【摘要】 目的:评估雾化吸入给药对重组人干扰素-α2b(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-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/m L、300万IU/m L和100万IU/m L rhIFN-α2b雾化后纯度分别为82.9%、90.1%和94.9%。600万IU/m L 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/m L。更多还原

【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’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 6millions IU / m L,3 millions IU / m L and 1 million IU / m L IFN-α2b were 82.9%,90.1% and 94.9% respectively. For 6 millions IU / m L 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 1million IU / m L in clinical treatment.更多还原