吡柔比星脂质体的制备及体内外评价  

Preparation and in vitro and vivo evaluation of pirarubicin liposomes

作　　者：姬浩洁 韩文豪 姜宇 郝贵周 张贵民 JI Hao-jie;HAN Wen-hao;JANG Yu;HAO Gui-zhou;ZHANG Gui-min(Xiangya School of Pharmaceutical Sciences,Central South University,Changsha 410000;Shandong Complex Injectable Engineering Research Center,Shandong New Times Pharmaceutical Co.,Ltd.,Linyi Shandong 276006;Lunan Pharmaceutical Group Co.,Ltd.,Linyi Shandong 276005)

机构地区：[1]中南大学湘雅药学院,长沙410000 [2]山东新时代药业有限公司,山东省复杂注射剂工程技术研究中心,山东临沂276006 [3]鲁南制药集团股份有限公司,山东临沂276005

出　　处：《中南药学》2025年第4期880-886,共7页Central South Pharmacy

基　　金：山东省重点研发计划(竞争性创新平台)(No.2022CXPT046)。

摘　　要：目的采用3种不同工艺制备吡柔比星脂质体(PL),并考察其药剂学性质及抗肿瘤效果。方法分别采用硫酸铵梯度法、葡糖醛酸铵梯度法和pH梯度法,制备PL-Sul、PL-Glu、PL-pH,建立HPLC法测定脂质体中吡柔比星的含量和包封率。以包封率为指标,优化沉淀剂浓度。考察3种PL的形态、粒径、Zeta电位、包封率、体外释放曲线、稳定性等药剂学特性。使用CCK-8法检测PL对小鼠结肠癌细胞(CT26)和小鼠B细胞淋巴瘤细胞(A20)的增殖抑制效果。以吡柔比星注射液为阳性对照,采用小鼠CT26皮下肿瘤模型,比较3种PL的抗肿瘤活性。结果3种PL的药物包封率均大于90%,平均粒径约为85mm,Zeta电位约为-26mV。4℃条件下放置30d,各样品的主要质量指标均未发生明显变化。冷冻透射电镜观察形态结果表明,在脂质体亲水内腔,PL-Glu中没有观察到药物与沉淀剂的沉淀晶体,而PL-Sul和PL-pH中可观察到棒状和片状结晶。体外释放结果表明,3种PL均缓慢释放药物,以PL-pH释放速度最慢。体外抗肿瘤结果显示PL-Sul和PL-Glu对细胞增殖的抑制作用更强。药效学研究结果表明,3种PL都显示出较高的抗肿瘤活性,但仅PL-Sul疗效显著优于吡柔比星注射液。结论使用硫酸铵梯度法制备的PL能显著提高药物的抗肿瘤效果,为PL药物的开发奠定了基础。Objective To prepare pirarubicin liposome(PL)with three methods and evaluate their properties and antitumor effects.Methods PL was prepared by ammonium sulfate gradient method(PL-Sul),ammonium glucuronate gradient method(PL-Glu)and pH gradient method(PL-pH).HPLC method was used to determine the drug loading and encapsulation efficiency of the PL.The concentration of the precipitant was optimized with the encapsulation efficiency.The morphology,size and Zeta potential of the PL were analyzed by cryo-transmission electron microscope(cryo-TEM)and nanometer particle size analyzer.The in vitro release rate and stability were also evaluated.The cell proliferation inhibitory effect of the PL on CT26 cells and A20 cells was detected with CCK-8.The antitumor effect of the PL was measured in mice bearing CT26 colon tumor and compared with that of pirarubicin injection.Results The encapsulation efficiencies of three PLs were all over 90%.The average particle sizes of the PL were about 85 mm,and the Zeta potentials were about-26 mV.The PL was stable,no obvious change was found in each index of the PL after storing at 4℃for 30 d.Furthermore,the cryo-TEM pictures showed that some rod-like or slice-like crystals were found inside PL-Sul and PL-pH,respectively,but no drug crystal was found in PL-Glu.The release rate of PL-pH was the slowest,which indicated that the drug crystal probably influenced the in vitro drug release.In vitro antitumor results showed that PL-Sul and PL-Glu had stronger inhibitory effect on the cell proliferation.The pharmacodynamic test showed that all the three PLs showed high antitumor effect.However,only the antitumor effect of the PL-Sul was much better than pirarubicin injection.Conclusion PL prepared by ammonium glucuronate gradient method can effectively improve the antitumor effect of drugs,providing a basis for further research on PL drugs.

关 键 词：吡柔比星 脂质体 硫酸铵梯度法 葡糖醛酸铵梯度法 PH梯度法 结肠癌 

分 类 号：R97[医药卫生—药品]