药学学报, 2018, 53(3): 453-459
季宇彬, 聂凡茹, 周欣欣, 王向涛. 藤黄酸纳米混悬剂的制备及抗肿瘤作用[J]. 药学学报, 2018, 53(3): 453-459.
JI Yu-bin, NIE Fan-ru, ZHOU Xin-xin, WANG Xiang-tao. Preparation of gambogic acid naonosuspensions and their anti-tumor efficacy[J]. Acta Pharmaceutica Sinica, 2018, 53(3): 453-459.

季宇彬1, 聂凡茹1,2, 周欣欣1,2, 王向涛1,2
1. 哈尔滨商业大学, 生命科学与环境科学研究中心, 黑龙江 哈尔滨 150076;
2. 中国医学科学院、北京协和医学院药用植物研究所, 北京 100193
藤黄酸(gambogic acid,GA)是藤黄科植物藤黄中提取的具有良好选择性和高效抗肿瘤活性的化合物,但其溶解性差,限制了进一步的体内研究和临床应用。本研究通过将其制备成纳米混悬剂来解决此问题。采用碱溶酸法制备藤黄酸纳米混悬剂(GA-NSps),以粒径和多分散指数(PDI)为指标选择稳定剂;采用动态光散射法、透射电镜考察GA-NSps粒径和形态,在生物介质中的稳定性;透析法测量GA-NSps体外释放;MTT比色法考察GA-NSps细胞毒性作用;建立H22荷瘤小鼠模型,考察GA-NSps体内抗肿瘤作用。结果表明,泊洛沙姆为GA-NSps的优良稳定剂;药载比7∶1时,所制备的GA-NSps近似规则球形,分布均匀,粒径为135.9 ±5.1 nm,zeta电位为-35.1 ±1.36 mV,PDI为0.26 ±0.01,在多种生物介质中均可以稳定存在;体外释放缓慢,312 h累计释放量为90.26%。MTT结果显示,藤黄酸制备成纳米混悬剂后,体外抗肿瘤活性显著提高,GA-NSps和藤黄酸的IC50值分别为0.851 8和2.104 μg·mL-1,两者相比具有显著性差异(P<0.05)。在荷瘤鼠药效学实验中,GA-NSps显示出良好的量效关系,高剂量组的肿瘤抑制率(72.35%)高于紫杉醇注射液组(66.80%,P<0.05)。综上,本研究制备的GA-NSps载药量高、稳定性良好,增强了藤黄酸的抗肿瘤效果,为藤黄酸的剂型研究奠定了基础。
Preparation of gambogic acid naonosuspensions and their anti-tumor efficacy
JI Yu-bin1, NIE Fan-ru1,2, ZHOU Xin-xin1,2, WANG Xiang-tao1,2
1. Life Sciences and Environmental Sciences Center, Harbin University of Commerce, Harbin 150076, China;
2. Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China
Gambogic acid (GA), the main active ingredient in gamboge, has been reported to have good anti-tumor activity with excellent selectivity. However, its clinical application is limited by the poor water solubility. GA nanosuspensions were designed in this study in order to solve this problem. GA nanosuspensions were prepared by microprecipitation method based on pH adjustment. Suitable stabilizer was screened according to the size and polydispersity index (PDI) of the resultant nanosuspensions. Dynamic light scattering method was used to measure the particle size and transmission electron microscopy was used to observe the morphology. The stability was studied in different medium. The drug release was evaluated using a dialysis method. MTT assay was used to assess their cytotoxicity in vitro against cancer cell line. Anti-tumor effect in vivo was investigated on H22-bearing mice. In result, Poloxamer (P188) was found to be a good stabilizer. The resultant GA nanosuspensions (GA-NSps) were 135.9 ±5.1 nm in diameter, with PDI value being 0.26 ±0.01 and the zeta potential being −35.1 ±1.36) mV. GA-NSps were nearly spherical. They were quite stable in various physiological media. GA-NSps exhibited a sustained drug release pattern, with the cumulative release reaching 90.26% within 312 h. In MTT assay, GA-NSps had a stronger cytotoxicity against HepG2 cells than the free drug (IC50, 0.851 8 μg·mL−1 vs 2.104 μg·mL−1, P < 0.05). The pharmacodynamics study suggest that the antitumor effect of GA-NSps was dose-dependent. The anti-tumor effect at the high dose is better than that of paclitaxel (72.35% vs 66.80%, P < 0.01). In summary, we prepared GA-NSps with high drug loading capacity, small particle size and good stability, and provided a solid basis for the effective dosage form of gambogic acid.
Key words:   
收稿日期: 2017-10-16
DOI: 10.16438/j.0513-4870.2017-1005
基金项目: 国家自然科学基金−;广东联合基金资助项目(U1401223).
通讯作者: 王向涛,Tel/Fax:86-10-57833264,E-mail:xtaowang@163.com
Email: xtaowang@163.com
PDF(434KB) Free

[1] Hou WJ, Xiao W. Advances in studies on gambogic acid[J]. Chin Tradit Herb Drugs (中草药), 2011, 42:617-620.
[2] Ou SP, Wang S, Yang QY, et al. Advances in studies on dried resin of Garcinia hanburyi processing in toxicity reducing and efficacy enhancing effects[J]. Chin Tradit Herb Drugs (中草药), 2011, 42:2560-2563.
[3] Li Ping. Study on Phytochemistry and Metabonomics of Garcinia (藤黄属植物的化学成分及其代谢组学研究)[D]. Beijing:Minzu University of China, 2016.
[4] Lv GB, Yang XX, Huang QS. Isolation and structure of neo-gambogic acid from gamboge (Garcinia hanburyi)[J]. Acta Pharm Sin (药学学报), 1984, 19:636-639.
[5] Kashyap D, Mondal R, Tuli HS, et al. Molecular targets of gambogic acid in cancer:recent trends and advancements[J]. Tumour Biol, 2016, 37:12915-12925.
[6] Li D, Yang H, Li RP, et al. Antitumor activity of gambogic acid on NCI-H1993 xenografts via MET signaling pathway downregulation[J]. Oncol Lett, 2015, 10:2802-2806.
[7] Chen J, Zhou M, Zhang Q, et al. Anticancer effect and apoptosis induction of gambogic acid in human leukemia cell line K562 in vitro[J]. Med Sci Monit, 2015, 21:1604- 1610.
[8] Zhang Z, Qian H, Yang M, et al. Gambogic acid-loaded biomimetic nanoparticles in colorectal cancer treatment[J]. Int J Nanomed, 2017, 12:1593-1605.
[9] Xia G, Wang H, Song Z, et al. Gambogic acid sensitizes gemcitabine efficacy in pancreatic cancer by reducing the expression of ribonucleotide reductase subunit-M2(RRM2)[J]. J Exp Clin Cancer Res, 2017, 36:107.
[10] Ye JL, Yu YJ, Wu AL et al. Sensitization of human colon cancer HT-29 cells to TRAIL-induced apoptosis by gambognic acid[J]. Acta Pharm Sin (药学学报), 2015, 50:1252-1257.
[11] Gu H, Rao S, Zhao J, et al. Gambogic acid reduced bcl-2 expression via p53 in human breast MCF-7 cancer cells[J]. J Cancer Res Clin Oncol, 2009, 135:1778-1782.
[12] Jang JH, Kim JY, Sung EG, et al. Gambogic acid induces apoptosis and sensitizes TRAIL-mediated apoptosis through downregulation of cFLIPl in renal carcinoma Caki cells[J]. Int J Oncol, 2016, 48:376-384.
[13] Zhu X, Zhang H, Lin Y, et al. Mechanisms of gambogic acid-induced apoptosis in non-small cell lung cancer cells in relation to transferrin receptors[J]. J Chemother, 2009, 21:666-672.
[14] Zhang W, Zhou H, Yu Y, et al. Combination of gambogic acid with cisplatin enhances the antitumor effects on cisplatin-resistant lung cancer cells by downregulating MRP2 and LRP expression[J]. OncoTargets Ther, 2016, 9:3359-3368.
[15] Hou WJ, Chen BL, Zhao YW, et al. Stability of gambogic acid[J]. Chin Tradit Herb Drugs (中草药), 2011, 42:1755- 1758.
[16] Zhou ZT. Phase I Trial of Human Tolerability and Pharma­cokinetics to Gambogic Acid (注射用藤黄酸Ⅰ期临床耐受性及药代动力学试验)[D]. Beijing:Peking Union Medical College, 2006.
[17] Zhu J, Wang M, Zhu Y. Quantitative cardiotoxicity assess­ment of gambogic acid using multiple cellular phenotype analysis[J]. Chin J Pharmacol Toxicol (中国药理与毒理学杂志), 2017, 31:73-79.
[18] Wang BN. Research on Extraction and Separation and Pharmaco-toxicology of Gambogic Acid and Neogambogic Acid in Garcinia hanburyi (藤黄中藤黄酸和新藤黄酸的提取分离及药理毒理研究)[D]. Changchun:Jilin Agricultural University, 2015.
[19] Au JL, Yeung BZ, Wientjes MG, et al. Delivery of cancer therapeutics to extracellular and intracellular targets:determi­nants, barriers, challenges and opportunities[J]. Adv Drug Deliv Rev, 2016, 97:280-301.
[20] Maeda H. Toward a full understanding of the EPR effect in primary and metastatic tumors as well as issues related to its heterogeneity[J]. Adv Drug Deliv Rev, 2015, 91:3-6.
[21] Wang LY, Liu YM, Wu LY, et al. Preparation and pharma­cokinetics of gambogic acid long-circulating liposomes[J]. Chin Tradit Herb Drugs (中草药), 2016, 47:1309-1314.
[22] Yan X, Yang Y, He L, et al. Gambogic acid grafted low molecular weight heparin micelles for targeted treatment in a hepatocellular carcinoma model with an enhanced anti-angiogenesis effect[J]. Int J Pharm, 2017, 522:110-118.
[23] Zhang D, Zou Z, Ren W, et al. Gambogic acid-loaded PEG-PCL nanoparticles act as an effective antitumor agent against gastric cancer[J]. Pharm Dev Technol, 2018, 23:33-40.
[24] Dai YJ, Wang F, Huang P, et al. Preparation and characteri­zation of magnetic nano Fe3O4 load neogambogic acid[J]. Guangzhou Chem Ind (广州化工), 2016, 44:71-73, 84.
[25] Liu CY, Wang W, Zhou JP, et al. Preparation and evaluation of gambogic acid-loaded reconstituted high density lipoprotein nanoparticles[J]. China Pharm Univ (中国药科大学学报), 2013, 44:311-315.
[26] Yang XF, Guo RQ, Su WJ, et al. Prescription screening and preparation process of 10-hydroxycamptothecin nanocrystals and industrialization research[J]. Her Med (医药导报), 2016, 35:1333-1340.
[27] Zhang CY, Li J, Gao JM, et al. The impurity profiling of paclitaxel and its injection by UPLC-MS/MS[J]. Acta Pharm Sin (药学学报), 2016, 51:965-971.