CAJ | 학술논문

目的：合成一种聚天冬酰胺衍生物并对其细胞水平的安全性进行评价，为其作为药物载体应用提供研究基础。方法通过L-天冬氨酸热缩聚合成聚琥珀酰亚胺，利用苯丙氨酸甲酯盐酸盐和乙醇胺对聚琥珀酰亚胺进行开环反应得到载体PSI-Phe-EA；利用1H NMR进行聚合物结构表征；采用内标法磁氢谱计算其开环率；通过水溶性的比较验证其亲水性变化；采用MTT法对聚合物的细胞增殖抑制进行研究，利用倒置显微镜观察聚合物对细胞微观形态的影响；利用碘化丙啶（PI）染色法通过流式细胞仪研究聚合物对细胞周期的影响。结果1H NMR确证了开环衍生物PSI-Phe-EA的结构，PSI的开环率为40%；乙醇胺开环后聚合物的亲水性得到了明显改善；MTT实验表明， PSI-Phe-EA在所检测浓度范围内（<100 mg/L），对NIH3T3和HepG2两种细胞的24 h细胞存活率均在80%以上；倒置显微镜观察表明，50 mg/L的PSI-Phe-EA孵育24 h后以上两种细胞的形态与对照组无明显差异；细胞周期分析表明PSI-Phe-EA处理与否对细胞周期分布的影响无明显差异。结论合成的聚天冬酰胺衍生物PSI-Phe-EA亲水性明显提高，且对细胞的存活、微观形态以及周期分布均无明显影响，是一种安全的高分子材料。
목적：합성일충취천동선알연생물병대기세포수평적안전성진행평개，위기작위약물재체응용제공연구기출。방법통과L-천동안산열축취합성취호박선아알，이용분병안산갑지염산염화을순알대취호박선아알진행개배반응득도재체PSI-Phe-EA；이용1H NMR진행취합물결구표정；채용내표법자경보계산기개배솔；통과수용성적비교험증기친수성변화；채용MTT법대취합물적세포증식억제진행연구，이용도치현미경관찰취합물대세포미관형태적영향；이용전화병정（PI）염색법통과류식세포의연구취합물대세포주기적영향。결과1H NMR학증료개배연생물PSI-Phe-EA적결구，PSI적개배솔위40%；을순알개배후취합물적친수성득도료명현개선；MTT실험표명， PSI-Phe-EA재소검측농도범위내（<100 mg/L），대NIH3T3화HepG2량충세포적24 h세포존활솔균재80%이상；도치현미경관찰표명，50 mg/L적PSI-Phe-EA부육24 h후이상량충세포적형태여대조조무명현차이；세포주기분석표명PSI-Phe-EA처리여부대세포주기분포적영향무명현차이。결론합성적취천동선알연생물PSI-Phe-EA친수성명현제고，차대세포적존활、미관형태이급주기분포균무명현영향，시일충안전적고분자재료。
Objective To synthesize poly asparagine derivatives and to evaluate its safety at the cellular level, which provide research platform for its potential application as drug carrier. Methods Polysuccinimide was synthesized by ther?mal polymerization of L-polyaspartic acid, and the target product of PSI-Phe-EA was obtained by the ring-opening reaction of polysuccinimide using L-phenylalanine methyl ester hydrochloride and ethanol amine. The structure of PSI-Phe-EA were characterized by 1H NMR. The rate of ring-opening of PSI was calculated by internal standard method of 1H NMR. The change of hydrophilicity was studied by the comparison of solubility. The cytotoxicity and morphology modification by PSI-Phe-EA at designate concentrations was investigated by MTT method and inverted microscopy respectively. The effects on cell cycles were analyzed by flow cytometry after propidium iodide (PI) staining. Results 1H NMR results confirmed the structure of PSI-Phe-EA and the ring-openning rate of PSI was 40%. The hydrophilicity of PSI-Phe-EA was greatly in?creased upon ring opening using ethanol amine. MTT test showed that the cell survival rates of NIH 3T3 and HepG2 cells were higher than 80%under the examined concentration (<100 mg/L). Inverted microscopy showed that 50 mg/L of PSI-Phe-EA treatment had no adverse effects on cell morphology. Cell cycle analysis indicated that PSI-Phe-EA treatment had no in?fluence on cell cycles of NIH 3T3 and HepG2 cell lines. Conclusion PSI-Phe-EA showed high hydrophilicity without sig?nificant effects on the cells survival, cells morphology and cell cycles. It is a kind of safe polymer material.