透膜肽修饰的载流感泰得PLGA纳米粒的制备与评价  

Preparation and evaluation of flutide-loaded PLGA nanoparticles modified with cell-penetrating peptides

作　　者：徐清波[1,2] 丁晓然[2] 杜丽娜[2] 鲁丹丹[2] 金义光[2] 王升启[2] 

机构地区：[1]安徽医科大学,合肥230031 [2]军事医学科学院放射与辐射医学研究所,北京100850

出　　处：《军事医学》2014年第7期518-522,共5页Military Medical Sciences

摘　　要：目的制备与评价包载反义核酸药物-流感泰得的透膜肽修饰的聚乳酸-羟基乙酸共聚物[poly(D,Llactic-co-glycolic acid),PLGA]纳米粒。方法通过固相多肽合成法合成透膜肽(trans-activating transcriptional activator,TAT),使用高效液相色谱及质谱仪检测其纯度及相对分子质量。用TAT修饰PLGA后,采用双重乳化挥发溶剂法制备包载流感泰得的新型纳米粒,并对纳米粒进行理化分析和体外缓释效果评价,主要指标有:粒径、分散系数、Zeta电位、表面形态、载药量及体外释放。Cell Count Kit-8(CCK-8)实验评价TAT-PLGA纳米粒的细胞毒性。结果合成的TAT纯度为95.6%,相对分子质量为1495.8,TAT修饰PLGA后所制备的纳米粒粒径为(159.5±2.1)nm,Zeta电位为-(1.87±0.6)mV,载药量为(5.75±0.17)μg/mg。在扫描电子显微镜下观察纳米粒形态为圆形,表面光滑,粒径分布均匀。体外释放实验结果显示TAT修饰的PLGA纳米粒包封的流感泰得具有缓释作用。细胞毒性实验结果显示,TAT修饰的PLGA纳米粒对细胞体外生长无不良影响,细胞相容性良好。结论通过固相多肽合成法成功合成了TAT,并成功使用双重乳化溶剂挥发法制备了TAT修饰的PLGA纳米粒,在流感预防与治疗方面具有潜在应用前景。Objective To prepare and evaluate flutide-loaded PLGA nanoparticles modified with cell-penetrating peptide-TAT.Methods The sequence of TAT was synthesized with florenl methyoxycarbonyl amino acids.The purity and molecular weight of TAT were determined using RP-HPLC and MALDI-TOF-MS.PLGA was modified with the TAT peptide and then prepared into flutide-loaded nanoparticles（TAT-PLGA NPs） with the double emulsion method.The physical and chemical properties were evaluated,including size distribution,Zeta potential,SEM of nanoparticles,loading ratio of drug content and release profiles of TAT-PLGA NPs in vitro.The cytotoxicity of TAT-PLGA NPs was evaluated by CCK-8methods.Results The purity of synthesized TAT was 95.6%,and molecular weight was 1495.8.The mean diameter,Zeta potential,drug loading ratio of TAT-PLGA nanoparticals were（159.5 ± 2.1） nm,-（1.87 ± 0.6） mV,and（5.75 ± 0.17）μg/mg,respectively.The nanoparticles observed by transmission electron microscopy（TEM） had a spherical shape and uniform size without aggregation.In vitro release test showed sustained release of flutide from TAT-PLGA nanoparticles.Cell proliferation assay revealed that the TAT-PLGA nanoparticles did not damage the cell growth in vitro and showed good compatibility.Conclusion TAT-PLGA nanoparticles are prepared successfully by double emulsion method,and have sustainedrelease effect and good compatibility in vitro.They have potential application prospect in prevention and treatment of influenza.

关 键 词：纳米粒 透膜肽 聚乳酸-羟基乙酸 反义寡核苷酸 缓释 

分 类 号：TQ460.6[化学工程—制药化工]