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GSK343

Name: GSK343
Brand: Selleck Chemicals
SKU: S7164
Rating: 5 (59 reviews)

Catalog No.S7164 Purity:99.95%

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.

GSK343 Chemical Structure

CAS No. 1346704-33-3

Purity & Quality Control

Batch: Purity: 99.95%

99.95

GSK343 Related Products

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Related Compound Libraries	Kinase Inhibitor Library FDA-approved Drug Library Natural Product Library Bioactive Compound Library-I Highly Selective Inhibitor Library	Click to Expand

Signaling Pathway

Histone Methyltransferase Signaling Pathway

Cell Data

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

Biological Activity

Description

Features

A chemical probe for the SGC epigenetics initiative. Potential use in a variety of solid tumors.

Targets

EZH2 ^[1] (Cell-free assay)	EZH1 ^[1] (Cell-free assay)
4 nM	240 nM

In vitro
In vitro	GSK343 inhibits trimethylation of H3K27 (H3K27me3) with IC50 of 174 nM in HCC1806 breast cancer cells. GSK343 potently inhibits cell proliferation in breast cancer cells and prostate cancer cells, and the prostate cancer cell line LNCaP is the most sensitive to GSK343, with IC50 of 2.9 μM. ^[1] GSK343 significantly suppresses the growth of EOC cells cultured in 3D matrigel extracellular matrix (ECM), which mimics the tumor microenvironment in vivo. In addition, GSK343 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. Compounds are 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 compound 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.
Cell Research	Cell lines	Breast cancer cell lines (HCC1806, Sk-Br-3, ZR-75-1), prostate cancer cell lines (DU145, PC3, LNCaP)
	Concentrations	~50 μM
	Incubation Time	6 days
	Method	To account for varying doubling rates among cancer cell lines, the optimal cell seeding is determined empirically for all cell lines by examining their growth in a 384-well plate over 6 days with a wide range of seeding densities. Cells are then plated at the optimal seeding density and allowed to adhere overnight. Cells are treated in duplicate with a 20-point 2-fold dilution series of compound or 0.147% DMSO (vehicle control) and incubated for 6 days at 37C in 5% CO2. Cells are then lysed with 25 μl CellTiter-Glo per well and chemiluminescence is quantified with a TECAN Safire2 microplate reader. In addition, an untreated plate of cells is harvested at the time of compound addition (T0) to quantify the starting number of cells. CTG values after 6 days of treatment were expressed as a percent of the T0 value and plotted against compound concentration. Data are fit with a 4-parameter equation to generate a concentration response curve and the concentration of compound required to inhibit 50% of growth (gIC50) is determined
Experimental Result Images	Methods	Biomarkers	Images	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

In Vivo
In vivo	GSK343, a selective EZH2 inhibitor, inhibits phagocytosis.

References

Chemical Information & Solubility

Molecular Weight	541.69	Formula	C₃₁H₃₉N₇O₂
CAS No.	1346704-33-3	SDF	Download GSK343 SDF
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
Storage (From the date of receipt)	3 years-20°Cpowder	Storage of Stock Solutions Aliquot the stock solutions to avoid repeated freeze-thaw cycles	1 year-80°Cin solvent
Storage (From the date of receipt)	3 years-20°Cpowder		1 month-20°Cin solvent

In vitro
Batch:

Ethanol : 4 mg/mL

DMSO : 1 mg/mL ( (1.84 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Molecular Weight Calculator

In vivo Batch: Add solvents to the product individually and in order.				In vivo Formulation Calculator
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.

Preparing Stock Solutions

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
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Dilution Calculator Molecular Weight Calculator

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.

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
If you have any other enquiries, please leave a message.

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