Search Site
Related Products
AZ20ATR inhibitor,potent and selective

AZ20

Catalog No. A3210
Size Price Stock Qty
10mM (in 1mL DMSO) $141.00 In stock
5mg $128.00 In stock
10mg $214.00 In stock
50mg $727.00 In stock

Tel: +1-832-696-8203

Email: sales@apexbt.com

Worldwide Distributors

Sample solution is provided at 25 µL, 10mM.

Product Citations

1. Shah, Govind A., and Clodagh C. O’Shea. "Viral and cellular genomes activate distinct DNA damage responses." Cell 162.5 (2015): 987-1002. PMID:26317467

Quality Control

Quality Control & MSDS

View current batch:

Chemical structure

AZ20

Related Biological Data

AZ20

Related Biological Data

AZ20

Biological Activity

Description AZ20 is a potent and selective inhibitor of ATR kinase with IC50 value of 5 nM.
Targets ATR kinase          
IC50 5 nM          

Protocol

Kinase experiment [1]:

Inhibitory activities

ATR for use in the vitro enzyme assay was obtained from HeLa nuclear extract by immunoprecipitation with rabbit polyclonal antiserum raised to amino acids 400-480 of ATR contained in the following buffer: 25 mM HEPES (pH 7.4), 2 mM MgCl2, 250 mM NaCl, 0.5 mM EDTA, 0.1 mM Na3VO4, 10% v/v glycerol, and 0.01% v/v Tween 20. ATR-antibody complexes were isolated from nuclear extract by incubating with protein A-Sepharose beads (Sigma, no. P3476) for 1 h and then through centrifugation to recover the beads. In the well of a 96-well plate, 10 μL ATR-containing Sepharose beads were incubated with 1 μg of substrate glutathione S-transferase-p53N66 in ATR assay buffer (50 mM HEPES (pH 7.4), 150 mM NaCl, 6 mM MgCl2, 4 mM MnCl2, 0.1 mM Na3VO4, 0.1 mM DTT, and 10% (v/v) glycerol) at 37℃ in the presence or absence of inhibitor. After 10 min with gentle shaking, ATP was added to a final concentration of 3 μM and the reaction continued at 37℃ for an additional 1 h. The reaction was stopped by addition of 100 μL of PBS, and the reaction was transferred to a white opaque glutathione coated 96-well plate and incubated overnight at 4℃. This plate was then washed with PBS/0.05% (v/v) Tween 20, blotted dry, and analyzed by a standard ELISA technique with a phosphoserine 15 p53 (16G78) antibody. The detection of phosphorylated glutathione S-transferase-p53N66 substrate was performed in combination with a goat anti-mouse horseradish peroxidase-conjugated secondary antibody.

Cell experiment [1]:

Cell lines

BT-474 cells; HT29 colorectal adenocarcinoma cells.

Preparation method

The solubility of this compound in DMSO is >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reacting condition

60 min

Applications

AZ20 inhibits pAKT T308 in BT-474 cells and inhibits pATM Ser-1981 and pDNA-PK Ser-2056 in HT29 cells. In LoVo colorectal adenocarcinoma cells, AZ20 induces growth inhibition with GI50 value of 0.20 μM.

Animal experiment [1]:

Animal models

Female nude mice bearing LoVo tumors.

Dosage form

25 mg/kg twice daily or 50 mg/kg once daily for 13 days; administered orally.

Preparation method

Dissolved in 10% DMSO/40% propylene glycol/50% water.

Application

AZ20 significantly inhibits tumor growth. Body weight loss is in an acceptable range and AZ20 is generally well tolerated.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

References:

[1]. Discovery of 4-{4-[(3R)-3-Methylmorpholin-4-yl]-6- [1-(methylsulfonyl)cyclopropyl]pyrimidin-2-yl}-1H-indole (AZ20): a potent and selective inhibitor of ATR protein kinase with monotherapy in vivo antitumor activity. J Med Chem, 2013, 56(5): 2125-2138.

AZ20 Dilution Calculator

Concentration (start)
x
Volume (start)
=
Concentration (final)
x
Volume (final)
 
 
 
C1
V1
C2
V2

calculate

AZ20 Molarity Calculator

Mass
=
Concentration
x
Volume
x
MW*
 
 
 
g/mol

calculate

Chemical Properties

Cas No. 1233339-22-4 SDF Download SDF
Synonyms AZ-20;AZ 20
Chemical Name (3R)-4-[2-(3H-indol-4-yl)-6-(1-methylsulfonylcyclopropyl)pyrimidin-4-yl]-3-methylmorpholine
Canonical SMILES CC1COCCN1C2=NC(=NC(=C2)C3(CC3)S(=O)(=O)C)C4=C5CC=NC5=CC=C4
Formula C21H24N4O3S M.Wt 412.51
Solubility Soluble in DMSO > 10 mM Storage Store at -20°C
General tips For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.
Shipping Condition Evaluation sample solution : ship with blue ice
All other available size: ship with RT , or blue ice upon request

Background

AZ20 is a potent and selective inhibitor of ATR. AZ20 inhibits ATR immunoprecipitated from HeLa nuclear extracts with an IC50 value of 5 nM and ATR mediated phosphorylation of Chk1 in HT29 colorectal adenocarcinoma tumor cells with an IC50 value of 50 nM.
ATR is an attractive new anticancer drug target whose inhibitors have potential as chemo- or radiation sensitizers or as monotherapy in tumors addicted to particular DNA-repair pathways.
AZ20 is the first reported inhibitor of ATR protein kinase demonstrating tumor growth inhibition in vivo and is therefore a useful probe molecule to aid further investigation of ATR tumor biology. AZ20 potently inhibited the growth of LoVo colorectal adenocarcinoma tumor cells in vitro. In the mTOR (pAKT) cell assay, AZ20 weak inhibited recombinant mTOR enzyme activity. AZ20 shows good selectivity against all of the PI3K isoforms together with ATM and DNA-PK, and when tested in a large panel of kinases, AZ20 shows very high general kinase selectivity.
Female nude mice bearing LoVo tumors were treated with AZ20 orally at a dose of 25 mg/kg twice daily or 50 mg/kg once daily for 13 days. AZ20 showed monotherapy in vivo antitumor efficacy in LoVo colorectal xenografts in nude mice.
References:
[1]. Foote KM, Blades K, Cronin A et al. Discovery of 4-{4-[(3R)-3-Methylmorpholin-4-yl]-6- [1-(methylsulfonyl)cyclopropyl]pyrimidin-2-yl}-1H-indole (AZ20): a potent and selective inhibitor of ATR protein kinase with monotherapy in vivo antitumor activity. J Med Chem. 2013 Mar 14;56(5):2125-38.