目的 制备共载高三尖杉酯碱（homoharringtonine，HHT）和雷公藤甲素（triptolide，TRL）-前列腺特异性膜抗原（prostate specific membrane antigen，PSMA）靶向肽修饰脂质-聚合物纳米粒（PSMA-targeting peptide-modified lipid-polymer nanoparticles co-loaded with HHT and TRL，HHT/TRL-PSMA-Lips），优化其制备工艺，并进行体外抗前列腺癌肿瘤作用评价。方法 测定HHT与TPL的溶解度及油水分配系数（lgP），合成PSMA靶向肽修饰PEG化磷脂。采用乳化-溶剂挥发法制备HHT/TPL-PSMA-Lips，采用中心复合设计-响应面法（central composite design-response surface methodology，CCD-RSM）优化处方，并考察其粒径、ζ电位、包封率等指标。考察制剂在不同介质中的体外释放行为。并采用MTT法、流式细胞术考察纳米粒对前列腺癌细胞的体外抑制及诱导凋亡作用，采用细胞摄取实验考察纳米粒的肿瘤细胞靶向效果。结果 HHT与TPL在不同介质中的溶解度均较低，亲脂性较强。合成PSMA靶向肽修饰PEG化磷脂的相对分子质量为4 643.8。采用最优处方的制备的HHT/TPL-PSMA-Lips粒径均一，平均粒径为（209.12±3.24）nm，ζ电位为（-34.4±2.5）mV，综合包封率为（81.20±0.40）%［HHT（80.73±0.86）%，TPL（82.00±0.40）%］，体外释放实验证明，HHT/TPL-PSMA-Lips可有效提高HHT和TPL的溶解度，并具有一定的pH响应性释放效果。细胞实验表明，HHT/TPL联合给药组显示出高于单一药物组的前列腺肿瘤细胞抑制作用，及诱导凋亡效果，HHT/TPL-Lips可提高抗肿瘤活性，靶向修饰的纳米粒HHT/TPL-PSMA-Lips显示出对前列腺肿瘤细胞最强的抑制效果。细胞摄取实验表明，PSMA靶向肽的修饰可有效提高纳米制剂在肿瘤细胞的摄取。结论 制备的HHT/TPL-PSMA-Lips不仅可解决药物的溶解性问题，并可显著提升肿瘤靶向作用以及抗肿瘤效果，为中药新制剂用于前列腺癌的治疗提供了新思路。

Objective To prepare prostate specific membrane antigen (PSMA)-targeting peptide-modified lipid-polymer nanoparticles co-loaded with homoharringtonine and triptolide (HHT/TPL-PSMA-Lips), optimize its preparation process, and evaluate its in vitro anti-prostate cancer activity. Methods The solubility and oil-water partition coefficient (lgP) of HHT and TPL were determined, and PSMA-targeting peptide-modified PEGylated phospholipids were synthesized. HHT/TPL-PSMA-Lips was prepared by the emulsification-solvent evaporation method, and the formulation was optimized using central composite design-response surface methodology (CCD-RSM). Indicators including particle size, ζ potential, and encapsulation efficiency were investigated. The in vitro release behavior of the formulation in different media was evaluated. MTT assay and flow cytometry were used to assess the in vitro inhibitory effect and apoptosis-inducing activity of the nanoparticles on prostate cancer cells, while cell uptake assay was performed to verify the tumor cell targeting efficiency of the nanoparticles. Results HHT and TPL showed low solubility in various media and strong lipophilicity. The molecular weight of the synthesized PSMA-targeting peptide-modified PEGylated phospholipids was 4 643.8. HHT/TPL-PSMA-Lips prepared with the optimal formulation exhibited uniform particle size, with an average particle size of (209.12 ± 3.24) nm, a ζ potential of (-34.4 ± 2.5) mV, and a comprehensive encapsulation efficiency of (81.20 ± 0.40)% [HHT (80.73 ± 0.86)%, TPL (82.00 ± 0.40)%]. In vitro release experiments demonstrated that HHT/TPL-PSMA-Lips could effectively improve the solubility of HHT and TPL, and exhibite a certain pH-responsive release property. Cell experiments showed that the HHT/TPL combined administration group displayed stronger inhibitory effect on prostate cancer cells and better apoptosis-inducing activity than the single drug groups. HHT/TPL-Lips enhanced the anti-tumor activity, and the targeted modified nanoparticles (HHT/TPL-PSMA-Lips) showed the strongest inhibitory effect on prostate cancer cells. Cell uptake assay indicated that modification with PSMA-targeting peptide could effectively enhance the cellular uptake of the nanoformulation in tumor cells. Conclusion The prepared HHT/TPL-PSMA-Lips not only solves the solubility problem of the drugs but also significantly enhances tumor targeting and anti-tumor efficacy, providing a new approach for the development of new traditional Chinese medicine formulations for the treatment of prostate cancer.

R283.6

国家自然科学基金资助项目（U1804181）；河南省中医药专项课题（2024ZY2134）；无锡市卫生健康委科研青年项目（Q202307）；无锡市卫生健康委中青年后备拔尖人才项目（HB2023117）；中国博士后科学基金第73批面上资助（2023M730572）