ETP-46464_DataSheet_MedChemExpress

Inhibitors, Agonists, Screening Libraries

https://www.wendangku.net/doc/3a8471893.html, Data Sheet

BIOLOGICAL ACTIVITY:

ETP–46464 is a cell–permeable quinoline–containing heterotricyclic compound that acts as a potent inhibitor against mTOR, ATR,DNA–PK, PI 3–Kα, and ATM (IC50= 0.6, 14, 36, 170, and 545 nM, respectively).

IC50 Value: 0.6 nM (mTOR); 14 nM (ATR) [1]

Target: ATR; mTOR

ETP–46464 preferentially suppresses radiation–induced cellular ATR activity (>90% at 500 nM) over ATM or DNA–PK activity (IC50 > 5uM) in U2OS cells, while exhibiting much reduced or little potency toward a panel of 26 other kinases (>55% inhibition at 5 uM).ETP–46464 is Shown to synergize with UCN–01 in replicative stress induction in S–phase replicating U2OS (1.5–, 61–, and 190–fold of control RS, respectively, with 1 uM ETP–46464, 50 nM UCN–01, or combined treatment).

PROTOCOL (Extracted from published papers and Only for reference)

Cell assay [1]

For G2/M checkpoint analysis cells were incubated with the various compounds 15 min before ionizing radiation. Concentrations were as indicated. One hour later cells were collected and fixed in suspension in ice–cold 70% (v/v) ethanol in PBS. After a wash in PBS, cells were incubated with an antibody recognizing Ser10–phosphorylated histone H3 in PBS containing 1% (w/v) BSA and 0.1% (v/v)

Tween–20 for 2 h at room temperature. After one wash with the same solution and incubation with Alexa 488–conjugated secondary antibody, cells were resuspended in a PBS solution containing 1% (w/v) BSA, propidium iodide (10 μg/ml) and RNase A (0.5 mg/ml)and were analyzed by flow cytometry in a Becton–Dickinson FACSCalibur machine. To monitor the hydroxyurea–induced reversible replication block, cells were incubated with EdU for 1 h at 37°C. After fixation with 4% (w/v) paraformaldehyde for 30min, cells were processed with a Click–iT Edu Flow Cytometry kit as recommended by the manufacturer. Finally, cells were resuspended in a PBS solution containing 1% (w/v) BSA, propidium iodide (10μg/ml) and RNase A (0.5 mg/ml) and were analyzed by flow cytometry in a Becton–Dickinson FACSCalibur machine.

References:

[1]. Toledo, Luis I.; Murga, Matilde; Zur, Rafal; et al. A cell–based screen identifies ATR inhibitors with synthetic lethal properties for cancer–associated mutations. Nature Structural & Molecular Biology (2011), 18(6), 721–727.

Product Name:

ETP–46464Cat. No.:

HY-15521CAS No.:

1345675-02-6Molecular Formula:

C 30H 22N 4O 2Molecular Weight:

470.52Target:

ATM/ATR; ATM/ATR; mTOR Pathway:

Cell Cycle/DNA Damage; PI3K/Akt/mTOR; PI3K/Akt/mTOR Solubility:

10 mM in DMSO

Caution: Product has not been fully validated for medical applications. For research use only.

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