For research use only.

Catalog No.S7050

28 publications

AZ20 Chemical Structure

CAS No. 1233339-22-4

AZ20 is a novel potent and selective inhibitor of ATR kinase with IC50 of 5 nM in a cell-free assay, 8-fold selectivity over mTOR.

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Selleck's AZ20 has been cited by 28 publications

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Biological Activity

Description AZ20 is a novel potent and selective inhibitor of ATR kinase with IC50 of 5 nM in a cell-free assay, 8-fold selectivity over mTOR.
Features ATR-selective inhibitor with high permeability and good stability.
ATR [2]
(Cell-free assay)
mTOR [2]
(Cell-free assay)
5 nM 38 nM
In vitro

AZ20 shows good selectivity against all of the PI3K isoforms together with ATM and DNA-PK. [2] In vitro, AZ20 decreases pChk1 Ser345, pChk1 Ser317 and pChk1 Ser296 levels in a concentration-dependent manner. Prolonged exposure with AZ20 increases γH2AX pan-nuclear staining, indicative of replication stress. This is associated with S-phase arrest and increase in phospho-histone H3. AZ20 induces growth inhibition and cell death in vitro and its profile of activity is distinct from other cytotoxic agents. The cytotoxic effect of AZ20 can be increased in combination with the selective ATM inhibitor KU-60019. [1]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
HT29 cells M4DJ[2Z2dmO2aX;uJIF{e2G7 Mn3wNUBp M1LUO:KhUW6qaXLpeIlwdiCxZjDBWHIudWWmaXH0[YQhS0iNMTDwbI9{eGixconsZZRqd25iYYSgd4VzcW6nIEO0OUBqdiCqdX3hckBJXDJ7IHPlcIx{KGGodHXyJFEhcHJiaX6gdJJme2WwY3Wgc4YhPC2waYTyc5F2cW6xbHnu[UAyNW:6aXTlMEBKSzVyPUCuNFUh|ryP MnXBNlM{QTR{MEW=
LoVo cells Mln5S5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? M3m4XFczKGh? NIXzbI5Iem:5dHigbY5pcWKrdHnvckBw\iCqdX3hckBNd1[xIHPlcIx{KGGodHXyJFczKGi{czDifUBOXFNiYYPzZZktKEeLNUC9NE4zKM7:TR?= Mlu5NlM{QTR{MEW=
human MDA-MB-468 cells Mn;ySpVv[3Srb36gZZN{[Xl? MmXDTY5pcWKrdHnvckBw\iCvVF;SMY1m\GmjdHXkJGFMXCCyaH;zdIhwenmuYYTpc44h[XRic3XybY5mKDR5MzDpckBpfW2jbjDNSGEuVUJvNE[4JINmdGy|LDDJR|UxRTJwNDFOwG0> M2\jNFI{Ozl2MkC1

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Methods Test Index PMID
Growth inhibition assay

PubMed: 28176818     

(a and b) AML cell lines and primary patient samples were treated with variable concentrations of AZ20 in 96-well plates for 72 h and viable cells were determined using MTT reagent. IC50 values were calculated as drug concentration necessary to inhibit 50% OD590 compared to vehicle control treated cells. AML cell line data are graphed as mean values ± SEM from three independent experiments (panel a). For the patient samples, the IC50values are mean values of duplicates from one experiment due to limited sample. The horizontal lines indicate the median. (c) AML cell lines and primary patient sample AML#53 were treated with AZ20 for 24 h and then subjected to annexin V-FITC/PI staining and flow cytometry analyses. Mean percent annexin V + cells ± SEM from one representative experiment performed in triplicates are shown. For cell lines, experiments were repeated three times, while patient sample experiments were performed once due to limited available sample.

Western blot
p-CDK1 / CDK1 / p-CDK2 / CDK2 ; 

PubMed: 28176818     

OCI-AML3 and THP-1 cells were treated with 0–8 μM AZ20 for 24 h. Whole cell lysates were subjected to Western blotting and probed with the indicated antibodies. Densitometry measurements normalized to β-actin and then compared to vehicle control are presented. Western blots were repeated at least three times and one representative cropped blot is shown.

γH2AX / RRM1 / RRM2 ; 

PubMed: 28176818     

OCI-AML3 cells were treated with 8 μM AZ20 for 0, 2, 4, 8, 12 or 24 h. Whole cell lysates were subjecte d to Western blotting and probed with the indicated antibodies (panel c).

In vivo Female nude mice bearing LoVo tumors are treated with AZ20 orally at a dose of 25 mg/kg twice daily or 50 mg/kg once daily for 13 days, led to significant tumor growth inhibition. [2] This is associated with a persistent elevation of γH2AX pan-nuclear staining in xenograft tissue, but a transient increase in mouse bone marrow at therapeutic doses, suggesting a favourable therapeutic index. [1] AZ20 is assessed for drug−drug interaction (DDI) potential specifically from inhibition of cytochrome P450 enzymes. AZ20 is found to inhibit the cytochrome 3A4-mediated metabolism of midazolam by 50% at 10 μM. AZ20 has respectable bioavailability in a low dose rat PK study. [2]


Animal Research:[2]
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  • Animal Models: LoVo colorectal adenocarcinoma xenografts
  • Dosages: 25 mg/kg twice daily and 50 mg/kg once daily
  • Administration: orally
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 83 mg/mL (201.2 mM)
Ethanol 3 mg/mL (7.27 mM)
Water 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 412.51


CAS No. 1233339-22-4
Storage powder
in solvent
Synonyms N/A
Smiles CC1COCCN1C2=NC(=NC(=C2)C3(CC3)S(=O)(=O)C)C4=C5C=CNC5=CC=C4

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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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Frequently Asked Questions

  • Question 1:

    If I want to completely block the kinase activity from the in vitro cell lines, how much concentration I should use?

  • Answer:

    IC50 5nM was quoted from a previous publication in which the author tested IC50 of AZ20 in cell free assay. In cell culture, many factors, such as membrane permeability and target protein concentration, may affect the efficiency. Each cell line responses to the same compound differently and it is very difficult to predict the optimized concentration simply based on cell free data. In cell culture experiment, the required concentration is usually higher. We recommend that you perform a pilot experiment and test different concentrations (50nM to 500uM) to get the optimized condition.

ATM/ATR Signaling Pathway Map

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID