人胸腺肽α_1的固相合成及体外活性研究  

Study on Fmoc solid-phase synthesis and activity of thymosin α_1

作　　者：韩香[1] 顾军[1] 苑庆兰[1] 甘一如[1] 

机构地区：[1]武警医学院化学教研室,天津300162

出　　处：《中国药物化学杂志》2004年第1期27-29,共3页Chinese Journal of Medicinal Chemistry

基　　金：天津市自然科学基金资助项目 (993 80 43 1 1 )

摘　　要：目的对胸腺肽α1的固相合成工艺进行研究 ,并对合成产物进行活性评价。方法采用对碱敏感的Nα 芴甲氧羰基 (Fmoc)作为α 氨基的保护基 ,以Fmoc Asn(Trt) WangResin为起点 ,逐个延伸固相合成法合成胸腺肽α1。与Nα Fmoc 基团配套的保护策略还有 :叔丁氧羰基 (Boc)保护Lys的侧链氨基 ,叔丁酯基 (OtBu)保护Asp、Glu的侧链羧基 ,叔丁氧基 (tBu)保护Ser、Thr的侧链羟基 ,三苯甲基 (Trt)保护Asn的侧链酰胺基。采用DCC HOBt缩合剂法进行接肽反应 ,茚三酮定性显色法和水杨醛定量自由氨基检测法控制反应进程 ,肽段最后用TFA DCM(V∶V=1∶1 )定量地从树脂上切除。结果胸腺肽α1总产率为 3 3 2 %。纯化后的产物 ,经SDS PAGE和RP HPLC鉴定 ,纯度为98 8%以上 ,活性与日达仙对照品相当。结论Nα 芴甲氧羰基保护策略的固相合成方法操作简便、条件温和、副反应少 ,产品纯度高、收率高。Aim To synthesize thymosin α _1 by the solid-phase method.Methods Peptide synthesis was performed manually by the stepwise solid-phase method using the base-labile Fmoc group for protecting the α -amino acid.The peptide was assembled on a p -alkoxybenzyl alcohol resin.And the peptide sequence was prepared employing Fmoc-amino acid and Boc-protected Lys,O t Bu-protected Asp,Glu,t Bu-protected Ser,Thr,Trt-protected Asn as building blocks.The Fmoc group was removed with piperidine-DCM( V∶V =1∶1).The peptide resin was treated with TFA-DCM( V∶V =1∶1),and purified by ion-exchange chromatography and prepared RP-HPLC.The analytical strategies included the reaction of ninhydrin reagents with small samples of resin peptide and salicylaldehyde reagents testing methods were used to monitor solid-phase reactions and determine the structures of resin-bound compounds.Results The yield of the total synthesis was 33.2%,based on the amino acid content of the starting Fmoc-Asn(Trt)-resin.The purity was range to above 98.8%,the activity is equal to that of Zadaxin.Conclusion A strategy for the synthesis of thymosin α _1 was presented.This orthogonal protection strategy has bright prospects for peptide synthesis.

关 键 词：药物化学 制备 固相多肽合成 胸腺肽Α1 N^α-芴甲氧羰基(Fmoc) 

分 类 号：R914[医药卫生—药物化学] R965[医药卫生—药学]