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JNK-IN-7

现货
Catalog No.
A3519
JNK抑制剂
组合的产品项目
规格价格库存 数量
10mM (in 1mL DMSO)
¥ 2,890.00
现货
5mg
¥ 2,790.00
现货
10mg
¥ 3,600.00
现货
50mg
¥ 8,800.00
现货
100mg
¥ 14,030.00
现货

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Background

JNK-IN-7 is a selective JNK inhibitor with IC50 values of 1.54 nM, 1.99 nM, 0.75 nM to JNK1, JNK2, JNK3, respectively. It also inhibits phosphorylation of c-Jun, which is a direct substrate of JNK kinase.

JNK-IN-8, an analog of JNK-IN-7 with an extra flag methyl, dramatically improved in selectivity and eliminated binding to IRAK1, PIK3C3, PIP4K2C and PIP5K3, comparing to JNK-IN-7. JNK-IN-7 and JNK-IN-8 require Cys116 for JNK2 inhibition. JNK-IN-7 can indeed inhibit IRAK-1 dependent E3 ligase activity of pellino, which plays an role in the Toll receptor signaling pathway in cells at relative high compound concentrations (1–10 mM).

The IRAK1 inhibitor JNK-IN-7 inhibited the IL-1-stimulated activation of Pellino 1 in IL-1R cells, but not the Pam3CSK4-stimulated activation of Pellino 1 in RAW264.7 macrophages. JNK-IN-7 also suppressed the phosphorylation of c-Jun in Pam3CSK4-stimulated RAW macrophages, but in contrast with IL-1R cells it did not affect the activation of Pellino 1.

References:
[1].  Zhang T, Inesta-Vaquera F, Niepel M et al. Discovery of potent and selective covalent inhibitors of JNK. Chem Biol. 2012 Jan 27;19(1):140-54.
[2].  Goh ET, Arthur JS, Cheung PC et al. Identification of the protein kinases that activate the E3 ubiquitin ligase Pellino 1 in the innate immune system. Biochem J. 2012 Jan 1;441(1):339-46.

Chemical Properties

Physical AppearanceA solid
StorageStore at -20°C
M.Wt493.56
Cas No.1408064-71-0
FormulaC28H27N7O2
SynonymsJNK inhibitor
Solubility≥24.7 mg/mL in DMSO, <2.65 mg/mL in EtOH, <2.54 mg/mL in H2O
Chemical Name3-[[(E)-4-(dimethylamino)but-2-enoyl]amino]-N-[4-[(4-pyridin-3-ylpyrimidin-2-yl)amino]phenyl]benzamid
SDFDownload SDF
Canonical SMILESCN(C)CC=CC(=O)NC1=CC=CC(=C1)C(=O)NC2=CC=C(C=C2)NC3=NC=CC(=N3)C4=CN=CC=C4
运输条件试用装:蓝冰运输。 其他可选规格:常温运输或根据您的要求用蓝冰运输。
一般建议为了使其更好的溶解,请用37℃加热试管并在超声波水浴中震动片刻。不同厂家不同批次产品溶解度各有差异,仅做参考。若实验所需浓度过大至产品溶解极限,请添加助溶剂助溶或自行调整浓度。

试验操作

激酶实验 [1]:

基于细胞的c-Jun磷酸化测定

使用分别稳定表达GFP-c-Jun 1 ~ 79和GFP-ATF2 19 ~ 106的LanthaScreen c-Jun (1 ~ 79) HeLa细胞系进行基于细胞的c-Jun磷酸化激酶实验。通过测定铽标记的磷酸化c-Jun特异性抗体与GFP之间的TR-FRET以检测磷酸化程度。将细胞加入含白色组织培养液的384孔板中,每孔含10,000个细胞和32 μL测定液(Opti-MEM,含0.5%经过活性炭/葡聚糖处理的FBS,100 U/mL Penicillin,100 μg/mL Streptomycin,0.1 mM非必需氨基酸,1 mM丙酮酸钠,25 mM HEPES [pH 7.3]和少量酚红)。孵育过夜后,使用4 μL测定缓冲液将JNK-IN-7稀释到指定浓度,再将其加入到细胞中预处理90分钟,加入4 μL含5 ng/mL of TNF-α的测定缓冲液(最终体积为40 μL)孵育30分钟以激活细胞。吸除培养液,加入20 μL裂解缓冲液(20 mM Tris-HCl [pH 7.6],5 mM EDTA,1% Nonidet P-40 替代物,5 mM NaF,150 mM NaCl和1:100蛋白酶和磷酸酶抑制剂混合物,分别为P8340和P2850)裂解细胞。裂解液包括2 nM铽标记的抗c-Jun (pSer73) 测定抗体。在室温下,实验平衡60分钟,使用BMG Pherastar荧光板读取器测定TR-FRET发射率。测定参数为:在340 nm处激发,在520和490 nm处发射;100毫秒滞后时间;200微秒积分时间;发射率 = Em 520/Em 490。使用GraphPad Prism 4分析所有数据并绘图。

细胞实验 [2]:

细胞系

人IL-1R细胞和RAW264.7巨噬细胞

制备方法

溶于DMSO。若配制更高浓度的溶液,一般步骤如下:请将试管置于37℃加热10分钟和/或将其置于超声波浴中震荡一段时间。原液于-20℃可放置数月。

反应条件

0.1、1和10 mM;1小时

实验结果

在人IL-1R细胞中,JNK-IN-7抑制IL-1β诱导的c-Jun磷酸化和Pellino1活化。在Pam3CSK4诱导的RAW巨噬细胞中,JNK-IN-7也抑制c-Jun磷酸化。

References:

[1]. Zhang T, Inesta-Vaquera F, Niepel M, et al. Discovery of potent and selective covalent inhibitors of JNK. Chem Biol, 2012, 19(1): 140-154.

[2]. Goh ET, Arthur JS, Cheung PC, et al. Identification of the protein kinases that activate the E3 ubiquitin ligase Pellino 1 in the innate immune system. Biochem J, 2012, 441(1): 339-346.

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