半乳糖化氟尿嘧啶前体脂质体的制备及体外细胞毒性初步研究
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摘要
目的建立半乳糖化氟尿嘧啶前体脂质体的制备方法,并对该制剂体外细胞毒性进行初步研究。方法采用逆向蒸发-冷冻干燥法制备前体脂质体,CCK-8法分别考察半乳糖化氟尿嘧啶脂质体对SMMC-7721和MCF7的细胞毒性,Annexin V-FITC/PI流式细胞分析法分别检测原药和半乳糖化氟尿嘧啶前体脂质体对SMMC-7721细胞株和MCF7细胞株的细胞凋亡率。结果该方法制得的前体脂质体呈球形或近球形,粒径为(188. 2±2. 4) nm,包封率为(71. 2±1. 0)%,在4℃贮120 d渗漏率(4. 1±0. 9)%,对于SMMC-7721细胞,半乳糖化氟尿嘧啶前体脂质体的抑制率最高,对于MCF7细胞,半乳糖化氟尿嘧啶前体脂质体和氟尿嘧啶前体脂质体的抑制率无明显差别,但均高于原药。原药对MCF7细胞有较强的诱导细胞凋亡能力,半乳糖化氟尿嘧啶前体脂质体对SMMC-7721细胞有较强的诱导细胞凋亡能力。结论该方法制得的前体脂质体包封率高,稳定性好,初步判定半乳糖化氟尿嘧啶前体脂质体有一定的主动肝靶向性。
Objective To establish a preparation method of galactosylated fluorouracil precursor liposome and to study the cytotoxicity of the preparation in vitro. Methods Precursor liposomes were prepared by reverse evaporating-freezing-drying method. The cytotoxicity of lactosylated fluorouracil liposomes against SMMC-7721 and MCF7 was investigated by CCK-8 method. The apoptosis rate of the prodrug and galactosylated fluorouracil precursor liposome on SMMC-7721 cell line and MCF7 cell line was detected by Annexin V-FITC/PI flow cytometry analysis. Results The precursor liposome prepared by the method was spherical or nearly spherical,and the particle size was( 188. 2 ± 2. 4) nm,the encapsulation efficiency was( 71. 2 ± 1. 0) %,and the leakage rate was( 4. 1 ±0. 9) % stored at 4 ℃ for 120 days. The inhibition rate of galactosylated fluorouracil precursor liposome was the highest for SMMC-7721 cells,and there was no significant difference in the inhibition rate of galactosylated fluorouracil precursor liposome and fluorouracil precursor liposome for MCF7 cells,but all of them were higher than the original drug. The original drug had strong ability to induce apoptosis of MCF7 cells,and galactosylated fluorouracil precursor liposome had strong ability to induce apoptosis of SMMC-7721 cells. Conclusion The precursor liposomes prepared by this method have high encapsulation efficiency and good stability. It is preliminarily determined that galactosylated fluorouracil precursor liposomes have certain active liver-targeting properties.
Objective To establish a preparation method of galactosylated fluorouracil precursor liposome and to study the cytotoxicity of the preparation in vitro. Methods Precursor liposomes were prepared by reverse evaporating-freezing-drying method. The cytotoxicity of lactosylated fluorouracil liposomes against SMMC-7721 and MCF7 was investigated by CCK-8 method. The apoptosis rate of the prodrug and galactosylated fluorouracil precursor liposome on SMMC-7721 cell line and MCF7 cell line was detected by Annexin V-FITC/PI flow cytometry analysis. Results The precursor liposome prepared by the method was spherical or nearly spherical,and the particle size was( 188. 2 ± 2. 4) nm,the encapsulation efficiency was( 71. 2 ± 1. 0) %,and the leakage rate was( 4. 1 ±0. 9) % stored at 4 ℃ for 120 days. The inhibition rate of galactosylated fluorouracil precursor liposome was the highest for SMMC-7721 cells,and there was no significant difference in the inhibition rate of galactosylated fluorouracil precursor liposome and fluorouracil precursor liposome for MCF7 cells,but all of them were higher than the original drug. The original drug had strong ability to induce apoptosis of MCF7 cells,and galactosylated fluorouracil precursor liposome had strong ability to induce apoptosis of SMMC-7721 cells. Conclusion The precursor liposomes prepared by this method have high encapsulation efficiency and good stability. It is preliminarily determined that galactosylated fluorouracil precursor liposomes have certain active liver-targeting properties.
引文
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[3]宋韶芳.李科,刘于飞.原发性肝癌患者生存质量及其影响因素的研究现状[J].华南预防医学,2018,44(1):71-73.
[4]史占萍,申世刚,岳志莲,等.靶向可控抗肿瘤药物递送体研究进展[J].中国新药杂志,2017,26(4):410-417.
[5]仲跻云,陈梁,杨肖斌.口服用前体脂质体的研究进展[J].药学与临床研究,2012,20(1):56-59.
[6] Zhou Q,Liu L,Zhang D,et al. Preparation and characterization of gemcitabine liposome injections[J]. Original Articles,2012,67(1):844-847.
[7]王小宁,贾慧婷,李伟泽,等.水飞蓟宾前体脂质体的制备及其质量评价[J].中草药,2017,48(7):1314-1320.
[8]李菲,王建筑,毕研平,等.叔丁醇-水共溶剂体系冻干法制备多西紫杉醇脂质体[J].中国药学杂志,2016,51(4):293-296.
[9]王金晶,张芹,肖晗,等.盐酸博安霉素脂质体的制备及表征[J].中国抗生素杂志,2014,36(3):220-223.
[10]陈奋,张玲,郑佳彤,等.星点设计-效应面法优化平阳霉素脂质体的处方及工艺[J].中国医院药学杂志,2017,37(22):2231-2235.
[11]王永利,王立华,张江伟,等.大黄酚冻干脂质体的制备工艺研究[J].中草药,2013,44(8):960-964.
[12]方晓旭,郭伟英.硬脂醇半乳糖苷修饰阿西替尼脂质体的制备及体外细胞毒性研究[J].中国新药杂志,2017,26(3):343-349.
[13]李建林,孟路华,赵玉玺,等.透明质酸磷脂酰衍生物修饰姜黄素脂质体的制备及细胞毒性[J].中国医药工业杂志,2013,44(5):471-474.
[14]刘宇欣,李红玉,严鹏.海龙蛋白质提取物诱导宫颈癌Hela细胞凋亡的机制[J].解放军医学院学报,2014,35(6):623-626.
[15]孟庆硕,张鹏程,尹琦,等.光敏感型盐酸多柔比星脂质体逆转乳腺癌耐药[J].药学学报,2017,52(5):809-820.
[16]孟路华,赵怡,潘黎军,等.维生素A-姜黄素脂质体的制备及细胞毒性研究[J].中国药学杂志,2011,46(14):1104-1107.
[17]邓意辉,徐晖,贺亮,等.脂质体[M].第2版.北京:化学工业出版社,2007:120-126.