CRF（促肾上腺皮质激素释放因子）是一种主要在下丘脑产生的 41 肽。肽激素刺激垂体前叶释放 ACTH。CRH 在内分泌、行为和对压力的免疫反应中起重要作用，并且可能在能量平衡的调节中也发挥着重要作用。人类序列 EEPPISLDLTFHLLREVLEMARAEQLAQQAHSNRKLMEII 酰胺也对应于犬、猫、鼠和猪 CRH 的序列。
编号：118180
CAS号：79804-71-0/121249-14-7/9015-71-8
单字母：H2N-SQEPPISLDLTFHLLREVLEMTKADQLAQQAHSNRKLLDIA-CONH2
CRF（促肾上腺皮质激素释放因子）是一种主要在下丘脑产生的 41 肽。肽激素刺激垂体前叶释放 ACTH。CRH 在内分泌、行为和对压力的免疫反应中起重要作用，并且可能在能量平衡的调节中也发挥着重要作用。人类序列 EEPPISLDLTFHLLREVLEMARAEQLAQQAHSNRKLMEII 酰胺也对应于犬、猫、鼠和猪 CRH 的序列。
CRF (corticotropin-releasing factor) is a 41-peptide produced mainly in the hypothalamus. The peptide hormone stimulates ACTH release from the anterior lobe of the pituitary gland. CRH plays an important role in the endocrine, behavioral, and immune response to stress and probably as well in the regulation of energy balance. The human sequence EEPPISLDLTFHLLREVLEMARAEQLAQQAHSNRKLMEII amide also corresponds to the sequence of canine, feline, murine, and porcine CRH.
Corticotropin-releasing factor (CRF)是一种内源性神经肽激素，与压力反应，抑郁，焦虑和其他情绪障碍有关。
| DOI | 名称 | |
|---|---|---|
| 10.1002/cbic.200300769 | Structure-activity studies on the corticotropin releasing factor antagonist astressin, leading to a minimal sequence necessary for antagonistic activity | 下载 |
| 10.1016/j.bcp.2004.07.013 | Cell-type specific calcium signaling by corticotropin-releasing factor type 1 (CRF1) and 2a (CRF2(a)) receptors: phospholipase C-mediated responses in human embryonic kidney 293 but not SK-N-MC neuroblastoma cells | 下载 |
| 10.1152/ajpregu.00099.2006 | Carboxyl-terminal and intracellular loop sites for CRF1 receptor phosphorylation and beta-arrestin-2 recruitment: a mechanism regulating stress and anxiety responses | 下载 |
| 10.1111/j.1471-4159.2007.05052.x | Expression, binding, and signaling properties of CRF2(a) receptors endogenously expressed in human retinoblastoma Y79 cells: passage-dependent regulation of functional receptors | 下载 |
| 10.1111/j.1476-5381.2011.01390.x | Quantitative pharmacological analysis of antagonist binding kinetics at CRF1 receptors in vitro and in vivo | 下载 |
| 10.1073/pnas.83.23.8824 | Identification and characterization of a vasopressin isoreceptor in porcine seminal vesicles | 下载 |
| 10.1126/science.6267699 | Characterization of a 41-residue ovine hypothalamic peptide that stimulates secretion of corticotropin and beta-endorphin | 下载 |
| 10.1113/jphysiol.2007.150078 | Corticotropin-releasing factor increases mouse ventral tegmental area dopamine neuron firing through a protein kinase C-dependent enhancement of Ih | 下载 |
多肽H2N-Ser-Gln-Glu-Pro-Pro-Ile-Ser-Leu-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Glu-Val-Leu-Glu-Met-Thr-Lys-Ala-Asp-Gln-Leu-Ala-Gln-Gln-Ala-His-Ser-Asn-Arg-Lys-Leu-Leu-Asp-Ile-Ala-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1.44g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1.3mmol Fmoc-Ala-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2.59mmol DIPEA，1.23mmol 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-Ala-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-Ala-Linker-MBHA Resin
Fmoc-Ile-Ala-Linker-MBHA Resin
H2N-Ile-Ala-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-His(Trt)-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-His(Trt)-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Leu-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Leu-Asp(OtBu)-Leu-Thr(tBu)-Phe-His(Trt)-Leu-Leu-Arg(Pbf)-Glu(OtBu)-Val-Leu-Glu(OtBu)-Met-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-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-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-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-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-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-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-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-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-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-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-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-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-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-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-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-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-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-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-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-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Gln(Trt)-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-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
H2N-Gln(Trt)-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-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-Linker-MBHA Resin
Fmoc-Ser(tBu)-Gln(Trt)-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-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-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-Ser(tBu)-Gln(Trt)-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-Thr(tBu)-Lys(Boc)-Ala-Asp(OtBu)-Gln(Trt)-Leu-Ala-Gln(Trt)-Gln(Trt)-Ala-His(Trt)-Ser(tBu)-Asn(Trt)-Arg(Pbf)-Lys(Boc)-Leu-Leu-Asp(OtBu)-Ile-Ala-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-Ser-Gln-Glu-Pro-Pro-Ile-Ser-Leu-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Glu-Val-Leu-Glu-Met-Thr-Lys-Ala-Asp-Gln-Leu-Ala-Gln-Gln-Ala-His-Ser-Asn-Arg-Lys-Leu-Leu-Asp-Ile-Ala-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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