重组人干扰素-α1b雾化后生物学活性与粒径及安全性分析
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摘要
目的研究雾化给药重组人干扰素-α1b(rhIFN-α1b)对微粒粒径、生物活性、分子结构的影响及安全性分析。方法应用Winner311XP激光粒度仪检测并记录药液雾化后微粒的粒径分布;细胞病变法测定雾化前后rhIFN-α1b的生物学活性;应用高效液相色谱法(HPLC)和十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)法测定雾化前后rhIFN-α1b的分子结构变化;用异常毒性试验检测rhIFN-α1b经雾化给药的安全性。结果 rhIFN-α1b雾化后,81%的微粒粒径分布在1～5μm区间;10μg、20μg、30μg、40μg、50μg、60μg rhIFN-α1b雾化后生物学活性保留率分别为95.3%、96.8%、96.2%、96.5%、95.85%和94.27%;小鼠肌注30μg/mL干扰素溶液0.5 mL(相当于人肌注15μg),安全无异常。结论雾化rhIFN-α1b可使药液到达肺泡,且生物活性基本保留,绝大部分分子结构保持完整,二聚体含量略有增加,且安全无毒性。
Objective To study the effect of aerosolized recombinant human interferon-α1b(rhIFN-α1b on the particle size, biological activity, and molecular structure, and the safety analysis.Methods Winner 311 XP laser particle size analyzer was used to detect and record the particle size distribution of aerosolized particles; the cytopathogenic assay was used to detect the change of biological activity of rhIFN-α1b after aerosolization; high performance liquid chromatography(HPLC) and dodecane sodium sulphate-polyacrylamide gel electrophoresis(SDS-PAGE) was used to check the molecular structure changes of rhIFN-α1b after nebulization. The safety of rhIFN-α1b after atomization was detected by abnormal toxicity test.Results After atomization of rhIFN-α1b, 81% of the microparticles were distributed in the range of 1-5μm;the biological activity retention rates of 10 μg, 20μg,30μg,40 μg,50 μg,60 μg rhIFN-α1b, after atomization were 95.3% and 96.8 respectively. %, 96.2%, 96.5%, 95.85%,and 94.27%;No abnormality was found in mice after intramuscular injection of 30 μg/mL interferon solution 0.5 mL(equivalent to 15 μg of human intramuscular injection).Conclusion The aerosolized rhIFN-α1b can reach the alveoli, and the biological activity remains. Most of the molecular structure remains intact, the dimer content is slightly increased, and it is safe and non-toxic.
Objective To study the effect of aerosolized recombinant human interferon-α1b(rhIFN-α1b on the particle size, biological activity, and molecular structure, and the safety analysis.Methods Winner 311 XP laser particle size analyzer was used to detect and record the particle size distribution of aerosolized particles; the cytopathogenic assay was used to detect the change of biological activity of rhIFN-α1b after aerosolization; high performance liquid chromatography(HPLC) and dodecane sodium sulphate-polyacrylamide gel electrophoresis(SDS-PAGE) was used to check the molecular structure changes of rhIFN-α1b after nebulization. The safety of rhIFN-α1b after atomization was detected by abnormal toxicity test.Results After atomization of rhIFN-α1b, 81% of the microparticles were distributed in the range of 1-5μm;the biological activity retention rates of 10 μg, 20μg,30μg,40 μg,50 μg,60 μg rhIFN-α1b, after atomization were 95.3% and 96.8 respectively. %, 96.2%, 96.5%, 95.85%,and 94.27%;No abnormality was found in mice after intramuscular injection of 30 μg/mL interferon solution 0.5 mL(equivalent to 15 μg of human intramuscular injection).Conclusion The aerosolized rhIFN-α1b can reach the alveoli, and the biological activity remains. Most of the molecular structure remains intact, the dimer content is slightly increased, and it is safe and non-toxic.
引文
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[2]陈伟卓,高向东,徐晨,牛春.2'-5'寡聚腺苷酸合成酶(OAS)及其临床意义的研究进展.[J].现代生物医学进展,2013,36(018):7165-7170.
[3]JACKSON D J.Inhaled interferon:a novel treatment for virusinduced asthma?[J].Am J Respir Crit Care Med,2014,190(Iss2):123-124.
[4]尚云晓,黄英,刘恩梅,等.雾化吸入重组人干扰素α1b治疗小儿急性毛细支气管炎多中心研究[J].中华实用临床儿科杂志,201429(11):840-844.
[5]RATKOD,TIMH,SEBASTIAN L J,et al.The effect of inhaled IFN-on worsening of asthma symptoms caused by viral infections:a randomized trial[J].Am J Respir Crit Care Med,2017,Vol.23,N0.3(Iss2):145-154.
[6]申昆玲,张国成,尚云晓等.重组人干扰素α1b在儿科的临床应用专家共识[J].中国实用儿科杂志,2015,30(16):1214-1218.
[7]《中华人民共和国药典》2015版三部3523第一法.
[8]杨丽华.重组人干扰素α1b注射液治疗小儿病毒性疾病的安全性及对下呼吸道感染患儿的远期影响[D].第四军医大学,2017.
[9]欧宝莲.重组人干扰素α_(1b)治疗小儿病毒性呼吸道感染的效果及安全性[J].中国当代医药,2015,22(11):128-129+132.
[10]杨永弘,马香.小儿呼吸道感染的细菌病原[J].实用儿科临床杂志,2011,26(04):229-232.
[11]刘鉴峰,刘金剑,褚丽萍,王德芝,于佳,刘金毅.雾化吸入干扰素α1b在兔体内的分布及代谢途径[J].医药导报,2013,32(01):1-5.
[12]周莉.雾化吸入疗法临床应用新进展[J].医学信息(中旬刊),2010,5(02):147-149.