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甘草次酸纳米粒的制备、表征及其抗肿瘤活性研究 Δ

1＊
张丽娟 ，喻红梅 ，张 勇 ，龚宁波 （1.首都医科大学燕京医学院临床医学学系，北京 101300；2.北京协和
2，3 #
2
3
医学院/中国医学科学院药物研究所药物晶型研究中心，北京 100050；3.海南医学院基础医学与生命科学学院
药理教研室，海口 571199）

中图分类号 R943；R932 文献标志码 A 文章编号 1001-0408（2020）13-1589-06
DOI 10.6039/j.issn.1001-0408.2020.13.09

摘 要 目的：制备和表征甘草次酸（GA）纳米粒，并评价其体外抗肿瘤活性。方法：以聚乙烯吡咯烷酮K30为载体，使用反溶剂
沉淀-冷冻干燥法制备GA纳米粒。采用X射线衍射分析、红外光谱分析、差示扫描量热分析、粒度分析等方法对所制纳米粒进行
表征；采用高效液相色谱法测定纳米粒中 GA 的溶解度和载药量；采用 MTT 法考察 GA 原料药及纳米粒（GA 剂量均为 12.5、25、
50、100、200 μmol/L）对人肝癌细胞HepG2的体外抑制活性并计算半数抑制浓度（IC50 ）。结果：所制纳米粒中GA的X射线衍射特
征峰和红外特征吸收峰均消失，吸热峰发生改变。纳米粒的粒径为（194.88±23.52）nm，低于原料药的（2 592.33±207.51）nm；分
散指数为 0.24±0.04，高于原料药的 0.15±0.03；纳米粒的平均载药量为 15.99％；溶解度由原料药的（1.05±0.01）μg/mL 升至
（250.00±0.15）μg/mL。体外抗肿瘤试验结果显示，GA原料药200 μmol/L组和纳米粒各剂量组的细胞存活率均较空白对照组显
著降低，且GA纳米粒各剂量组（除12.5 μmol/L组外）的细胞存活率均显著低于同剂量原料药组（P＜0.01）；GA纳米粒的IC50值为86.3
μmol/L，低于原料药的364.4 μmol/L。结论：成功制得GA纳米粒；所制纳米粒粒径小且分布均匀，溶解度增大且体外抗肿瘤活性增强。
关键词 甘草次酸纳米粒；反溶剂沉淀-冷冻干燥法；制备；表征；抗肿瘤活性；HepG2细胞

Preparation，Characterization and Anti-tumor Activity Study of Glycyrrhetinic Acid Nanoparticles
ZHANG Lijuan ，YU Hongmei ，ZHANG Yong ，GONG Ningbo （1. Dept. of Clinical Medicine，Yanjing Medical
3
2，3
1
2
College，Capital Medical University，Beijing 101300，China；2. Drug Crystal Form Research Center，Institute
of Materia Medica，Chinese Academy of Medical Sciences/Peking Union Medical College，Beijing 100050，
China；3. Dept. of Pharmacology，College of Basic Medicine and Life Sciences，Hainan Medical University，
Haikou 571199，China）

ABSTRACT OBJECTIVE：To prepare and characterize glycyrrhetinic acid （GA） nanoparticles，and to evaluate its in vitro
anti-tumor activity. METHODS：Using PVP K30 as carrier，GA nanoparticles were prepared by anti-solvent precipitation and
freeze-drying method. X-ray diffraction，infrared spectrum，differential scanning calorimetry and granularity analysis were used to
characterize the nanoparticles； HPLC method was used to measure the solubility and drug-loading amount of GA in the
nanoparticles. MTT method was used to assay the in vitro inhibition activity of GA raw material and nanoparticles（GA doses were
12.5，25，50，100，200 μmol/L）on human liver cancer cell HepG2 and calculate its IC50. RESULTS：The characteristic peaks of
X-ray diffraction and infrared absorption of GA disappeared in the nanoparticles and the endothermic peak changed. The particle
size of the nanoparticles was（194.88±23.52）nm，which was lower than（2 592.33±207.51）nm of raw material. The dispersion
index was 0.24±0.04，which was higher than 0.15±0.03 of raw material. The average drug-loading amount of GA was 15.99％.
Moreover，the solubility of nanoparticles increased from（1.05±0.01）μg/mL to（250.00±0.15）μg/mL. The results of antitumor
test in vitro showed that the cell survival rates in the group of GA raw material 200 μmol/L and GA nanoparticles groups were
significantly lower than that in blank control group，and the cell survival rates of GA nanoparticles groups（except for 12.5 μmol/L
group） were significantly lower than that of same dose group of raw material （P＜0.01）. IC50 of GA nanoparticles was 86.3
μmol/L，which was lower than 364.4 μmol/L of raw material. CONCLUSIONS：GA nanoparticles are prepared successfully；the
prepared nanoparticles have small size and uniform distribution，and the solubility are increased and antitumor activity in vitro are
enhanced.
KEYWORDS Glycyrrhetinic acid nanoparticle；Anti-solvent
Δ 基金项目：国家科技重大专项（民口）课题（No.2018ZX09711-
precipitation and freeze-drying method；Preparation；Charac-
001）；中国医学科学院医学与健康科技创新工程项目（No.2017-I2M-
terization；Anti-tumor activity；HepG2 cells
1-010）；首都医科大学燕京医学院科研培育基金（No.18qdky02）
＊副教授，硕士。研究方向：天然产物的作用机理、分子生物学。
电话：010-81476189。E-mail：zhanglijuan@ccmu.edu.cn
# 通信作者：副研究员，硕士生导师，博士。研究方向：药物分析、 甘草次酸（Glycyrrhetinic acid，GA）是一种由从豆科
晶型药物研发。电话：010-63030566。E-mail：gnb@imm.ac.cn 植物甘草（Glycyrrhiza uralensis Fisch.）、胀果甘草（G. in-

中国药房 2020年第31卷第13期 China Pharmacy 2020 Vol. 31 No. 13 ·1589 ·