中文名称: JIB-04
英文名称: JIB-04
CAS No: 199596-05-9
分子式: C17H13ClN4
分子量: 308.76
EINEC: 693627
J10024 JIB-04 ≥98%(HPLC) (psaitong)
包装规格:
5mg 25mg 100mg in glass bottle
溶解性:
溶于DMSO(20mg/mL 超声 加热)
产品描述:

基本信息

产品编号:J10024

产品名称:JIB-04

CAS:

199596-05-9

 

储存条件

粉末

-20℃

四年

 

 

分子式:

C17H13ClN4

溶于液体

-80℃

六个月

分子量:

308.76

-20℃

一个月

化学名: 

 

 

Solubility (25°C)

 

体外

DMSO

62mg/mL warmed with 50ºC water bath (200.8mM)

Ethanol

Insoluble

Water

Insoluble

体内(现配现用)

4% DMSO+corn oil

3mg/mL

1mg/ml表示微溶或不溶。

普西唐提供的所有化合物浓度为内部测试所得,实际溶液度可能与公布值有所偏差,属于正常的批间细微差异现象。

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液;⼀旦配成溶液,请分装保存,避免反复冻融造成的产品失效。

 

制备储备液

 

浓度

 

溶液体积

质量

 

1mg

 

5mg

 

10mg

1mM

3.2388mL

16.1938mL

32.3876mL

5mM

0.6478mL

3.2388mL

6.4775mL

10mM

0.3239mL

1.6194mL

3.2388mL

50mM

0.0648mL

0.3239mL

0.6478mL

 

生物活性

产品描述

一种Jumonji组蛋白脱甲基酶抑制剂,能抑制JARID1A、JMJD2E、JMJD3、JMJD2A、JMJD2B、JMJD2C和JMJD2D的活性,IC50值分别为 230、340、855、445、435、1100和290nM。

靶点/IC50

JARID1A
(Cell-free assay)

JMJD2D
(Cell-free assay)

JMJD2E
(Cell-free assay)

JMJD2B
(Cell-free assay)

JMJD2A
(Cell-free assay)

230nM

290nM

340nM

435nM

445nM

 

体外研究

JIB-04 is consistently selective for cancer vs. normal cells, demonstrated by the higher sensitivity of lung and prostate cancer lines (with IC50 as low as 10nM) compared to HBECs and PrSCs/PrECs. JIB-04 inhibits cellular Jumonji demethylase activity,and Jumonji levels affect JIB-04 action in cells. JIB-04 significantly inhibits the proliferation of GB cell lines and stemenriched cultures. JIB-04 exerts its maximal inhibitory activity against KDM5A, and modulates the expression of genes involved in the control of cancer cell growth and leads to hypermethylation of H3K4. Furthermore, JIB-04 (2500nM) activates the autophagy and apoptotic pathways and inactivates PI3K. JIB-04 also cooperates with TMZ in killing GB cells.

体内研究

JIB-04 results in a significant reduction in cancer-induced death rates in mice,prolonging survival.JIB-04 (60, 40 and 20mg/kg, i.p.) reaches bioactive concentration in the brain of the mice.The orthotopic GB xenograft model shows a trend toward longer survival in JIB-04-treated mice with an Hazard Ratio of 0.5.

 

推荐实验方法(仅供参考)

激酶实验:

Jumonji脱甲基酶/脯氨酰羟化酶/LSD1活性检测

活性JMJD2E氨基酸1-350从E.coli中纯化,体外环境中与α-酮戊二酸,5-10μM铁离子,抗坏血酸,组蛋白肽段基质一起使用,偶联反应时加入甲醛脱氢酶和NAD+来定量NADH的生成量,或使用Epigentek kit P-3081。组蛋白脱甲基反应结果通过Western分析来定量,构建His-tagged hJMJ2D氨基酸1-350表达体系,E.coli表达产物通过Qiagen Ni-NTA琼脂糖手册进行纯化。蛋白用50mM TrisHCl pH 7.8+0.3M NaCl + 10%甘油+200mM immidazol洗脱,后用20mM TrisHCl pH 7.4,0.15 M NaCl,0.2mM PMSF,0.5mM DTT,8%甘油透析。蛋白分装,速冻、储存在-80°C。Western blot中,~1.5μg酶与0.3 μg H3K9me3基质(Active Motif #31213),10μM (NH4)2Fe(SO4)2,1mM α-酮戊二酸,2mM L-抗坏血酸钠在50mM pH 7.9溶液中混合,与溶媒或药物在37°C孵育30min-2hrs。加入SDS loading buffer后,煮沸,样品在NuPage 4-12% Bis-Tris胶上电泳,转膜,用Upstate #07-523处理硝化纤维素膜以检测H3K9me3。H3总信号是通过Active Motif #39763和连接IRDye 680的驴抗兔IgG(二抗, LI-COR # 926-32223),在Odyssey Infrared成像系统中获取图像。体外IC50检测和竞争性试验,100-200ng纯化蛋白与溶媒、JIB-04或类似物共孵育,通过ELISA(H3K9me3去甲基作用:Epigentek kit P-3081,H3K4me3去甲基作用:P-3083,H3K27me3去甲基作用:P-3085)检测活性,反应液包含50mM Hepes pH 7.5,0.01% Tween 20,120nM (NH4)2Fe(SO4)2,1mMα-酮戊二酸,2mM L-抗坏血酸钠,50ng肽段基质。最终酶浓度分别为:206nM JMJD2A,12nM JMJD2B,60nM JMJD2C,90nM JMJD2D E.coli,30nM JMJD2D Sf9,30nM JMJD2E,30nM Jarid1a,35nM JMJD3。E.coli表达的hJMJD2A(氨基酸1-350)纯化后以每孔400ng的量进行检测。通过GraphPad Prism软件计算IC50和进行曲线拟合。需要注意的是在基质竞争性试验中, 应该使分析处于线性范围并且不超出ELISA板的结合能力。细胞裂解物的H3K9me3脱甲基酶活性的直接测定:超声处理细胞(细胞接种浓度为2x10E+06/10cm板)或肿瘤PBS匀浆(3x4 sec),等量的蛋白与组蛋白H3K9me3基质在反应液中37°C孵育2h后,用Epigentek kit P-3081进行免疫检测。500ng E.coli表达的PHD2纯化蛋白,溶于40mM Tris pH 7.4,100mM NaCl,20% glycerol,5mM β-mercapto-ethanol,10mM 麦芽糖备用。从HIF-1 ODD分离出的生物素化的肽段固定在亲和素包裹的96孔板中。酶与被试药物在反应液(20mM Tris-Cl pH 7.5,5mM KCl,1.5mM MgCl2,2mM DTT,0.12μM硫酸亚铁,0.5mM戊邻酮二酸盐,1mM抗坏血酸)中室温孵育45min。α-酮戊二酸的竞争性类似物DMOG作为抑制作用的阳性对照。肽段羟基化是通过对羟基化HIF肽表位使用多克隆兔抗体,再加入连接羊抗兔HPR的二抗。EnVision上读取发光值。LSD1重组蛋白活性通过使用Epigentek kit P-3075检测。

 

细胞实验:

 

For cell viability assays, cells are plated at 1500-3000 cells/well in 96 well plates and treated the next day with increasing doses of compound over 4 days and their viability assessed by standard MTS assays using Promega’s Cell Titer or Cell TiterGlo reagents. Absorbance at 490nm and 650nm or luminescence is measured by a Spectra Max or a FlouroStar Omega plate reader. Data are normalized to the untreated controls (100% viability). Each cell line is tested in 2-5 independent assays,each containing 4-8 replicates. IC50 values are calculated using DIVISA, a high-throughput software, developed in house, for storing and analyzing drug sensitivity assays.Dose-response curves are plotted using a non-linear regression model and IC50s are determined from the fitted curves. The average IC50 derived from 2-5 independent assays, each containing 4-8 replicates is reported.

 

动物实验:

 

For xenografts, 4-6 week old female nude mice are housed under standard conditions in a clean facility at UTSW. Two million H358 cells or five million A549 cells are injected subcutaneously and allowed to grow for 2-3 weeks with monitoring.When tumors reach appr 200mm3,therapy is started in weight and tumor volume matched pairs (n=7 for each treatment group for each cell line). Drug or vehicle is administered by inter-peritoneal injection in 10% DMSO 90% sesame oil 2 to 3 times weekly for 5 weeks at 110mg/kg to all mice harboring H358 xenografts or 3 times per week by gavage in 12.5% Cremophor EL, 12.5% DMSO as an aqueous suspension at 55mg/kg to

mice harboring A549 xenografts. Tumor volumes are monitored twice weekly by caliper measurements. Animals are weighed and observed during the five weeks of treatment. At the end point, mice are euthanized by CO2 asphyxiation and cervical dislocation, and blood, tumors and major organs collected and weighed. Paired, unequal variance, one-tailed t-tests are performed across treatment groups using Excel software.

保存条件:
-20℃
UN码:
HazardClass:
危害声明:
安全说明:
搜索质检报告(COA)
参考文献 & 客户发表文献

本计算器可帮助您计算出特定溶液中溶质的质量、溶液浓度和体积之间的关系,公式为:
质量 (g) = 浓度 (mol/L) x 体积 (L) x 分子量 (g/mol)

摩尔浓度计算公式

  • =
    *
    *
    *选择对应的单位 *空出希望得到的变量,填写另外两个变量

用本工具协助配置特定浓度的溶液,使用的计算公式为:
开始浓度 x 开始体积 = 最终浓度 x 最终体积

稀释公式

稀释公式一般简略地表示为:C1V1 = C2V2 ( 输入 输出 )

  • * = *

连续稀释计算器方程

  • 连续稀释

  • 初始浓度:
  • 稀释倍数:
  • 计算结果

  • C1=C0/X C1: LOG(C1):
    C2=C1/X C2: LOG(C2):
    C3=C2/X C3: LOG(C3):
    C4=C3/X C4: LOG(C4):
    C5=C4/X C5: LOG(C5):
    C6=C5/X C6: LOG(C6):
    C7=C6/X C7: LOG(C7):
    C8=C7/X C8: LOG(C8):