一种新型hPTH(1-34)基因相关肽的制备及其应用的初步研究-东南大学学报医学版

一种新型hPTH(1-34)基因相关肽的制备及其应用的初步研究

单位：1.中国药科大学生命科学与技术学院,微基因药学实验室,江苏,南京,210009； 2.中国药科大学生命科学与技术学院,微基因药学实验室,江苏,南京,210009;上海水产大学食品学院,海洋生物制药学科点,上海,200090

分类号：R587.02

出版年·卷·期（页码）：2008·27·第一期（31-36）

摘要：

目的:对hPTH(1-34)进行结构改造,提高蛋白水解酶的稳定性,增强酶活性,使之更适合制成滴鼻剂,并验证治疗骨质疏松症的效果.方法:确定将hPTH(1-34)改造成为Pro-Pro-[Arg11]hPTH(1-34)-Pro-Pro后,构建表达质粒pED-4PhPTH(1-34),转化E.coli BL21(DE3)LysE,乳糖诱导表达,经分离纯化获得目的多肽.将目的多肽与二甲基-β-环糊精、聚丙烯酸、EDTA制成滴鼻剂,治疗卵巢摘除大鼠(OVXed Rats)的骨质疏松症.结果:获得纯度较高的Pro-Pro-[Arg11]hPTH(1-34)-Pro-Pro,将制成的滴鼻剂用于卵巢摘除大鼠,与生理盐水滴鼻治疗组相比能显著增加大鼠椎骨骨密度(P<0.01)及股骨头骨小梁厚度(P<0.001),对鼻黏膜没有损伤.结论:Pro-Pro-[Arg11]hPTH(1-34)-Pro-Pro制成滴鼻剂治疗卵巢摘除大鼠的骨质疏松症效果明显,有进一步开发的前景.

Objectives To find a novel analog of hPTH(1-34),which has more activity for osteoporosis and more stability against degradation of proteases before absorption and is suitable for preparing nasal drops and to evaluate the effectiveness of intranasal administration of the novel analog for osteoroporsis.Methods The recombinant Pro-Pro-hPTH(1-34)-Pro-Pro was obtained through combining fusion expression of gene,cell disruption,inclusion body washing,ethanol fraction precipitation,acid hydrolysis,and CM-52 ion exchange column chromatography.The purified Pro-Pro-hPTH(1-34)-Pro-Pro was mixed up with dimethyl-cyclodextrin(DM-β-CD),polyacrylic acid,and EDTA-Na to prepare nasal drops and the nasal drops were administrated to the ovariectomized rats through nasal mucosa daily for 12 weeks.Then the bone material densities(BMD)of the vertebrae and the thickness of trabecular bone of femoral head were measured.Results The average BMDs of these groups treated with the nasal drops of the peptide were significantly higher than that of the group treated with normal saline(NS)(P<0.01).And the trabecular thicknesses of these groups were all decreased,significantly different to that of the group treated with NS(P<0.001).Conclusion The nasal drops of Pro-Pro-hPTH(1-34)-Pro-Pro are effective for osteoporosis.

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