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氨基酸构型对多肽纳米纤维体内分布的影响

徐宏艳1,张玉民1,杨翠红1,刘金剑2,褚丽萍2,刘鉴峰2,宋娜玲1   

  1. 1. 北京协和医学院＆中国医学科学学院放射医学研究所
    2. 中国医学科学院放射医学研究所
  • 收稿日期:2013-10-31 修回日期:2013-11-27 出版日期:2014-02-15 发布日期:2014-02-15
  • 通讯作者: 宋娜玲

The effect of amino acid configuration on the biodistribution of peptide nanofiber

  • Received:2013-10-31 Revised:2013-11-27 Published:2014-02-15 Online:2014-02-15

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摘要： 【摘要】 目的 比较由L构型和D构型氨基酸自组装形成的纳米纤维在体内分布的差异，为不同构型氨基酸自组装纳米多肽的体内应用提供指导。方法 固相合成法合成多肽Nap-GFFYGRGD（L-肽）和Nap-GDFDFDYGRGD（D-肽），利用核磁和质谱对多肽分子进行结构表征。多肽溶液通过煮沸、冷却、自组装形成纳米纤维（L-纤维和D-纤维），透射电镜观察纳米纤维的微观形貌。125I标记多肽分子后，由其自组装形成的纳米纤维通过尾静脉注射入小鼠体内，分别在1、3、6和12 h采血并处死小鼠，取心、肝、脾、肺、肾、胃、大肠、小肠、肌肉、脑等主要器官，用γ计数仪测量其放射性强度。结果 L-肽和D-肽均可自组装形成纳米纤维，纤维直径约为10~20 nm，且两者微观形貌差异无统计学意义。两种纳米纤维在体内的分布差异有统计学意义。D-纤维在注射后1 h的血液浓度为（8.17±0.32）%ID/g，但较迅速地从血液中清除；L-纤维浓度为（5.96±0.30）%ID/g，在注射后6 h基本保持不变。D-纤维主要分布于肝中而L纤维主要分布在胃中。结论 氨基酸构型（D/L）对多肽纳米纤维在体内的分布影响显著，在未来的医学应用中考虑氨基酸构型对体内分布的影响有可能更好地指导多肽纳米纤维的应用。

关键词: 肽类, 氨基酸序列, 同位素标记, 组织分布, 多肽纳米纤维

Abstract: [Abstract] Objective To compare the biodistribution difference of peptide nanofibers which were self-assembled by peptide composed of L- or D-amino acids, respectively. And provide guidance for the in vivo applications of peptide nanofibers. Methods The Nap-GFFYGRGD (L-peptide) and Nap-GDFDFDYGRGD (D-peptide, F and Y were D-configuration) were synthesized with 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 was observed with transmission electron microscope (TEM). The peptides were radiolabled with iodine-125 and self-assembled into nanofibers, then administrated into BALB/c mice via tail vein. The blood was collected and then the mice were sacrificed at 1, 3, 6 and 12 hours. The main organs (heart, liver, spleen, lung, kidney, stomach, large intestine, small intestine, muscle and brain) were isolated and weighed. The radioactivity of organs was detected with a gamma counter. Results The two peptides could self-assemble into nanofibers with diameter of 10-20 nanometers. The diameter and morphology of the two naofibers had no statistically significant difference. Biodistribution of the two nanofibers had statistically significant difference. The concentration of L-fiber in blood at one hour after injection was 5.9±0.30 %ID/g and maintained at a stable level during six hours after injection. While the conterpart of D-fiber was 8.17±0.32 %ID/g and then cleared rapidly from the blood. The L-fiber mainly accumulated in stomach while the D-fiber mainly accumulated in liver. Conculsion The configuration of amino acids (D/L) could affect the biodistribution of peptide nanofibers dramatically, this results may provide guidance for the medical applications of peptide nanofibers.

Key words: peptides, amino acid sequence, isotope labeling, tissue distribution, peptide nanofiber