专肽生物_定制多肽合成服务_多肽合成定制服务_多肽合成厂家_多肽合成公司

NEP(1-40) TFA 是 Nogo-66 受体 (NgR) 的肽类拮抗剂，通过限制髓蛋白抑制逆转损伤引起的小胶质细胞形态分布的改变。
NEP(1-40) TFA is a Nogo-66 receptor (NgR) antagonist peptide, reversing the injury-induced shift in distribution of microglia morphologies by limiting myelin-based inhibition[1].

Jenna M Ziebell, et al. Nogo Presence Is Inversely Associated With Shifts in Cortical Microglial Morphology Following Experimental Diffuse Brain Injury. Neuroscience. 2017 Sep 17;359:209-223. : https://pubmed.ncbi.nlm.nih.gov/28736137

多肽H2N-Arg-Ile-Tyr-Lys-Gly-Val-Ile-Gln-Ala-Ile-Gln-Lys-Ser-Asp-Glu-Gly-His-Pro-Phe-Arg-Ala-Tyr-Leu-Glu-Ser-Glu-Val-Ala-Ile-Ser-Glu-Glu-Leu-Val-Gln-Lys-Tyr-Ser-Asn-Ser-NH2的合成步骤：

1、合成MBHA树脂：取若干克的MBHA树脂（如初始取代度为0.5mmol/g）和1倍树脂摩尔量的Fmoc-Linker-OH加入到反应器中，加入DMF，搅拌使氨基酸完全溶解。再加入树脂2倍量的DIEPA，搅拌混合均匀。再加入树脂0.95倍量的HBTU，搅拌混合均匀。反应3-4小时后，用DMF洗涤3次。用2倍树脂体积的10%乙酸酐/DMF 进行封端30分钟。然后再用DMF洗涤3次，甲醇洗涤2次，DCM洗涤2次，再用甲醇洗涤2次。真空干燥12小时以上，得到干燥的树脂{Fmoc-Linker-MHBA Resin}，测定取代度。这里测得取代度为 0.3mmol/g。结构如下图：

2、脱Fmoc：取1.28g的上述树脂，用DCM或DMF溶胀20分钟。用DMF洗涤2遍。加3倍树脂体积的20%Pip/DMF溶液，鼓氮气30分钟，然后2倍树脂体积的DMF 洗涤5次。得到 H2N-Linker-MBHA Resin 。（此步骤脱除Fmoc基团，茚三酮检测为蓝色，Pip为哌啶）。结构图如下：

3、缩合：取1.15mmol Fmoc-Ser(tBu)-OH 氨基酸，加入到上述树脂里，加适当DMF溶解氨基酸，再依次加入2.3mmol DIPEA，1.09mmol HBTU。反应30分钟后，取小样洗涤，茚三酮检测为无色。用2倍树脂体积的DMF 洗涤3次树脂。（洗涤树脂，去掉残留溶剂，为下一步反应做准备）。得到Fmoc-Ser(tBu)-Linker-MBHA Resin。氨基酸：DIPEA：HBTU：树脂=3：6：2.85：1（摩尔比）。结构图如下：

4、依次循环步骤二、步骤三，依次得到

H2N-Ser(tBu)-Linker-MBHA Resin

Fmoc-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Ile-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Ile-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Val-Ile-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Val-Ile-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Gly-Val-Ile-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Gly-Val-Ile-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Lys(Boc)-Gly-Val-Ile-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Lys(Boc)-Gly-Val-Ile-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Tyr(tBu)-Lys(Boc)-Gly-Val-Ile-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Tyr(tBu)-Lys(Boc)-Gly-Val-Ile-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Ile-Tyr(tBu)-Lys(Boc)-Gly-Val-Ile-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

H2N-Ile-Tyr(tBu)-Lys(Boc)-Gly-Val-Ile-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

Fmoc-Arg(Pbf)-Ile-Tyr(tBu)-Lys(Boc)-Gly-Val-Ile-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin

以上中间结构，均可在专肽生物多肽计算器-多肽结构计算器中，一键画出。

最后再经过步骤二得到 H2N-Arg(Pbf)-Ile-Tyr(tBu)-Lys(Boc)-Gly-Val-Ile-Gln(Trt)-Ala-Ile-Gln(Trt)-Lys(Boc)-Ser(tBu)-Asp(OtBu)-Glu(OtBu)-Gly-His(Trt)-Pro-Phe-Arg(Pbf)-Ala-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Glu(OtBu)-Val-Ala-Ile-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Leu-Val-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-Asn(Trt)-Ser(tBu)-Linker-MBHA Resin，结构如下：

5、切割：6倍树脂体积的切割液（或每1g树脂加8ml左右的切割液），摇床摇晃 2小时，过滤掉树脂，用冰无水乙醚沉淀滤液，并用冰无水乙醚洗涤沉淀物3次，最后将沉淀物放真空干燥釜中，常温干燥24小试，得到粗品H2N-Arg-Ile-Tyr-Lys-Gly-Val-Ile-Gln-Ala-Ile-Gln-Lys-Ser-Asp-Glu-Gly-His-Pro-Phe-Arg-Ala-Tyr-Leu-Glu-Ser-Glu-Val-Ala-Ile-Ser-Glu-Glu-Leu-Val-Gln-Lys-Tyr-Ser-Asn-Ser-NH2。结构图见产品结构图。

切割液选择：1）TFA：H2O=95%：5%

2）TFA：H2O：TIS=95%：2.5%：2.5%

3）三氟乙酸：茴香硫醚：1，2-乙二硫醇：苯酚：水=87.5%：5%：2.5%：2.5%：2.5%

（前两种适合没有容易氧化的氨基酸，例如Trp、Cys、Met。第三种适合几乎所有的序列。）

6、纯化冻干：使用液相色谱纯化，收集目标峰液体，进行冻干，获得蓬松的粉末状固体多肽。不过这时要取小样复测下纯度 是否目标纯度。

7、最后总结：

杭州专肽生物技术有限公司（ALLPEPTIDE https://www.allpeptide.com）主营定制多肽合成业务，提供各类长肽，短肽，环肽，提供各类修饰肽，如：荧光标记修饰（CY3、CY5、CY5.5、CY7、FAM、FITC、Rhodamine B、TAMRA等），功能基团修饰肽（叠氮、炔基、DBCO、DOTA、NOTA等），同位素标记肽（N15、C13），订书肽（Stapled Peptide）,脂肪酸修饰肽（Pal、Myr、Ste），磷酸化修饰肽（P-Ser、P-Thr、P-Tyr），环肽（酰胺键环肽、一对或者多对二硫键环），生物素标记肽，PEG修饰肽，甲基化修饰肽等。

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