ACTH的定義
促腎上腺皮質(zhì)激素（ACTH、Adrenocorticotropic hormone）或促腎上腺皮質(zhì)激素是垂體前葉產(chǎn)生的一種激素，它刺激腎上腺皮質(zhì)。

促腎上腺皮質(zhì)激素（ACTH），也稱為促腎上腺皮質(zhì)激素，是一種由垂體前葉腺產(chǎn)生和分泌的多肽促性激素。它是下丘腦-垂體-腎上腺軸的重要組成部分，通常是在對(duì)生物壓力的反應(yīng)中產(chǎn)生的（連同下丘腦的促腎上腺皮質(zhì)激素釋放激素）。它的主要作用是增加皮質(zhì)類固醇的產(chǎn)生和釋放，以及顧名思義，是腎上腺皮質(zhì)的皮質(zhì)醇。 

ACTH是由前opiomelanocortin（pre-POMC）合成的。翻譯過(guò)程中信號(hào)肽的去除產(chǎn)生了241個(gè)氨基酸的多肽POMC，在被內(nèi)肽酶進(jìn)行蛋白水解切割之前，它會(huì)經(jīng)歷一系列的翻譯后修飾，例如磷酸化和糖基化，以產(chǎn)生具有不同生理活性的各種多肽片段。這些片段包括NPP，促黑素Gamma（γ-MSH），潛在肽，促腎上腺皮質(zhì)激素（促腎上腺皮質(zhì)激素或ACTH），促黑素Alpha（促黑素細(xì)胞激素或α-MSH），促腎上腺皮質(zhì)激素樣中間肽（CLIP），促腎上腺皮質(zhì)激素Beta （β-LPH），脂蛋白Gamma（γ-LPH），促黑素β（β-MSH），β-內(nèi)啡肽和Met-腦啡肽。POMC，

為了調(diào)節(jié)ACTH的分泌，該軸內(nèi)分泌的許多物質(zhì)表現(xiàn)出緩慢/中間和快速的反饋環(huán)活性。腎上腺皮質(zhì)分泌的糖皮質(zhì)激素起抑制下丘腦CRH分泌的作用，進(jìn)而降低了ACTH的垂體前葉分泌。糖皮質(zhì)激素也可能抑制POMC基因轉(zhuǎn)錄和肽合成的速率。后者是一個(gè)緩慢的反饋循環(huán)的例子，它的工作時(shí)間從幾小時(shí)到幾天不等，而前者的工作時(shí)間則在幾分鐘到幾分鐘。
ACTH還與許多生物體的晝夜節(jié)律有關(guān)。ACTH在人血中的半衰期約為十分鐘。
相關(guān)肽
六個(gè)相關(guān)肽包括較小的生物活性片段（激素），這些片段是通過(guò)對(duì)前opiomelanocortin多蛋白（POMC）進(jìn)行差分加工而從共同的前體衍生而來(lái)的。ACTH，促腎上腺皮質(zhì)激素樣中間葉蛋白[CLIP]，β-內(nèi)啡肽，γ-脂蛋白[yLPH]，腦啡肽和α-黑素蛋白[aMSH] 1。

ACTH的發(fā)現(xiàn)
1930年代首次研究了ACTH的性質(zhì)。1933年，由詹姆斯·科利普（James Collip），赫伯特·埃文斯（Herbert Evans）和貝瑪爾多·侯賽（Bemardo Houssay）領(lǐng)導(dǎo)的研究小組使用垂體提取物刺激腎上腺皮質(zhì)。美國(guó)生物化學(xué)家Choh Hao Li是1943年分離ACTH并在1963年對(duì)其進(jìn)行合成的幾位科學(xué)家之一。

ACTH的結(jié)構(gòu)特征
ACTH的分子量為4541.3 K Da 2。它是由39個(gè)氨基組成的直鏈肽分子。前24個(gè)氨基酸和后7個(gè)氨基酸相同，而第25至32個(gè)氨基酸略有不同。只有前20個(gè)氨基酸才能發(fā)揮全部活性，稱為活性中心。

ACTH的作用機(jī)制
ACTH進(jìn)入全身循環(huán)并與位于腎上腺皮質(zhì)細(xì)胞和皮膚表面的特定高親和力受體結(jié)合。ACTH受體是七種跨膜的G蛋白偶聯(lián)受體3，在配體結(jié)合后會(huì)經(jīng)歷構(gòu)象變化，從而刺激腺苷酸環(huán)化酶，從而導(dǎo)致細(xì)胞內(nèi)cAMP的增加和蛋白激酶A的激活。最終導(dǎo)致類固醇生成的刺激。

ACTH的功能
ACTH的主要功能是刺激腎上腺皮質(zhì)分泌一組類固醇激素，稱為糖皮質(zhì)激素，鹽皮質(zhì)激素和雄激素類固醇。糖皮質(zhì)激素可控制人體對(duì)糖的使用，并在壓力時(shí)刻幫助調(diào)節(jié)生物功能。它刺激膽固醇轉(zhuǎn)化為孕烯醇酮，這是所有類固醇激素的前體。它用于治療類風(fēng)濕關(guān)節(jié)炎，潰瘍性結(jié)腸炎，肝炎和減輕疼痛。它們通過(guò)促進(jìn)必需的蛋白質(zhì)在特定于記憶的位點(diǎn)4合成所需的蛋白質(zhì)，在記憶處理中起主要作用。

ACTH的的文獻(xiàn)參考

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-Glu-Glu-Pro-Pro-Ile-Ser-Leu-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Glu-Val-Leu-Glu-Met-Ala-Arg-Ala-Glu-Gln-Leu-Ala-Gln-Gln-Ala-His-Ser-Asn-Arg-Lys-Leu-Met-Glu-Asn-Phe-NH2的合成步驟：

1、合成MBHA樹脂：取若干克的MBHA樹脂（如初始取代度為0.5mmol/g）和1倍樹脂摩爾量的Fmoc-Linker-OH加入到反應(yīng)器中，加入DMF，攪拌使氨基酸完全溶解。再加入樹脂2倍量的DIEPA，攪拌混合均勻。再加入樹脂0.95倍量的HBTU，攪拌混合均勻。反應(yīng)3-4小時(shí)后，用DMF洗滌3次。用2倍樹脂體積的10%乙酸酐/DMF 進(jìn)行封端30分鐘。然后再用DMF洗滌3次，甲醇洗滌2次，DCM洗滌2次，再用甲醇洗滌2次。真空干燥12小時(shí)以上，得到干燥的樹脂{Fmoc-Linker-MHBA Resin}，測(cè)定取代度。這里測(cè)得取代度為 0.3mmol/g。結(jié)構(gòu)如下圖：

2、脫Fmoc：取2.34g的上述樹脂，用DCM或DMF溶脹20分鐘。用DMF洗滌2遍。加3倍樹脂體積的20%Pip/DMF溶液，鼓氮?dú)?0分鐘，然后2倍樹脂體積的DMF 洗滌5次。得到 H2N-Linker-MBHA 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2.11mmol Fmoc-Phe-OH 氨基酸，加入到上述樹脂里，加適當(dāng)DMF溶解氨基酸，再依次加入4.21mmol DIPEA，2.0mmol 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倍樹脂體積的DMF 洗滌3次樹脂。（洗滌樹脂，去掉殘留溶劑，為下一步反應(yīng)做準(zhǔn)備）。得到Fmoc-Phe-Linker-MBHA Resin。氨基酸：DIPEA：HBTU：樹脂=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-Phe-Linker-MBHA Resin

Fmoc-Asn(Trt)-Phe-Linker-MBHA Resin

H2N-Asn(Trt)-Phe-Linker-MBHA Resin

Fmoc-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

H2N-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

Fmoc-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

H2N-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

Fmoc-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

H2N-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

Fmoc-Lys(Boc)-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

H2N-Lys(Boc)-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

Fmoc-Arg(Pbf)-Lys(Boc)-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

H2N-Arg(Pbf)-Lys(Boc)-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

Fmoc-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

H2N-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

Fmoc-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

H2N-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

Fmoc-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

H2N-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

Fmoc-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

H2N-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

以上中間結(jié)構(gòu)，均可在專肽生物多肽計(jì)算器-多肽結(jié)構(gòu)計(jì)算器中，一鍵畫出。

最后再經(jīng)過(guò)步驟二得到 H2N-Ser(tBu)-Glu(OtBu)-Glu(OtBu)-Pro-Pro-Ile-Ser(tBu)-Leu-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Met-Glu(OtBu)-Asn(Trt)-Phe-Linker-MBHA Resin，結(jié)構(gòu)如下：

5、切割：6倍樹脂體積的切割液（或每1g樹脂加8ml左右的切割液），搖床搖晃 2小時(shí)，過(guò)濾掉樹脂，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-Ser-Glu-Glu-Pro-Pro-Ile-Ser-Leu-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Glu-Val-Leu-Glu-Met-Ala-Arg-Ala-Glu-Gln-Leu-Ala-Gln-Gln-Ala-His-Ser-Asn-Arg-Lys-Leu-Met-Glu-Asn-Phe-NH2。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)，提供各類長(zhǎng)肽，短肽，環(huán)肽，提供各類修飾肽，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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