ASV-30 是一种构象受限的 sauvagine 类似物,是一种针对小鼠 CRF 受体 2β (mCRFR2β) 的高效选择性拮抗剂。它已被证明可抑制表达 mCRFR2β 的人胚胎肾 (HEK) 293 细胞中激动剂诱导的腺苷酸环化酶活性。
编号:136970
CAS号:220673-95-0
单字母:H2N-fHLLRKMIEIEKQEKEKQQAANNRLLLDTI-NH2
编号: | 136970 |
中文名称: | Antisauvagine-30 |
英文名: | Antisauvagine-30 |
英文同义词: | (D-Phe11,His12)-Sauvagine (11-40);11-D-Phenylalanine-12-L-histidine-11-40-sauvagine (11-40) |
CAS号: | 220673-95-0 |
单字母: | H2N-fHLLRKMIEIEKQEKEKQQAANNRLLLDTI-NH2 |
三字母: | H2N N端氨基 -DPheD型笨丙氨酸 -His组氨酸 -Leu亮氨酸 -Leu亮氨酸 -Arg精氨酸 -Lys赖氨酸 -Met甲硫氨酸 -Ile异亮氨酸 -Glu谷氨酸 -Ile异亮氨酸 -Glu谷氨酸 -Lys赖氨酸 -Gln谷氨酰胺 -Glu谷氨酸 -Lys赖氨酸 -Glu谷氨酸 -Lys赖氨酸 -Gln谷氨酰胺 -Gln谷氨酰胺 -Ala丙氨酸 -Ala丙氨酸 -Asn天冬酰胺 -Asn天冬酰胺 -Arg精氨酸 -Leu亮氨酸 -Leu亮氨酸 -Leu亮氨酸 -Asp天冬氨酸 -Thr苏氨酸 -Ile异亮氨酸 -NH2C端酰胺化 |
氨基酸个数: | 30 |
分子式: | C161H274N48O46S1 |
平均分子量: | 3650.26 |
精确分子量: | 3648.03 |
等电点(PI): | 10.33 |
pH=7.0时的净电荷数: | 2.22 |
平均亲水性: | 0.46333333333333 |
疏水性值: | -0.76 |
外观与性状: | 白色粉末状固体 |
消光系数: | - |
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 |
储存条件: | 负80℃至负20℃ |
标签: | D型氨基酸肽 |
ASV-30, a conformationally constrained analog of sauvagine, is a highly potent and selective antagonist directed against mouse CRF receptor type 2β (mCRFR2β). It has been shown to suppress agonist-induced adenylate cyclase activity in human embryonic kidney (HEK) 293 cells expressing mCRFR2β.
Antisauvagine-30是一种选择性、竞争性和强效的促肾上腺皮质激素释放因子CRF2受体拮抗剂,与大鼠CRF1和小鼠CRF2β受体结合的Kd值分别为153.6和1.4nM。它可以防止小鼠体内应激增强的恐惧条件和ERK1/2的MEK1/2依赖性激活。它抑制阴道刺激的cAMP在HEK-mCRF2β细胞中的积累。
Antisauvagine-30 is a selective, competitive and potent corticotropin-releasing factor CRF2 receptor antagonist with Kd values of 153.6 and 1.4 nM for binding to rat CRF1 and mouse CRF2β receptors respectively. It prevents stress-enhanced fear conditioning and MEK 1/2-dependent activation of ERK1/2 in mice in vivo. It inhibits sauvagine-stimulated cAMP accumulation in HEK-mCRF2β cells.
很多蛋白在细胞中非常容易被降解,或被标记,进而被选择性地破坏。但含有部分D型氨基酸的多肽则显示了很强的抵抗蛋白酶降解能力。
DOI | 名称 | |
---|---|---|
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 |
10.1016/j.bbr.2011.03.026 | Wake-promoting effects of orexin: Its independent actions against the background of an impaired corticotropine-releasing hormone receptor system | 下载 |
10.1073/pnas.95.26.15264 | Structural requirements for peptidic antagonists of the corticotropin-releasing factor receptor (CRFR): development of CRFR2beta-selective antisauvagine-30 | 下载 |
多肽H2N-DPhe-His-Leu-Leu-Arg-Lys-Met-Ile-Glu-Ile-Glu-Lys-Gln-Glu-Lys-Glu-Lys-Gln-Gln-Ala-Ala-Asn-Asn-Arg-Leu-Leu-Leu-Asp-Thr-Ile-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1.72g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1.55mmol Fmoc-Ile-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3.1mmol DIPEA,1.47mmol 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-Ile-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-Ile-Linker-MBHA Resin
Fmoc-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Met-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Met-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Lys(Boc)-Met-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Lys(Boc)-Met-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Lys(Boc)-Met-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Arg(Pbf)-Lys(Boc)-Met-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Leu-Arg(Pbf)-Lys(Boc)-Met-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Leu-Arg(Pbf)-Lys(Boc)-Met-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-Leu-Leu-Arg(Pbf)-Lys(Boc)-Met-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-Leu-Leu-Arg(Pbf)-Lys(Boc)-Met-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-His(Trt)-Leu-Leu-Arg(Pbf)-Lys(Boc)-Met-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
H2N-His(Trt)-Leu-Leu-Arg(Pbf)-Lys(Boc)-Met-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-Linker-MBHA Resin
Fmoc-DPhe-His(Trt)-Leu-Leu-Arg(Pbf)-Lys(Boc)-Met-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-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-DPhe-His(Trt)-Leu-Leu-Arg(Pbf)-Lys(Boc)-Met-Ile-Glu(OtBu)-Ile-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Gln(Trt)-Ala-Ala-Asn(Trt)-Asn(Trt)-Arg(Pbf)-Leu-Leu-Leu-Asp(OtBu)-Thr(tBu)-Ile-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-DPhe-His-Leu-Leu-Arg-Lys-Met-Ile-Glu-Ile-Glu-Lys-Gln-Glu-Lys-Glu-Lys-Gln-Gln-Ala-Ala-Asn-Asn-Arg-Leu-Leu-Leu-Asp-Thr-Ile-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。
以上所有内容,为专肽生物原创内容,请勿发布到其他网站上。