苗口服接种新载体：荆豆凝集素化壳聚糖纳石米粒传输系统的研究.docVIP

PAGE PAGE 1 苗口服接种新载体：荆豆凝集素化壳聚糖纳米粒传输系统的研究 李凤前，费轶博，刘继勇，朱全刚，胡晋红? ?基金项目：国家自然科学基金资助项目；上海市科委纳米专项课题（0552nm040）. ?通讯作者 Tel / Fax: 86-21 E-mail: HYPERLINK mailto:hujh@smmu.edu.cn hujh@smmu.edu.cn 第二军医大学长海医院药学部，上海 200433 摘要 本文依据肠黏膜及派伊尔集合淋巴结（peyer’s patches, PP）的免疫学特点，以乙型肝炎疫苗为模型药物，将能同肠道微皱褶细胞上岩藻糖特异性结合的荆豆凝集素，对壳聚糖进行修饰，构建并研究PP定位的疫苗微粒口服传输系统。 离子交联-高压匀化工艺制备的壳聚糖纳米粒形态较规则。用激光散射粒度仪测定纳米粒平均粒径为366.8nm，多分散系数为0.122，粒度分布较窄。以低温高速离心法测得乙肝疫苗纳米粒平均包封率在90%以上，通过SDS电泳表明，乙肝疫苗在反应前后没有发生分子量的变化。用戊二醛活化法制备凝集素修饰的壳聚糖纳米粒，结合率可达85.3±3.2%。岩藻糖的存在会让BSM与凝集素化纳米粒的结合受到抑制，凝集素在结合到纳米粒表面后糖结合活性的保持。荧光显微镜观察发现纳米粒在淋巴结中有一定富集效果。与市售制剂肌注组、口服乙肝疫苗未凝集素化纳米粒组、口服乙肝疫苗组、阴性对照组（生理盐水）比较，口服凝集素化乙肝疫苗纳米粒组在Balb/c小鼠，能在一定程度上引发免疫应答。 本研究表明，凝集素化壳聚糖纳米粒可以作为疫苗类药物有效的载体，用于肠PP的定向给药，为疫苗口服免疫开辟了新思路。 关键词：离子交联-高压匀化法；多聚磷酸钠；凝集素化壳聚糖纳米粒；岩藻糖；牛颌下腺粘蛋白；疫苗口服接种；派伊尔集合淋巴结 New particular carrier of oral vaccination: Ulex europaeus agglutinin Anchored chitosan nanoparticles delivery system Feng-Qian Li, Yi-Bo Fei, Ji-Yong Liu, Quan-Gang Zhu, Jin-Hong Hu? Department of Pharmaceutical Sciences, Changhai Hospital, Second Military Medical University, Shanghai, 200433, China Abstract This paper is proposed upon the immunologic characteristics of enteric-mucosa and Peyers patches (PP). Biodegradable material of chitosan was modified with the lectin of ulex europaeus agglutinin (UEA), which could be specific to the rhodeose within the microfold cell under intestinal tract, and then used for nanoencapsulation. The model hepatitis B vaccine was then entrapped within the lectin anchored nanoparticles for oral delivery. The uniform-shaped chitosan nanoparticles were prepared by inonotropic gelation-homogenization process with tripolyphosphate as gelatinizer. The mean diameter of the chitosan nanoparticles was 366.8nm with the polydispersity index of 0.122. The vaccine entrapment efficiency of nanoparticles which is separated by centrifugalization was above 90%. The lectin anchored chitosan nanoparticles were prepared with the glutaraldehyde activation method. And t