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| 雾化给药对重组人干扰素-α2b生物活性和分子结构的影响 |
| 王兵1,2,程婷1,卢晨1,周乐春1,倪晓燕1,李增礼1,王荣海1,2,宋礼华1,2 |
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| (1. 安徽安科生物工程(集团)股份有限公司,安徽合肥 230088;2. 安徽省生物研究所,安徽合肥 230032) |
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| 摘要: |
| 目的:评估雾化吸入给药对重组人干扰素-琢2b(rhIFN-α2b) 生物活性和分子结构的影响。方法:采用病毒抑制法检测 rhIFN-α2b 的生物活性,通过比较雾化前后rhIFN-α2b 生物活性的变化分析空气射流对其生物活性的影响。采用高效液相色谱(HPLC)法和十二烷基磺酸钠鄄聚丙烯酰胺凝胶电泳(SDS-PAGE) 法分析干扰素分子结构的改变。结果:雾化给药不会造成rhIFN-α2b 蛋白质肽键断裂,但是空气射流会造成二硫键断裂,断裂比例与雾化前rhIFN-α2b 浓度有关,600 万IU/mL、300 万IU/mL 和100 万IU/mL rhIFN-α2b雾化后纯度分别为82.9%、90.1%和94.9%。600 万IU/mL rhIFN-α2b 溶液雾化后17.1%的rhIFN-α2b分子出现二硫键的断裂,其中0.5%的rhIFN-α2b分子出现双二硫键断裂,雾化后喷雾壶内未完全汽化的剩余液中rhIFN-α2b 的生物活性保留率为98.2%,雾化后收集的凝集液中rhIFN-α2b的生物活性保留率为96.1%。结论:rhIFN-α2b能以雾化状态稳定传输给药,生物活性基本保留。雾化后绝大部分rhIFN-α2b分子结构保持完整,少数分子出现单个或双二硫键断裂,为了尽量减少rhIFN-α2b分子的损耗,临床上雾化吸入rhIFN-α2b的浓度不应超过100 万IU/mL。 |
| 关键词: 重组人干扰素-α2b 雾化给药 生物活性 分子结构 二硫键 |
| DOI：doi:10.13407/j.cnki.jpp.1672-108X.2017.03.001 |
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| 基金项目: |
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| The Effects of Atomization Inhalation on Biological Activity and Molecular Structure of Human Recombinant Interferon Alpha 2b |
| Wang Bing1,2 , Cheng Ting1 , Lu Chen1 , Zhou Lechun1 , Ni Xiaoyan1 , Li Zengli1 , Wang Ronghai1,2 , Song Lihua1,2 |
| (1. Anhui Anke Biotechnology (Group) Co., Ltd. Anhui Hefei 230088, China; 2. Anhui Biological Institute, Anhui Hefei 230032, China) |
| Abstract: |
| Objective: To evaluate the effects of atomization inhalation of administration with recombinant human interferon alpha 2b (rhIFN-α2b) on its biological activity and molecular structure. Methods: The rhIFN-α2b biological activity was detected by virus inhibition method. The influence of the air jet on IFN-α2b biological activity was studied by comparing its biological activity before and after atomization. The changes of IFN-α2b molecular structure were analyzed by high performance liquid chromatography (HPLC) method and sodium dodecyl sulfate-polyacrylamide gel electrophoresis ( SDS-PAGE) method. Results: Atomization inhalation of administration didn爷t cause the peptide bond of IFN-α2b protein to break, but air jet could cause disulfide bond to break. The ratio of IFN-α2b molecular with disulfide bond breaking was related to its concentration before atomization. After atomization the purity of 6 millions IU/mL, 3 millions IU/mL and 1 million IU/mL IFN-α2b were 82.9%, 90.1% and 94.9% respectively. For 6 millions IU/mL concentration rhIFN-α2b solution, after atomization would cause disulfide bond of 17.1% rhIFN-α2b molecular to break. Therein a pair
of two disulfide bonds breaking occured in 0.5% of rhIFN-α2b molecules. And the retention rate of rhIFN-α2b biological activity after atomization in remainder fluid was about 98.2% and in condensed fluid was 96.1%. Conclusion: Recombinant human interferon α2b can be transmitted stably in spray pattern. Its biological activity can be retained basic completely. The molecular structure of most of IFN-α2b keeps perfectly after the atomization. Single or double disulfide bond breaking in a few molecules of rhIFN-α2b is caused by air jet. In order to minimize the loss of rhIFN-α2b molecules, it is suggested atomization rhIFN-α2b concentration should be controlled under 1 million IU/ mL in clinical treatment. |
| Key words: recombinant human interferon alpha 2b atomization inhalation of administration biological activity molecular structure disulfide bond |
