Page 42 - 《中国药房》2021年24期

白屈菜红碱纳米粒的制备及体外抗黑色素瘤活性评价 Δ

1＊
1
2，3
1
1
1 #
1
杨 锦 ，韩 伟 ，张永萍 ，陈晓兰 ，李 哲 ，刘 杰 ，吴静澜 （1.贵州中医药大学药学院，贵阳 550025；
2.贵州中医药大学基础医学院，贵阳 550025；3.贵州中医药大学兽用中药制剂创新研发中心，贵阳 550025）
中图分类号 R283.6；R979.1 文献标志码 A 文章编号 1001-0408（2021）24-2980-07
DOI 10.6039/j.issn.1001-0408.2021.24.07
摘 要 目的：制备白屈菜红碱纳米粒（CHE-NPs），对其处方进行优化，并评价其体外释药行为及对黑色素瘤的抑制作用。方法：
以甲氧基聚乙二醇-聚乳酸-羟基乙酸共聚物（mPEG-PLGA）为载体材料，采用纳米沉淀法制备CHE-NPs，采用高效液相色谱法和
透析袋法测定包封率、载药量。以二者的总评归一（OD）值为因变量，以CHE投药量、mPEG-PLGA质量浓度、泊洛沙姆188（F68）
浓度为自变量，采用Box-Behnken响应面设计优化CHE-NPs处方。检测最优处方所制CHE-NPs的粒径和Zeta电位，考察其体外
释药特征，比较CHE原料药和CHE-NPs对小鼠B16黑色素瘤细胞存活率的影响并计算二者的半数抑制浓度。结果：最优处方为
CHE 投药量 2 mg、mPEG-PLGA 质量浓度 13 mg/mL、F68 浓度 1.8％。以此所制 CHE-NPs 的平均包封率为（80.18±1.11）％，平均
载药量为（11.36±0.28）％，平均OD值为0.96±0.04[与OD预测值（0.90）的相对偏差为6.67％]，粒径为（113.1±1.40）nm，Zeta电位
为（－21.6±0.29）mV，多分散性指数为 0.07±0.01（n＝3）。CHE 对照品、CHE-NPs 在孵育 8 h 时的累积释放率分别为 90.87％、
68.68％，后者的体外释药行为符合 Weibull 动力学模型。CHE-NPs 对 B16 黑色素瘤细胞的抑制作用显著强于 CHE 原料药，
CHE-NPs 和 CHE 原料药的 24 h 半数抑制浓度分别为 69.35、107.36 μg/mL。结论：所制 CHE-NPs 具有缓释作用和较强的载药能
力，同时增强了CHE对黑色素瘤的体外抑制作用。
关键词 白屈菜红碱；黑色素瘤；抑瘤作用；甲氧基聚乙二醇-聚乳酸-羟基乙酸共聚物；纳米粒；Box-Behnken响应面设计

Preparation of Chelerythrine Nanoparticles and Evaluation of Anti-melanoma Activity in vitro
YANG Jin ，HAN Wei ，ZHANG Yongping ，CHEN Xiaolan ，LI Zhe ，LIU Jie ，WU Jinglan（1. College of
1
1
2，3
1
1
1
1
Pharmacy，Guizhou University of Traditional Chinese Medicine，Guiyang 550025，China；2. College of Basic
Medicine， Guizhou University of Traditional Chinese Medicine， Guiyang 550025， China； 3. Innovation
Research and Development Center of Veterinary Traditional Chinese Medicine Preparations，Guizhou University
of Traditional Chinese Medicine，Guiyang 550025，China）
ABSTRACT OBJECTIVE：To prepare chelerythrine nanoparticles（CHE-NPs），optimize their formulation，and evaluate its drug
release behavior in vitro and its inhibitory effect on melanoma. METHODS：Using methoxy polyethylene glycol-poly（lactic-co-
glycolic acid）（mPEG-PLGA）as carrier，CHE-NPs were prepared by the nano-precipitation method. HPLC method and dialysis
bag method were used to determine entrapment efficiency and drug loading. The formulation of CHE-NPs was optimized by
Box-Behnken response surface design using overall desirability（OD）of them as dependent variables，CHE dosage，mPEG-PLGA
concentration and poloxamer 188（F68）concentration as independent variables. The particle size and Zeta potential of CHE-NPs
prepared by the optimal formulation were detected；the characteristics of drug release in vitro were investigated；the effects of CHE
and CHE-NPs on survival rate of mice B16 melanoma cells were compared，and median inhibition concentrations（IC50 ）of them
were calculated. RESULTS：The optimal formulation included CHE of 2 mg，mPEG-PLGA of 13 mg/mL，F68 of 1.8％. Average
entrapment efficiency rate of CHE-NPs prepared by the optimal formulation was （80.18 ± 1.11）％ ，average drug loading was
（11.36±0.28）％，average OD value was 0.96±0.04 [the relative deviation from predicted value（0.90）of OD was 6.67％]；
particle size was（113.1±1.40）nm，and Zeta potential was（－21.6±0.29）mV；polydispersity index was 0.07±0.01（n＝3）；
accumulative release rates of CHE control and CHE-NPs were 90.87％ and 68.68％ within 8 h，and drug release behavior in vitro
of the latter was in accordance with Weibull kinetic model. Inhibitory effect of CHE-NPs on B16 melanoma cells was significantly
stronger than that of CHE；the 24 h IC 50 of CHE-NPs and CHE
Δ 基金项目：国家苗药工程技术研究中心组建项目（No.2014FU- were 69.35 and 107.36 μg/mL，respectively. CONCLUSIONS：The
125Q09）；贵州省科技厅学术新苗培养及创新探索专项项目（No.黔科合
prepared CHE-NPs show good sustained-effect and high
平台人才〔2018〕5766号-13）；地方病与少数民族性疾病教育部重点实验
capacity of drug loading，and strengthen the inhibitory effect
室（贵州医科大学）开放课题基金资助项目（No.黔教合KY字〔2019〕047）
＊硕士研究生。研究方向：中药及民族药物新制剂与新剂型。 of CHE on melanoma.
E-mail：1510428733@qq.com KEYWORDS Chelerythrine； Melanoma；Anti-tumor effect；
# 通信作者：教授，硕士生导师，硕士。研究方向：中药及民族药 Methoxy polyethylene glycolpolylactic acid-hydroxyacetic acid
物新制剂与新剂型。E-mail：1259803072@qq.com copolymer；Nanoparticles；Box-Behnken response surface design

·2980 · China Pharmacy 2021 Vol. 32 No. 24 中国药房 2021年第32卷第24期