A-769662

Catalog No.S2697

For research use only.

A-769662 is a potent, reversible AMPK activator with EC50 of 0.8 μM in cell-free assays, little effect on GPPase/FBPase activity.

A-769662 Chemical Structure

CAS No. 844499-71-4

Selleck's A-769662 has been cited by 60 publications

Purity & Quality Control

Choose Selective AMPK Inhibitors

Other AMPK Products

Biological Activity

Description A-769662 is a potent, reversible AMPK activator with EC50 of 0.8 μM in cell-free assays, little effect on GPPase/FBPase activity.
Targets
AMPK [1]
(Cell-free assay)
Fatty acid synthesis [1]
(in primary rat hepatocytes)
0.8 μM(EC50) 3.2 μM
In vitro

A-769662 stimulates partially purified rat liver AMPK with EC50 with 0.8 μM. A-769662 activates AMPK purified from multiple tissues and species in a dose-responsive manner with modest variations in observed EC50s. EC50s determined for A-769662 using partially purified AMPK extracts from rat heart, rat muscle, or human embryonic kidney cells (HEKs) are 2.2 mM, 1.9 mM, or 1.1 mM, respectively. A 4 hours treatment of primary rat hepatocytes with A-769662 dose-dependently increases ACC phosphorylation, which correlated inhibition of fatty acid synthesis with IC50 of 3.2 μM. A-769662 also inhibits fatty acid sythesis in mouse hepatocytes with IC50 with 3.6 μM [1] A-769662 activates AMPK both allosterically and by inhibiting dephosphorylation of AMPK on Thr-172, similar to the effects of AMP. [2] A-769662 inhibits proteasomal function by an AMPK-independent mechanism. A-769662 affects the in vitro activity of purified 26S proteasomes but not the in vitro activity of purified 20S proteasomes. A-769662 has toxic effects on MEF cells. [3] A recent research shows A-769662 inhibited cell proliferation and DNA synthesis. [4]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
mouse hepatocytes MVTGeY5kfGmxbjDhd5NigQ>? M3HMeVEhdU1? NH;6d2pFVVOR MnrQbY5pcWKrdIOg[oF1fHliYXPp[EB{gW62aHXzbZMhf2m2aDDJR|UxKG:oIEOuOkDPxE1? MUK8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yPjd3M{W3Okc,OTZ5NUO1O|Y9N2F-
rat hepatocytes NWPiV|FNTnWwY4Tpc44h[XO|YYm= M2XxNVEhdU1? NVnjdVlsTE2VTx?= M3L4WIlvcGmkaYTzJIZifHS7IHHjbYQhe3mwdHjld4l{KHerdHigTWM2OCCxZjCzMlYh|ryP NF;2cos9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zNke1N|U4Pid-MU[3OVM2PzZ:L3G+
HEK293 NHm4dVBMcW6jc3WgZZN{[Xl? NXHiRolPOjByIN88US=> M3jxV2ROW09? NFy2S29i[3SrdnH0[ZMh\W6mb3flco92eyCDTWDL MUm8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yPzd{OEK0NUc,OTd5MkiyOFE9N2F-
CCL13 MWPLbY5ie2ViYYPzZZk> NGHwVlEzODBizszN NFGwWolFVVOR NH7LU29i[3SrdnH0[ZMh\W6mb3flco92eyCDTWDL MVm8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yPzd{OEK0NUc,OTd5MkiyOFE9N2F-
MEFs M{n4O2Z2dmO2aX;uJIF{e2G7 M3Kw[lMxOCEQvF2= NULL[nFMTE2VTx?= NHz6TllqdmirYnn0d{Bxem:2ZXHzc41idCCodX7jeIlwdiCkeTDhckBCVVCNLXnu[IVx\W6mZX70JI1m[2ijbnnzcS=> M3HleFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF6NUmzOVg1Lz5zOEW5N|U5PDxxYU6=
epididymal clear cells MV;GeY5kfGmxbjDhd5NigQ>? NUD2dnB1OjByIN88US=> NEntdlFFVVOR NG[xRnBqdmirYnn0d{B1cGVicFitcYVlcWG2ZXSgWk1CXFCjc3WgZYNkfW23bHH0bY9vKGG2IITo[UBieGmlYXygcYVu[nKjbnW= NE\VSVE9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zOUKxNVkyQCd-MUmyNVE6OTh:L3G+
3T3-L1 MmLaSpVv[3Srb36gZZN{[Xl? M17vfVEvOiCvTR?= MlzpSG1UVw>? NFLER4dqdmirYnn0d{A{XDNvTEGgRYRqeG:pZX7ld4l{ MWi8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yQTR6M{OwOEc,OTl2OEOzNFQ9N2F-
3T3-L1 NXX0NXZ2TnWwY4Tpc44h[XO|YYm= MUKxMlIhdU1? NEj0O4ZFVVOR NHHwPHlqdmirYnn0d{B1cGViRYjwdoV{e2mxbjDv[kBC\Gmyb3flcoV{cXOUZXzheIVlKFS{YX7zZ5JqeHSrb36gSoFkfG:{czDhcoQhVWG{a3Xydy=> NX7hRVV[RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMUm0PFM{ODRpPkG5OFg{OzB2PD;hQi=>
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3T3-L1 NV3tPHI{[3m2b4TvfIlkcXS7IHHzd4F6 NULxW|JSOS5{IH3N MXfEUXNQ NH\NdlBl\WO{ZXHz[ZMhS2WubDDWbYFjcWyrdIm= MWW8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yQTR6M{OwOEc,OTl2OEOzNFQ9N2F-
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L6 skeletal muscle cells MXrGeY5kfGmxbjDhd5NigQ>? NYn0XFI{OjVyIN88US=> NV7yblJZTE2VTx?= M3;YTolvcGmkaYTzJJRp\SCQYTutT{suSVSSYYPlJJRz[W6|cH;yeEBi[3Srdnn0fUBidmRiY3XscEB{fXKoYXPlJIFjfW6mYX7j[S=> NFy4W489[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zOUiyPFg{Pid-MUm4Nlg5OzZ:L3G+
MDA-MB231 M2LkO2Fxd3C2b4Ppd{Bie3OjeR?= MlfPOFAxKM7:TR?= MUjEUXNQ MlfBd4Vve2m2aYrld{BpfW2jbjDidoVie3RiY3HuZ4VzKGOnbHygcIlv\XNidH:gWHJCUUxvaX7keYNm\CCjcH;weI9{cXN? M2\GXFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF7OEm2OFY6Lz5zOUi5OlQ3QTxxYU6=
BT474 MnSwRZBweHSxc3nzJIF{e2G7 NY[1fIpGPDByIN88US=> NWjDd|Q5TE2VTx?= NGHzfm5{\W6|aYTpfoV{KGi3bXHuJIJz\WG|dDDjZY5k\XJiY3XscEBtcW6nczD0c{BVWkGLTD3pcoR2[2WmIHHwc5B1d3Orcx?= NFHVe5I9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zOUi5OlQ3QSd-MUm4PVY1Pjl:L3G+
MCF7 M3;FW2Fxd3C2b4Ppd{Bie3OjeR?= NWTafFhsPDByIN88US=> MlXmSG1UVw>? NGjnZXR{\W6|aYTpfoV{KGi3bXHuJIJz\WG|dDDjZY5k\XJiY3XscEBtcW6nczD0c{BVWkGLTD3pcoR2[2WmIHHwc5B1d3Orcx?= M4DLT|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF7OEm2OFY6Lz5zOUi5OlQ3QTxxYU6=
Mesenchymal stem cells MVzLbY5ie2ViYYPzZZk> NUjQd5MyOTBiwsXN Mle1SG1UVw>? M{LjPIlv\HWlZYOgZUBzd2K3c4SgZY5lKHO3c4ThbY5m\CCDTWDLJIFkfGm4YYTpc44> M4LSSFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2MUC0PFc6Lz5{NEGwOFg4QTxxYU6=
Mesenchymal stem cells MY\jfZRwfG:6aXPpeJkh[XO|YYm= MnTxNVAxKML3TR?= MWTEUXNQ NGXsfGZl\WO{ZXHz[ZMhfGinIF3TR{Bxem:uaX\ldoF1cW:w M3TN[|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2MUC0PFc6Lz5{NEGwOFg4QTxxYU6=
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MG-63 NXP6U4VQSXCxcITvd4l{KGG|c3H5 NEO3[VEyOCEEtV2= MUTEUXNQ M3fEeZN2eHC{ZYPz[ZMhUDKRMj3JcoR2[2WmIF;zeIVw[myjc4SgR4VtdCCDcH;weI9{cXN? MlS4QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjR7NkCzOlIoRjJ2OU[wN|YzRC:jPh?=
MC3T3-E1 NUfnW|FsSXCxcITvd4l{KGG|c3H5 NIjITJAyOCEEtV2= MVPEUXNQ MkLvd5VxeHKnc4Pld{BJOk9{LVnu[JVk\WRiT4P0[Y9jdGG|dDDD[YxtKEGyb4D0c5Nqew>? NWHzb3hSRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS5OlA{PjJpPkK0PVYxOzZ{PD;hQi=>
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MC3T3-E1 NHXzSXlHfW6ldHnvckBie3OjeR?= NFXOeFEyOCEEtV2= Mnz4SG1UVw>? Mn\6ZYxt\X[rYYTld{BTV1NiYXPjeY12dGG2aX;uJIFv\CCDVGCg[IVxdGW2aX;uJINifXOnZDDifUBJOk9{ MmDLQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjR7NkCzOlIoRjJ2OU[wN|YzRC:jPh?=
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MC3T3-E1 NVHQNWtTTnWwY4Tpc44h[XO|YYm= NVzpZXFVOTBiwsXN MX\EUXNQ NU\1R4wx\mGlaXzpeIF1\XNiSELPNk1qdmS3Y3XkJIF2fG:yaHHnfUBi[3SrdnH0bY9v MXy8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDl4MEO2Nkc,OjR7NkCzOlI9N2F-
PC3 M3nQ[mtqdmG|ZTDhd5NigQ>? M2TybVExOCEEtV2= MYjEUXNQ MoDPeZBz\We3bHH0[ZMhfGinIHzleoVteyCxZjDBUXBMKGGwZDDBR2MheGixc4Doc5J6dGG2aX;u MoK4QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjV3OUSwOFMoRjJ3NUm0NFQ{RC:jPh?=
PC3M MULLbY5ie2ViYYPzZZk> MVWxNFAhyrWP MlX5SG1UVw>? NHjYXGF2eHKnZ4XsZZRmeyC2aHWgcIV3\Wy|IH;mJGFOWEtiYX7kJGFESyCyaH;zdIhwenmuYYTpc44> NVfDN2pPRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkW1PVQxPDNpPkK1OVk1ODR|PD;hQi=>
PC3 Ml35SpVv[3Srb36gZZN{[Xl? MX2xNFAhyrWP M1zwb2ROW09? NXXhd|JucW6mdXPld{BRUTONL33UU3IheGG2aIfhfZM> NX\SeG43RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkW1PVQxPDNpPkK1OVk1ODR|PD;hQi=>
PC3M NFu2N3ZHfW6ldHnvckBie3OjeR?= NWn0elh5OTByINM1US=> MWHEUXNQ M135cIlv\HWlZYOgVGk{Uy:vVF;SJJBifGi5YYnz NYPGZ2NXRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkW1PVQxPDNpPkK1OVk1ODR|PD;hQi=>
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T47D MkTMS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? MoXYTY5pcWKrdHnvckBw\iCqdX3hckBVPDeGIHPlcIwh\3Kxd4ToJIlvKGFiY3XscEB3cWGkaXzpeJkh[XO|YYmsJGlEPTBiPTC0PU44ODB5IN88UU4> M2HvbVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFUyOjVyND:nQnNCVkeHUkyvZV4>
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Assay
Methods Test Index PMID
Western blot pCofilin / Cofilin / detyrosinated alpha tubulin / acetylated alpha tubulin ; p-AMPK / AMPK / p-S6K / S6K / p-ACC 26431377 22456226
In vivo Short-term treatment of normal Sprague Dawley rats with A-769662 decreases liver malonyl CoA levels and the respiratory exchange ratio, VCO2/VO2, indicating an increased rate of whole-body fatty acid oxidation. Treatment of ob/ob mice with 30 mg/kg b.i.d. A-769662 decreases hepatic expression of PEPCK, G6Pase, and FAS, lowers plasma glucose by 40%, reduced body weight gain and significantly decreases both plasma and liver triglyceride levels. [1]

Protocol (from reference)

Kinase Assay:

[1]

  • 96-well AMPK assay:

    AMPK activity is measured by monitoring phosphorylation of the SAMS peptide substrate (20 mM in standard assays and 100 mM in additivity assays) following a previously described protocol (Anderson et al., 2004). To determine whether A-769662-induced AMPK activation occurs in a reversible manner, AMP or A-769662 are preincubated with rat liver AMPK for 10 minutes at 20 times standard assay concentrations prior to dilution and measurement of AMPK activity.

Cell Research:

[3]

  • Cell lines: MEF cells
  • Concentrations: 300 μM
  • Incubation Time: 24 hours
  • Method:

    Cell viability of MEF cells treated or not with A-769662 is performed as follows: cells are harvested by trypsinization and incubated with 0.5 mg/mL RNase and 50 μg/mL propidium iodine at room temperature in the dark; cell viability is analyzed by flow cytometry using a FACScanto flow cytometer, using an excitation laser at 488 nm and a propidium iodine fluorescence detection at 600 nm. To determine the proportion of cells in each phase of the cell cycle, cells are harvested by trypsinization, collected by centrifugation, washed in PBS and fixed overnight in 80% ethanol at -20 °C. Subsequently, these fixed cells are centrifuged to remove the fixative and incubated for 20 minutes in the dark at room temperature in PBS containing 0.5 mg/mL RNase and 50 μg/mL propidium iodine. Flow cytometry analysis is performed as above. The proportion of cells in G1, S, and G2 is determined using the MODFIT program. Cell culture pictures are taken at the indicated times using a camera coupled to an inverted microscope with a 20 × objective.

Animal Research:

[1]

  • Animal Models: Sprague Dawley rats
  • Dosages: 30 mg/kg
  • Administration: Administered via i.p.

Solubility (25°C)

In vitro

DMSO 72 mg/mL
(199.78 mM)
Water Insoluble
Ethanol Insoluble

In vivo

Add solvents to the product individually and in order
(Data is from Selleck tests instead of citations):
5% DMSO+40% PEG 300+5% Tween 80+50% ddH2O
For best results, use promptly after mixing.

5 mg/ml

Chemical Information

Molecular Weight 360.39
Formula

C20H12N2O3S

CAS No. 844499-71-4
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles C1=CC=C(C(=C1)C2=CC=C(C=C2)C3=CSC4=C3C(=C(C(=O)N4)C#N)O)O

In vivo Formulation Calculator (Clear solution)

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

mg/kg g μL

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.

Molarity Calculator

Mass Concentration Volume Molecular Weight

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

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