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(1)
中圖分类号 R943 文献标志码 A 文章编号 1001-0408(2018)05-0611-04
DOI 10.6039/j.issn.1001-0408.2018.05.09
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 mL 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/mL(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。
关键词 麦胚凝集素；长春瑞滨；阳离子脂质体；处方优选；星点设计-响应面法；含量测定
ABSTRACT 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， 3β-[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 mL 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/mL(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.
KEYWORDS Wheat germ agglutinin； Vinorelbine； Cationic liposomes； Formulation optimization； Central composite design- response surface methodology； Content determination, 百拇医药(肖瑶 王鑫 符敏 刘晶晶 李学涛 程岚)
DOI 10.6039/j.issn.1001-0408.2018.05.09
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 mL 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/mL(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。
关键词 麦胚凝集素；长春瑞滨；阳离子脂质体；处方优选；星点设计-响应面法；含量测定
ABSTRACT 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， 3β-[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 mL 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/mL(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.
KEYWORDS Wheat germ agglutinin； Vinorelbine； Cationic liposomes； Formulation optimization； Central composite design- response surface methodology； Content determination, 百拇医药(肖瑶 王鑫 符敏 刘晶晶 李学涛 程岚)