新型阿霉素抗耐药性隐形脂质体的体外细胞毒和体内毒性研究  

The in vitro cytotoxicity and in vivo toxicity of doxorubicin antiresistant stealth liposomes

作　　者：王坚成[1] 刘晓岩[2] 吕万良[1] How-Sung Lee Boon-Cher Goh 张强[1] 

机构地区：[1]北京大学药学院药剂系,北京100083 [2]北京大学药学院细胞与分子药理系,北京100083 [3]新加坡国立大学医学院药理系,新加坡117597 [4]新加坡国立大学医院肿瘤研究所,新加坡117597

出　　处：《药学学报》2005年第5期475-480,共6页Acta Pharmaceutica Sinica

摘　　要：目的　多药耐药(multidrugresistance,MDR)是目前临床肿瘤治疗的主要障碍。本文研制了新型阿霉素抗耐药性隐形脂质体(DARSLs),并对其体外细胞毒和体内毒性进行评价。方法　采用硫酸铵梯度法将阿霉素(DOX)和维拉帕米(VER)药物同时包载到隐形脂质体内,制备成DARSLs;采用耐药性鼠前列腺肿瘤细胞株MLLB2和人子宫肉瘤细胞株MES SA/DX5进行体外细胞毒性评价;采用SD大鼠对阿霉素抗耐药性隐形脂质体进行体内毒性评价。结果　在药脂比(DOX/VER/Lipid,w/w/w)为1∶0 11∶10时,阿霉素包封率大于90%,维拉帕米包封率约为70%。平均粒径为(118 .1±22. 3)nm。体外细胞毒性实验证实该脂质体能够在体外有效地逆转肿瘤细胞耐药性,并导致耐药肿瘤细胞生长抑制。体内系统毒性及心脏毒性实验结果显示,该脂质体能够明显改善游离阿霉素单独使用或与维拉帕米联合使用时产生的全身毒性,尤其是心脏毒性。结论　DARSLs具有相对较低的毒性,且能有效抑制耐药肿瘤的生长。Aim Multidrug resistance (MDR) as a major obstacle to successful clinical cancer chemotherapy, searching a novel effective antiresistant drug would be necessary. Methods A novel doxorubicin anti-resistant stealth liposomes (DARSLs) was prepared by co-encapsulating doxorubicin (DOX) and verapamil (VER) into stealth liposomes with ammonium sulfate gradient remote loading approach. In vitro cytotoxity of various DOX formulations and in vivo toxicity of DARSLs were evaluated using DOX-resistant rat prostate cancer cell line (MLLB2), human uterus sarcoma cell line (MES-SA/DX5) and normal SD rats, separately. Results The DARSLs liposome suspensions mainly consisted of homogeneous large unilamellar vesicles (LUV) with average particle size of (118.1±22.3) nm. Encapsulation efficiencies of DOX and VER in DARSLs were more than 90% and about 70%, respectively, when the ratio of DOX/VER/Lipid was 1∶0.11∶10 (w/w/w). In vitro cytotoxicity tests of the DARSLs using rat prostate cancer cell line (MLLB2) and human uterus sarcoma cell line (MES-SA/DX5) showed that 5 μmol·L^(-1) VER significantly reversed DOX-resistance of these 2 cell lines and DARSLs was the most effective on inhibition of DOX-resistant cell growth. Besides, compared to FDFV, much slower DOX distribution (confocal microscopy) to nuclei and cytoplasm in MLLB2 cells for DARSLs suggested that it might possess distinct mechanism of cytotoxicity. Systemic and cardiac toxicity evaluations in normal SD rats suggested that liposomal encapsulation could significantly improve the severe cardiotoxicity arising from simultanous administration of DOX and VER. Conclusion DARSLs is a novel anticancer liposome formulation with lower cardiotoxicity, effective drug-resistance reversal and intravenous injection.

关 键 词：阿霉素 维拉帕米 隐形脂质体 多药耐药性 毒性 

分 类 号：R943.4[医药卫生—药剂学] R965.3[医药卫生—药学]