group significantly inhibited tumor growth in nude mice. Conclusion In this experiment, the pH sensitive C-NP micelle was successfully constructed, which has good physicochemical properties, pH sensitivity, and strong inhibitory effect on NSCLC preliminarily confirmed by a variety of models.

R283.6

2021年度江苏省高校基础科学（自然科学）面上项目（21KJD350004）；2020年度江苏省高校基础科学（自然科学）面上项目（20KJD350007）；江苏省大学生创新创业训练计划项目（202113980003Y）；南京医科大学康达学院2021年度科研发展基金（KD2021KYJJZD010）；南京医科大学康达学院第二期“科研创新团队项目”（KD2023KYCXTD003）