AZ20

SKU: orb1306978

Description

AZ20 is a potent and selective ATR kinase inhibitor with an IC50 of 5 nM, demonstrating 8-fold selectivity over mTOR. This small molecule is a valuable chemical probe for investigating DNA damage response pathways in both cellular and biochemical assays.

Research Area

Molecular Biology, Signal Transduction

Key Properties

• CAS Number: 1233339-22-4
• MW: 412.51
• Purity: >99.99% (May vary between batches)
• Formula: C₂₁H₂₄N₄O₃S
• SMILES: C[C@@H]1COCCN1c1cc(nc(n1)-c1cccc2[nH]ccc12)C1(CC1)S(C)(=O)=O
• Target: mTOR,ATM/ATR
• Solubility: H2O:< 1 mg/mL (insoluble or slightly soluble);Ethanol:3 mg/mL (7.27 mM);DMSO:50 mg/mL (121.21 mM);10% DMSO+40% PEG300+5% Tween 80+45% Saline:2 mg/mL (4.85 mM)

Bioactivity

• Target IC50:
  mTOR:38 nM|ATR:5 nM|PI3Kα:13000 nM
• In Vivo:
  In vitro, the concentration of AZ20 correspondingly decreases the levels of pChk1 Ser345/317/296. Extending the treatment duration with AZ20 intensifies γH2AX nuclear staining due to replication stress, associated with S phase arrest and an increase in histone H3 phosphorylation levels. The ability of AZ20 to inhibit cellular growth and induce cell death is entirely distinct from other cytotoxic drugs. The cytotoxic effects are enhanced when AZ20 is used in combination with the ATM inhibitor KU-60019.
• In Vitro:
  AZ20 (10 μM) inhibits 50% of the metabolism of midazolam mediated by CYP3A4. In female nude mice with LoVo tumors, treatment with AZ20 (25 mg/kg, twice daily, orally; or 50 mg/kg/day, orally) for 13 days significantly suppresses tumor growth. This effect is associated with a sustained and extensive increase in γH2AX nuclear staining in xenografted tissues.
• Cell Research:
  AZ20 is dissolved in 100% DMSO. Compound dose ranges are created by diluting in 100% DMSO and then further into assay medium (EMEM, 10% FCS, 1% glutamine) using a Labcyte Echo acoustic dispensing instrument. Cells are plated in 384-well Costar plates at 9×104 cells per mL in 40 μL of EMEM, 10% FCS, 1% glutamine and grown for 24 h. Following addition of compound the cells are incubated for 60 min. A final concentration of 3 μM 4NQO (prepared in 100% DMSO) is then added using the Labcyte Echo, and the cells are incubated for a further 60 min. The cells are fixed by adding 40 μL of 3.7% v/v formaldehyde solution for 20 min. After removal of fix, cells are washed with PBS and permeabilized in 40 μL of PBS containing 0.1% Triton X-100. The cells are then washed, and 15 μL primary antibody solution (pChk1 Ser345) is added. The plates are incubated at 4°C overnight. The primary antibody is then washed off, and 20 μL of secondary antibody solution and 1 μM Hoechst 33258 added for 90 min at room temperature. The plates are washed and left in 40 μL of PBS. Plates are then read on an ArrayScan VTI instrument to determine staining intensities, and dose responses are obtained and used to determine the IC50 values for the compounds.

Storage & Handling

• Storage: Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice/Shipping at ambient temperature.
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

ATR, AZ 20, Ataxia telangiectasia mutated, ATM, ATM and RAD3 related, ATM/ATR, AZ20, AZ-20, Inhibitor, inhibit, mTOR

Compound Identifiers

PubChem CID

46244454