郭凝,杨仲璠,王俊玲,吴彦卓,徐明波,张景海
GUO Ning1,2,YANG Zhong-fan2,WANG Jun-ling2,WU Yan-zhuo2,XU Ming-bo2

• 1:沈阳药科大学生命科学与生物制药学院
• 2:北京双鹭药业股份有限公司

摘要(Abstract)：

目的制备一种新型聚乙二醇化重组人生长激素(PEG-(rhGH)2),并对其理化性质及药理作用进行研究。方法以分子质量为2×104u的聚乙二醇二丙醛(PEG-(ALD)2)对重组人生长激素(rhGH)进行修饰,采用Sephacryl S-200凝胶过滤色谱、Q Sepharose FF离子交换色谱对修饰产物进行分离纯化,SDS-PAGE鉴定各组分,通过SEC-HPLC、IEF、MALDI-TOF-MS等方法对目标产物进行分析,并采用去脑垂体大鼠模型研究其体内生物活性与长效作用。结果获得了一种新型聚乙二醇化重组人生长激素(PEG-(rhGH)2)。其纯度质量分数达到97%以上,等电点为5.27,分子质量为6.6×104u。与未修饰的rhGH相比,PEG-(rhGH)2具有高效、长效的促进机体生长的药理作用。结论新型聚乙二醇化重组人生长激素PEG-(rhGH)2是一种具有开发前景的长效蛋白质药物,值得进一步深入研究。
Objective To prepare a novel PEGylated recombinant human growth hormone(PEG-(rhGH)2),and study the characterization and pharmacological effects of(PEG-(rhGH)2).Methods rhGH was modified with(PEG-(ALD)2)(mw 2.0×104 u),the products were purified by Sephacryl S-200 gel filtration chromatography,Q Sepharose FF ion exchange chromatography and identified with SDS-PAGE.SEC-HPLC,IEF,MALDI-TOF-MS were applied to analyze the final derivative.The long-term pharmacological effects and in vivo bioactivities in hypophysectomized(Hpx)male rats treated with PEG-(rhGH)2 were analyzed.Results A novel PEGylated recombinant human growth hormone(PEG-(rhGH)2)was obtained.The purity of PEG-(rhGH)2 was higher than 97%,the isoelectric point was 5.27,the molecular weight was 6.6×104 u.Compared with unmodified rhGH,PEG-(rhGH)2′s effects on body and organ growth was stronger and lasted longer.Conclusions The novel PEGylated recombinant human growth hormone PEG-(rhGH)2 is a long-acting protein drug with good developing perspective and is worthy of further investigation.

关键词(KeyWords)： 重组人生长激素;聚乙二醇二丙醛;化学修饰;纯化;长效
recombinant human growth hormone;PEG-(ALD)2;chemical modification;purification;long-acting

Abstract:

Keywords:

基金项目(Foundation): 国家重大科技专项基金资助项目(2008ZX09401)

作者(Author): 郭凝,杨仲璠,王俊玲,吴彦卓,徐明波,张景海
GUO Ning1,2,YANG Zhong-fan2,WANG Jun-ling2,WU Yan-zhuo2,XU Ming-bo2

DOI: 10.14066/j.cnki.cn21-1349/r.2010.10.012

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