de Jonge Jørgen, Amorij Jean-Pierre, Hinrichs Wouter L J, Wilschut Jan, Huckriede Anke, Frijlink Henderik W
Department of Medical Microbiology, Molecular Virology Section, University Medical Center Groningen and University of Groningen, Groningen, The Netherlands.
Eur J Pharm Sci. 2007 Sep;32(1):33-44. doi: 10.1016/j.ejps.2007.05.112. Epub 2007 Jun 2.
Influenza virosomes are reconstituted influenza virus envelopes that may be used as vaccines or as carrier systems for cellular delivery of therapeutic molecules. Here we present a procedure to generate influenza virosomes as a stable dry-powder formulation by freeze-drying (lyophilization) using an amorphous inulin matrix as a stabilizer. In the presence of inulin the structural integrity and fusogenic activity of virosomes were fully preserved during freeze-drying. For example, the immunological properties of the virosomes, i.e. the HA potency in vitro and the immunogenic potential in vivo, were maintained during lyophilization in the presence of inulin. In addition, compared to virosomes dispersed in buffer, inulin-formulated virosomes showed substantially prolonged preservation of the HA potency upon storage. Also the capacity of virosomes to mediate cellular delivery of macromolecules was maintained during lyophilization in the presence of inulin and upon subsequent storage. Specifically, when dispersed in buffer, virosomes with encapsulated plasmid DNA lost their transfection activity completely within 6 weeks, whereas their transfection activity was fully preserved for at least 12 weeks after incorporation in an inulin matrix. Thus, in the presence of inulin as a stabilizing agent, the shelf-life of influenza virosomes with and without encapsulated macromolecules was considerably prolonged. Formulation of influenza virosomes as a dry-powder is advantageous for storage and transport and offers the possibility to develop needle-free dosage forms, e.g. for oral, nasal, pulmonal, or dermal delivery.
流感病毒体是重组的流感病毒包膜，可作为疫苗或作为治疗分子细胞递送的载体系统。在此，我们展示了一种通过冷冻干燥（冻干）使用无定形菊粉基质作为稳定剂来生成流感病毒体稳定干粉制剂的方法。在菊粉存在下，病毒体的结构完整性和融合活性在冷冻干燥过程中得到充分保留。例如，在菊粉存在下冻干过程中，病毒体的免疫学特性，即体外血凝素（HA）效力和体内免疫原性潜力得以维持。此外，与分散在缓冲液中的病毒体相比，用菊粉配制的病毒体在储存时HA效力的保存时间显著延长。在菊粉存在下冻干过程中以及随后储存时，病毒体介导大分子细胞递送的能力也得以维持。具体而言，当分散在缓冲液中时，包裹有质粒DNA的病毒体在6周内完全丧失其转染活性，而在掺入菊粉基质后，其转染活性至少可完全保留12周。因此，在菊粉作为稳定剂存在的情况下，有无包裹大分子的流感病毒体的保质期都显著延长。将流感病毒体制备成干粉有利于储存和运输，并为开发无针剂型提供了可能性，例如用于口服、鼻腔、肺部或皮肤给药。
