KU-55933 (ATM Kinase Inhibitor)

Catalog No.S1092

For research use only.

KU-55933 (ATM Kinase Inhibitor) is a potent and specific ATM inhibitor with IC50/Ki of 12.9 nM/2.2 nM in cell-free assays, and is highly selective for ATM as compared to DNA-PK, PI3K/PI4K, ATR and mTOR. KU‑55933 (ATM Kinase Inhibitor) inhibits the activation of autophagy‑initiating kinase ULK1 and results in a significant decrease of autophagy.

KU-55933 (ATM Kinase Inhibitor) Chemical Structure

CAS No. 587871-26-9

Selleck's KU-55933 (ATM Kinase Inhibitor) has been cited by 282 publications

Purity & Quality Control

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Biological Activity

Description KU-55933 (ATM Kinase Inhibitor) is a potent and specific ATM inhibitor with IC50/Ki of 12.9 nM/2.2 nM in cell-free assays, and is highly selective for ATM as compared to DNA-PK, PI3K/PI4K, ATR and mTOR. KU‑55933 (ATM Kinase Inhibitor) inhibits the activation of autophagy‑initiating kinase ULK1 and results in a significant decrease of autophagy.
Targets
ATM [1]
(Cell-free assay)
12.9 nM
In vitro

KU-55933 inhibits DNA-PK and PI3K with IC50 of 2.5 μM and 16.6 μM, respectively. Besides, KU-55933 also prevents the activity of mTOR with IC50 of 9.3 μM. KU-55933 is active at the cellular level in ablating a well-characterized ATM-dependent phosphorylation event. KU-55933 has a dose-dependent effect in inhibiting this ATM-dependent phosphorylation event with IC50 of 300 nM. KU-58050 does not prevent the ATM-dependent phosphorylation of p53 serine 15 until a dose of 30 μM. Addition of KU-55933 has no appreciable effects on UV-induced phosphorylation of H2AX on serine 139, NBS1 on serine 343, CHK1 on serine 345, and SMC1 on serine 966. In stark contrast to the UV responses, KU-55933 ablates the ionizing radiation-induced phosphorylation of these ATM substrates. KU-55933 sensitizes HeLa cells to a range of ionizing radiation doses. [1] KU-55933 inhibits the phosphorylation of Akt induced by growth factors in cancer cells. KU-55933 suppresses the proliferation of cancer cells. Furthermore, suppression of ATM by KU-55933 improves survival, probably via prevention of downstream activation of TAp63α. [2]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
DU-145 M3LUZWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NF3oXYJKSzVyPUOuNlc{PTJizszN NFHWR3c9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmwzOjJzMEKvK|5USU6JRWK8M4E,
HuO-3N1 NXj5WopMT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NEHVdI5KSzVyPUSuNVcyPDJizszN NX7lbGFtRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNOjJ{MUCyM{c,W0GQR1XSQE9iRg>?
LAMA-84 MXTHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MW\JR|UxRTRwNUi0OlUh|ryP Mnn5QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMNlIzOTB{Lze+V2FPT0WUPD;hQi=>
CAL-72 NXnCfFBDT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MWTJR|UxRTVwNEiwPFQh|ryP NF74Omo9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmwzOjJzMEKvK|5USU6JRWK8M4E,
LoVo MVjHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NYDZelNDUUN3ME22Mlk{OjN7IN88US=> NWq2TYdbRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNOjJ{MUCyM{c,W0GQR1XSQE9iRg>?
HH M{nzcGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MmDJTWM2OD16LkK3OlcyKM7:TR?= NWTtO5J3RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNOjJ{MUCyM{c,W0GQR1XSQE9iRg>?
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KM12 NU\CWHlZT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MluxTWM2OD17LkKxNVQzKM7:TR?= M2\rTFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFIzOjFyMj:nQnNCVkeHUkyvZV4>
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LB2241-RCC M4nBVmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NHOzTXFKSzVyPUGxMlcyQDZizszN MX[8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyyNlIyODJxJ{7TRW5ITVJ:L3G+
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CAL-12T NUjz[XQ{T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MorMTWM2OD1zMz62NVch|ryP M33VWlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFIzOjFyMj:nQnNCVkeHUkyvZV4>
COLO-684 M4rsdWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 M3ThUmlEPTB;MUSuNVU3QSEQvF2= MnrjQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMNlIzOTB{Lze+V2FPT0WUPD;hQi=>
DOK M3fId2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MX7JR|UxRTF3LkOzNlkh|ryP NVmzXG1ZRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNOjJ{MUCyM{c,W0GQR1XSQE9iRg>?
Hs-578-T MVTHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? M3HYRmlEPTB;MUWuOFE5OiEQvF2= NU\OeWlERGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNOjJ{MUCyM{c,W0GQR1XSQE9iRg>?
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KG-1 NHv1UZBIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NUC1WJZMUUN3ME2xOk4xQTl4IN88US=> MU[8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyyNlIyODJxJ{7TRW5ITVJ:L3G+
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ChaGo-K-1 MV3Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MlnKTWM2OD1zNj62OVY5KM7:TR?= M1z0O|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFIzOjFyMj:nQnNCVkeHUkyvZV4>
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DMS-273 M{Dab2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NUXNendwUUN3ME20OE44PTZ5IN88US=> MWG8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyyNlIyODJxJ{7TRW5ITVJ:L3G+
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BALL-1 NFzUOFZIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NIfvV|hKSzVyPUS3Mlg{OiEQvF2= NGTuU|I9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmwzOjJzMEKvK|5USU6JRWK8M4E,
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MFE-280 MV3Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MkLkTWM2OD12OT60OlE4KM7:TR?= M{DXblxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFIzOjFyMj:nQnNCVkeHUkyvZV4>
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BB30-HNC NHPNbnJIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MlfuTWM2OD12OT65OFk5KM7:TR?= M{jHWVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFIzOjFyMj:nQnNCVkeHUkyvZV4>
SK-N-DZ NEHYW3dIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= M1G2UGlEPTB;NUCuNFQ5OSEQvF2= NH;NUJk9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmwzOjJzMEKvK|5USU6JRWK8M4E,
HepG2  MXzHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NXrxR4JSOTEEoN88US=> MUeyOEBp NULmS5Fs[myxY3vzJHNENUmLSUOtbY5lfWOnZDDTJJBp[XOnIHHydoV{fA>? M3PZSlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ3NUK3NVI{Lz5{NUWyO|EzOzxxYU6=
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HeLa  MXjHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? M3;lN|EuOTByzszN MWKyOEBp MnTwSmJU NV;WVFBIcW6qaXLpeJMhfGinIHPlcIwheHKxbHnm[ZJifGmxbh?= NHPqS4U9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{M{G4OVM1Pyd-MkOxPFU{PDd:L3G+
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IMR-32 M4\m[2Z2dmO2aX;uJGF{e2G7 NX;VSod4OTEEoN88UeKh MoK3NlQhcA>? M37tT4lvcGmkaYTzJINtcW:zdXnuc4wucW6mdXPl[EBxcG:|cHjvdplt[XSrb36gc4YheDV| NWDWcY9mRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkK2NlczQTRpPkKyOlI4Ojl2PD;hQi=>
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T47D  NHWyc3BHfW6ldHnvckBCe3OjeR?= NWTiPHM{OjEEoH3N MWWyOEBp Mnq2SG1UVw>? M1jxO5Bz\X[nboTzJGlTNWmwZIXj[YQh\GWpcnHkZZRqd25ib3[gTe67Ss7z NGH0UGw9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MUG0OFgxPSd-MkGxOFQ5ODV:L3G+
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MDA-MB-453  MUnHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MljPOU01OCEQvF2= MYe3NkBp NEXmVndKSzVyIH;mJFExKM7:TR?= MnTKQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjByNUO3PFEoRjJyMEWzO|gyRC:jPh?=
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MCF7 MULGeY5kfGmxbjDhd5NigQ>? Mn;NNUBpeg>? MX3Jcohq[mm2aX;uJI9nKEGWTTDrbY5ie2ViaX6gbJVu[W5iTVPGO{Bk\WyuczDh[pRmeiBzIHjyJIJ6KGmvbYXuc4ZtfW:{ZYPj[Y5k\SCjc4PhfUwhUUN3MDC9JFAvOyEQvF2u MoDUQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjZ4M{K5OlUoRjJ4NkOyPVY2RC:jPh?=
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MCF7 NUDaOFV6TnWwY4Tpc44h[XO|YYm= MVmxNEB2VQ>? MUCxNEBucW6| NFXnT2NU\W6|aYTpfoF1cW:wIH;mJIlv\nKjcnXkMYlv\HWlZXSgSG5CKGSjbXHn[UBqdiCqdX3hckBOS0Z5IHPlcIx{KGG|c3Xzd4VlKGG|IILl[JVkfGmxbjDpckBkd2yxbomg[o9zdWG2aX;uJIF1KDFyIIXNJJBz\XS{ZXH0[YQh\m:{IEGwJI1qdnNiZn;scI94\WRiYomgbZJz[WSrYYTpc44h\m:{IESgbJJ{KG2nYYP1doVlKGGodHXyJFExKGSjeYOgZpkh[3K7c4ThcEB3cW:uZYSgd5RicW6rbnegZY5idHm|aYO= MVq8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPjZ|Mkm2OUc,OjZ4M{K5OlU9N2F-
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Assay
Methods Test Index PMID
Western blot p-AKT(Ser473) / p-AKT(Thr308) ; PARP / Cleaved PARP / Caspase-3 / Cleaved caspase-3 ; ATM-S1981 / ATM / p-p53(S15) / p53 ; p21 / p27 / p53 22739265 20639198 20177072
Immunofluorescence p-S824 KAP1 / ZEBRA 28249048
In vivo Suppression of ATM-dependent STAT3 activation by KU-55933 enhances TRAIL-mediated apoptosis through up-regulation of surface DR5 expression, whereas suppression of both STAT3 and NF-κB appeares to be involved in down-regulation of cFLIP accompanied by an additional increase in apoptotic levels. The ATM inhibitor KU-55933 affectes TRAIL-mediated apoptosis more strongly than the JAK2 inhibitor, AG490, or overexpression of STAT3β. [3]

Protocol (from reference)

Kinase Assay:

[1]

  • Purified enzyme assays:

    ATM for use in the in vitro assay is obtained from HeLa nuclear extract by immunoprecipitation with rabbit polyclonal antiserum raised to the COOH-terminal 400 amino acids of ATM in buffer containing 25 mM HEPES (pH 7.4), 2 mM MgCl2, 250 mM KCl, 500 μM EDTA, 100 μM Na3VO4, 10% v/v glycerol, and 0.1% v/v Igepal. ATM-antibody complexes are isolated from nuclear extract by incubating with protein A-Sepharose beads for 1 hour and then through centrifugation to recover the beads. In the well of a 96-well plate, ATM-containing Sepharose beads are incubated with 1 μg of substrate glutathione S-transferase–p53N66 (NH2-terminal 66 amino acids of p53 fused to glutathione S-transferase) in ATM assay buffer [25 mM HEPES (pH 7.4), 75 mM NaCl, 3 mM MgCl2, 2 mM MnCl2, 50 μM Na3VO4, 500 μM DTT, and 5% v/v glycerol] at 37 °C in the presence or absence of inhibitor. After 10 minutes with gentle shaking, ATP is added to a final concentration of 50 μM and the reaction continued at 37 °C for an additional 1 hour. The plate is centrifuged at 250 × g for 10 minutes (4 °C) to remove the ATM-containing beads, and the supernatant is removed and transferred to a white opaque 96-well plate and incubated at room temperature for 1.5 hours to allow glutathione S-transferase-p53N66 binding. This plate is then washed with PBS, blotted dry, and analyzed by a standard ELISA technique with a phospho-serine 15 p53 antibody. The detection of phosphorylated glutathione S-transferase-p53N66 substrate is performed in combination with a goat antimouse horseradish peroxidase-conjugated secondary antibody. Enhanced chemiluminescence solution is used to produce a signal and chemiluminescent detection is carried out.

Cell Research:

[1]

  • Cell lines: U2OS cells
  • Concentrations: 10 μM
  • Incubation Time: 2 hours
  • Method:

    U2OS cells are exposed to ionizing radiation (3, 5, or 15 Gy) or UV (5 or 50 J/m2) and the ATM response determined by Western blot analysis of p53 serine 15 phosphorylation and stabilization of wild-type p53. Whole cell extracts are obtained from each time point, proteins separated by SDS-PAGE, and the ATM-specific increase in phosphorylated serine 15 measured with a p53 phospho-serine 15 specific antibody. Overall p53 stabilization with time is also observed with a p53-specific antibody (DO-1). Similarly, for studying ATM-dependent phosphorylations on H2AX, CHK1, NBS1, and SMC1, the following antibodies are used: CHK1 phospho-serine 345 and NBS1 phospho-serine 343 antibodies. Histone H2A (H-124) and CHK1 antibodies are also used, as well as SMC1 and SMC1 phospho-serine 966 antibodies. For determination of a cellular IC50 for KU-55933, the peak response time for p53 serine 15 phosphorylation of 2 hours is used to monitor inhibition of ATM. KU-55933 is titrated onto cells and preincubated for 1 hour before ionizing radiation. Using scanning densitometry, the percentage inhibition relative to vehicle control is calculated, and the IC50 value is calculated as for the in vitro determinations.

Animal Research:

[3]

  • Animal Models: BALB/c nu/nu nude mice bearing LU1205 cells
  • Dosages: 10 μM
  • Administration: --

Solubility (25°C)

In vitro

In vivo

Add solvents to the product individually and in order
(Data is from Selleck tests instead of citations):
5%DMSO+40%PEG300+5%Tween80+50%ddH2O
For best results, use promptly after mixing.

3.95 mg/mL

Chemical Information

Molecular Weight 395.49
Formula

C21H17NO3S2

CAS No. 587871-26-9
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles C1COCCN1C2=CC(=O)C=C(O2)C3=C4C(=CC=C3)SC5=CC=CC=C5S4

In vivo Formulation Calculator (Clear solution)

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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
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Molarity Calculator

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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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