高等学校化学学报 ›› 2016, Vol. 37 ›› Issue (4): 661.doi: 10.7503/cjcu20150941

• 有机化学 • 上一篇    下一篇

嗜神经病毒衍生肽RVG29介导的靶向纳米载药胶束的合成及抗肿瘤活性

韩海玲1,2, 金顺子1, 苗壮3, 陈平3, 王占峰3, 谢志刚4   

  1. 1. 吉林大学公共卫生学院, 卫生部放射生物学重点实验室, 长春 130021;
    2. 北华大学附属医院心内三科, 吉林 132011;
    3. 吉林大学中日联谊医院神经外二科, 长春 130033;
    4. 中国科学院长春应用化学研究所, 高分子物理与化学国家重点实验室, 长春 130022
  • 收稿日期:2015-12-09 出版日期:2016-04-10 发布日期:2016-03-04
  • 通讯作者: 王占峰,男,博士,副主任医师,主要从事脑肿瘤和脑血管病的纳米药物治疗和基础研究.E-mail:1206hhl@sina.com E-mail:1206hhl@sina.com
  • 基金资助:

    国家自然科学基金(批准号: 20904002)、教育部新世纪优秀人才计划项目(批准号: NCET-10-0171)、吉林省科技厅青年项目(批准号: 20130522020JH)、吉林大学白求恩医学科研支持计划(批准号2013107027)、吉林省教育厅十二五重大课题(批准号: 2013107027)和吉林省高技术产业发展专项项目(批准号: JF2012C005-2)资助.

Synthesis and Antitumor Activity of Targeting and Nano Drug-Carrying Micelles Mediated by Rabies-virus-derived Peptide RVG29

HAN Hailing1,2, JIN Shunzi1, MIAO Zhuang3, CHEN Ping3, WANG Zhanfeng3, XIE Zhigang4   

  1. 1. Ministry of Health Key Laboratory of Radiobiology, School of Public Health, Jilin University, Changchun 130021, China;
    2. Department of Cardiac, Affiliated Hospital of Beihua University, Jilin 132011, China;
    3. Department of Neurosurgery, China Japan Union Hospital, Jilin University, Changchun 130033, China;
    4. State Key Lab of Polymer Physics and Chemistry, Changchun Institute of Applied Chemistry, Changchun 130022, China
  • Received:2015-12-09 Online:2016-04-10 Published:2016-03-04
  • Supported by:

    Supported by the National Natural Science Foundation of China(No.20904002), the Program for New Century Excellent Talents in University of Ministry of Education of China(No.NCET-10-0171), the Young Scholars Programs of Science and Technology Commission Foundation of Jilin Province, China(No. 20130522020JH), the Programs of Educational Commission of Jilin Province of China During the 12th Five-Year Plan Period(No.2013107027) and the High-Technology Research and Development Programs of Jilin Province of China(No.JF2012C005-2).

摘要:

将羧基化的水溶性葡聚糖(Dex)与紫杉醇(PTX)化学偶联, 制得载药纳米胶束M(PTX), 再将M(PTX)与嗜神经性病毒衍生肽(RVG29)化学偶联, 得到RVG29靶向的载药纳米胶束M(RVG,PTX). 采用核磁共振氢谱(1H NMR)测定了Dex-PTX及RVG-Dex-PTX键合物的分子量, 并对2种胶束进行了表征, 考察了2种胶束对肿瘤细胞的抑制效果及细胞凋亡情况, 观察了C6细胞对荧光标记M(RVG,PTX)和M(PTX)的摄取情况. 结果表明, 羧基化葡聚糖-紫杉醇键合物的分子量约为16500, 紫杉醇的质量约为葡聚糖的20%, RVG29的质量约为葡聚糖的10%. 2种胶束的粒径在45~60 nm之间; M(RVG,PTX)胶束对C6细胞的抑制作用具有浓度和时间依赖性, 细胞抑制率随着作用时间和药物浓度增加而增加, 且M(RVG,PTX)胶束对C6细胞的抑制作用强于M(PTX)胶束. 细胞摄取实验结果表明, 与M(PTX)相比, C6细胞摄取了更多的M(RVG,PTX)胶束. 如果先用游离的RVG29处理C6细胞, 再进行细胞实验, 则M(RVG,PTX)胶束对C6细胞生长的抑制作用及被C6细胞摄取的比率显著降低, 与 M(PTX)相当. 表明靶向载药胶束M(RVG,PTX)中的RVG29保留了游离RVG29的活性, 对C6细胞依然具有靶向效应, 从而介导了M(RVG,PTX)被C6细胞的摄取, 增强了对C6细胞的生长抑制作用. 由于M(RVG,PTX)胶束只使用水溶性葡聚糖作载体, 不涉及疏水高分子链段, 不需要分别制备载药高分子和靶向高分子然后再共组装, 因而制备过程比较简单, 同时具有载药和靶向功能.

关键词: 嗜神经性病毒衍生肽, 脑胶质瘤, 肿瘤靶向, 紫杉醇, 纳米载药胶束

Abstract:

Starting with water soluble dextran(Dex), paclitaxel(PTX)-carrying micelles M(PTX) and Rabies-virus-derived peptide(RVG29)-targeted and PTX-carrying micelles M(RVG,PTX) were prepared, by carboxylation of dextran and subsequently by conjugating PTX and RVG29 onto the carboxylated dextran. The products were characterized with nuclear magnetic resonance(1H NMR) spectroscopy, transmission electron microscopy(TEM), and dynamic light scattering(DLS). The uptakes of the micelles by C6 cells and the inhibition rates of C6 cell growth by the micelles were measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide(MTT) assay, by confocal laser scanning microscopy(CLSM), and by flow cytometry(FCM). The results showed that molecular weight of Dex-PTX was about 16500 and that of RVG-Dex-PTX was about 19800. Micelles M(PTX) and M(RVG, PTX) assumed a shape of spheres with a diameter of 45-60 nm. The mass ratio of Dex, PTX and RVG29 was about 100: 20: 10. M(RVG, PTX) exhibited higher cell growth inhibition rate against C6 cells than M(PTX) and C6 cells took in more M(RVG,PTX) than M(PTX) under the same experimental conditions. When C6 cells were treated with free RVG29 prior to cellular experiments, the inhibition rate against C6 cells and the uptake by C6 cells of M(RVG,PTX) were significantly decreased and came to a level of M(PTX). This implied that the enhanced inhibition of C6 cell growth by M(RVG,PTX) and the enhanced uptake of M(RVG,PTX) by C6 cells were mediated by RVG29, and the RVG29 species after conjugation with Dex reserved the targeting ability of RVG29 itself to a reasonable extent. Furthermore, in the preparation of M(RVG, PTX) micelles, only Dex was used as a carrier polymer, no hydrophobic polymer segments were involvd and there was no need of preparing drug-carrying copolymers and targeting copolymers, separately, and then doing co-assembling of them. In short, the manufacturing of M(RVG,PTX) micelles is relatively simple but they have the functions of drug-carrying and C6-cell targeting, therefore, their practical application is expected.

Key words: Rabies-virus-derived peptide, Glioma, Tumor targeting, Paclitaxel, Nano drug-carrying micelle

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