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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 poly (D, Llactic-co-glycolic acid) (PLGA) nanoparticle with encapsulated antitumor drug-flutadet. Methods Trans-activating transcriptional activator (TAT) was synthesized by solid-phase peptide synthesis. The purity and relative molecular mass of TAT were determined by high performance liquid chromatography and mass spectrometry. After modification of PLGA by TAT, the new emulsifiable nanoparticles loaded with fluconazole were prepared by dual emulsification of volatile solvents. The physicochemical properties and in vitro release of nanoparticles were evaluated. The main indexes of the nanoparticles were particle size, dispersion coefficient, Zeta potential, Surface morphology, drug loading and in vitro release. Cell Count Kit-8 (CCK-8) assay was used to evaluate the cytotoxicity of TAT-PLGA nanoparticles. Results The purity of TAT was 95.6% and the relative molecular mass was 1495.8. The diameter of the prepared nanoparticles after TAT modified PLGA was (159.5 ± 2.1) nm and the Zeta potential was (-1.87 ± 0.6) mV. The drug loading was ( 5.75 ± 0.17) μg / mg. Under the scanning electron microscope, the morphology of the nanoparticles was round, with a smooth surface and uniform particle size distribution. In vitro release experiments showed that TAT-modified PLGA nanoparticle encapsulated influenza Taide has a sustained release effect. Cytotoxicity test results showed that TAT modified PLGA nanoparticles on cell growth in vitro without adverse effects, good cell compatibility. Conclusion TAT was successfully synthesized by solid-phase peptide synthesis and TAT-modified PLGA nanoparticles were successfully prepared by dual emulsion solvent evaporation method. It has potential applications in the prevention and treatment of influenza.