Inhibitory activities

VE-821 (2 μM) was screened against the indicated human (h), rat (r), mouse (m) and fission yeast (y) kinases using the Millipore KinaseProfiler service, at ATP concentrations equal to each enzyme’s ATP Km.

Cell lines

HFL1 cells; HCT116 cancer cells; H23 cancer cell line.

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.

Reaction Conditions

10 μM; 24, 48 or 96 h.

Applications

HFL1 cells were pretreated with 10 μM VE-821 or DMSO before addition of 200 μM cisplatin (Cis), 1 μM gemcitabine (Gem), 100 μM etoposide (Etop) or 5 Gy ionizing radiation (IR), VE-821 blocks Chk1 Ser345 phosphorylation under all conditions and inhibits H2AX phosphorylation in treatment with cisplatin and gemcitabine. In the H23 cancer cell line, VE-821 shows marked synergy with cisplatin in growth arrest.

VE-821 is a potent, highly-selective, and ATP-competitive DNA damage response (DDR) kinase ATR inhibitor with Ki value of 13nM. VE-821 specifically inhibits ATR, revealing low cross-reactivity against the mammalian target of rapamycin (mTOR), DNA-dependent protein kinase (DNA-PK), phosphoinositol 3-kinase-γ (PI3K) and the related PIKKs ATM [1].

HL-60 cells treated with VE-821 (10μM) showed reduction of phosphorylatin of Chk1 (Ser 345), inhibition of cell growth, and a radiosensitizing effect after Gamma-ray irradiation [2].

VE-821 has also been demonstrated to down-regulate the phosphorylated Chk1 (Ser 345) but it does not inhibit the phosphorylation of Chk2 (Thr68) and ATM (Ser1981) in pancreatic cancer cell lines, including PSN-1 and MiaPaCa-2 cells that are treated with gemcitabine or radiation. VE-821 combined with gemcitabine (a nucleoside analog) has caused a remarkable increase of cytotoxic effect of gemcitabine against hypoxia [3].

References:
[1] Reaper PM1, Griffiths MR, Long JM, Charrier JD, Maccormick S, Charlton PA, Golec JM, Pollard JR. Selective killing of ATM- or p53-deficient cancer cells through inhibition of ATR. Nat Chem Biol. 2011 Apr 13;7(7):428-30.
[2] Vávrová J1, Zárybnická L, Lukásová E, Rezácová M, Novotná E, Sinkorová Z, Tichy A, Pejchal J, Durisová K. Inhibition of ATR kinase with the selective inhibitor VE-821 results in radiosensitization of cells of promyelocytic leukaemia (HL-60). Radiat Environ Biophys. 2013 Nov;52(4):471-9.
[3] Prevo R1, Fokas E, Reaper PM, Charlton PA, Pollard JR, McKenna WG, Muschel RJ, Brunner TB.The novel ATR inhibitor VE-821 increases sensitivity of pancreatic cancer cells to radiation and chemotherapy. Cancer Biol Ther. 2012 Sep;13(11):1072-81.