Home PI3K/Akt/mTOR ATM/ATR inhibitor AZ20

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AZ20 ATM/ATR inhibitor

Cat.No.S7050

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.
AZ20 ATM/ATR inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 412.51

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Quality Control

Cell Culture, Treatment & Working Concentration

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
HT29 Function assay 1 hr Inhibition of ATR-mediated CHK1 phosphorylation at serine 345 in human HT29 cells after 1 hr in presence of 4-nitroquinoline 1-oxide, IC50=0.05μM 23394205
LoVo Growth inhibition assay 72 hrs Growth inhibition of human LoVo cells after 72 hrs by MTS assay, GI50=0.2μM 23394205
MDA-MB-468 Function assay Inhibition of mTOR-mediated AKT phosphorylation at serine 473 in human MDA-MB-468 cells, IC50=2.4μM 23394205
LoVo Antitumor assay 50 mg/kg 13 days Antitumor activity against human LoVo cells xenografted in Swiss nu/nu mouse assessed as tumor growth inhibition at 50 mg/kg, po qd for 13 days relative to control 23394205
LoVo Antitumor assay 25 mg/kg 13 days Antitumor activity against human LoVo cells xenografted in Swiss nu/nu mouse assessed as tumor growth inhibition at 25 mg/kg, po bid for 13 days relative to control 23394205
HT29 Function assay 60 mins Inhibition of ATR in human HT29 cells after 60 mins by Hoechst 33258 staining-based assay, IC50=0.061μM 30346772
LoVo Cytotoxicity assay 72 hrs Cytotoxicity against human LoVo cells after 72 hrs by MTS assay, GI50=0.2μM 30346772
MDA-MB-468 Function assay Inhibition of mTOR in human MDA-MB-468 cells assessed as decrease in 70S6K S235/236 phosphorylation, IC50=0.72μM 30346772
HT-29 Cytotoxicity assay 72 hrs Cytotoxicity against human HT-29 cells after 72 hrs by MTS assay, GI50=0.97μM 30346772
LoVo Function assay 25 mg/kg 8 hrs Plasma concentration in Swiss nu/nu mouse xenografted with human LoVo cells at 25 mg/kg, po bid after 8 hrs, Cp=1.8μM 30346772
MDA-MB-468 Function assay Inhibition of mTOR in human MDA-MB-468 cells assessed as decrease in AKT phosphorylation at S473 residue, IC50=2.4μM 30346772
LoVo Function assay 50 mg/kg 8 hrs Plasma concentration in Swiss nu/nu mouse xenografted with human LoVo cells at 50 mg/kg, po qd after 8 hrs, Cp=3.5μM 30346772
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Chemical Information, Storage & Stability

Molecular Weight 412.51 Formula

C21H24N4O3S

 Storage (From the date of receipt)
CAS No. 1233339-22-4 Download SDF Storage of Stock Solutions

Synonyms N/A Smiles CC1COCCN1C2=NC(=NC(=C2)C3(CC3)S(=O)(=O)C)C4=C5C=CNC5=CC=C4

Solubility

In vitro
Batch:

DMSO : 83 mg/mL (201.2 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : 3 mg/mL

Water : Insoluble

Molarity Calculator

Mass Concentration Volume Molecular Weight

In vivo
Batch:

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.
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Mechanism of Action

Features
ATR-selective inhibitor with high permeability and good stability.
Targets/IC50/Ki
ATR [2]
(Cell-free assay)
5 nM
mTOR [2]
(Cell-free assay)
38 nM
In vitro
AZ20 shows good selectivity against all of the PI3K isoforms together with ATM and DNA-PK. [2] In vitro, this compound decreases pChk1 Ser345, pChk1 Ser317 and pChk1 Ser296 levels in a concentration-dependent manner. Prolonged exposure with this chemical increases γH2AX pan-nuclear staining, indicative of replication stress. This is associated with S-phase arrest and increase in phospho-histone H3. It induces growth inhibition and cell death in vitro and its profile of activity is distinct from other cytotoxic agents. The cytotoxic effect of this agent can be increased in combination with the selective ATM inhibitor KU-60019. [1]
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] This compound is assessed for drug−drug interaction (DDI) potential specifically from inhibition of cytochrome P450 enzymes. It is found to inhibit the cytochrome 3A4-mediated metabolism of midazolam by 50% at 10 μM. This chemical has respectable bioavailability in a low dose rat PK study. [2]
References

Applications

Methods Biomarkers Images PMID
Growth inhibition assay IC50 S7050-viability1 28176818
Western blot p-CDK1 / CDK1 / p-CDK2 / CDK2 γH2AX / RRM1 / RRM2 S7050-WB1 28176818

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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 of it should I use?

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

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