CAJ | 학술논문

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，L-lactic-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 acti-vator，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。
목적：제비여평개포재반의핵산약물-류감태득적투막태수식적취유산-간기을산공취물[poly（D，L-lactic-co-glycolic acid ），PLGA]납미립。방법통과고상다태합성법합성투막태（ trans-activating transcriptional acti-vator，TAT），사용고효액상색보급질보의검측기순도급상대분자질량。용TAT수식PLGA후，채용쌍중유화휘발용제법제비포재류감태득적신형납미립，병대납미립진행이화분석화체외완석효과평개，주요지표유：립경、분산계수、Zeta전위、표면형태、재약량급체외석방。 Cell Count Kit-8（CCK-8）실험평개TAT-PLGA납미립적세포독성。결과합성적TAT순도위95．6％，상대분자질량위1495．8，TAT수식PLGA후소제비적납미립립경위（159．5±2．1） nm，Zeta전위위-（1．87±0．6）mV，재약량위（5．75±0．17）μg/mg。재소묘전자현미경하관찰납미립형태위원형，표면광활，립경분포균균。체외석방실험결과현시TAT수식적PLGA납미립포봉적류감태득구유완석작용。세포독성실험결과현시，TAT수식적PLGA납미립대세포체외생장무불량영향，세포상용성량호。결론통과고상다태합성법성공합성료TAT，병성공사용쌍중유화용제휘발법제비료TAT수식적PLGA납미립，재류감예방여치료방면구유잠재응용전경。
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-8 methods.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 sustained-release effect and good compatibility in vitro.They have potential application prospect in prevention and treatment of influenza .