浙江省科技型企業(yè)---加速您的多肽研究
400-998-5282
專(zhuān)注多肽 服務(wù)科研
400-998-5282
專(zhuān)注多肽 服務(wù)科研
這種增強(qiáng)胰島素的 hGH 片段已在人血清中檢測(cè)到。用 hGH (1-43) 處理肥胖的黃色 Avy/A 小鼠可增強(qiáng)其脂肪組織對(duì)胰島素的體外敏感性。
編號(hào)：199421
CAS號(hào)：96827-07-5
單字母：H2N-FPTIPLSRLFDNAMLRAHRLHQLAFDTYQEFEEAYIPKEQKYS-OH
This insulin-potentiating hGH-fragment has been detected in human serum. Treatment of obese yellow Avy/A mice with hGH (1-43) enhanced the in vitro sensitivity of their adipose tissue to insulin.
| DOI | 名稱(chēng) | |
|---|---|---|
| 10.3181/00379727-191-42942 | Effects of hypophysectomy and the insulin-like and anti-insulin pituitary peptides on carbohydrate metabolism in yellow Avy/A (BALB/c x VY)F1 hybrid mice | 下載 |
| 10.1210/endo-122-6-2940 | Increased sensitivity of adipose tissue to insulin after in vivo treatment of yellow Avy/A obese mice with amino-terminal peptides of human growth hormone | 下載 |
多肽H2N-Phe-Pro-Thr-Ile-Pro-Leu-Ser-Arg-Leu-Phe-Asp-Asn-Ala-Met-Leu-Arg-Ala-His-Arg-Leu-His-Gln-Leu-Ala-Phe-Asp-Thr-Tyr-Gln-Glu-Phe-Glu-Glu-Ala-Tyr-Ile-Pro-Lys-Glu-Gln-Lys-Tyr-Ser-COOH的合成步驟：
1、合成CTC樹(shù)脂：稱(chēng)取0.87g CTC Resin（如初始取代度約為0.91mmol/g）和0.95mmol Fmoc-Ser(tBu)-OH于反應(yīng)器中，加入適量DCM溶解氨基酸（需要注意，此時(shí)CTC樹(shù)脂體積會(huì)增大好幾倍，避免DCM溶液過(guò)少），再加入2.38mmol DIPEA（Mw：129.1,d：0.740g/ml），反應(yīng)2-3小時(shí)后，可不抽濾溶液，直接加入1ml的HPLC級(jí)甲醇，封端半小時(shí)。依次用DMF洗滌2次，甲醇洗滌1次，DCM洗滌一次，甲醇洗滌一次，DCM洗滌一次，DMF洗滌2次（這里使用甲醇和DCM交替洗滌，是為了更好地去除其他溶質(zhì)，有利于后續(xù)反應(yīng)）。得到 Fmoc-Ser(tBu)-CTC Resin。結(jié)構(gòu)圖如下：
2、脫Fmoc：加3倍樹(shù)脂體積的20%Pip/DMF溶液，鼓氮?dú)?0分鐘，然后2倍樹(shù)脂體積的DMF 洗滌5次。得到 H2N-Ser(tBu)-CTC Resin 。（此步驟脫除Fmoc基團(tuán)，茚三酮檢測(cè)為藍(lán)色，Pip為哌啶）。結(jié)構(gòu)圖如下：
3、縮合：取2.38mmol Fmoc-Tyr(tBu)-OH 氨基酸，加入到上述樹(shù)脂里，加適當(dāng)DMF溶解氨基酸，再依次加入4.75mmol DIPEA，2.26mmol HBTU。反應(yīng)30分鐘后，取小樣洗滌，茚三酮檢測(cè)為無(wú)色。用2倍樹(shù)脂體積的DMF 洗滌3次樹(shù)脂。（洗滌樹(shù)脂，去掉殘留溶劑，為下一步反應(yīng)做準(zhǔn)備）。得到Fmoc-Tyr(tBu)-Ser(tBu)-CTC Resin。氨基酸：DIPEA：HBTU：樹(shù)脂=3：6：2.85：1（摩爾比）。結(jié)構(gòu)圖如下：
4、依次循環(huán)步驟二、步驟三，依次得到
H2N-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Ser(tBu)-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Ser(tBu)-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Leu-Ser(tBu)-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Leu-Ser(tBu)-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Pro-Leu-Ser(tBu)-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Pro-Leu-Ser(tBu)-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Ile-Pro-Leu-Ser(tBu)-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Ile-Pro-Leu-Ser(tBu)-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Thr(tBu)-Ile-Pro-Leu-Ser(tBu)-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Thr(tBu)-Ile-Pro-Leu-Ser(tBu)-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Pro-Thr(tBu)-Ile-Pro-Leu-Ser(tBu)-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
H2N-Pro-Thr(tBu)-Ile-Pro-Leu-Ser(tBu)-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
Fmoc-Phe-Pro-Thr(tBu)-Ile-Pro-Leu-Ser(tBu)-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin
以上中間結(jié)構(gòu)，均可在專(zhuān)肽生物多肽計(jì)算器-多肽結(jié)構(gòu)計(jì)算器中，一鍵畫(huà)出。
最后再經(jīng)過(guò)步驟二得到 H2N-Phe-Pro-Thr(tBu)-Ile-Pro-Leu-Ser(tBu)-Arg(Pbf)-Leu-Phe-Asp(OtBu)-Asn(Trt)-Ala-Met-Leu-Arg(Pbf)-Ala-His(Trt)-Arg(Pbf)-Leu-His(Trt)-Gln(Trt)-Leu-Ala-Phe-Asp(OtBu)-Thr(tBu)-Tyr(tBu)-Gln(Trt)-Glu(OtBu)-Phe-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Ile-Pro-Lys(Boc)-Glu(OtBu)-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Ser(tBu)-CTC Resin，結(jié)構(gòu)如下：
5、切割：6倍樹(shù)脂體積的切割液（或每1g樹(shù)脂加8ml左右的切割液），搖床搖晃 2小時(shí)，過(guò)濾掉樹(shù)脂，用冰無(wú)水乙醚沉淀濾液，并用冰無(wú)水乙醚洗滌沉淀物3次，最后將沉淀物放真空干燥釜中，常溫干燥24小試，得到粗品H2N-Phe-Pro-Thr-Ile-Pro-Leu-Ser-Arg-Leu-Phe-Asp-Asn-Ala-Met-Leu-Arg-Ala-His-Arg-Leu-His-Gln-Leu-Ala-Phe-Asp-Thr-Tyr-Gln-Glu-Phe-Glu-Glu-Ala-Tyr-Ile-Pro-Lys-Glu-Gln-Lys-Tyr-Ser-COOH。結(jié)構(gòu)圖見(jiàn)產(chǎn)品結(jié)構(gòu)圖。
切割液選擇：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%
（前兩種適合沒(méi)有容易氧化的氨基酸，例如Trp、Cys、Met。第三種適合幾乎所有的序列。）
6、純化凍干：使用液相色譜純化，收集目標(biāo)峰液體，進(jìn)行凍干，獲得蓬松的粉末狀固體多肽。不過(guò)這時(shí)要取小樣復(fù)測(cè)下純度 是否目標(biāo)純度。
7、最后總結(jié)：
杭州專(zhuān)肽生物技術(shù)有限公司（ALLPEPTIDE http://tsjxdd.com）主營(yíng)定制多肽合成業(yè)務(wù)，提供各類(lèi)長(zhǎng)肽，短肽，環(huán)肽，提供各類(lèi)修飾肽，如：熒光標(biāo)記修飾（CY3、CY5、CY5.5、CY7、FAM、FITC、Rhodamine B、TAMRA等），功能基團(tuán)修飾肽（疊氮、炔基、DBCO、DOTA、NOTA等），同位素標(biāo)記肽（N15、C13），訂書(shū)肽（Stapled Peptide）,脂肪酸修飾肽（Pal、Myr、Ste），磷酸化修飾肽（P-Ser、P-Thr、P-Tyr），環(huán)肽（酰胺鍵環(huán)肽、一對(duì)或者多對(duì)二硫鍵環(huán)），生物素標(biāo)記肽，PEG修飾肽，甲基化修飾肽等。
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