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GW4869 N-SMase inhibitor

Name: GW4869
Brand: Selleck Chemicals
SKU: S7609
Availability: InStock
Rating: 5 (89 reviews)

Cat.No.S7609

GW4869 (GW69A, GW554869A) is a neutral, noncompetitive inhibitor of sphingomyelinase (SMase) with an IC50 of 1 μM. It is selective for N-SMase, and does not inhibit acid SMase at up to at least 150 μM, also is a commonly used exosome inhibitor.

Chemical Structure

Molecular Weight: 577.5

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

Quality Control

Batch: Purity: >97%

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Related Targets	Apoptosis related Ras STAT TNF-alpha Bcl-2 Caspase PD-1/PD-L1 Ferroptosis p53 RIP kinase c-RET MDM2/MDMX Myc IAP OXPHOS PERK FKBP Thymidylate Synthase ABC Transporter Heme Oxygenase PFKFB PKD Survivin Pyroptosis ASK Bcl-6 Nur77 DAPK TRADD TACE
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Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	Activity Description	PMID
MCF7	Function assay	10-20 μM		the addition of 10 μM GW4869 significantly inhibited TNF-induced SM hydrolysis, whereas 20 μM of the compound protected completely from the loss of sphingomyelin	12154098
OPM2	Proliferation assay	0, 1.25, 2.5, 5, 10, 20, 40 μmol/L	24 h	the cell proliferation rate was increased after GW4869 treatment. The inhibition of cell proliferation was only significant when the concentration of GW4869 was 40 μmol/L or higher.	28858294
RPMI-8226	Cell viability assay		72 h	cytotoxic to a panel of MM cell lines	28211573
NCI-H929	Cell viability assay		72 h	cytotoxic to a panel of MM cell lines	28211573
JJN3	Cell viability assay		72 h	cytotoxic to a panel of MM cell lines	28211573
U266	Cell viability assay		72 h	cytotoxic to a panel of MM cell lines	28211573
Daudi	Cell viability assay		72 h	not cytotoxic to non-MM cell lines or PBMCs	28211573
Jurkat	Cell viability assay		72 h	not cytotoxic to non-MM cell lines or PBMCs	28211573
K562	Cell viability assay		72 h	not cytotoxic to non-MM cell lines or PBMCs	28211573
PBMCs	Cell viability assay		72 h	not cytotoxic to non-MM cell lines or PBMCs	28211573
Click to View More Cell Line Experimental Data

Chemical Information, Storage & Stability

Molecular Weight	577.5	Formula	C₃₀H₂₈N₆O₂.₂HCl.XH₂O	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	6823-69-4	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	GW69A, GW554869A			Smiles	C1CN=C(N1)C2=CC=C(C=C2)NC(=O)C=CC3=CC=C(C=C3)C=CC(=O)NC4=CC=C(C=C4)C5=NCCN5.Cl.Cl

Solubility

In vitro
Batch:

DMSO : 1 mg/mL (1.73 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

DMSO : 3.25 mg/mL (5.62 mM) Warmed with 50°C water bath; Ultrasonicated;
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
=	×	×

Dilution Calculator Molecular Weight Calculator

In vivo Batch:
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.	0.050mg/ml (0.09mM)	Taking the 1 mL working solution as an example, add 50 μL of 1 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.
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.	0.050mg/ml (0.09mM)	Taking the 1 mL working solution as an example, add 50 μL of 1 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.
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 95% Corn oil Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	0.162mg/ml (0.28mM)	Taking the 1 mL working solution as an example, add 50 μL of 3.25 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.
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.	0.250mg/ml (0.43mM)	Taking the 1 mL working solution as an example, add 50 μL of clarified DMSO stock solution of 5 mg/ml to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify it; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
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.	0.100mg/ml (0.17mM)	Taking the 1 mL working solution as an example, add 50 μL of 2 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
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.

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

Targets/IC₅₀/Ki	SMase ^[1] (Cell-free assay) 1 μM
In vitro	GW4869 inhibits N-SMase not only in vitro but also in a cellular model. This compound does not significantly impair TNF-induced NF-κB translocation to nuclei. Therefore, it does not interfere with other key TNF-mediated signaling effects. It is able, in a dose-dependent manner, to significantly protect from cell death as measured by nuclear condensation, caspase activation, PARP degradation, and trypan blue uptake. These protective effects are accompanied by significant inhibition of cytochrome c release from mitochondria and caspase 9 activation, therefore localizing N-SMase activation upstream of mitochondrial dysfunction. At up to 150 μM, this chemical does not inhibit the cloned human A-SMase. It shows no or minor inhibitory activity versus other hydrolytic enzymes, such as bacterial phosphatidylcholine-PLC and bovine protein phosphatase 2A, and it shows significantly higher activity versus the rat brain enzyme compared with the human lyso-PAF PLC^[1].
In vivo	Systemic administration of GW4869 does not alter the ceramide or sphingomylein content of liver, heart or skeletal muscle but does decrease the ceramide content and increase the sphingomyelin content in brain. Inhibition of nSMase2 with this compound slowed learning. Mice administered this chemical do not progressively decrease latency to locate the hidden platform with repeated training trials, suggesting that they has difficulty learning to use spatial cues to navigate the pool^[2]. Intraperitoneal injection of this agent reduces the levels of brain and serum exosomes, brain ceramide, and Aβ1-42 plaque load. It reduces amyloid plaque formation in vivo by preventing exosome secretion. This inhibitor may have a low toxicity at levels needed to achieve a biological effect from nSMase2 inhibition^[3].
References	[1] https://pubmed.ncbi.nlm.nih.gov/12154098/ [2] https://pubmed.ncbi.nlm.nih.gov/20629193/ [3] https://pubmed.ncbi.nlm.nih.gov/24650793/

Applications

Methods	Biomarkers	Images	PMID
Immunofluorescence	Nrf2		29794115

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