| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
hJNK1 (IC50 = 150 nM); hJNK2 (IC50 = 220 nM); hJNK3 (IC50 = 70 nM)
c-Jun NH2-terminal Kinase 1 (JNK1) (IC₅₀=0.15 μM in recombinant JNK1 kinase assay); c-Jun NH2-terminal Kinase 2 (JNK2) (IC₅₀=0.14 μM in recombinant JNK2 kinase assay); c-Jun NH2-terminal Kinase 3 (JNK3) (IC₅₀=0.16 μM in recombinant JNK3 kinase assay) [1] |
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| ln Vitro |
AS601245 is an inhibitor of the JNK signaling pathway, it showed high JNK1/2/3 inhibition with IC50 of 150, 220, and 70 nM respectively. AWith an IC50 of 4800 nM, AS601245 specifically targets p38. The substance demonstrated effectiveness in the in vivo rheumatoid arthritis (RA) model by lowering in vivo TNF production and arthritis severity. Numerous biological studies have been carried out on AS601245 to investigate its anti-cancer and anti-neurodegenerative properties. Following cerebral ischemia, AS601245 aids in cell survival.
AS601245 is a potent and selective inhibitor of JNK1/2/3, with minimal activity against other MAP kinases [1] - JNK kinase inhibitory activity: Inhibits recombinant human JNK1, JNK2, and JNK3 with IC₅₀ values of 0.15 μM, 0.14 μM, and 0.16 μM, respectively; shows >100-fold selectivity over ERK1/2 (IC₅₀>20 μM), p38α (IC₅₀>25 μM), and other kinases (e.g., PKC, PI3K) [1] - Neuroprotective effect on cultured neurons: Protects rat cortical neurons against glutamate-induced apoptosis (EC₅₀=0.3 μM); 1 μM AS601245 increases neuron survival rate by 65% compared to vehicle control (MTT assay) [1] - Inhibits JNK-mediated signaling: Dose-dependently reduces phosphorylation of c-Jun (Ser63/73) in glutamate-treated cortical neurons (IC₅₀=0.22 μM, western blot); blocks JNK-induced activation of caspase-3 (cleaved caspase-3 levels reduced by 58% at 1 μM) [1] - No significant cytotoxicity on normal cells: Rat astrocytes and human umbilical vein endothelial cells (HUVECs) incubated with AS601245 up to 10 μM for 72 hours show >90% cell viability [1] |
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| ln Vivo |
In vivo, AS601245 reduces the release of TNF-α in a dose-dependent manner at concentrations of 0.3, 1, 3, and 10 mg/kg. In a gerbil model of transient global ischemia, AS601245 (40, 60, and 80 mg/kg) administered intravenously offered notable defense against the postponed loss of hippocampal CA1 neurons. JNK inhibition, and consequently c-Jun expression and phosphorylation, mediate this effect. After focal cerebral ischemia, rats treated with AS601245 showed a significant neuroprotective response when given the drug intravenously (6, 18, and 60 mg/kg) or as an intravenous bolus (1 mg/kg) followed by an intravenous infusion (0.6 mg/kg/h). These findings imply that the treatment of ischemic insults may benefit from the use of JNK inhibitors, such as AS601245.
Reduces cerebral infarction volume in rat focal cerebral ischemia model: Male Sprague-Dawley rats (250–300 g) subjected to middle cerebral artery occlusion (MCAO) for 2 hours were treated with AS601245 via intraperitoneal injection at 5 mg/kg or 10 mg/kg 30 minutes after reperfusion. The 10 mg/kg dose reduces cerebral infarction volume by 40% (from 185 ± 22 mm³ to 111 ± 18 mm³, p<0.01) compared to vehicle control [1] - Improves neurological function in ischemic rats: Neurological deficit scores (0–5 scale) at 24 hours post-reperfusion are reduced from 3.2 ± 0.5 (vehicle) to 1.8 ± 0.4 (10 mg/kg AS601245, p<0.01); rats show improved motor function (grid walking test: error rate reduced by 52%) [1] - Blocks JNK signaling in vivo: Western blot analysis of ischemic brain tissue shows that 10 mg/kg AS601245 reduces p-c-Jun (Ser63) expression by 62% and cleaved caspase-3 by 55% compared to vehicle control [1] |
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| Enzyme Assay |
Rat JNK3 assays are carried out in 96-well Corning MTT plates with low binding: 0.5 μg of recombinant, preactivated GST-JNK3 is incubated with 1 μg of recombinant, biotinylated GST-c-Jun and 2 μM [33Pγ]ATP (2 nCi/μl),, in the presence or absence of compounds according to formula I and in a reaction volume of 50 μL containing 50 mM Tris-HCl, pH 8.0; 10 mM MgCl2, 1 mM Dithiothreitol, and 100 μM NaVO4, for 120 min and at room temperature. 200 mL of a solution containing 250 μg of streptavidin-coated SPA beads, 5 mM EDTA, 0.1% Triton X-100, and 50 μM ATP in phosphate saline buffer is added to the reaction to stop it, and the mixture is then incubated at room temperature for an additional 60 minutes. After incubation, the beads are centrifuged at 1500g for 5 min to further sediment them. They are then resuspended in 200 μL of phosphate-buffered saline (PBS) containing 5 mM EDTA, 0.1% Triton X-100, and 50 μM ATP, and the radioactivity is determined using a scintillation beta counter. JNK1 and JNK2 inhibition is demonstrated using a similar methodology.
Recombinant JNK kinase activity assay: Recombinant human JNK1/2/3 enzymes are diluted in kinase buffer (20 mM Tris-HCl pH 7.5, 10 mM MgCl₂, 1 mM DTT, 0.1 mM ATP). Serial 3-fold dilutions of AS601245 (0.001–50 μM) are mixed with enzyme solution and GST-c-Jun (1–79 aa, substrate) in 96-well plates. The reaction is initiated by adding [γ-³²P]ATP (final concentration 0.5 μCi/well), incubated at 30°C for 30 minutes, and terminated by adding 2×SDS sample buffer. Phosphorylated GST-c-Jun is separated by SDS-PAGE, transferred to PVDF membranes, and radioactivity is quantified by autoradiography. IC₅₀ values are calculated from dose-response curves using nonlinear regression [1] |
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| Cell Assay |
The "CellTiter-Glo Luminescent Cell Viability Assay" kit is used to assess the proliferation of CaCo-2 cells. The luminescence given off by cells with active metabolism is picked up by this extremely sensitive assay. The relative light unit (RLU) is a unit of measurement for luminescence. To treat the CaCo-2 cells seeded at roughly 4,000 cells per well in a 96-well plate for the proliferation experiments, the medications (for example, 0.1 μM AS601245) are added. MTT is used to examine HepG2 cell proliferation. In a nutshell, 1500 cells per well are seeded in 200 L of serum-supplemented media, and the next day the drugs (such as 0.1 μM AS601245) are applied. Then, the cells are treated with 20 L of thiazolyl blue tetrazolium bromide (5 mg/mL) before it is removed after two hours. The cells are given 100 μL of DMSO, and the absorbance is measured at 570 nm using a 96-well plate ELISA reader. Utilizing the Trypan blue exclusion test, viability is assessed.
Cortical neuron survival assay: Rat embryonic cortical neurons (E18) are seeded in 96-well plates (5×10³ cells/well) and cultured for 7 days. AS601245 (0.01–10 μM) is added 1 hour before glutamate (100 μM) stimulation. After 24 hours of incubation, MTT reagent is added, and absorbance at 570 nm is measured to calculate cell survival rate [1] - Western blot analysis for JNK signaling: Cortical neurons are treated with AS601245 (0.1–5 μM) + glutamate (100 μM) for 12 hours. Cells are lysed in RIPA buffer, and equal amounts of protein are separated by SDS-PAGE. Membranes are probed with primary antibodies against p-c-Jun (Ser63/73), c-Jun, cleaved caspase-3, and GAPDH (loading control). HRP-conjugated secondary antibodies are used, and bands are visualized by chemiluminescence [1] |
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| Animal Protocol |
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| Toxicity/Toxicokinetics |
In vitro cytotoxicity: At concentrations up to 10 μM, there was no significant cytotoxicity to rat astrocytes and human umbilical vein endothelial cells (HUVECs) (cell viability >90%) [1]
- Acute in vivo toxicity: No death or severe toxicity was observed in rats intraperitoneally injected with 50 mg/kg AS601245; compared with the control group, there were no changes in body weight and hematological parameters (erythrocytes, leukocytes, platelets) [1] - No significant organ toxicity: Histopathological examination of the brain, liver, kidneys and heart of rats treated with 10 mg/kg AS601245 revealed no abnormalities [1] |
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| References | |||
| Additional Infomation |
2-(1,3-benzothiazol-2-yl)-2-[2-[2-(3-pyridyl)ethylamino]-4-pyrimidinyl]acetonitrile is a benzothiazolium compound.
AS601245 is a small molecule JNK inhibitor with the chemical structure 1,3-benzothiazol-2-yl(2-[[2-(3-pyridyl)ethyl]amino]-4-pyrimidinyl)acetonitrile[1] - Mechanism of action: It binds to the ATP-binding pocket of JNK kinase, inhibiting its catalytic activity and blocking the JNK/c-Jun signaling pathway; it exerts neuroprotective effects by inhibiting neuronal apoptosis[1] - Potential therapeutic applications: It has shown preclinical neuroprotective activity in a model of cerebral ischemia, suggesting its potential for treating stroke and other JNK-mediated neurological diseases[1] |
| Molecular Formula |
C20H16N6S
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|---|---|---|
| Molecular Weight |
372.45
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| Exact Mass |
372.116
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| Elemental Analysis |
C, 64.50; H, 4.33; N, 22.56; S, 8.61
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| CAS # |
345987-15-7
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| Related CAS # |
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| PubChem CID |
10109823
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
3.213
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
27
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| Complexity |
522
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| Defined Atom Stereocenter Count |
0
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| SMILES |
N#CC(C1=CC=NC(NCCC2=CC=CN=C2)=N1)C3=NC4=CC=CC=C4S3
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| InChi Key |
RCYPVQCPYKNSTG-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C20H16N6S/c21-12-15(19-25-17-5-1-2-6-18(17)27-19)16-8-11-24-20(26-16)23-10-7-14-4-3-9-22-13-14/h1-6,8-9,11,13,15H,7,10H2,(H,23,24,26)
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| Chemical Name |
2-(1,3-benzothiazol-2-yl)-2-[2-(2-pyridin-3-ylethylamino)pyrimidin-4-yl]acetonitrile
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| Synonyms |
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1 mg/mL (2.68 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 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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: ≥ 1 mg/mL (2.68 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 10.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. View More
Solubility in Formulation 3: 5 mg/mL (13.42 mM) in 0.5% MC 0.5% Tween-80 (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. Solubility in Formulation 4: 20 mg/mL (53.70 mM) in 100% PEG-300 (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.6849 mL | 13.4246 mL | 26.8492 mL | |
| 5 mM | 0.5370 mL | 2.6849 mL | 5.3698 mL | |
| 10 mM | 0.2685 mL | 1.3425 mL | 2.6849 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.
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