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| 25mg |
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| 50mg |
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| 100mg |
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Purity: ≥98%
Chk2 Inhibitor II (also known as BML-277) is an ATP-competitive inhibitor of Chk2 (checkpoint kinase 2) with an IC50 of 15 nM. The selectivity towards Chk2 is 1000 times higher than that of Chk1 and Cdk1/B kinases. Human CD4+ and CD8+ T-cells are protected from ionizing radiation-induced apoptosis by BML-277 in a dose-dependent manner. With an observed EC50 of 3−7.6 μM, it can effectively rescue both T-cell populations from radiation-induced apoptosis in a dose-dependent manner. The biochemical measurement of chk2 inhibition is consistent with the concentration of BML-277 needed for radioprotection.
| Targets |
Chk2 (IC50 = 15 nM)
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| ln Vitro |
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| ln Vivo |
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| Enzyme Assay |
In order to ascertain the activity of chk2 inhibitors, the following conditions must be met: 5 nM recombinant human Chk2, 50 mM HEPES (pH 7.4), 100 mM NaCl, 10 mM MgCl2, 25 μM synthetic peptide substrate (biotin-SGLYRSPSMPENLNRPR), 1 μM ATP, 50 μCi/mL [γ-33P] ATP, and a combination of protease inhibitors. The peptide substrate is bound to streptavidin-conjugated agarose beads by the reaction mixtures after three hours of incubation at 37°C. A 0.1% Tween-20 solution in phosphate-buffered saline (pH 7.4) is used to repeatedly wash the agarose beads. The amount of radioactive phosphate bound to the substrate peptide at various concentrations of BML-277 (6.25, 12.5, 25, 50, 100, and 200 nM) is measured by scintillation counting, which is used to determine the enzyme activity. Kinetic experiments involve varying the concentration of ATP while maintaining a constant ratio between unlabeled and [γ-33P] labeled ATP. Samples are kept on ice for additional processing and reactions are halted at various times by adding 50 mM cold ATP[1].
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| Cell Assay |
Purified T-cells are incubated at 100,000 cells per well in BML-277 (102.5 nM, 1 μM, 100.5 μM, 10 μM, and 101.5 μM) or vehicle (DMSO) at different concentrations in 96-well stripwells for one hour in order to assess the radioprotective effect of Chk2 inhibitors. The cells are then placed back into the incubator for an additional 24 hours after being exposed to a dose of 0 or 10 Gy gamma irradiation from a 137Cs source at a dose rate of 3.65 Gy/min. Propidium iodide and Annexin V-FITC are used to stain cells in accordance with the manufacturer's instructions. Using a FACSCalibur FACS instrument, apoptotic and surviving cells are quantified. The number of survivors from treatment groups less the number of cells surviving in the irradiated control group divided by the number of surviving cells in the untreated control groups is the percentage of recovery used to report data[1].
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| Animal Protocol |
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| References |
| Molecular Formula |
C20H14CLN3O2
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| Molecular Weight |
363.80
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| Exact Mass |
363.077
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| Elemental Analysis |
C, 66.03; H, 3.88; Cl, 9.74; N, 11.55; O, 8.80
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| CAS # |
516480-79-8
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| Related CAS # |
516480-79-8; 516480-80-1;
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| PubChem CID |
9969021
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| Appearance |
White to off-white solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
637.1±63.0 °C at 760 mmHg
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| Flash Point |
339.1±33.7 °C
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| Vapour Pressure |
0.0±1.9 mmHg at 25°C
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| Index of Refraction |
1.703
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| LogP |
4.61
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
26
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| Complexity |
489
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| Defined Atom Stereocenter Count |
0
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| SMILES |
ClC(C=C1)=CC=C1OC(C=C2)=CC=C2C3=NC4=CC(C(N)=O)=CC=C4N3
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| InChi Key |
UXGJAOIJSROTTN-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C20H14ClN3O2/c21-14-4-8-16(9-5-14)26-15-6-1-12(2-7-15)20-23-17-10-3-13(19(22)25)11-18(17)24-20/h1-11H,(H2,22,25)(H,23,24)
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| Chemical Name |
2-[4-(4-chlorophenoxy)phenyl]-3H-benzimidazole-5-carboxamide
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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: ≥ 1.67 mg/mL (4.59 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 16.7 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: ≥ 1.67 mg/mL (4.59 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 16.7 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. View More
Solubility in Formulation 3: ≥ 1.67 mg/mL (4.59 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.7488 mL | 13.7438 mL | 27.4876 mL | |
| 5 mM | 0.5498 mL | 2.7488 mL | 5.4975 mL | |
| 10 mM | 0.2749 mL | 1.3744 mL | 2.7488 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.
ERK1/2 and CHK2 protein levels are elevated in human DLBCLs.Nat Commun.2011 Jul 19;2:402. |
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 Inhibition of ERK and CHK2 increases cell apoptosis.Nat Commun.2011 Jul 19;2:402. |
 ERK1/2 and CHK2 physically interact with each other.Nat Commun.2011 Jul 19;2:402. |
 CHK2 Thr68 is required for the physical interaction between ERK1/2 and CHK2.Nat Commun.2011 Jul 19;2:402. |
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 Inhibition of CHK2 activity increases ERK1/2 phosphorylation.Nat Commun.2011 Jul 19;2:402. |
 Cotreatment with ERK inhibitor and CHK2 inhibitor II results in decreased tumour growth in a xenograft model.Nat Commun.2011 Jul 19;2:402. |
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