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GSK343 EZH2 Inhibitor

Name: GSK343
Brand: Selleck Chemicals
SKU: S7164
Availability: InStock
Rating: 5 (60 reviews)

Cat.No.S7164

GSK343 is a potent and selective EZH2 inhibitor with IC50 of 4 nM in a cell-free assay, showing 60 fold selectivity against EZH1, and >1000 fold selectivity against other histone methyltransferases. GSK343 induces autophagy.

Chemical Structure

Molecular Weight: 541.69

Jump to Mechanism of Action Cell Culture & Working Concentration Solubility Chemical Information, Storage & Stability Specificity

Quality Control

Batch: Purity: 99.95%

99.95

Related Targets	Epigenetic Reader Domain HDAC JAK Histone Methyltransferase AMPK PKC PARP HIF Histone Acetyltransferase Aurora Kinase Sirtuin PRMT Histone Demethylase DNA Methyltransferase Pim LSD1 MLL JMJD G9a/GLP NSD FTO MicroRNA PAD SETD SMYD DOT1 COMT WDR5 HNMT SAM MTase
Related Products	Valemetostat (DS-3201) SHR2554 EZH2/HSP90-IN-29 Ezh2 Antibody [M10N8] PF-06821497 EBI-2511 Tulmimetostat (CPI-0209)

Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	Formulation	Activity Description	PMID
2D10	Function Assay	0.25-2.0 μM	96 h		reduces total cellular trimethylated H3K27 in a time- and dose-dependent manner	26041287
MDA-MB-231	Growth Inhibition Assay	0-20 μM	72 h		shows cytotoxicity in a dose-dependent manner	25203626
MDA-MB-231	Function Assay	10 μM	72 h		reduces the level of H3K27-me3	25203626
MDA-MB-231	Function Assay	10 μM	72 h		induces LC3-II accumulation	25203626
MDA-MB-231	Function Assay	10 μM	24 h		induces autophagy	25203626
HepG2	Growth Inhibition Assay	0-20 μM	72 h		shows cytotoxicity in a dose-dependent manner	25203626
A549	Growth Inhibition Assay	0-20 μM	72 h		shows cytotoxicity in a dose-dependent manner	25203626
HepG2	Function Assay	10 μM	24 h		induces autophagy	25203626
A549	Function Assay	10 μM	24 h		induces autophagy	25203626
OVCAR10	Growth Inhibition Assay	1 μM	0-12 d	DMSO	inhibits cell growth time dependently	23759589
UPN289	Growth Inhibition Assay	1 μM	0-12 d	DMSO	inhibits cell growth time dependently	23759589
SKOV3	Growth Inhibition Assay	1 μM	0-12 d	DMSO	inhibits cell growth time dependently	23759589
SKOV3	Apoptosis Assay	1 μM	4 d	DMSO	induces apoptosis cultured in 3D conditions	23759589
HCC1806	Function assay		72 hrs		Inhibition of EZH2-mediated nuclear H3K27 methylation in human HCC1806 cells after 72 hrs by immunofluorescence analysis, IC50 = 0.174 μM.	24900432
LNCaP	Function assay		6 days		Inhibition of EZH2-mediated proliferation of human LNCaP cells after 6 days by chemiluminescence analysis, IC50 = 2.9 μM.	24900432
DU145	Function assay		6 days		Inhibition of EZH2-mediated proliferation of human DU145 cells after 6 days by chemiluminescence analysis	24900432
SKBR3	Function assay		6 days		Inhibition of EZH2-mediated proliferation of human SKBR3 cells after 6 days by chemiluminescence analysis	24900432
PC3	Function assay		6 days		Inhibition of EZH2-mediated proliferation of human PC3 cells after 6 days by chemiluminescence analysis	24900432
ZR-75-1	Function assay		6 days		Inhibition of EZH2-mediated proliferation of human ZR-75-1 cells after 6 days by chemiluminescence analysis	24900432
HCC180	Function assay		6 days		Inhibition of EZH2-mediated proliferation of human HCC180 cells after 6 days by chemiluminescence analysis	24900432
Click to View More Cell Line Experimental Data

Chemical Information, Storage & Stability

Molecular Weight	541.69	Formula	C₃₁H₃₉N₇O₂	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	1346704-33-3	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	N/A			Smiles	CCCC1=C(C(=O)NC(=C1)C)CNC(=O)C2=C3C=NN(C3=CC(=C2)C4=CC(=NC=C4)N5CCN(CC5)C)C(C)C

Solubility

In vitro
Batch:

Ethanol : 4 mg/mL

DMSO : 1 mg/mL (1.84 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
=	×	×

Dilution Calculator Molecular Weight Calculator

In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	5.000mg/ml (9.23mM)	Taking the 1 mL working solution as an example, add 50 μL of 100 mg/ml clarified DMSO stock solution to 400 μL PEG300, mix evenly to clarify it; add 50 μL Tween80 to the above system, mix evenly to clarify it; then continue to add 500 μL ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	5% DMSO 1 95% corn oil Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	0.500mg/ml (0.92mM)	Taking the 1 mL working solution as an example, add 50 μL of 10 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

Dosage: mg/kg Average weight of animals: g Dosing volume per animal:μL Number of animals:

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Mechanism of Action

Features	A chemical probe for the SGC epigenetics initiative. Potential use in a variety of solid tumors.
Targets/IC₅₀/Ki	EZH2 ^[1] (Cell-free assay) 4 nM EZH1 ^[1] (Cell-free assay) 240 nM
In vitro	GSK343 inhibits trimethylation of H3K27 (H3K27me3) with IC50 of 174 nM in HCC1806 breast cancer cells. This compound potently inhibits cell proliferation in breast cancer cells and prostate cancer cells, and the prostate cancer cell line LNCaP is the most sensitive to it, with IC50 of 2.9 μM. ^[1] This chemical significantly suppresses the growth of EOC cells cultured in 3D matrigel extracellular matrix (ECM), which mimics the tumor microenvironment in vivo. In addition, it also induces apoptosis of EOC cells in 3D and significantly inhibits the invasion of EOC cells. ^[2]
Kinase Assay	In vitro biochemical assays against histone methyltransferases
Kinase Assay	Activity against EZH2 is assessed using 5 member PRC2 complex (Flag-EZH2, EED, SUZ12, AEBP2, RbAp48). The assay protocol may be summarized as follows: 10 mM stocks of compounds are prepared from solid in 100% DMSO. An 11 point serial dilution master plate is prepared in 384 well format (1:3 dilution, columns 6 and 18 were equal volume DMSO controls) and dispensed to assay ready plates using acoustic dispensing technology to create a 100 nL stamp of compound and DMSO controls. The assay additions consisted of equal volume additions of 10 nM EZH2 and the substrate solution (5 µg/mL HeLa nucleosomes and 0.25 µM [³H]-SAM) dispensed into assay plates using a multi-drop combi dispense. Reaction plates are incubated for 1 hr and quenched with an equal volume addition of 0.5 mg/mL PS-PEI Imaging Beads (RPNQ0098) containing 0.1 mM unlabeled SAM. The plates are sealed, dark adapted for 30 minutes, and a 5 minute endpoint luminescence image is acquired using a Viewlux imager. Plate statistics such as Z’ and signal to background as well as dose response curves are analyzed using Activity BaseXE. The in vitro biochemical activity of EZH1 is assessed as part of a 5 member PRC2 complex using a 384 well SPA assay identical to EZH2. Buffer components, reagent dispensing, compound plate preparation, quench conditions and data analysis are identical for EZH1 and EZH2 with final assay concentrations of 20 nM EZH1, 5 μM/mL HeLa nucleosomes and 0.25 μM [³H]-SAM. Further data analysis, pIC50 pivots and visualizations are enabled by TIBCO Spotfire. This compound is profiled at Reaction Biology Corp. (Malvern, PA) to assess inhibition in their panel of histone methyltransferase assays. Methyltransferase activity is assessed using HotSpot technology, a miniaturized radioisotope-based filter binding assay. Inhibitors are dissolved in dimethyl sulfoxide (DMSO) and tested at concentrations up to 100 uM with a final DMSO concentration of 2%. Buffer containing the methyltrasferase at the listed concentration and its preferred substrate as shown in the accompanying table is preincubated with this chemical for 10 min. Reactions are initiated by the addition of 1 uM S-adenosyl-L-[methyl-³H]methionine (SAM), allowed to incubate for 60 min at 30C followed by transfer to P81 filter-paper and PBS wash before detection.
In vivo	GSK343, a selective EZH2 inhibitor, inhibits phagocytosis.
References	[1] http://pubs.acs.org/doi/abs/10.1021/ml3003346 [2] https://pubmed.ncbi.nlm.nih.gov/23759589/ [3] https://pubmed.ncbi.nlm.nih.gov/35265199/

Applications

Methods	Biomarkers	PMID
Western blot	H3K27me3 EZH2 / EED / SUZ12 N-cadherin / Vimentin / Snail / Slug / MMP2 / MMP9	29228694
Immunofluorescence	N-cadherin / H3K27me3 / Vimentin	29228694
Growth inhibition assay	Cell viability	26973856

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