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尿紧张素UrotensinI

一种类 CRF 样肽,可作为 CRF 受体激动剂。

编号:160553

CAS号:83930-33-0

单字母:H2N-NDDPPISIDLTFHLLRNMIEMARIENEREQAGLNRKYLDEV-NH2

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  • 编号:160553
    中文名称:尿紧张素UrotensinI
    英文名:UrotensinI
    英文同义词:Catostomus urotensin I TFA)
    CAS号:83930-33-0
    单字母:H2N-NDDPPISIDLTFHLLRNMIEMARIENEREQAGLNRKYLDEV-NH2
    三字母:H2N

    N端氨基

    -Asn

    天冬酰胺

    -Asp

    天冬氨酸

    -Asp

    天冬氨酸

    -Pro

    脯氨酸

    -Pro

    脯氨酸

    -Ile

    异亮氨酸

    -Ser

    丝氨酸

    -Ile

    异亮氨酸

    -Asp

    天冬氨酸

    -Leu

    亮氨酸

    -Thr

    苏氨酸

    -Phe

    苯丙氨酸

    -His

    组氨酸

    -Leu

    亮氨酸

    -Leu

    亮氨酸

    -Arg

    精氨酸

    -Asn

    天冬酰胺

    -Met

    甲硫氨酸

    -Ile

    异亮氨酸

    -Glu

    谷氨酸

    -Met

    甲硫氨酸

    -Ala

    丙氨酸

    -Arg

    精氨酸

    -Ile

    异亮氨酸

    -Glu

    谷氨酸

    -Asn

    天冬酰胺

    -Glu

    谷氨酸

    -Arg

    精氨酸

    -Glu

    谷氨酸

    -Gln

    谷氨酰胺

    -Ala

    丙氨酸

    -Gly

    甘氨酸

    -Leu

    亮氨酸

    -Asn

    天冬酰胺

    -Arg

    精氨酸

    -Lys

    赖氨酸

    -Tyr

    酪氨酸

    -Leu

    亮氨酸

    -Asp

    天冬氨酸

    -Glu

    谷氨酸

    -Val

    缬氨酸

    -NH2

    C端酰胺化

    氨基酸个数:41
    分子式:C210H340N62O67S2
    平均分子量:4869.45
    精确分子量:4866.45
    等电点(PI):4.92
    pH=7.0时的净电荷数:-1.78
    平均亲水性:0.4
    疏水性值:-0.7
    外观与性状:白色粉末状固体
    消光系数:1490
    来源:人工化学合成,仅限科学研究使用,不得用于人体。
    纯度:95%、98%
    盐体系:可选TFA、HAc、HCl或其它
    生成周期:2-3周
    储存条件:负80℃至负20℃
    标签:激动剂多肽(Agonist Peptide)    尿紧张素   

  • Urotensin I (Catostomus urotensin I) TFA 是一种类 CRF 样肽,可作为 CRF 受体激动剂,在细胞实验中,对人 CRF1,CRF2 和大鼠 CRF2β 受体的 pEC50 值分别为 11.46,9.36 和 9.85,对 hCRF1,rCRF2β 和 mCRF2β 受体的 Ki 值分别为 0.4,1.8 和 5.7 nM。
    Urotensin I (Catostomus urotensin I) TFA, a CRF-like neuropeptide, acts as an agonist of CRF receptor with pEC50s of 11.46, 9.36 and 9.85 for human CRF1, human CRF2 and rat CRF2β receptors in CHO cells, and Kis of 0.4, 1.8, and 5.7 nM for hCRF1, rCRF2β and mCRF2β receptors, respectively[1][2].

  • Smart D, et al. Characterisation using microphysiometry of CRF receptor pharmacology. Eur J Pharmacol. 1999 Aug 27;379(2-3):229-35. : https://pubmed.ncbi.nlm.nih.gov/10497910/
    Reul JM, et al. Corticotropin-releasing factor receptors 1 and 2 in anxiety and depression. Curr Opin Pharmacol. 2002 Feb;2(1):23-33. : https://pubmed.ncbi.nlm.nih.gov/11786305/
    Fryer J, et al. Urotensin I, a CRF-like neuropeptide, stimulates acth release from the teleost pituitary. Endocrinology. 1983;113(6):2308-2310. : https://pubmed.ncbi.nlm.nih.gov/6315348/
    Gerritsen ME, et al. Urotensin I effects on intracellular content of cyclic AMP in the rat tail artery. Eur J Pharmacol. 1979;60(2-3):211-220. : https://pubmed.ncbi.nlm.nih.gov/230978/

  • 多肽H2N-Asn-Asp-Asp-Pro-Pro-Ile-Ser-Ile-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Asn-Met-Ile-Glu-Met-Ala-Arg-Ile-Glu-Asn-Glu-Arg-Glu-Gln-Ala-Gly-Leu-Asn-Arg-Lys-Tyr-Leu-Asp-Glu-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1.94g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1.75mmol 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3.49mmol DIPEA,1.66mmol 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-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Ile-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Ile-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Met-Ile-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Met-Ile-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Asn(Trt)-Met-Ile-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Asn(Trt)-Met-Ile-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Arg(Pbf)-Asn(Trt)-Met-Ile-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Arg(Pbf)-Asn(Trt)-Met-Ile-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Leu-Arg(Pbf)-Asn(Trt)-Met-Ile-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Leu-Arg(Pbf)-Asn(Trt)-Met-Ile-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    Fmoc-Leu-Leu-Arg(Pbf)-Asn(Trt)-Met-Ile-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

    H2N-Leu-Leu-Arg(Pbf)-Asn(Trt)-Met-Ile-Glu(OtBu)-Met-Ala-Arg(Pbf)-Ile-Glu(OtBu)-Asn(Trt)-Glu(OtBu)-Arg(Pbf)-Glu(OtBu)-Gln(Trt)-Ala-Gly-Leu-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(OtBu)-Val-Linker-MBHA Resin

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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