| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| 500mg |
|
||
| Other Sizes |
|
Purity: ≥98%
| Targets |
JNK (IC50 = 280 nM)
|
||
|---|---|---|---|
| ln Vitro |
BI-78D3, dose-dependently inhibits the phosphorylation of JNK substrates both in vitro and in cell. For JNK1 binding, BI-78D3 can outcompete the D-domain of JIP1 (amino acids 153–163; pepJIP1) (IC50=500 nM). BI-78D3 is found to be 100 times less active vs. p38α, a member of the MAPK family with a high structural similarity to JNK, and completely inactive against mTOR and PI3-kinase (-isoform), both unrelated protein kinases, using the same in vitro LanthaScreen kinase assay and the same ATF2 substrate. Furthermore, Lineweaver-Burk analysis clearly shows that BI-78D3, with an apparent Ki value of 200 nM, is competitive with ATF2 for binding to JNK1. The cell-based LanthaScreen kinase assay is utilized to profile the characteristics of BI-78D3 in the context of a complex cellular milieu. In this test, BI-78D3 can prevent the cellular phosphorylation of c-Jun that is induced by TNF-α (EC50=12.4 μM)[1].
|
||
| ln Vivo |
Studies using JNK1-/- and JNK2-/- mice have demonstrated a relationship between ConA-induced liver failure, TNF receptor signaling, and JNK function. In order to conduct this study, 25 mg/kg of BI-78D3 is administered to insulin-insensitive mice just once, 30 minutes prior to the administration of insulin. The impact of insulin is then evaluated on blood sugar levels. In comparison to the vehicle control, BI-78D3 causes a statistically significant drop in blood glucose levels. Therefore, BI-78D3's capacity to reverse ConA-induced liver damage and improve insulin sensitivity is consistent with its proposed role as a powerful JNK inhibitor. Liquid chromatography/mass spectrometry bio-availability analysis reveals that BI-78D3 has good microsome and plasma stability (T1/2=54 min)[1].
|
||
| Enzyme Assay |
The LanthaScreen c-Jun (1-79) HeLa and LanthaScreen ATF2 (19-106) A549 cell lines, which stably express GFP-c-Jun 1-79 and GFP-ATF2 19-106, respectively, are used for the cell-based kinase assays for c-Jun and ATF2 phosphorylation. By measuring the time resolved FRET (TR-FRET) between a terbium labeled phospho-specific antibody and the GFP-fusion protein, phosphorylation is identified. 10,000 cells per well in 32 μl of assay medium (supplemented with 1% charcoal/dextran-treated FBS, 100 U/mL Penicillin, 100 μg/mL Streptomycin, 0.1 mM nonessential amino acids, 1 mM sodium pyruvate, 25 mM Hepes, pH 7.3, and devoid of phenol red) are plated on white tissue culture-treated 384-well plates. After overnight incubation, cells are pretreated for 60 minutes with BI-78D3 (0.001, 0.01, 0.1, 1, 10, and 100 μM), and then stimulated for 30 minutes with TNF-α, which activates both JNK and p38. After aspirating the medium out of the cells, 20 μL of lysis buffer (20 mM Tris•HCl, pH 7.6, 5 mM EDTA, 1% Nonidet P-40 substitute, 5 mM NaF, 150 mM NaCl, and 1:100 protease and phosphatase inhibitor mix, SIGMA P8340 and P2850, respectively) is added. Anti-pc-Jun (pSer73) or anti-pATF2 (pThr71) detection antibodies that have been terbium-labeled are present in the lysis buffer at a concentration of 2 nM. TR-FRET emission ratios are calculated on a BMG Pherastar fluorescence plate reader (excitation at 340 nm, emission at 520 and 490 nm; 100 s lag time, 200 s integration time, emission ratio=Em520/Em 490)[1] after allowing the assay to equilibrate for 1 h at room temperature.
|
||
| Cell Assay |
Mice
Male BL/6 mice aged 6 to 8 weeks are given a 10 mg/kg i.v. injection of ConA and BI-78D3. Mice are given isofluorane anesthesia before undergoing midventral laparotomy and having the left lateral and median lobes of the liver removed. Animals are put to death, their livers are surgically removed, and blood is obtained by puncturing their hearts. The concentration of alanine-aminotranferase in serum is determined[1]. Male, eleven weeks old OlaHsd db/db BKS.Cg-+Leprdb/+Leprdb Based on blood glucose levels adjusted three days prior to drug administration, mice are randomly assigned. A portable glucose meter is used to measure blood sugar levels. Mice are fasted for 6 hours prior to receiving 25 mg/kg of BI-78D3 intravenously (i.p.). Bovine Insulin (I-0516) is injected intravenously for 30 minutes following the administration of the test substance at a dose of 0.75 mg/kg. At predetermined intervals, blood samples are drawn, and blood glucose levels are measured as specified. Three hours after the test article is administered, food is returned[1]. |
||
| Animal Protocol |
|
||
| References | |||
| Additional Infomation |
4-(2,3-dihydro-1,4-benzodioxin-6-yl)-3-[(5-nitro-2-thiazolyl)thio]-1H-1,2,4-triazol-5-one is an aryl sulfide.
|
| Molecular Formula |
C13H9N5O5S2
|
|
|---|---|---|
| Molecular Weight |
379.37
|
|
| Exact Mass |
379.005
|
|
| Elemental Analysis |
C, 41.16; H, 2.39; N, 18.46; O, 21.09; S, 16.90
|
|
| CAS # |
883065-90-5
|
|
| Related CAS # |
|
|
| PubChem CID |
2747117
|
|
| Appearance |
White to off-white solid powder
|
|
| Density |
1.922g/cm3
|
|
| Boiling Point |
705.721ºC at 760 mmHg
|
|
| Flash Point |
380.606ºC
|
|
| Index of Refraction |
1.879
|
|
| LogP |
2.37
|
|
| Hydrogen Bond Donor Count |
1
|
|
| Hydrogen Bond Acceptor Count |
9
|
|
| Rotatable Bond Count |
3
|
|
| Heavy Atom Count |
25
|
|
| Complexity |
588
|
|
| Defined Atom Stereocenter Count |
0
|
|
| SMILES |
O=C1N(C2C=C3C(OCCO3)=CC=2)C(SC2SC([N+](=O)[O-])=CN=2)=NN1
|
|
| InChi Key |
QFRLDZGQEZCCJZ-UHFFFAOYSA-N
|
|
| InChi Code |
InChI=1S/C13H9N5O5S2/c19-11-15-16-12(25-13-14-6-10(24-13)18(20)21)17(11)7-1-2-8-9(5-7)23-4-3-22-8/h1-2,5-6H,3-4H2,(H,15,19)
|
|
| Chemical Name |
4-(2,3-dihydro-1,4-benzodioxin-6-yl)-3-[(5-nitro-1,3-thiazol-2-yl)sulfanyl]-1H-1,2,4-triazol-5-one
|
|
| Synonyms |
|
|
| HS Tariff Code |
2934.99.9001
|
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
|
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
|
|||
|---|---|---|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.59 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.59 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.6359 mL | 13.1797 mL | 26.3595 mL | |
| 5 mM | 0.5272 mL | 2.6359 mL | 5.2719 mL | |
| 10 mM | 0.2636 mL | 1.3180 mL | 2.6359 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
 In vitrocharacterization of pepJIP1 and BI-78D3.
Binding affinity data.Proc Natl Acad Sci U S A.2008 Oct 28;105(43):16809-13. |
|---|
 Bioevaluation studies with BI-78D3.Proc Natl Acad Sci U S A.2008 Oct 28;105(43):16809-13. |
 Docking studies and NMR analysis.Proc Natl Acad Sci U S A.2008 Oct 28;105(43):16809-1 |
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA
