400-998-5282
专注多肽 服务科研
400-998-5282
专注多肽 服务科研
RFamide 肽家族的成员，具有促食欲作用，是小鼠孤儿受体 GPR103 的同源配体。
编号：200776
CAS号：600171-70-8
单字母：H2N-ASGPLGTLAEELSSYSRRKGGFSFRF-CONH2
| 参考文献（References）: | N.Chartrel et al., Proc. Natl. Acad. Sci. USA, 100, 15247 (2003) Y.Jiang et al., J. Biol. Chem., 278, 27652 (2003) |
Orphan GPCR SP9155 agonist P550 (mouse, rat) (26RFa (mouse, rat))，RFamide 肽家族的成员，具有促食欲作用，是小鼠孤儿受体 GPR103 的同源配体，也命名为 SP9155 或 AQ27。
Orphan GPCR SP9155 agonist P550 (mouse, rat) (26RFa (mouse, rat)), a member of the RFamide peptide family with orexigenic effect, is the cognate ligand of the mouse orphan receptor GPR103, also designated SP9155 or AQ27.
26Rfa是属于RFamide肽家族的神经肽。它具有促食欲活性，并与肥胖有关。人类、大鼠和青蛙26RFa的一级结构具有80%的同一性，C端八肽从两栖动物到哺乳动物都是完全保守的。
26Rfa is a neuropeptide belonging to the RFamide peptide family. It exhibits orexigenic activity and has been associated to obesity. The primary structures of human, rat, and frog 26RFa exhibit 80% identity, and the C-terminal octapeptide is fully conserved from amphibians to mammals.
P550（小鼠）是P518（人类）的小鼠同源物。已显示它对人类孤儿G蛋白偶联受体SP9155具有与P518（人类）相似的激动活性（EC50=6 nM）。
P550 (mouse) is the mouse homolog of P518 (human). It has been shown to exhibit similar agonistic activity towards the human orphan G-protein-coupled receptor SP9155 as P518 (human) (EC50 = 6 nM).
26Rfa 下丘脑肽（大鼠）是大鼠来源的酰胺化神经肽，含有影响结构动力学的芳香族和碱性残基。其序列支持神经内分泌信号传导和肽 - 受体接触的研究。柔性主链能够进行环境响应性构象变化。应用包括结构作图、配体结合分析和比较神经肽研究。
26Rfa Hypothalamic Peptide rat is a rat-derived amidated neuropeptide containing aromatic and basic residues that influence structural dynamics. Its sequence supports studies of neuroendocrine signaling and peptide-receptor contacts. The flexible backbone enables environment-responsive conformational changes. Applications include structural mapping, ligand-binding analysis, and comparative neuropeptide research.
孤儿 G 蛋白偶联受体 SP9155 激动剂 P550（小鼠）是一种多肽配体，用于激活小鼠系统中未表征的 G 蛋白偶联受体。其序列适用于受体去孤化和基序识别研究。研究人员将其用于鉴定细胞内信号结果。该多肽有助于发现新型 G 蛋白偶联受体通路。
Orphan GPCR SP9155 Agonist P550 (mouse) is a peptide ligand designed to activate an uncharacterized GPCR in murine systems. Its sequence supports studies of receptor deorphanization and motif recognition. Researchers employ it to identify intracellular signaling outcomes. The peptide aids discovery of novel GPCR pathways.
Orphan GPCR SP9155 Agonist P550 (mouse) 别名 P550 (mouse), Orphan G-Protein Coupled Receptor SP9155 Agonist Peptide P550 (mouse)，是P518 (human) 的鼠同源物。它对人孤儿 G-蛋白-偶联的受体 SP9155 (orphan G-protein-coupled receptor SP9155) 显示出和P518 (human) 相似的竞争活性。
Chartrel N, et al. Identification of 26RFa, a hypothalamic neuropeptide of the RFamide peptide family with orexigenic activity. Proc Natl Acad Sci U S A. 2003;100(25):15247-15252. : https://pubmed.ncbi.nlm.nih.gov/14657341/
Chartrel N, et al. The Neuropeptide 26RFa (QRFP) and Its Role in the Regulation of Energy Homeostasis: A Mini-Review. Front Neurosci. 2016;10:549. Published 2016 Nov 29. : https://pubmed.ncbi.nlm.nih.gov/27965532/
Egido EM, et al. 26RFa, a novel orexigenic neuropeptide, inhibits insulin secretion in the rat pancreas. Peptides. 2007;28(4):725-730. : https://pubmed.ncbi.nlm.nih.gov/16777265/
多肽H2N-Ala-Ser-Gly-Pro-Leu-Gly-Thr-Leu-Ala-Glu-Glu-Leu-Ser-Ser-Tyr-Ser-Arg-Arg-Lys-Gly-Gly-Phe-Ser-Phe-Arg-Phe-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1.28g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1.15mmol Fmoc-Phe-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2.3mmol DIPEA，1.09mmol 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-Phe-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-Phe-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Thr(tBu)-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Thr(tBu)-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Gly-Thr(tBu)-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Gly-Thr(tBu)-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Leu-Gly-Thr(tBu)-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Leu-Gly-Thr(tBu)-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Pro-Leu-Gly-Thr(tBu)-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Pro-Leu-Gly-Thr(tBu)-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Gly-Pro-Leu-Gly-Thr(tBu)-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Gly-Pro-Leu-Gly-Thr(tBu)-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Ser(tBu)-Gly-Pro-Leu-Gly-Thr(tBu)-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
H2N-Ser(tBu)-Gly-Pro-Leu-Gly-Thr(tBu)-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-Linker-MBHA Resin
Fmoc-Ala-Ser(tBu)-Gly-Pro-Leu-Gly-Thr(tBu)-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-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-Ala-Ser(tBu)-Gly-Pro-Leu-Gly-Thr(tBu)-Leu-Ala-Glu(OtBu)-Glu(OtBu)-Leu-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Ser(tBu)-Arg(Pbf)-Arg(Pbf)-Lys(Boc)-Gly-Gly-Phe-Ser(tBu)-Phe-Arg(Pbf)-Phe-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-Ala-Ser-Gly-Pro-Leu-Gly-Thr-Leu-Ala-Glu-Glu-Leu-Ser-Ser-Tyr-Ser-Arg-Arg-Lys-Gly-Gly-Phe-Ser-Phe-Arg-Phe-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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