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Protease-Activated Receptor-2, amide (SLIGKV-NH2) 是一种高效的蛋白酶活化受体-2 (PAR2) 活化肽。

Protease-Activated Receptor-2, amide (SLIGKV-NH2) is a highly potent protease-activated receptor-2 (PAR2) activating peptide.

H-Ser-Leu-Ile-Gly-Lys-Val-NH2是一种合成肽，是一种环化酶抑制剂。它与神经激肽1的受体结合，神经激肽1已被证明可抑制大鼠的泪腺功能。该化合物还抑制表皮生长因子的合成，并已显示在小鼠模型中具有抗炎作用。H-Ser-Leu-Ile-Gly-Lys-Val-NH2也被证明可以抑制大鼠前列腺细胞中的蛋白酶活性，可以用作蛋白酶的化学抑制剂。

H-Ser-Leu-Ile-Gly-Lys-Val-NH2 is a synthetic peptide that is a cyclase inhibitor. It binds to the receptor for neurokinin 1, which has been shown to inhibit lacrimal gland function in rats. The compound also inhibits the synthesis of epidermal growth factor and has been shown to have anti-inflammatory effects in mouse models. H-Ser-Leu-Ile-Gly-Lys-Val-NH2 has also been shown to inhibit protease activity in rat prostate cells and can be used as a chemical inhibitor for proteases.

这种蛋白酶激活的受体激活肽（PAR-2-AP）对应于PAR2束缚的配体。它也可以用于研究受体的功能。

This protease-activated receptor activating peptide (PAR-2-AP) corresponds to the PAR2 tethered ligand. It can also be utlized to study the functions of receptors.

人蛋白酶激活受体 2（1‑6）酰胺为蛋白酶触发受体研究提供稳定的最小激活基序。序列组成适用于切割事件动力学分析。折叠柔性有助于早期受体结合定位。该多肽常用于信号与多肽‑酶相互作用研究。

PAR-2 (1-6) amide (human) provides a stabilized minimal activation motif for protease-triggered receptor studies. Sequence composition supports kinetic profiling of cleavage events. Folding flexibility aids mapping of early receptor engagement. The peptide is commonly utilized in signaling and peptide–enzyme interaction research.

Protease-Activated Receptor-2, amide 蛋白酶活化的受体-2 Protease-Activated Receptor-2, amide (SLIGKV-NH2) 是一种高效的蛋白酶活化受体-2 (PAR2) 活化肽。 体外研究： The PAR2-activating peptides used are: SLIGKV-OH, SLIGRL-OH, SLIGKV-NH2, SLIGRL-NH2. The synthetic agonist peptides mimicking the tethered ligand of PAR2, Ser-Leu-Ile-Gly-Lys-Val (SLIGKV-OH), Ser-Leu-Ile-Gly-Arg-Leu (SLIGRL-OH) and their amidated forms Ser-Leu-Ile-Gly-Lys-Val-amide (SLIGKV-NH2) Ser-Leu-Ile-Gly-Arg-Leu-amide (SLIGRL-NH2) have also been demonstrated being able to activate the receptor without enzymatic cleavage, therefore, have been utilised as biological tools to examine physiological functions of PAR2. Protease-Activated Receptor-2, amide is one of a four family subgroup of G-protein-coupled receptors (GPCRs), called PARs. Protease-activated receptors are distinguished from other GPCRs through their unique proteolytic mechanism of activation. For PAR2, activating proteases, such as trypsin, tryptase and coagulation factors VIIa and Xa, cleave a specific extracellular amino-terminal domain of the receptor to reveal a "tethered ligand", SLIGKV- and SLIGRL- for human and mouse/rat PAR2, respectively, which subsequently interacts with the activation domain of the receptor, initiating intracellular signaling pathways[1]. The protease-activated receptor-2 (PAR2) has been implicated in the pathogenesis of several inflammatory and autoimmune disorders, and is expressed in a wide variety of human tissues and cells. PAR2 belongs to a family of seven transmembrane domain receptor proteins that are activated by proteolysis. Enzymatic digestion exposes an N-terminus ligand sequence that binds intramolecularly to the activation site on the extracellular loop II, initiating a G-protein-mediated cell-signalling cascade and nuclear factor-kappa B (NF-κB)-regulated gene transcription。
 

多肽H2N-Ser-Leu-Ile-Gly-Lys-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2.22g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2.0mmol 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4.0mmol DIPEA，1.9mmol 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-Lys(Boc)-Val-Linker-MBHA Resin

H2N-Lys(Boc)-Val-Linker-MBHA Resin

Fmoc-Gly-Lys(Boc)-Val-Linker-MBHA Resin

H2N-Gly-Lys(Boc)-Val-Linker-MBHA Resin

Fmoc-Ile-Gly-Lys(Boc)-Val-Linker-MBHA Resin

H2N-Ile-Gly-Lys(Boc)-Val-Linker-MBHA Resin

Fmoc-Leu-Ile-Gly-Lys(Boc)-Val-Linker-MBHA Resin

H2N-Leu-Ile-Gly-Lys(Boc)-Val-Linker-MBHA Resin

Fmoc-Ser(tBu)-Leu-Ile-Gly-Lys(Boc)-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-Ser(tBu)-Leu-Ile-Gly-Lys(Boc)-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-Ser-Leu-Ile-Gly-Lys-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

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