目的 通过接枝反应将油酸（OA）修饰于羟丙基壳聚糖（HPCS）分子链，合成HPCS/OA聚合物载体，制备大麻二酚纳米胶束（CBD-HPCS/OA-NMs），并研究其抗肿瘤活性。方法 以粒径、多分散系数（PDI）、Zeta电位为评价指标，采用单因素实验筛选HPCS/OA空白纳米胶束的制备方法，通过核磁共振氢谱（1H-NMR）、傅里叶红外光谱（FT-IR）表征其结构，并测定其临界胶束浓度（CMC）。采用透析法制备CBD-HPCS/OA-NMs，测定其粒径、PDI、Zeta电位、包封率、载药量、溶解度等关键理化性质；通过透射电镜（TEM）观察其形貌；评价胶束在模拟生理环境与细胞培养环境中的体外稳定性，以及体外释药行为；通过溶血性实验考察生物安全性，CCK-8法评估CBD-HPCS/OA-NMs对SKOV3卵巢癌细胞的增殖抑制作用；借助荧光显微镜分析SKOV3细胞对CBD-HPCS/OA-NMs的摄取情况；通过细胞划痕实验考察CBD-HPCS/OA-NMs对SKOV3细胞迁移能力的影响，初步阐明其抗肿瘤作用。结果 成功合成HPCS/OA聚合物载体，其最优处方工艺为： OA用量0.6 mL、反应时间6 h、反应温度60℃，此条件下制备的HPCS/OA空白纳米胶束CMC为0.010 2 mg·mL^-1。透射电镜下CBD-HPCS/OA-NMs呈规则球形，粒径、PDI、Zeta电位分别为（194.80±1.2） nm、0.27±0.01、（39.13±3.46） mV，包封率为（72.85±0.37）%，载药量为（29.14±0.21）%，其在水中的溶解度为290 μg·mL^-1，相较于游离CBD提高了近743倍；稳定性实验结果表明，CBD-HPCS/OA-NMs在模拟生理与细胞培养环境中储存168 h粒径均无显著变化（P＞0.05），提示胶束稳定性良好；溶血性实验结果表明CBD-HPCS/OA-NMs有较小的溶血率，生物相容性好； CCK-8结果显示，HPCS/OA空白纳米胶束无细胞毒性，给药24 h后CBD-HPCS/OA-NMs对SKOV3细胞的半数抑制浓度（IC50）为8.51 μg·mL^-1；荧光显微镜观察证实，SKOV3细胞对CBD-HPCS/OA-NMs的摄取效率显著强于游离CBD；细胞划痕实验表明，CBD-HPCS/OA-NMs对SKOV3细胞迁移的抑制效果显著优于游离CBD（P＜0.05）。结论 制备的CBDHPCS/OA-NMs能有效改善CBD的水溶性与稳定性，兼具良好生物安全性与体外抗肿瘤活性。

Objective Oleic acid (OA) was modified onto the hydroxypropyl chitosan (HPCS) molecular chain through grafting reaction to synthesize the HPCS/OA polymer carrier. Cannabidiol nanomicelles (CBD-HPCS/OA-NMs) were prepared and their antitumor activities were studied. Methods Taking particle size, polydispersity index (PDI), and Zeta potential as evaluation indicators, the preparation method of blank nanomicelles of HPCS/OA was screened by single-factor experiments. Its structure was characterized by nuclear magnetic resonance hydrogen spectroscopy (1H-NMR) and fourier transform infrared spectroscopy (FT-IR), and its critical micelle concentration (CMC) was determined. CBD-HPCS/OA-NMs was prepared by dialysis method, and its key physicochemical properties such as particle size, PDI, Zeta potential, encapsulation efficiency, drug loading and solubility were determined. The morphology was observed by transmission electron microscope (TEM). The in vitro stability and drug release of micelles in simulated physiological and cell culture environments were evaluated. The biological safety was investigated through hemolytic experiments. The inhibitory effect of CBD-HPCS/OA-NMs on the SKOV3 ovarian cancer cells was evaluated by the CCK-8 method. The uptake of CBD-HPCS/OA-NMs by SKOV3 cells was analyzed by fluorescence microscopy. The effect of CBD-HPCS/OA-NMs on the migration ability of SKOV3 cells was investigated through cell scratch assay, and its anti-tumor effect was preliminarily clarified. Results The HPCS/OA polymer carrier was successfully synthesized. The optimal prescription process is as follows: OA dosage 0.6 mL, reaction time 6 h, and reaction temperature 60 ℃.The CMC of HPCS/OA blank nanomicelles prepared under this condition was 0.010 2 mg·mL^-1.The CBD-HPCS/OA-NMs showed a regular spherical shape under TEM. The particle size, PDI and Zeta potential were (194.80 ±1.2) nm, 0.27 ±0.01 and (39.13 ±3.46) mV, respectively. The encapsulation efficiency was (72.85 ±0.37) %, and the drug loading was (29.14 ±0.21) %. Its solubility in water is 290 μg·mL^-1, which is 743 times higher than that of free CBD. Stability test results showed that CBD-HPCS/OA-NMs had no significant change in particle size after 168 h storage in simulated physiological and cell culture environment (P > 0.05), indicating good stability of micelles. The hemolytic test results indicated that CBD-HPCS/OA-NMs had a relatively low hemolysis rate and good biocompatibility. The CCK-8 results showed that the blank nanomicelles of HPCS/OA had no cytotoxicity. The median inhibitory concentration (IC50) of CBD-HPCS/OA-NMs on SKOV3 cells was 8.51 μg·mL^-1 after 24 h of administration. Fluorescence microscopy observation confirmed that the uptake efficiency of CBDHPCS/OA-NMs by SKOV3 cells was significantly stronger than that of free CBD. The cell scratch assay indicated that the inhibitory effect of CBD-HPCS/OA-NMs on the migration of SKOV3 cells was significantly better than that of free CBD (P < 0.05). Conclusion The CBD-HPCS/OA-NMs prepared in this study can effectively improve the water solubility and stability of CBD, and has both good biological safety and in vitro anti-tumor activity.

R979.1

山西省应用基础研究计划项目（201801D221278）；企业横向项目（2022-05）；中药药剂学学科经费项目（2025XK35）