人甲状旁腺素(1-34)的合成与聚乙二醇化修饰

目的合成人甲状旁腺素(1 34) hPTH(1-34) ]并进行聚乙二醇化修饰。方法应用固相多肽合成方法合成并经高效液相色谱纯化得hPTH(1-34) ,Cys-hPTH(1- 34)和hPTH(1-34)-Cys-NH2 。Cys hPTH(1-34)和hPTH(1-34)-Cys-NH2 在水溶液(pH 7～8)中分别与平均相对分子质量为5 0 0 0的单甲氧基马来酰亚胺基聚乙二醇(mPEG50 0 0 MAL)反应,经高效液相色谱纯化得Cys(mPEG50 0 0 MAL)- hPTH(1-34)和hPTH(1-34)-Cys(mPEG50 0 0- MAL) -NH2 。结果hPTH(1-34) ,Cys(mPEG50 0 0 MAL) hPTH(1 34)和hPTH(1 34) Cys(mPEG50 0 0 MAL) NH2 的质谱和氨基酸组成分析结果均与理论值一致。hPTH(1-34) -Cys(mPEG50 0 0-MAL) NH2 保持了较好的体外活性,Cys(mPEG50 0 0- MAL) hPTH(1-34)保持了较好的体内活性。结论采用一种简便的方法成功实现了对hPTH(1-34)的N端或C端的聚乙二醇化修饰

Synthesis and pegylation of human parathyroid hormones (1-34)

PurposeTo synthesize human parathyroid hormone(1-34)hPTH(1-34)] and pegylate it.MethodshPTH(1-34) and Cys-hPTH(1-34) were assembled on Wang resin with DCC/HOBt as coupling reagents and under cleavaged with TFA/m-cresol/TMBS/thiolanisol/EDT. hPTH(1-34)-Cys- NH_2 was assembled on Rink AM Amide resin with the same couple and under cleavage condition. hPTH(1-34),Cys-hPTH((1-34)) and hPTH(1-34)-Cys-NH_2 were purified by PR-HPLC. Cys-hPTH(1-34) and hPTH(1-34)-Cys-NH_2 reacted with mPEG_(5000)-MAL in aqueous solution(pH 7～8) for about 2h to get Cys(mPEG_(5000)-MAL)-hPTH((1-34)) and hPTH(1-34)-Cys(mPEG_(5000)-MAL)-NH_2.ResultsThe structures of hPTH(1-34),Cys(mPEG_(5000)-MAL)-hPTH(1-34) and hPTH(1-34)-Cys(mPEG_(5000)-MAL)-NH_2 were confirmed by MALDI-TOF-MS and amino acid analysis. In comparison with hPTH(1-34), hPTH(1-34)-Cys(mPEG_(5000)-MAL)-NH_2 showed some bioactivity in vitro and Cys(mPEG_(5000)-MAL)-hPTH(1-34) exhibited better effects in vivo.ConclusionhPTH(1-34) was successfully pegylated at N-or C-terminal by a simple method.