KU-55933 (ATM Kinase Inhibitor)

For research use only.

Catalog No.S1092

234 publications

KU-55933 (ATM Kinase Inhibitor) Chemical Structure

CAS No. 587871-26-9

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.

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Selleck's KU-55933 (ATM Kinase Inhibitor) has been cited by 234 publications

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Choose Selective ATM/ATR Inhibitors

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
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T47D  M2fHSWZ2dmO2aX;uJGF{e2G7 MX:yNOKhdU1? M4\FOlI1KGh? M{fTUmROW09? MmXYdJJmfmWwdIOgTXIucW6mdXPl[EBl\We{YXTheIlwdiCxZjDJ{tpD|rF? M2LkdlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJzMUS0PFA2Lz5{MUG0OFgxPTxxYU6=
A29 MEF M4r1bWZ2dmO2aX;uJGF{e2G7 MYOxNOKh|ryPwrC= M1LVPFFp Ml\DZoxw[2u|wrD0bIUheGixc4Doc5J6dGG2aX;uJI9nKEGtdDDheEBU\XJ2N{RCpC=> NXyxWY1WRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkCwOVM4QDFpPkKwNFU{PzhzPD;hQi=>
MDA-MB-453  NE\3TYlIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= Mom0OU01OCEQvF2= NYi2eJVvPzJiaB?= NFXac3pKSzVyIH;mJFExKM7:TR?= MnPRQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjByNUO3PFEoRjJyMEWzO|gyRC:jPh?=
PC-3 NHfKSGRIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NIm3RWU2NTRyIN88US=> MonjO|IhcA>? NF\JS5pKSzVyIH;mJFExKM7:TR?= NVrpbZZRRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkCwOVM4QDFpPkKwNFU{PzhzPD;hQi=>
U2OS MYTGeY5kfGmxbjDhd5NigQ>? NIi0V2JKdmirYnn0bY9vKG:oIFHUUUBqdiCqdX3hckBWOk:VIHPlcIx{KGG|c3Xzd4VlKGG|IHnubIljcXSrb36gc4YheDV|IIDoc5NxcG:{eXzheIlwdiCjdDDT[ZIyPSC{ZYPp[JVmNCCLQ{WwJF0hOC5{NTFOwG0v M1HM[VxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ4NkOyPVY2Lz5{Nk[zNlk3PTxxYU6=
MCF7 M3i2V2Z2dmO2aX;uJIF{e2G7 MofLNUBpeg>? MU\Jcohq[mm2aX;uJI9nKEGWTTDrbY5ie2ViaX6gbJVu[W5iTVPGO{Bk\WyuczDh[pRmeiBzIHjyJIJ6KGmvbYXuc4ZtfW:{ZYPj[Y5k\SCjc4PhfUwhUUN3MDC9JFAvOyEQvF2u MojQQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjZ4M{K5OlUoRjJ4NkOyPVY2RC:jPh?=
BJ MlLqSpVv[3Srb36gZZN{[Xl? MUKxNEB2VQ>? MlLkNVAh\GG7cx?= M{fY[HN2eHC{ZYPzbY9vKG:oIIPlcoV{[2WwY3WgbY4hcHWvYX6gRmoh[2WubIOgZZN{\XO|ZXSgZZMhcW6lcnXhd4UhcW5iY3XscEBvfW2kZYKgZZQhOTBidV2gZYZ1\XJiMUCg[IF6eyCkeTDz[Y5me2OnbnPlJJJmfmW{c3HsJIF{e2G7 NILpWFk9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zNke2O|A5PSd-MU[3OlcxQDV:L3G+
BJ MoT5SpVv[3Srb36gZZN{[Xl? MmHtNVAhfU1? Ml;INVAh\GG7cx?= NYrnSHZIUW6qaXLpeIlwdiCxZjDheIF5cWFidHXsZY5ocWWldHHzbYEudXW2YYTl[EBqdiCqdX3hckBDUiClZXzsd{Bie3Onc4Pl[EBieyCrbnPy[YF{\SCrbjDj[YxtKG63bXLldkBifCBzMDD1UUBi\nSncjCxNEBl[Xm|IHL5JJNmdmW|Y3XuZ4UhemW4ZYLzZYwh[XO|YYm= Mn3FQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTZ5NkewPFUoRjF4N{[3NFg2RC:jPh?=
MCF7 NHrGclBHfW6ldHnvckBie3OjeR?= Mlz4NVAhfU1? M2eyXVExKG2rboO= NWfIXlN1W2Wwc3n0bZpifGmxbjDv[kBqdm[{YYLl[E1qdmS3Y3XkJGRPSSCmYX3h[4UhcW5iaIXtZY4hVUOINzDj[YxteyCjc4Pld5Nm\CCjczDy[YR2[3Srb36gbY4h[2:ub375JIZwem2jdHnvckBifCBzMDD1UUBxemW2cnXheIVlKG[xcjCxNEBucW6|IH\vcIxwf2WmIHL5JIlzemGmaXH0bY9vKG[xcjC0JIhzeyCvZXHzeZJm\CCjZoTldkAyOCCmYYnzJIJ6KGO{eYP0ZYwhfmmxbHX0JJN1[WmwaX7nJIFv[Wy7c3nz MkLJQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjZ4M{K5OlUoRjJ4NkOyPVY2RC:jPh?=
SK-N-MC MWDxTHRUKGG|c3H5 NGS5dHhyUFSVIH;mJJBm\GmjdILpZ{Bk[W6lZYKgZ4VtdCCuaX7ld{B1dyCrZHXueIlngSCvdXz0bZBt\SCxcIDvdpR2dmm2aXXzJIZweiCmcoXnJJJmeHW{cH;zbY5oQiCScnntZZJ6KHOlcnXlckBnd3JiU1utUk1OSyClZXzsdy=> NYHnfGNxRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkm0N|UyOzlpPkK5OFM2OTN7PD;hQi=>

... Click to View More Cell Line Experimental Data

Assay
Methods Test Index PMID
Western blot
p-AKT(Ser473) / p-AKT(Thr308); 

PubMed: 22739265     


Cells were serum-starved for 6 h and treated with KU-55933 (10 µmol/L) for 1 h before treatment with insulin (100 nmol/L) for 45 min. Cells were lysed, and cell lysates were subjected to SDS-PAGE and Western blotting for the detection of phospho-Akt at Se䲧疝Ỵ疞㧀疜膉痘 瘿�෋ᾰƌ෋à 㺣痖

PARP / Cleaved PARP / Caspase-3 / Cleaved caspase-3 ; 

PubMed: 22739265     


ATM inhibition by KU-55933 induces apoptosis in differentiated but not undifferentiated SH-SY5Y cells. Cells were starved for 6 h in serum-free medium. KU-55933 (10 µmol/L) was added 1 h before insulin (100 nmol/L) treatment. After 3 h of insulin stimulat䲧疝Ỵ疞㧀疜膉痘 瘿�෋ᾰƌ෋à 㺣痖帉痖Ѐ瑖堘𢡄빢᎒෋à鑸᎒彿堙奋堙巫堙᎒ﻺ᎒彿堙ﻮ᎒塚堙ﻺ᎒ꍈ堞빢᎒學堙漸堞圔堙빢᎒圞堙圭堙𢡄玚Wᾰƌ ᾰƌ戤

ATM-S1981 / ATM / p-p53(S15) / p53 ; 

PubMed: 20639198     


Antioxidant and ATM inhibitors blocked phosphorylation of ATM-S1981, Akt-S473, and p53-S15 as stimulated by H2O2 exposure. Cells were treated with 100 μm H2O2 for 30 min in the absence or presence of KU-55933. Whole cell lysates were subjected to Western 䲧疝Ỵ疞㧀疜膉痘 瘿뾠ՂᾰƌՂĀ 㺣痖

p21 / p27 / p53 ; 

PubMed: 20177072     


HMGE cells were incubated for the indicated times with 1 μm KU-55933 or DMSO. Total protein extracts were analyzed by Western blotting for p21WAF1/CIP1, p27KIP1, p53, or β-actin.

22739265 20639198 20177072
Immunofluorescence
p-S824 KAP1 / ZEBRA ; 

PubMed: 28249048     


HH514-16 cells were treated with NaB (upper panel) or NaB plus KU-55933 (1μM; middle panel) or NaB plus Torin1 (0.5μM; lower panel) for 24 hours and stained with anti-phospho KAP1 (S824) plus anti-ZEBRA antibodies and visualized at 1000X magnification.

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

Kinase Assay:

[1]

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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]

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  • 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.


    (Only for Reference)
Animal Research:

[3]

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  • Animal Models: BALB/c nu/nu nude mice bearing LU1205 cells
  • Dosages: 10 μM
  • Administration: --
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 33 mg/mL (83.44 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%PEG300+5%Tween80+50%ddH2O
For best results, use promptly after mixing.
3.95 mg/mL

* 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 395.49
Formula

C21H17NO3S2

CAS No. 587871-26-9
Storage powder
in solvent
Synonyms N/A
Smiles C1COCCN1C2=CC(=O)C=C(O2)C3=C4C(=CC=C3)SC5=CC=CC=C5S4

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID