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Helospectin I 是血管活性肠肽 (VIP) 家族的神经肽。Helospectin I 具有血管扩张和抗高血压活性，并可降低血压。Helospectin I 最初是从蜥蜴 Heloderma suspectum 的唾液腺毒液中分离出来的。

Helospectin I和II是分泌素/胰高血糖素/血管活性肠肽（VIP）超家族中两种亲缘关系密切的哺乳动物神经肽，它们与VIP共同定位于血管平滑肌周围的神经纤维中。然而，VIP受体在完整外周微循环中对Helospectin I和II的血管舒张效应的转导作用是否起作用，目前尚不明确。

通过活体显微镜观察发现：当Helospectin I和II（各1.0 nmol)灌注时，在完整仓鼠颊囊微循环中可诱发显著的血管舒张反应（P值< 0.05)。而0.1 nmol剂量的螺旋素I和II对小动脉直径无明显影响。预先使用VPAC1/VPAC2受体拮抗剂VIP10-28或PAC1/VPAC2受体拮抗剂PACAP6-38处理后，Helospectin I和II诱导的反应均未见显著改变。此外，indomethacin预处理对Helospectin I和II引发的血管舒张也无明显影响。

展示了Helospectin I和II对颊囊内小动脉直径的影响。当分别注射0.1 nmol剂量的螺旋素I和II时，未观察到显著影响（各组n = 4只动物，与基线值相比P>0.5）。然而，当以1.0 nmol浓度持续输注这两种螺旋素时，引发了显著且持久的血管舒张效应（各组n = 4只动物，与基线值相比P < 0.05；组间比较P>0.5）。实验数据显示，螺旋素I和II开始输注后2分钟内即可观察到小动脉直径增大，随后在∼20分钟内恢复至基线水平。

综合数据表明，研究人员发现helospectin I和II在完整的外周微循环中引起强效的血管舒张，这既不是通过VIP或PACAP受体，也不是通过花生四烯酸代谢的环氧合酶产物。

参考文献
Tsueshita T, et al. Helospectin I and II evoke vasodilation in the intact peripheral microcirculation. Peptides. 2004 Jan;25(1):65-9

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

1、合成CTC树脂：称取1.97g CTC Resin（如初始取代度约为1.01mmol/g）和2.39mmol Fmoc-Ser(tBu)-OH于反应器中，加入适量DCM溶解氨基酸（需要注意，此时CTC树脂体积会增大好几倍，避免DCM溶液过少），再加入5.97mmol DIPEA（Mw：129.1,d：0.740g/ml），反应2-3小时后，可不抽滤溶液，直接加入1ml的HPLC级甲醇，封端半小时。依次用DMF洗涤2次，甲醇洗涤1次，DCM洗涤一次，甲醇洗涤一次，DCM洗涤一次，DMF洗涤2次（这里使用甲醇和DCM交替洗涤，是为了更好地去除其他溶质，有利于后续反应）。得到  Fmoc-Ser(tBu)-CTC Resin。结构图如下：

2、脱Fmoc：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-Ser(tBu)-CTC 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5.97mmol 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11.94mmol DIPEA，5.67mmol 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)-Ser(tBu)-CTC 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)-Ser(tBu)-CTC Resin

Fmoc-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Thr(tBu)-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Thr(tBu)-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Phe-Thr(tBu)-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Phe-Thr(tBu)-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Thr(tBu)-Phe-Thr(tBu)-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Thr(tBu)-Phe-Thr(tBu)-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Ala-Thr(tBu)-Phe-Thr(tBu)-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Ala-Thr(tBu)-Phe-Thr(tBu)-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Asp(OtBu)-Ala-Thr(tBu)-Phe-Thr(tBu)-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Asp(OtBu)-Ala-Thr(tBu)-Phe-Thr(tBu)-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-Ser(tBu)-Asp(OtBu)-Ala-Thr(tBu)-Phe-Thr(tBu)-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

H2N-Ser(tBu)-Asp(OtBu)-Ala-Thr(tBu)-Phe-Thr(tBu)-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC Resin

Fmoc-His(Trt)-Ser(tBu)-Asp(OtBu)-Ala-Thr(tBu)-Phe-Thr(tBu)-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC 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-His(Trt)-Ser(tBu)-Asp(OtBu)-Ala-Thr(tBu)-Phe-Thr(tBu)-Ala-Glu(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Leu-Leu-Ala-Lys(Boc)-Leu-Ala-Leu-Gln(Trt)-Lys(Boc)-Tyr(tBu)-Leu-Glu(OtBu)-Ser(tBu)-Ile-Leu-Gly-Ser(tBu)-Ser(tBu)-Thr(tBu)-Ser(tBu)-Pro-Arg(Pbf)-Pro-Pro-Ser(tBu)-Ser(tBu)-CTC 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-His-Ser-Asp-Ala-Thr-Phe-Thr-Ala-Glu-Tyr-Ser-Lys-Leu-Leu-Ala-Lys-Leu-Ala-Leu-Gln-Lys-Tyr-Leu-Glu-Ser-Ile-Leu-Gly-Ser-Ser-Thr-Ser-Pro-Arg-Pro-Pro-Ser-Ser-COOH。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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