For research use only.

Catalog No.S7609 Synonyms: GW69A, GW554869A

12 publications

GW4869 Chemical Structure

CAS No. 6823-69-4

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.

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Selleck's GW4869 has been cited by 12 publications

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  • U87 and SG2 cells cultured in the indicated conditions for 48 h were exposed to TMZ for 12 h and then were stained with γ-H2AX immediately. Scale bar: 5 μm

    Cancer Lett, 2018, 433:210-220. GW4869 purchased from Selleck.

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Biological Activity

Description 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.
SMase [1]
(Cell-free assay)
1 μM
In vitro

GW4869 inhibits N-SMase not only in vitro but also in a cellular model. GW4869 does not significantly impair TNF-induced NF-κB translocation to nuclei. Therefore, GW4869 does not interfere with other key TNF-mediated signaling effects. GW4869 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, GW4869 does not inhibit the cloned human A-SMase. GW4869 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].

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
MCF7 NYf1ZmtVTnWwY4Tpc44h[XO|YYm= MUCxNE0zOCEQvF2= MUD0bIUh[WSmaYTpc44hd2ZiMUCg{txOKEeZNEi2PUB{cWewaX\pZ4FvfGy7IHnubIljcXSnZDDUUmYucW6mdXPl[EBUVSCqeXTyc4x6e2m|LDD3bIVz\WG|IEKwJO69VSCxZjD0bIUh[2:vcH;1coQheHKxdHXjeIVlKGOxbYDs[ZRmdHliZoLvcUB1cGVibH;zd{Bw\iC|cHjpcodwdXmnbHnu MWqxNlE2PDB7OB?=
OPM2 NFy1VWVRem:uaX\ldoF1cW:wIHHzd4F6 MmL6NEwhOS5{NTygNk42NCB3LDCxNEwhOjBuIESwJO69dW:uL1y= M{HYOFI1KGh? NGXsdId1cGViY3XscEBxem:uaX\ldoF1cW:wIILheIUhf2G|IHnuZ5Jm[XOnZDDh[pRmeiCJV{S4OlkhfHKnYYTt[Y51NiCWaHWgbY5pcWKrdHnvckBw\iClZXzsJJBzd2yrZnXyZZRqd25id3HzJI9vdHlic3nncolncWOjboSge4hmdiC2aHWgZ49v[2WwdILheIlwdiCxZjDHW|Q5Pjlid3HzJFQxKM7:bX;sM2whd3JiaHnnbIVzNg>? M2PLNFI5QDV6Mkm0
RPMI-8226 MlT6R4VtdCC4aXHibYxqfHliYYPzZZk> NYPzU4R{PzJiaB?= NFHse21kgXSxdH;4bYMhfG9iYTDwZY5mdCCxZjDNUUBk\WyuIHzpcoV{ MoTaNlgzOTF3N{O=
NCI-H929 MmrjR4VtdCC4aXHibYxqfHliYYPzZZk> MY[3NkBp MYnjfZRwfG:6aXOgeI8h[SCyYX7lcEBw\iCPTTDj[YxtKGyrbnXz NVTscJpjOjh{MUG1O|M>
JJN3 M1TpcGNmdGxidnnhZoltcXS7IHHzd4F6 M1jmTlczKGh? M{C4S4N6fG:2b4jpZ{B1dyCjIIDhcoVtKG:oIF3NJINmdGxibHnu[ZM> MUeyPFIyOTV5Mx?=
U266 M2ezWmNmdGxidnnhZoltcXS7IHHzd4F6 NFXiT4I4OiCq NUe0W2db[3m2b4TvfIlkKHSxIHGgdIFv\Wxib3[gUW0h[2WubDDsbY5mew>? MmrvNlgzOTF3N{O=
Daudi NFXsOo5E\WyuII\pZYJqdGm2eTDhd5NigQ>? MmL2O|IhcA>? MoXVco91KGO7dH;0c5hq[yC2bzDuc44uVU1iY3XscEBtcW6nczDvdkBRSk2Fcx?= MXuyPFIyOTV5Mx?=
Jurkat NXXzc2tJS2WubDD2bYFjcWyrdImgZZN{[Xl? Ml[0O|IhcA>? NHnEeIdvd3RiY4n0c5RwgGmlIITvJI5wdi2PTTDj[YxtKGyrbnXzJI9zKFCETVPz MUWyPFIyOTV5Mx?=
K562 NHzUNYhE\WyuII\pZYJqdGm2eTDhd5NigQ>? MlPWO|IhcA>? NHG2[Ilvd3RiY4n0c5RwgGmlIITvJI5wdi2PTTDj[YxtKGyrbnXzJI9zKFCETVPz MX:yPFIyOTV5Mx?=
PBMCs M1:w[WNmdGxidnnhZoltcXS7IHHzd4F6 M1nHNVczKGh? NXvZcoNXdm:2IHP5eI91d3irYzD0c{Bvd25vTV2gZ4VtdCCuaX7ld{BweiCSQl3Ddy=> MX2yPFIyOTV5Mx?=

... Click to View More Cell Line Experimental Data

Methods Test Index PMID

PubMed: 29794115     

Confocal microscopy of RAW264.7 stimulated or not with GW4869 (2 h).

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 GW4869 slowed learning. Mice administered GW4869 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 GW4869 reduces the levels of brain and serum exosomes, brain ceramide, and Aβ1-42 plaque load. GW4869 reduces amyloid plaque formation in vivo by preventing exosome secretion. GW4869 may have a low toxicity at levels needed to achieve a biological effect from nSMase2 inhibition[3].


Cell Research:


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  • Cell lines: MCF7 cells
  • Concentrations: 10-20 μM
  • Incubation Time: 30 min
  • Method:

    MCF7 human breast cancer cells are routinely cultured in RPMI 1640 containing 10% FBS at 37 °C in 5% CO2. Unless otherwise indicated, for treatment, cells are seeded at 1.7 × 106 cells/10-cm culture dish in 10 ml of complete medium; after 24 h, the medium is replaced with 7 ml of RPMI 1640 containing 2% FBS and 25 mM Hepes, pH 7.5, and the cells are rested for 2 h prior to treatment. GW4869 is routinely stored at −80 °C as a 1.5 mM stock suspension in Me2SO. Right before use, the suspension is solubilized by the addition of 5% methane sulfonic acid (MSA) (2.5 μl of 5% MSA in sterile double-distilled H2O are added to 50 μl of GW4869 stock suspension; therefore, the concentration of the GW4869 stock solution at the time of the experiments is 1.43 mM). The suspension is mixed and warmed up at 37 °C until clear. Cells are preincubated with the inhibitor for 30 min prior to treatment with TNF. Control cells are treated with Me2SO containing 5% MSA, similarly to the samples receiving the GW4869 solution. When different doses of GW4869 are tested, amounts of vehicle solution are added in order to equal the volume of GW4869 used for the highest dose.

    (Only for Reference)
Animal Research:


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  • Animal Models: Mice with a complete loss of nSMase2 activity (background: C57BL/6J mice)
  • Dosages: 1.25 mg/kg
  • Administration: i.p.
    (Only for Reference)

Solubility (25°C)

In vitro DMSO Insoluble . Please powderise before use.
Water Insoluble
Ethanol Insoluble

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information

Molecular Weight 577.5


CAS No. 6823-69-4
Storage powder
in 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

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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.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID