| Size | Price | Stock | Qty |
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| 5mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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Purity: ≥98%
MLS2052 (also known as BIO, GSK3 Inhibitor IX, 6-Bromoindirubin-3'-oxime) is a novel, potent, cell-permeable bis-indolo and specific inhibitor of GSK-3 (glycogen synthase kinase-3) with potential antitumor activity. In a cell-free assay, it inhibits GSK-3 with an IC50 of 5 nM for GSK-3α/β and displays >16-fold selectivity over CDK5. In a mouse xenograft model, it inhibited the growth of the melanoma tumor and demonstrated strong antiproliferative activity.
| Targets |
GSK-3α (IC50 = 5 nM); GSK-3β (IC50 = 5 nM); CDK5/p35 (IC50 = 80 nM); Cdk1/cyclin B (IC50 = 320 nM); cdk2/cyclin A (IC50 = 300 nM); Cdk4/cyclin D1 (IC50 = 10 μM); MAPKK (IC50 = 10 μM); protein kinase Cα (IC50 = 12 μM)
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| ln Vitro |
BIO (6-bromoindirubin-3'-oxime) is a specific inhibitor of glycogen synthase kinase-3 (GSK-3), with IC50 of 5 nM for GSK-3α/β, shows >16-fold selectivity over CDK5. In cellular models, BIO reduces β-catenin phosphorylation on a GSK-3-specific site and closely resembles Wnt signaling in Xenopus embryos. It also interacts with these kinases' ATP binding pockets. [1] The transcription factors Oct-3/4, Rex-1, and Nanog, which are specific to the pluripotent state, are maintained in the undifferentiated phenotype in human and mouse embryonic stem cells by BIO. Withholding the compound causes normal multidifferentiation programs in both human and mouse embryonic stem cells, demonstrating that BIO-mediated Wnt activation is functionally reversible.[2] 6BIO is also a pan-JAK inhibitor, with IC50 values for TYK2, JAK1, JAK2, and JAK3 of 0.03, 1.5, 8.0, and 0.5 μM.
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| ln Vivo |
BIO suppresses melanoma tumor growth in a mouse xenograft model.[4]
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| Enzyme Assay |
In Buffer A or C, kinase activities are measured at 30 °C with a final ATP concentration of 15 M. Activities are calculated as pmoles of phosphate incorporated during a 10 minute incubation after subtracting blank values. Dimethylsulfoxide is diluted appropriately for controls to be performed. After SDS-PAGE, phosphorylation of the substrate is occasionally measured by autoradiography. Purification of GSK-3α/β is accomplished in porcine brain via affinity chromatography using immobilized axin. It is assayed, following a 1/100 dilution in 1 mg BSA/ml 10 mM DTT, with 5 μl 40 μM GS-1 peptide, a specific GSK-3 substrate, (YRRAAVPPSPSLSRHSSPHQSpEDEEE), in buffer A, in the presence of 15 μM [γ-32P] ATP (3,000 Ci/mmol; 1 mCi/ml) in a final volume of 30 μl. After being incubated for 30 minutes at 30 degrees Celsius, 25 μl aliquots of the supernatant are spotted onto pieces of Whatman P81 phosphocellulose paper that are 2.5 × 3 cm in size. The filters are then washed five times (for at least five minutes each time) in a solution of 10 ml phosphoric acid/liter of water. A 1 ml sample of ACS scintillation fluid is used to count the wet filters.
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| Cell Assay |
In Dulbecco's Modified Medium (DMEM) containing 10% fetal bovine serum, COS1, Hepa (wild-type, CEM/LM AhR deficient, and ELB1 ARNT deficient) or SH-SY5Y cells are grown in 6 cm culture dishes. When cell density reaches about ~70% confluence, IO (5 μM), BIO (5 or 10 μM), MeBIO (5 or 50 μM), LiCl (20 or 40 mM), or a mock solution (DMSO, 0.5% final concentration) are added to the medium for treatment. After 12 (SH-SY5Y) or 24 hours, the cells are lysed with lysis buffer (1% SDS, 1 mM sodium orthovanadate, 10 mM Tris [pH 7.4]), still in the plate. The lysate is passed through a 26G needle several times, centrifuged at 10,000 × g for five minutes, and then adjusted to the same protein concentration. Each sample is loaded with about μg for immunoblotting. The detection method is enhanced chemiluminescence. The following primary antibodies are used: rabbit anti-AhR (Aryl hydrocarbon receptor), rabbit anti-actin, mouse anti-phospho-catenin (Upstate Biotechnologies, Clone 8E7, recognizes dephosphorylated -catenin), mouse anti-GSK-3, mouse anti-GSK-3 phosphoTyr216, and rabbit anti-actin.
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| Animal Protocol |
10 mg/kg for mice, 5 mg/kg for rats, 2 mg/kg for dogs and monkeys
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| References |
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| Additional Infomation |
6-bromoindirubin-3'-oxime is a member of the class of biindoles that is indirubin substituted at position 6 by a bromo group and in which the keto group at position 3' has undergone condensation with hydroxylamine to form the corresponding oxime. It has a role as an EC 2.7.11.1 (non-specific serine/threonine protein kinase) inhibitor and an EC 2.7.11.26 (tau-protein kinase) inhibitor. It is a ketoxime, an organobromine compound, a member of oxindoles and a biindole.
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| Molecular Formula |
C16H10BRN3O2
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|---|---|
| Molecular Weight |
356.1735
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| Exact Mass |
354.995
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| Elemental Analysis |
C, 53.95; H, 2.83; Br, 22.43; N, 11.80; O, 8.98
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| CAS # |
667463-62-9
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| Related CAS # |
667463-62-9
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| PubChem CID |
448949
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| Appearance |
Brown to red solid powder
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| Density |
1.8±0.1 g/cm3
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| Boiling Point |
554.3±50.0 °C at 760 mmHg
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| Melting Point |
300°C(lit.)
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| Flash Point |
289.0±30.1 °C
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| Vapour Pressure |
0.0±1.6 mmHg at 25°C
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| Index of Refraction |
1.802
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| LogP |
2.41
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
22
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| Complexity |
438
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| Defined Atom Stereocenter Count |
0
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| SMILES |
BrC1C([H])=C([H])C2=C(C=1[H])N([H])C(=C2C1=C(C2=C([H])C([H])=C([H])C([H])=C2N1[H])N=O)O[H]
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| InChi Key |
SAQUSDSPQYQNBG-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C16H10BrN3O2/c17-8-5-6-9-12(7-8)19-16(21)13(9)15-14(20-22)10-3-1-2-4-11(10)18-15/h1-7,18-19,21H
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| Chemical Name |
6-bromo-3-(3-nitroso-1H-indol-2-yl)-1H-indol-2-ol
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| Synonyms |
GSK-3 Inhibitor IX; 6-bromoindirubin-3-oxime; BIO; GSK 3 Inhibitor IX; MLS 2052; MLS-2052; MLS2052
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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 |
| 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) |
DMSO: ~71 mg/mL (~199.3 mM)
Water: <1 mg/mL Ethanol: ~21 mg/mL (~59.0 mM) |
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| Solubility (In Vivo) |
30% PEG400+0.5% Tween80+5% propylene glycol: 30 mg/mL (Please use freshly prepared in vivo formulations for optimal results.)
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| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.8076 mL | 14.0382 mL | 28.0765 mL | |
| 5 mM | 0.5615 mL | 2.8076 mL | 5.6153 mL | |
| 10 mM | 0.2808 mL | 1.4038 mL | 2.8076 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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