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
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.
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。
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。
