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ACTH的定义
促肾上腺皮质激素（ACTH、Adrenocorticotropic hormone）或促肾上腺皮质激素是垂体前叶产生的一种激素，它刺激肾上腺皮质。

促肾上腺皮质激素（ACTH），也称为促肾上腺皮质激素，是一种由垂体前叶腺产生和分泌的多肽促性激素。它是下丘脑-垂体-肾上腺轴的重要组成部分，通常是在对生物压力的反应中产生的（连同下丘脑的促肾上腺皮质激素释放激素）。它的主要作用是增加皮质类固醇的产生和释放，以及顾名思义，是肾上腺皮质的皮质醇。 

ACTH是由前opiomelanocortin（pre-POMC）合成的。翻译过程中信号肽的去除产生了241个氨基酸的多肽POMC，在被内肽酶进行蛋白水解切割之前，它会经历一系列的翻译后修饰，例如磷酸化和糖基化，以产生具有不同生理活性的各种多肽片段。这些片段包括NPP，促黑素Gamma（γ-MSH），潜在肽，促肾上腺皮质激素（促肾上腺皮质激素或ACTH），促黑素Alpha（促黑素细胞激素或α-MSH），促肾上腺皮质激素样中间肽（CLIP），促肾上腺皮质激素Beta （β-LPH），脂蛋白Gamma（γ-LPH），促黑素β（β-MSH），β-内啡肽和Met-脑啡肽。POMC，

为了调节ACTH的分泌，该轴内分泌的许多物质表现出缓慢/中间和快速的反馈环活性。肾上腺皮质分泌的糖皮质激素起抑制下丘脑CRH分泌的作用，进而降低了ACTH的垂体前叶分泌。糖皮质激素也可能抑制POMC基因转录和肽合成的速率。后者是一个缓慢的反馈循环的例子，它的工作时间从几小时到几天不等，而前者的工作时间则在几分钟到几分钟。
ACTH还与许多生物体的昼夜节律有关。ACTH在人血中的半衰期约为十分钟。
相关肽
六个相关肽包括较小的生物活性片段（激素），这些片段是通过对前opiomelanocortin多蛋白（POMC）进行差分加工而从共同的前体衍生而来的。ACTH，促肾上腺皮质激素样中间叶蛋白[CLIP]，β-内啡肽，γ-脂蛋白[yLPH]，脑啡肽和α-黑素蛋白[aMSH] 1。

ACTH的发现
1930年代首次研究了ACTH的性质。1933年，由詹姆斯·科利普（James Collip），赫伯特·埃文斯（Herbert Evans）和贝玛尔多·侯赛（Bemardo Houssay）领导的研究小组使用垂体提取物刺激肾上腺皮质。美国生物化学家Choh Hao Li是1943年分离ACTH并在1963年对其进行合成的几位科学家之一。

ACTH的结构特征
ACTH的分子量为4541.3 K Da 2。它是由39个氨基组成的直链肽分子。前24个氨基酸和后7个氨基酸相同，而第25至32个氨基酸略有不同。只有前20个氨基酸才能发挥全部活性，称为活性中心。

ACTH的作用机制
ACTH进入全身循环并与位于肾上腺皮质细胞和皮肤表面的特定高亲和力受体结合。ACTH受体是七种跨膜的G蛋白偶联受体3，在配体结合后会经历构象变化，从而刺激腺苷酸环化酶，从而导致细胞内cAMP的增加和蛋白激酶A的激活。最终导致类固醇生成的刺激。

ACTH的功能
ACTH的主要功能是刺激肾上腺皮质分泌一组类固醇激素，称为糖皮质激素，盐皮质激素和雄激素类固醇。糖皮质激素可控制人体对糖的使用，并在压力时刻帮助调节生物功能。它刺激胆固醇转化为孕烯醇酮，这是所有类固醇激素的前体。它用于治疗类风湿关节炎，溃疡性结肠炎，肝炎和减轻疼痛。它们通过促进必需的蛋白质在特定于记忆的位点4合成所需的蛋白质，在记忆处理中起主要作用。

ACTH的的文献参考

1、Funkelstein L, Toneff T, Mosier C, Hwang SR, Beuschlein F, Lichtenauer UD, Reinheckel T, Peters C, Hook V (2008). Major role of cathepsin L for producing the peptide hormones ACTH, beta-endorphin, and alpha-MSH, illustrated by protease gene knockout and expression. J Biol Chem., 283(51):35652-35659.

2、Lee TH, Lerner AB, Buettner-Janusch V (1961). On the structure of human corticotropin (adrenocorticotropic hormone). J. Biol. Chem., 236:2970-2974.

3、Mountjoy KG, Robbins LS, Mortrud MT, Cone RD (1992). The cloning of a family of genes that encode the melanocortin receptors. Science, 257:248–1251.

4、Flood JF, Jarvik ME, Bennett EL, Orme AE (1976). Effects of ACTH peptide fragments on memory formation. Pharmacol Biochem Behav., 5:41-51.

多肽H2N-Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Lys-Pro-Val-Gly-Lys-Lys-Arg-Arg-Pro-Val-Lys-Val-Tyr-Ala-Asn-Gly-Ala-Glu-Glu-Glu-Ser-Ala-Glu-Ala-Phe-Pro-Leu-Glu-Phe-COOH的合成步骤：

1、合成CTC树脂：称取1.56g CTC Resin（如初始取代度约为0.8mmol/g）和1.5mmol Fmoc-Phe-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3.74mmol 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-Phe-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-Phe-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3.74mmol Fmoc-Glu(OtBu)-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7.49mmol DIPEA，3.56mmol 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-Glu(OtBu)-Phe-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-Glu(OtBu)-Phe-CTC Resin

Fmoc-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Fmoc-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Phe-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Phe-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-His(Trt)-Phe-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-His(Trt)-Phe-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Glu(OtBu)-His(Trt)-Phe-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Glu(OtBu)-His(Trt)-Phe-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Met-Glu(OtBu)-His(Trt)-Phe-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Met-Glu(OtBu)-His(Trt)-Phe-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Ser(tBu)-Met-Glu(OtBu)-His(Trt)-Phe-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Ser(tBu)-Met-Glu(OtBu)-His(Trt)-Phe-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Tyr(tBu)-Ser(tBu)-Met-Glu(OtBu)-His(Trt)-Phe-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

H2N-Tyr(tBu)-Ser(tBu)-Met-Glu(OtBu)-His(Trt)-Phe-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-CTC Resin

Fmoc-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Met-Glu(OtBu)-His(Trt)-Phe-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-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-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Met-Glu(OtBu)-His(Trt)-Phe-Arg(Pbf)-Trp(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(Pbf)-Arg(Pbf)-Pro-Val-Lys(Boc)-Val-Tyr(tBu)-Ala-Asn(Trt)-Gly-Ala-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ser(tBu)-Ala-Glu(OtBu)-Ala-Phe-Pro-Leu-Glu(OtBu)-Phe-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-Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Lys-Pro-Val-Gly-Lys-Lys-Arg-Arg-Pro-Val-Lys-Val-Tyr-Ala-Asn-Gly-Ala-Glu-Glu-Glu-Ser-Ala-Glu-Ala-Phe-Pro-Leu-Glu-Phe-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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