麦胚凝集素修饰的长春瑞滨阳离子脂质体的处方优选及细胞毒性试验
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摘要
目的:制备麦胚凝集素(WGA)修饰的长春瑞滨(VRB)阳离子脂质体(WGA-VRB阳离子脂质体),优化处方并进行细胞毒性试验。方法:以磷脂、胆固醇为辅料,以3β-[N-(N′-N′-二甲基氨基乙烷)-氨基甲酰基]胆固醇盐酸盐(DC-Chol)为阳离子材料,以二硬脂酰磷脂酰乙醇胺-聚乙二醇2000(DSPE-PEG2000)为长循环链,采用薄膜分散法及硫酸铵梯度法制备WGA-VRB阳离子脂质体。以包封率为指标,采用星点设计-响应面法优化处方中DC-Chol、胆固醇和VRB用量。测定所制VRB脂质体和WGA-VRB阳离子脂质体中VRB含量,比较二者和空白阳离子脂质体作用于人乳腺癌细胞MCF-7和人非小细胞肺癌细胞A549后对细胞存活率的影响。结果:5 m L WGA-VRB阳离子脂质体的最优处方为磷脂22 mg、胆固醇12 mg、DC-Chol 8 mg、VRB 0.5 mg,其包封率为(92.24±1.21)%(n=3),与预测值的相对误差为5.3%。VRB脂质体和WGA-VRB阳离子脂质体中VRB含量分别为(96.01±3.26)、(93.39±1.59)μg/m L(n=3);与空白阳离子脂质体和VRB脂质体比较,WGA-VRB阳离子脂质体可明显降低MCF-7、A549细胞的存活率(P<0.05)。结论:成功制得WGA-VRB阳离子脂质体,其对MCF-7、A549细胞存活的抑制作用强于VRB脂质体。
OBJECTIVE: To prepare Wheat germ agglutinin(WGA) modified vinorelbine(VRB) cationic liposomes(WGA-VRB cationic liposomes),and to optimize the formulation and conduct cytotoxicity test. METHODS:Thin-film diffusion and ammonium sulfate gradient method were used to prepare WGA-VRB cationic liposomes using phospholipid and cholesterol as excipient,3 β-[N-(N′-N′-dimethyl aminoethane)-carbamoyl] cholesterol hydrochloride(DC-Chol) as cationic material,distearoyl phosphatidylethanolamine-polyethylene glycol 2000(DSPE-PEG2000) as long cycle chain. Using encapsulation rate as index,central composite design-response surface methodology was used to optimize the amount of DC-Chol,cholesterol and VRB. The contents of VRB in VRB liposomes and WGA-VRB cationic liposomes were determined. The effects of them and blank cationic liposomes on survival rates of human breast cancer cell MCF-7 and human non-small cell lung cancer cells A549 were compared.RESULTS:The optimal formulation of 5 m L WGA-VRB cationic liposomes was as follows as phospholipid 22 mg,cholesterol 12 mg,DC-Chol 8 mg,VRB 0.5 mg. Encapsulation rate of the liposomes was(92.24 ± 1.21)%(n=3),relative error of which to predicted value was 5.3%. The contents of VRB in VRB liposomes and WGA-VRB cationic liposomes were(96.01 ± 3.26),(93.39±1.59)μg/m L(n=3). Compared with blank cationic liposomes and VRB liposomes,WGA-VRB cationic liposomes could significantly reduce survival rate of MCF-7 and A549. CONCLUSIONS:WGA-VRB cationic liposomes are prepared successfully.Inhibitory effect of WGA-VRB cationic liposomes on MCF-7 and A549 cell survival is stronger than that of VRB liposomes.
OBJECTIVE: To prepare Wheat germ agglutinin(WGA) modified vinorelbine(VRB) cationic liposomes(WGA-VRB cationic liposomes),and to optimize the formulation and conduct cytotoxicity test. METHODS:Thin-film diffusion and ammonium sulfate gradient method were used to prepare WGA-VRB cationic liposomes using phospholipid and cholesterol as excipient,3 β-[N-(N′-N′-dimethyl aminoethane)-carbamoyl] cholesterol hydrochloride(DC-Chol) as cationic material,distearoyl phosphatidylethanolamine-polyethylene glycol 2000(DSPE-PEG2000) as long cycle chain. Using encapsulation rate as index,central composite design-response surface methodology was used to optimize the amount of DC-Chol,cholesterol and VRB. The contents of VRB in VRB liposomes and WGA-VRB cationic liposomes were determined. The effects of them and blank cationic liposomes on survival rates of human breast cancer cell MCF-7 and human non-small cell lung cancer cells A549 were compared.RESULTS:The optimal formulation of 5 m L WGA-VRB cationic liposomes was as follows as phospholipid 22 mg,cholesterol 12 mg,DC-Chol 8 mg,VRB 0.5 mg. Encapsulation rate of the liposomes was(92.24 ± 1.21)%(n=3),relative error of which to predicted value was 5.3%. The contents of VRB in VRB liposomes and WGA-VRB cationic liposomes were(96.01 ± 3.26),(93.39±1.59)μg/m L(n=3). Compared with blank cationic liposomes and VRB liposomes,WGA-VRB cationic liposomes could significantly reduce survival rate of MCF-7 and A549. CONCLUSIONS:WGA-VRB cationic liposomes are prepared successfully.Inhibitory effect of WGA-VRB cationic liposomes on MCF-7 and A549 cell survival is stronger than that of VRB liposomes.
引文
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[2]何文,胡翠苹,郭咸希.N-三甲基壳聚糖包覆水飞蓟宾脂质体在小鼠体内的药动学及靶向性[J].中国医院药学杂志,2016,36(3):196-200.
[3]喻樊,杨锦明,李金娟.PEG修饰β-谷甾醇制备新型绞股蓝总皂苷长循环脂质体的研究[J].中国中药杂志,2014,39(6):997-1001.
[4]袁端锋,宗太丽,高会乐.穿膜肽TAT和脑肿瘤靶向肽T7双修饰脂质体的制备和体外靶向性评价[J].药学学报,2015,50(1):104-110.
[5]吴迪,赵颖,任会丹,等.阳离子抗癌肽Temporin-1CEa脂质体的构建及其体外抗乳腺癌活性评价[J].中国生化药物杂志,2016,36(11):22-26.
[6]陈伟光,刘源岗,王士斌,等.阳离子脂质体共载si RNA与紫杉醇的制备与性能表征[J].科学通报,2013,58(11):1014-1020.
[7]马一平.异长春花碱抑制人乳腺癌细胞转移侵袭的作用及其机制[J].中草药,2013,44(13):1786-1792.
[8]王艳红,王晓敏,刘爽.星点设计-响应面法优化WGA修饰柔红霉素-粉防己碱脂质体处方[J].食品与药品,2016,18(4):229-234.
[9]杨雯姝,邱利焱,金一.RP-HPLC法测定脂质体中重酒石酸长春瑞滨的含量[J].药物分析杂志,2006,26(5):646-648.
[10]姜英,王晓敏,王艳红.星点设计-效应面法优化转铁蛋白修饰粉防己碱与硫酸长春新碱脂质体的处方[J].中国药房,2015,26(31):4399-4401.
[11]SHI JF,SUN MG,LI XY,et al.A combination of targeted sunitinib liposomes and targeted vinorelbine liposomes for treating invasive breast cancer[J].Journal of Biomedical Nanotechnology,2015,11(9):1568-1582.
[12]国家药典委员会.中华人民共和国药典:四部[S].2015年版.北京:中国医药科技出版社,2015:371-374.
[13]赵轶男,刘安,付莹,等.阳离子脂质体基因载体的稳定性研究[J].大连民族大学学报,2017,19(3):202-206.
[14]陈伟光,刘源岗,王士斌,等.阳离子脂质体共载基因与化疗药物用于癌症治疗的研究进展[J].药学学报,2012,47(8):986-992.