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(P<0.01)。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。
Objective To study the biodistribution and in vivo tumor-targeting ability of the RGD-functionalized peptide nanofibers and provide foundation for its in vivo applications as anti-tumor drug carriers. Methods The Nap-GFFYGRGD(RGD-peptide) and Nap-GFFYGRGD(RGE-peptide)were synthesized by solid phase peptide synthesis(SPPS). The structure of the two peptides was identified by nuclear magnetic resonance spectroscopy(1H NMR) and high-resolution mass spectrometry(HR-MS). The two peptides could self-assemble into nanofibers during the cooling process after being boiled and the morphology of the nanofibers were observed by transmission electron microscope(TEM). The peptides were radiolabeled with iodine-125 and purified by radioactive HPLC. The mouse xenograft tumor models of 4T1 was established in BALB /c mice,the iodine-125 labeled nanofibers was administrated into BALB /c mice via the tail vein. The blood were collected and then the mice were sacrificed at 1,3,6 and 12 hours,respectively. Tumors and main organs(heart,liver,spleen,lung,kidney,stomach,large intestine,small intestine,muscle and brain) were harvested and weighted. The radioactivity of organs was detected with a gamma counter. Results The RGD and RGE peptides could self-assemble into nanofibers with diameter of 10-20 nanometers( RGD-fiber and RGE fiber,respectively). Biodistribution of the two nanofibers were silimar,which mainly accumulated in stomach followed by intestine. But the distribution of RGD-fiber and RGE-fiber in tumors were significantly difference. There was a gradual accumulation process in the first 6 hours for RGD-fiber. The max difference of tumor accumulation was observed at 6 h between RGD-fiber and RGE-fiber(with concentration of 6.25% ID/g and 2.79% ID/g,respectively,P < 0.01). Conclusion The tumor-targeting tripeptide RGD could significantly improve the tumor accmulation of peptide nanofibers in vivo,these results may provide powerful experimental evidence for the application of this pepide nanofibers as drug carriers in tumor-targe
出处
《中国生化药物杂志》
CAS
北大核心
2014年第5期5-8,共4页
Chinese Journal of Biochemical Pharmaceutics
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)
