RGD功能化多肽纳米纤维的制备及其体内肿瘤靶向性研究

发布时间：2018-03-26 09:53

本文选题：RGD　切入点：同位素标记　出处：《中国生化药物杂志》2014年05期

【摘要】：目的制备肿瘤靶向RGD多肽纳米纤维,研究其体内分布和肿瘤靶向性。方法通过多肽固相合成法制备靶向多肽Nap-GFFYGRGD(RGD-肽)和对照多肽Nap-GFFYGRGE(RGE-肽),利用核磁和质谱对多肽的分子结构进行表征。多肽溶液经煮沸后冷却可自组装形成纳米纤维(RGD-纤维和RGE-纤维),通过透射电镜(TEM)对纳米纤维的微观形貌进行观察。采用氯胺-T法对多肽进行125I标记,利用放射性HPLC对标记多肽进行分离纯化。建立BALB/c小鼠皮下乳腺癌肿瘤模型,125I标记的多肽自组装形成纳米纤维后经尾静脉注射,分别于注射后1、3、6、12 h进行眼球取血并处死小鼠,取肿瘤、心、肝、脾、肺、肾、胃、大肠、小肠、肌肉和脑等,用γ计数仪测量各组织放射性信号强度。结果 RGD-肽和RGE-肽均可自组装形成直径约为10~20 nm的纳米纤维。RGD-纤维和RGE-纤维在注射后各个时间点在体内主要脏器的分布规律相似,主要分布于胃中,其次是肠。但2者在肿瘤组织的分布规律存在显著性差异,RGD-纤维注射后6 h内在肿瘤组织中呈现出一个逐渐积累的过程,而RGE-纤维在3 h时达最高浓度,在6 h 2者差异达到最大,分别为6.25%ID/g和2.79%ID/g(P0.01)。结论肿瘤靶向肽RGD能明显提高多肽纳米纤维在体内的肿瘤靶向分布,为该载体作为抗肿瘤药物载体用于肿瘤靶向治疗提供了强有力的数据支持。
[Abstract]:Objective to prepare tumor targeting RGD polypeptide nanofibers. Methods the target peptide Nap-GFFYGRGDG (RGD- peptide) and the control peptide Nap-GFFYGRGERGE-peptide (Nap-GFFYGRGERGE-peptide) were prepared by solid phase peptide synthesis method. The molecular structure of the peptide was characterized by NMR and mass spectrometry. The polypeptide solution was boiled. The nanofibers RGD- and RGE-fiber can be self-assembled by cooling. The micromorphology of the nanofibers was observed by transmission electron microscopy (TEM). The polypeptides were labeled with 125i by chloramine-T method. The labeled polypeptides were isolated and purified by radioactive HPLC. A tumor model of subcutaneous breast cancer in BALB/c mice was established. The peptides were self-assembled to form nanofibers and injected via tail vein. The blood was taken from the eyeball at 12 h after injection, and the mice were killed. Tumor, heart, liver, spleen, lung, kidney, stomach, large intestine, small intestine, muscle and brain, Results RGD- peptide and RGE-peptide were self-assembled to form nanofibers, RGD- and RGE-fibers with diameter of about 1020 nm. The distribution of RGD- and RGE-fibers in the main organs was similar at each time point after injection. But there was a significant difference in the distribution of RGD- fibers in the tumor tissues within 6 h after RGD- fiber injection, and the RGE- fibers reached the highest concentration at 3 h after the injection of RGD- fibers, which were mainly distributed in the stomach, followed by the intestine, but there was a significant difference in the distribution of RGD- fibers between the two groups within 6 h after the injection of RGD- fibers. At 6 h 2, the difference was the greatest, 6.25%ID/g and 2.79 IDP / G P 0.01, respectively. Conclusion tumor targeting peptide RGD can significantly improve the tumor targeting distribution of polypeptide nanofibers in vivo. It provides a powerful data support for the use of the vector as an antitumor drug carrier for tumor targeting therapy.
【作者单位】：北京协和医学院&中国医学科学院放射医学研究所
【基金】：国家自然科学基金资助项目(51303213) 协和青年科研基金滚动支持项目(3332014003) 中国医学科学院放射医学研究所所基金项目(SK1425,SF1416,SF1417)
【分类号】：R318.08

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