400-998-5282
专注多肽 服务科研
400-998-5282
专注多肽 服务科研
一种神经肽，也是一种有效的内源性 CRFR 激动剂。
编号：194578
CAS号：171543-83-2
单字母：H2N-DDPPLSIDLTFHLLRTLLELARTQSQRERAEQNRIIFDSV-CONH2
| 参考文献（References）: | J. Vaughan et al, Nature, 378, 287(1995) 1. Vaughan, J., Donaldson, C., Bittencourt, J., et al . Urocortin, a mammalian neuropeptide related to fish urotensin I and to corticotropin- releasing factor . Nature 378(6554), 287-292 (1995) . <ALLPEPTIDE>2. Martinez, V., Wang, L., Million, M., et al . Urocortins and the regulation of gastrointestinal motor function and visceral pain . Peptides 25(10), 1733-1744 (2004) . <ALLPEPTIDE>3. Sumii, K., Miyake, H., Enatsu, N., et al . Characterization of urocortin as an anti- apoptotic protein in experimental ischemia- reperfusion model of the rat testis . Biochem. Biophys. Res. Commun. 479(2), 387-392 (2016) . |
| 溶解度: | Formic Acid: 1 mg/ml |
| SMILES: | NC([C@H](C(C)C)NC([C@H](CO)NC([C@@H](NC([C@@H](NC([C@@]([C@H](CC)C)([H])NC([C@@]([C@H](CC)C)([H])NC([C@H](CCCNC(N)=N)NC([C@H](CC(N)=O)NC([C@H](CCC(N)=O)NC([C@H](CCC(O)=O)NC([C@@H](NC([C@H](CCCNC(N)=N)NC([C@H](CCC(O)=O)NC([C@H](CCCNC(N)=N)NC([C@H](CCC(N)=O)NC([C@H](CO)NC([C@H](CCC(N)=O)NC([C@@]([C@@H](C)O)([H])NC([C@H](CCCNC(N)=N)NC([C@@H](NC([C@H](CC(C)C)NC([C@H](CCC(O)=O)NC([C@H](CC(C)C)NC([C@H](CC(C)C)NC([C@@]([C@@H](C)O)([H])NC([C@H](CCCNC(N)=N)NC([C@H](CC(C)C)NC([C@H](CC(C)C)NC([C@@H](NC([C@@H](NC([C@@]([C@@H](C)O)([H])NC([C@H](CC(C)C)NC([C@@H](NC([C@@]([C@H](CC)C)([H])NC([C@H](CO)NC([C@H](CC(C)C)NC([C@@H]1CCCN1C([C@@H]2CCCN2C([C@@H](NC([C@@H](N[H])CC(O)=O)=O)CC(O)=O)=O)=O)=O)=O)=O)=O)CC(O)=O)=O)=O)=O)CC3=CC=CC=C3)=O)CC4=CN=CN4)=O)=O)=O)=O)=O)=O)=O)=O)=O)C)=O)=O)=O)=O)=O)=O)=O)=O)=O)C)=O)=O)=O)=O)=O)=O)=O)CC5=CC=CC=C5)=O)CC(O)=O)=O)=O)=O.FC(F)(C(O)=O)F |
| InChI/InChI Code: | InChI=1S/C206H338N62O64.C2HF3O2/c1-28-102(20)156(262-192(323)141(92-271)258-182(313)129(78-98(12)13)251-193(324)142-53-42-72-267(142)201(332)143-54-43-73-268(143)200(331)138(88-154(291)292)256-165(296)113(207)84-151(285)286)194(325)255-137(87-153(289)290)187(318)247-131(80-100(16)17)189(320)266-161(109(27)274)199(330)254-133(82-111-46-35-32-36-47-111)183(314)248-134(83-112-89-223-93-229-112)185(316)246-128(77-97(10)11)180(311)244-126(75-95(6)7)178(309)236-118(52-41-71-228-206(221)222)176(307)265-160(108(26)273)198(329)252-130(79-99(14)15)181(312)245-127(76-96(8)9)179(310)240-124(60-66-150(283)284)172(303)243-125(74-94(4)5)177(308)231-106(24)163(294)232-116(50-39-69-226-204(217)218)175(306)264-159(107(25)272)197(328)242-121(57-63-146(210)277)173(304)257-139(90-269)190(321)241-119(55-61-144(208)275)168(299)235-115(49-38-68-225-203(215)216)167(298)239-123(59-65-149(281)282)170(301)234-114(48-37-67-224-202(213)214)166(297)230-105(23)164(295)233-122(58-64-148(279)280)169(300)238-120(56-62-145(209)276)171(302)249-135(85-147(211)278)186(317)237-117(51-40-70-227-205(219)220)174(305)261-158(104(22)30-3)196(327)263-157(103(21)29-2)195(326)253-132(81-110-44-33-31-34-45-110)184(315)250-136(86-152(287)288)188(319)259-140(91-270)191(322)260-155(101(18)19)162(212)293;3-2(4,5)1(6)7/h31-36,44-47,89,93-109,113-143,155-161,269-274H,28-30,37-43,48-88,90-92,207H2,1-27H3,(H2,208,275)(H2,209,276)(H2,210,277)(H2,211,278)(H2,212,293)(H,223,229)(H,230,297)(H,231,308)(H,232,294)(H,233,295)(H,234,301)(H,235,299)(H,236,309)(H,237,317)(H,238,300)(H,239,298)(H,240,310)(H,241,321)(H,242,328)(H,243,303)(H,244,311)(H,245,312)(H,246,316)(H,247,318)(H,248,314)(H,249,302)(H,250,315)(H,251,324)(H,252,329)(H,253,326)(H,254,330)(H,255,325)(H,256,296)(H,257,304)(H,258,313)(H,259,319)(H,260,322)(H,261,305)(H,262,323)(H,263,327)(H,264,306)(H,265,307)(H,266,320)(H,279,280)(H,281,282)(H,283,284)(H,285,286)(H,287,288)(H,289,290)(H,291,292)(H4,213,214,224)(H4,215,216,225)(H4,217,218,226)(H4,219,220,227)(H4,221,2 |
| InChI Key: | BTOGVDGOXGIAEB-FJAJBMFWSA-N |
| Formal Name: | L-α-aspartyl-L-α-aspartyl-L-prolyl-L-prolyl-L-leucyl-L-seryl-L-isoleucyl-L-α-aspartyl-L-leucyl-L-threonyl-L-phenylalanyl-L-histidyl-L-leucyl-L-leucyl-L-arginyl-L-threonyl-L-leucyl-L-leucyl-L-α-glutamyl-L-leucyl-L-alanyl-L-arginyl-L-threonyl-L-glutaminyl-L-seryl-L-glutaminyl-L-arginyl-L-α-glutamyl-L-arginyl-L-alanyl-L-α-glutamyl-L-glutaminyl-L-asparaginyl-L-arginyl-L-isoleucyl-L-isoleucyl-L-phenylalanyl-L-α-aspartyl-L-seryl-L-valinamide,trifluoroacetatesalt |
Urocortin, rat (Urocortin (Rattus norvegicus)) 是一种神经肽，也是一种有效的内源性 CRFR 激动剂，对人 CRF1，大鼠 CRF2α 和小鼠 CRF2β 的 Ki 分别为 13 nM，1.5 nM 和 0.97 nM。
Urocortin, rat (Urocortin (Rattus norvegicus)) is a neuropeptide and a potent endogenous CRFR agonist with Kis of 13 nM, 1.5 nM, and 0.97 nM for human CRF1, rat CRF2α and mouse CRF2β, respectively.
尿皮质素是一种肽，也称为促肾上腺皮质激素释放因子（CRF）。它是蛋白磷酸酶1和蛋白磷酸酶2A的抑制剂，蛋白磷酸酶1和蛋白磷酸酶2A是调节其他蛋白质活性的酶。尿皮质素还具有与糖皮质激素受体（如GRP和MR）结合的能力，从而激活它们。该肽用于研究离子通道，神经元兴奋性和细胞生物学的其他方面。尿皮质素已被证明是一种高纯度的试剂，没有可检测的杂质。
Urocortin is a peptide that is also known as corticotropin-releasing factor (CRF). It is an inhibitor of the protein phosphatase 1 and protein phosphatase 2A, which are enzymes that regulate the activity of other proteins. Urocortin also has the ability to bind to receptors for glucocorticoid hormones, such as GRP and MR, in a manner that activates them. This peptide is used in research studies to study ion channels, neuronal excitability, and other aspects of cell biology. Urocortin has been shown to be a high purity reagent with no detectable impurities.
在结合和激活2型CRF受体方面比CRF更有效的肽，并且与人类版本共享约95%的序列。作为与鱼尾加压素和促肾上腺皮质激素释放因子相关的哺乳动物神经肽，它还在富含2型CRF受体的区域中充当c-Fos的诱导剂。
Peptide that is more potent than CRF at binding and activating type-2 CRF receptors and shares ~95% sequence with human version. As a mammalian neuropeptide related to fish urotensin and corticotropin-releasing factor, it also serves as an inducer of c-Fos in regions enriched in type-2 CRF receptors.
尿皮质素属于促肾上腺皮质激素释放因子家族神经肽激素。大鼠尿皮质素与硬骨鱼尾紧张素、促肾上腺皮质激素释放因子序列同源性分别为 63% 和 45%，主要表达于成年大鼠脑部特定区域，在外周组织也有分布。它可结合一型、二型促肾上腺皮质激素释放因子受体及促肾上腺皮质激素释放因子结合蛋白，能够诱导相关细胞产生环磷酸腺苷，刺激大鼠垂体前叶细胞分泌促肾上腺皮质激素。体内给药可降低大鼠平均动脉压，提升血浆促肾上腺皮质激素水平，抑制大小鼠固体食物胃排空，阻断迷走神经兴奋引发的胃部蠕动。睾丸扭转致大鼠睾丸缺血再灌注损伤后，睾丸组织内尿皮质素信使核糖核酸表达量上升。
Urocortin is a neuropeptide hormone and member of the corticotropin-releasing factor (CRF) family which includes mammalian CRF, urocortin II (Item Nos. 24746 | 24747 ), urocortin III (Item Nos. 24748 | 24749 | 24750 ), frog sauvagine, and piscine urotensin I. Rat urocortin shares 63 and 45% sequence identity to urotensin and CRF, respectively. It is primarilly expressed in the Edinger-Westphal nucleus and lateral superior olive regions in the adult rat brain but can also be expressed in the periphery. Urocortin binds to type 1 and 2 CRF receptors as well as CRF binding protein (CRF-BP) with K values of 0.16, 0.41, and 0.10 nM for CRF , CRF , and CRF-BP, respectively. It induces cAMP production in CHO cells expressing CRF and CRF (EC s = 0.8 and 0.18 nM, respectively) and secretion of adrenocorticotropic hormone (ACTH) by rat anterior pituitary cells (EC = 0.006 nM). In vivo , urocortin (0.94-18.85 μg/kg) decreases mean arterial pressure and increases plasma levels of ACTH in rats. Urocortin inhibits gastric emptying of a solid meal in mice and rats and inhibits gastric motility and transit induced by vagal stimulation in rats. Levels of urocortin mRNA increase in testis in response to ischemia-reperfusion injury in a rat model of testicular torsion.
| DOI | 名称 | |
|---|---|---|
| 10.1111/j.1749-6632.1999.tb08687.x | Corticotropin releasing factor receptors and their ligand family | 下载 |
| 10.1016/s0167-0115(00)00180-4 | The neurobiology of urocortin | 下载 |
| 10.1016/s1471-4892(01)00117-5 | Corticotropin-releasing factor receptors 1 and 2 in anxiety and depression | 下载 |
多肽H2N-Asp-Asp-Pro-Pro-Leu-Ser-Ile-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Thr-Leu-Leu-Glu-Leu-Ala-Arg-Thr-Gln-Ser-Gln-Arg-Glu-Arg-Ala-Glu-Gln-Asn-Arg-Ile-Ile-Phe-Asp-Ser-Val-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2.35g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2.11mmol Fmoc-Val-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4.23mmol DIPEA，2.01mmol 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-Val-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-Val-Linker-MBHA Resin
Fmoc-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Ile-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Ile-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Ser(tBu)-Ile-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Ser(tBu)-Ile-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Leu-Ser(tBu)-Ile-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Leu-Ser(tBu)-Ile-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Pro-Leu-Ser(tBu)-Ile-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Pro-Leu-Ser(tBu)-Ile-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Pro-Pro-Leu-Ser(tBu)-Ile-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Pro-Pro-Leu-Ser(tBu)-Ile-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Pro-Pro-Leu-Ser(tBu)-Ile-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
H2N-Asp(OtBu)-Pro-Pro-Leu-Ser(tBu)-Ile-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Asp(OtBu)-Pro-Pro-Leu-Ser(tBu)-Ile-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-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-Asp(OtBu)-Asp(OtBu)-Pro-Pro-Leu-Ser(tBu)-Ile-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Thr(tBu)-Leu-Leu-Glu(OtBu)-Leu-Ala-Arg(Pbf)-Thr(tBu)-Gln(Trt)-Ser(tBu)-Gln(Trt)-Arg(Pbf)-Glu(OtBu)-Arg(Pbf)-Ala-Glu(OtBu)-Gln(Trt)-Asn(Trt)-Arg(Pbf)-Ile-Ile-Phe-Asp(OtBu)-Ser(tBu)-Val-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-Asp-Asp-Pro-Pro-Leu-Ser-Ile-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Thr-Leu-Leu-Glu-Leu-Ala-Arg-Thr-Gln-Ser-Gln-Arg-Glu-Arg-Ala-Glu-Gln-Asn-Arg-Ile-Ile-Phe-Asp-Ser-Val-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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