目的 优化核酸适配体（APS613-1）和还原响应长循环链（mal-PEG-ss-DSPE）修饰的去甲斑蝥素脂质体 （Apt-ss-NCTD-Lip）的处方工艺，并探讨其体外抗肝肿瘤作用。方法 采用乙醇注入法制备 Apt-ss-NCTDLip，以磷脂种类、水合温度、水合时间、胆脂比等为考察因素，以去甲斑蝥素-脂质体（NCTD-Lip）包封率、 粒径、多分散指数（PDI）和 Zeta 电位为评价指标。采用单因素考察和 Box-Behnken 设计-响应面法优化 NCTDLip 的处方工艺，并考察 APS613-1 与 mal-PEG-ss-DSPE 的配比。采用透射电子显微镜观察脂质体形态；激光 纳米粒度测定仪测定粒径、Zeta 电位及 PDI。通过体外释放、还原响应性测定及 CCK-8 法进行体外抗肝癌研 究。结果 Apt-ss-NCTD-Lip 的最佳制备工艺：以大豆卵磷脂、胆固醇为膜材，药脂比为 1∶15，胆脂比为 1∶9，mal-PEG-ss-DSPE 用量为 10% 磷脂膜材量，APS613-1 与 mal-PEG-ss-NCTD-Lip 比例为 1∶150，水合温 度为 65 ℃，水合时间为 90 min。Apt-ss-NCTD-Lip 的粒径约 130 nm，包封率约 80%。Apt-ss-NCTD-Lip 溶液 外观显示为澄清透明带淡蓝色乳光；透射电镜下显示其形态圆整、大小均一、无粘连现象。体外实验结果显示 为 Apt-ss-NCTD-Lip 具有较好的还原响应性能，能有效抑制 HepG2 细胞增殖，具有良好的肝肿瘤靶向性。 结论 Apt-ss-NCTD-Lip 是一种具有开发潜力的抗肝肿瘤纳米递送系统。

Objective To optimize the prescription process of nucleic acid aptamer（APS613-1） and reductionresponsive long-circulating chain （mal-PEG-ss-DSPE）-modified norcantharidin liposomes （Apt-ss-NCTD-Lip） and initially investigate the physicochemical properties and in vitro anti-liver tumor effects. Methods Apt-ss-NCTD-Lip was prepared by ethanol injection method. The phospholipid species， hydration temperature， hydration time and cholesterol/lipid ratio were used as the investigation factors， and the encapsulation efficiency， particle size， polydispersity index （PDI） and Zeta potential of norcantharidin-liposomes （NCTD-Lip）were used as the evaluation indexes. The prescription process of NCTD-Lip was optimized by single factor investigation and Box-Behnken designresponse surface method， and the ratio of APS613-1 to mal-PEG-ss-DSPE was investigated. The morphology of liposomes was observed by transmission electron microscopy. The particle size，Zeta potential and PDI were measured by laser nano particle size analyzer. The in vitro anti-hepatocellular carcinoma study was carried out by in vitro release，reduction response determination and CCK-8 method. Results The optimum preparation process of Apt-ss-NCTD-Lip： soybean lecithin and cholesterol were used as membrane materials，the ratio of drug to lipid was 1∶15，the ratio of bile to lipid was 1∶9， the amount of mal-PEG-ss-DSPE was 10% phospholipid membrane material ， the ratio of APS613-1 to mal-PEG-ss-NCTD-Lip was 1∶150，the hydration temperature was 65 ℃，and the hydration time was 90 minutes. The particle size of Apt-ss-NCTD-Lip was about 130 nm，and the encapsulation efficiency was about 80%. The appearance of the liposome solution showed that the clear transparent zone was light blue opalescent； transmission electron microscopy showed its round shape， uniform size， no adhesion phenomenon. The in vitro evaluation results indicated that Apt-ss-NCTD-Lip exhibits a favorable reducibility response，effectively inhibits the proliferation of HepG2 cells，and demonstrates strong targeting capabilities towards liver tumors. Conclusion Apt-ssNCTD-Lip represents a promising nanodelivery system for the treatment of hepatic tumors.

R284.1

国家自然科学基金项目（81573621）；湖南省科技创新计划资助项目（2021RC4064）；湖南省自然科学基金项目（2022JJ40320）；湖南省 教育厅优秀青年项目（23B0362）；湖南省中医药管理局一般课题项目（B2024015）； 湖南中医药大学本科生科研创新基金项目（2024BKS064）。