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BIO GSK-3 inhibitor

Cat.No.S7198

BIO (GSK-3 Inhibitor IX, 6-bromoindirubin-3-oxime, 6-Bromoindirubin-3'-oxime, MLS 2052) is a specific inhibitor of GSK-3 with IC50 of 5 nM for GSK-3α/β in a cell-free assay, shows >16-fold selectivity over CDK5, also a pan-JAK inhibitor with IC50 of 30 nM for Tyk2. This compound induces apoptosis in human melanoma cells.
BIO GSK-3 inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 356.17

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Quality Control

Batch: Purity: 99.93%
99.93

Cell Culture, Treatment & Working Concentration

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
HEI-OC1 Function assay Protection against cisplatin-induced cell death in neonatal mouse HEI-OC1 cells assessed as reduction in caspase-3/7 activity, EC50 = 0.192 μM. 30091915
SH-SY5Y Function assay Inhibition of GSK3-mediated beta-casein phosphorylation in human SH-SY5Y cells in presence of MG132 by Western blot analysis, IC50 = 0.29 μM. 18816110
K562 Antiproliferative assay 72 hrs Antiproliferative activity against human K562 cells after 72 hrs by MTT assay, IC50 = 1.3 μM. 19783149
IMR90 Antiproliferative assay 72 hrs Antiproliferative activity against human IMR90 cells after 72 hrs by MTT assay, IC50 = 1.9 μM. 19783149
HCT116 Antiproliferative assay 72 hrs Antiproliferative activity against human HCT116 cells after 72 hrs by MTT assay, IC50 = 5.2 μM. 19783149
HepG2 Cytotoxicity assay 24 hrs Cytotoxicity against human HepG2 cells assessed as cell growth inhibition after 24 hrs by alamar blue assay, IC50 = 5.3 μM. 28743492
HL60 Antiproliferative assay 5 days Antiproliferative activity against human HL60 cells after 5 days by MTT assay, IC50 = 5.4 μM. 19783149
HuH7 Antiproliferative assay 72 hrs Antiproliferative activity against human HuH7 cells after 72 hrs by MTT assay, IC50 = 6.2 μM. 19783149
SH-SY5Y Cytotoxicity assay 48 hrs Cytotoxicity against human SH-SY5Y cells after 48 hrs by MTS reduction assay, IC50 = 9 μM. 18816110
SH-SY5Y Function assay 48 hrs Survival of human SH-SY5Y cells after 48 hrs by MTS reduction assay, IC50 = 9.5 μM. 16854069
SH-SY5Y Function assay Death of human SH-SY5Y cells in absence of 20 uM Q-VD-OPh by MTS reduction assay, IC50 = 10 μM. 16854069
SH-SY5Y Function assay Death of human SH-SY5Y cells in presence of 20 uM Q-VD-OPh by MTS reduction assay, IC50 = 13 μM. 16854069
SH-SY5Y Function assay 24 hrs Survival of human SH-SY5Y cells after 24 hrs by MTS reduction assay, IC50 = 18 μM. 16854069
IMR32 Cytotoxicity assay 48 hrs Cytotoxicity against human IMR32 cells assessed as cell viability after 48 hrs by MTT assay 21802947
SK-N-SH Cytotoxicity assay 48 hrs Cytotoxicity against human SK-N-SH cells assessed as cell viability after 48 hrs by MTT assay 21802947
NB39 Cytotoxicity assay 48 hrs Cytotoxicity against human NB39 cells assessed as cell viability after 48 hrs by MTT assay 21802947
A673 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells 29435139
DAOY qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for DAOY cells 29435139
Saos-2 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Saos-2 cells 29435139
BT-37 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-37 cells 29435139
RD qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for RD cells 29435139
SK-N-SH qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-SH cells 29435139
BT-12 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-12 cells 29435139
MG 63 (6-TG R) qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for MG 63 (6-TG R) cells 29435139
NB1643 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB1643 cells 29435139
OHS-50 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for OHS-50 cells 29435139
Rh41 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh41 cells 29435139
Rh30 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh30 cells 29435139
SJ-GBM2 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SJ-GBM2 cells 29435139
SK-N-MC qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells 29435139
NB-EBc1 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB-EBc1 cells 29435139
LAN-5 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for LAN-5 cells 29435139
Rh18 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh18 cells 29435139
A549 Function assay 10 uM 15 mins Inhibition of human mPGES1 from human A549 cells assessed as PGE2 at 10 uM after 15 mins by HPLC 24697244
HEK293 Function assay 0.5 to 1 uM Inhibition of human GSK3 activity in HEK293 cells containing the Wnt/beta-catenin activated reporter pSuperTOPFLASH (STF293 cells) at 0.5 to 1 uM by Wnt reporter gene assay 24697244
sf9 Function assay 1 uM Inhibition of recombinant human N-terminal GST-tagged CDK4 (S4 to E303 residues)/Cyclin D1 (Q4 to I295 residues) expressed in sf9 cells at 1 uM using RB-CTF as substrate by filter binding assay 28557430
sf9 Function assay 1 uM Inhibition of human N-terminal GST/His6-tagged GSK3beta (M1 to T420 residues) expressed in baculovirus infected sf9 cells at 1 uM using RBER-IRStide as substrate by filter binding assay 28557430
sf9 Function assay 1 uM Inhibition of recombinant human N-terminal GST-tagged CDK2 (M1 to L298 residues)/Cyclin A2 (M1 to L432 residues) expressed in baculovirus infected sf9 cells at 1 uM using Histone H1 as substrate by filter binding assay 28557430
sf9 Function assay 1 uM Inhibition of recombinant human N-terminal GST-tagged CDK5 (M1 to P292 residues)/p35NCK (M1 to R307 residues) expressed in baculovirus infected sf9 cells at 1 uM using RB-CTF as substrate by filter binding assay 28557430
sf9 Function assay 1 uM Inhibition of human N-terminal GST/His6-tagged Aurora B (A2 to A344 residues) expressed in sf9 cells at 1 uM using tetra(LRRLSLG) as substrate by filter binding assay 28557430
sf9 Function assay 1 uM Inhibition of human N-terminal GST/His6-tagged FGFR1 (G400 to R820 residues) expressed in baculovirus infected sf9 cells at 1 uM using Poly(Glu,Tyr)4:1 as substrate by filter binding assay 28557430
sf9 Function assay 1 uM Inhibition of recombinant human N-terminal GST/His6-tagged CDK1 (M1 to M297 residues)/Cyclin B1 (M1 to V433 residues) expressed in sf9 cells at 1 uM using RB-CTF as substrate by filter binding assay 28557430
sf9 Function assay 1 uM Inhibition of recombinant human N-terminal GST-tagged CDK2 (M1 to L298 residues)/Cyclin E1 (M1 to A395 residues) expressed in sf9 cells at 1 uM using RB-CTF as substrate by filter binding assay 28557430
sf9 Function assay 1 uM Inhibition of human N-terminal GST/His6-tagged Aurora A (M1 to S403 residues) expressed in baculovirus infected sf9 cells at 1 uM using tetra(LRRLSLG) as substrate by filter binding assay 28557430
sf9 Function assay 1 uM Inhibition of human N-terminal GST-tagged Aurora B (M1 to S27 residues) expressed in sf9 cells at 1 uM using CDC25C-derived peptide as substrate by filter binding assay 28557430
HT22 Function assay 10 uM 24 hrs Inhibition of GSK3-mediated beta casein phosphorylation in mouse HT22 cells at 10 uM after 24 hrs by Western blot analysis in presence of MG132 22998443
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Solubility

In vitro
Batch:

DMSO : 71 mg/mL (199.34 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : 7 mg/mL

Water : Insoluble

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Dilution Calculator Molecular Weight Calculator

In vivo
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Chemical Information, Storage & Stability

Molecular Weight 356.17 Formula

C16H10BrN3O2

 Storage (From the date of receipt)
CAS No. 667463-62-9 Download SDF Storage of Stock Solutions

Synonyms GSK-3 Inhibitor IX, 6-bromoindirubin-3-oxime, 6-Bromoindirubin-3'-oxime, MLS 2052 Smiles C1=CC=C2C(=C1)C(=C(N2)C3=C(NC4=C3C=CC(=C4)Br)O)N=O

Mechanism of Action

Features
The first pharmacological agent shown to maintain self-renewal in human and mouse embryonic stem cells.
Targets/IC50/Ki
GSK-3
(Cell-free assay)
5 nM
TYK2
(Cell-free assay)
30 nM
CDK5/p35
(Cell-free assay)
0.08 μM
CDK2/CyclinA
(Cell-free assay)
0.30 μM
CDK1/CyclinB
(Cell-free assay)
0.32 μM
JAK3
(Cell-free assay)
0.5 μM
In 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. This compound interacts within the ATP binding pocket of these kinases, reduces β-catenin phosphorylation on a GSK-3-specific site in cellular models, closely mimicks Wnt signaling in Xenopus embryos. In human and mouse embryonic stem cells, this chemical maintains the undifferentiated phenotype and sustains expression of the pluripotent state-specific transcription factors Oct-3/4, Rex-1 and Nanog. This compound-mediated Wnt activation is functionally reversible, as withdrawal of the compound leads to normal multidifferentiation programs in both human and mouse embryonic stem cells. It promotes proliferation in mammalian cardiomyocytes. 6BIO is also a pan-JAK inhibitor, with IC50 values of 0.03, 1.5, 8.0, 0.5 μM for TYK2, JAK1, JAK2 and JAK3. This compound selectively inhibits phosphorylation of STAT3 and induces apoptosis of human melanoma cells.
Kinase Assay
Kinase assay
Kinase activities are assayed in Buffer A or C at 30°C, at a final ATP concentration of 15 μM. Blank values are subtracted and activities calculated as pmoles of phosphate incorporated during a 10 min incubation. Controls are performed with appropriate dilutions of dimethylsulfoxide. In a few cases phosphorylation of the substrate is assessed by autoradiography after SDS-PAGE. GSK-3α/β is purified from porcine brain by affinity chromatography on 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 30 min incubation at 30°C, 25 μl aliquots of supernatant are spotted onto 2.5 × 3 cm pieces of Whatman P81 phosphocellulose paper, and 20 seconds later, the filters are washed five times (for at least 5 min each time) in a solution of 10 ml phosphoric acid/liter of water. The wet filters are counted in the presence of 1 ml ACS scintillation fluid.
In vivo
BIO suppresses melanoma tumor growth in a mouse xenograft model.
References
  • [4] https://pubmed.ncbi.nlm.nih.gov/21610112/

Applications

Methods Biomarkers Images PMID
Western blot p-AKT / AKT / p21 / p27 p-β-catenin / β-catenin FoxO3a / FoxO1 / p-FoxO3a / p-FoxO1
S7198-WB1
27510556
Immunofluorescence pAKT / p21 / p27 TNF-α E-cadherin / Nanog Oct3/4
S7198-IF1
27510556
Growth inhibition assay Cell proliferation
S7198-viability1
27510556

Clinical Trial Information

(data from https://clinicaltrials.gov, updated on 2024-05-22)

NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT06337422 Not yet recruiting
Healthy Volunteer
International Bio service
 September 23 2024 Phase 1
NCT04276857 Not yet recruiting
Locally Advanced Pancreatic Cancer|Irreversible Electroporation
University of Saskatchewan
 September 1 2024 Not Applicable
NCT06359041 Not yet recruiting
Generalized Myasthenia Gravis (gMG)
Cabaletta Bio
 August 2024 Phase 1|Phase 2

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