结核多肽壳聚糖纳米粒的制备及表征  

Preparation and Characterization of Chitosan Nanoparticles Loaded with Mycobacterium tuberculosis Protective Antigen Polypeptide

作　　者：王梦娅 祖向阳 陈凡[2] 赵战勤[3] 薛云[1] WANG Mengya;ZU Xiangyang;CHEN Fan;ZHAO Zhanqin;XUE Yun(College of Medical Technology and Engineering,Henan University of Science and Technology,Luoyang 471023;School of Life Sciences,Hubei University,Wuhan 430062;College of Animal Science and Technology,Henan University of Science and Technology,Luoyang 471023)

机构地区：[1]河南科技大学医学技术与工程学院,河南洛阳471023 [2]湖北大学生命科学院,湖北武汉430062 [3]河南科技大学动物与科技学院,河南洛阳471023

出　　处：《中国医药工业杂志》2020年第4期505-510,共6页Chinese Journal of Pharmaceuticals

基　　金：国家自然科学基金(31672530);河南省基础与前沿技术研究计划项目(162300410091)。

摘　　要：采用离子交联法制备壳聚糖纳米粒(CS-NPs)载体,并以粒径和多分散系数(PDI)为主要评价指标优化了工艺参数。通过文献检索,筛选出结核分枝杆菌复合群(Mycobacterium tuberculosis complex,MTC)标准强菌株H37Rv的保护性抗原蛋白Rv0180c(GLDKNKFDIRVVSPDEARRLY),经异硫氰荧光素(FITC)标记后载入优化的空白CS-NPs中,获得结核多肽壳聚糖纳米粒(Pep-CS-NPs)。所得空白CS-NPs的平均粒径、PDI和ζ电位分别为(148.13±2.24)nm、0.197±0.013和(+18.00±0.89)mV;4℃放置28 d,粒径未见显著增大。Pep-CS-NPs的平均粒径、PDI和ζ电位分别为(186.93±8.80)nm、0.254±0.014和(+12.07±1.68)mV,包封率和载药量为(45.20±2.95)%和(12.92±1.12)%。体外释放结果表明,Pep-CS-NPs缓慢释放多肽,48 h累积释放率为56.6%。本试验表明,离子交联法制备载多肽CS-NPs的工艺简便稳定、条件温和,所得制品粒径大小适宜、分布均匀,且具有明显缓释作用,有望作为治疗结核病的新型疫苗。The chitosan nanoparticles(CS-NPs)were prepared by ion cross-linking method,and the preparation parameters were optimized with mean diameter and polydispersion index(PDI)as indexes.The protective antigen protein Rv0180c(GLDKNKFDIRVVSPDEARRLY)in Mycobacterium tuberculosis H37Rv was synthesized according to the results of literature search.Then,the fluorescein isothiocyanate(FITC)labelled protein was encapsulated into the optimal blank CS-NPs to obtain the title nanoparticles(Pep-CS-NPs).The mean diameter,PDI andζpotential of the blank CS-NPs were(148.13±2.24)nm,0.197±0.013 and(+18.00±0.89)mV,respectively.No significant changes in particle size and size distribution of blank CS-NPs stored at 4℃for 28 d were observed.The mean diameter,PDI,ζpotential,encapsulation efficiency and drug loading of the Pep-CS-NPs were(186.93±8.80)nm,0.254±0.014,(+12.07±1.68)mV,(45.20±2.95)%and(12.92±1.12)%,respectively.In vitro release results showed that Pep-CS-NPs slowly released peptides and the cumulative release at 48 h was 56.6%.It was concluded that the process of preparing peptide-loaded CS-NPs by ion cross-linking method was simple,stable and mild,and the product had suitable particle size,uniform distribution and sustained-release profile,which was expected to be a new vaccine for treating tuberculosis.

关 键 词：结核病 壳聚糖 结核多肽 保护性抗原多肽 纳米粒 体外释放 

分 类 号：R944.9[医药卫生—药剂学]