ATR/ATM kinases can phosphorylate a broad and overlapping catalogue of several thousand substrates in DNA damage signaling. The kinase activity of ATR, activated by single-stranded DNA (ssDNA) coated with replication protein A (RPA), is increased at damaged replication forks and resected DNA double-strand breaks (DSBs). AZD6738 is a highly selective and potent inhibitor of ATR kinase activity with IC50 value of 1nM in isolated enzyme assay and showed inhibitory effect on ATR kinase-dependent cellular Chk1 phosphorylation with IC50 value of 74nM. AZD6738 impaired viability of the Kras mutant cell line, H23, H460, A549 and H358, with the lowest GI50 and greatest maximal inhibition in H460 and H23 cells (1.05μM, 88.0% and 2.38μM, 86.2%, respectively). Meanwhile, induced activation of ATM, as evident by increased pATM-S1981, stabilized p53 and expression of p21 and p27 can be observed in p53-wildtype H460 and A549 cells treated with 1μM AZD6738. A marked increase in pH2A.X-ser139 and cleaved PARP, indicative of accumulation of DNA damage and induction of apoptosis with greater sensitivity in cell viability assay, can be observed in H23 and H460 cells treated with AZD6738. Combination of AZD6738 (1μM) and cisplatin caused accumulation of cells in early S-phase and at the G1/S border, as well as dramatic cell death of H23 and H460 cells, suggesting its potent synergy with cisplatin in ATM-deficient cells independent of the ATM-p53 signaling pathway. Consistent with the in vitro study, daily administration of AZD6738 at dose of 25mg/kg for 14 consecutive days enhanced the therapeutic efficacy of cisplatin (3mg/kg) and caused rapid regression of ATM-deficient H23 tumors[1].
To assess the sensitivity of ATM-proficient cells to Ceralasertib
Nucleic Acids Res. 2021 Sep 7;49(15):8665-8683.
ATM-deficient mouse embryonic stem cells
100 nM
6 days
To assess the sensitivity of ATM-deficient cells to Ceralasertib
Nucleic Acids Res. 2021 Sep 7;49(15):8665-8683.
CD3/CD28 stimulated T-cells
1 μM
4 days
To evaluate the effect of Ceralasertib on T-cell proliferation, the results showed that Ceralasertib significantly increased the expression of DNA damage markers.
Nat Commun. 2024 Feb 24;15(1):1700.
IMR-5
0.05 μM
48 h
To study the DNA damage effects of CX-5461 on neuroblastoma cells, results showed that CX-5461 induced DNA damage at low concentrations.
Nat Commun. 2021 Nov 9;12(1):6468.
CHP-134
0.04 μM
3 days
To study the cytotoxicity of CX-5461 on neuroblastoma cells, results showed that CX-5461 effectively induced apoptosis at low concentrations.
Nat Commun. 2021 Nov 9;12(1):6468.
HEK293 cells
>0.333 μM
24 h
To evaluate the inhibitory effect of Ceralasertib on HRR, results showed that Ceralasertib could inhibit HRR
Cancer Res. 2022 Mar 15;82(6):1140-1152.
A549 cells
>0.333 μM
72 h
To evaluate the inhibitory effect of Ceralasertib on BIR repair, results showed that Ceralasertib could inhibit BIR repair
Cancer Res. 2022 Mar 15;82(6):1140-1152.
FaDu WT and FaDu ATM −/− cells
1 µM
24, 48, 72 h
Investigated the sensitivity of ATM-deficient cells to Ceralasertib, showing that ATM −/− cells exhibited higher DNA damage, early apoptosis, and cell death after Ceralasertib treatment.
Oncoimmunology. 2022 Sep 13;11(1):2117321.
LL2-luc cells
1 µM
1 hour
To evaluate the effect of AZD6738 on the radiosensitivity of LL2-luc cells, results showed that AZD6738 enhanced the radiosensitivity of the cells.
Br J Cancer. 2020 Sep;123(5):762-771.
LL2-luc cells
0.6 µM
72 h
To evaluate the effect of AZD6738 on the proliferation of LL2-luc cells, results showed that AZD6738 has anti-proliferative effects.
Br J Cancer. 2020 Sep;123(5):762-771.
HPAF-II
2000 nM
24 h
To evaluate the effect of ATR inhibitor AZD6738 in combination with gemcitabine on DDR signaling pathways, results showed that DNA-PK-driven phosphorylation of Chk1 and Chk2 persisted.
Br J Cancer. 2020 Oct;123(9):1424-1436.
MIA PaCa-2
500 nM
24 h
To evaluate the effect of ATR inhibitor AZD6738 on the cell cycle, results showed that ATM-deficient cells exhibited increased intra-S phase accumulation and induced cell death.
Br J Cancer. 2020 Oct;123(9):1424-1436.
MiaPaCa-2 cells
2 µM
24 h
AZD6738 in combination with gemcitabine significantly induced phosphorylation of pRPA and γH2AX, indicating a strong induction of replication stress in these cells
Mol Cancer Ther. 2018 Aug;17(8):1670-1682.
K8484 cells
2 µM
7 h
AZD6738 completely prevented gemcitabine-induced Chk1 phosphorylation and restored Cdk1 activity
Mol Cancer Ther. 2018 Aug;17(8):1670-1682.
Ceralasertib 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Mouse
CT26 tumor model
Oral gavage
75 mg/kg
Once daily for 3 days
To evaluate the impact of ATR inhibitor on CD8+ T cell activation and proliferation. ATR inhibitor reduced the number of proliferating CD8+ T cells.
Cell Rep. 2022 Sep 20;40(12):111371.
Mice
CT26, MC38, 4T1, and A20 tumor models
Oral
25 mg/kg
Twice daily, 7 days on/7 days off
To evaluate the antitumor effect of Ceralasertib under an intermittent dosing schedule, the results showed that Ceralasertib significantly inhibited tumor growth, and its antitumor effect was dependent on CD8+ T cells.
Nat Commun. 2024 Feb 24;15(1):1700.
Mice
Neuroblastoma xenograft model
Intraperitoneal injection
5 mg/kg
Once daily for 5 days, starting a second 5-day course on day 8, repeated every 21 days
To study the anti-tumor activity of CX-5461 in vivo, results showed that CX-5461 significantly improved survival when combined with TOP1 inhibitors in orthotopic PDX mouse models.
Nat Commun. 2021 Nov 9;12(1):6468.
Nude mice
Granta-519 mantle cell lymphoma model
Oral
50 mg/kg
Once daily for 21 days
To evaluate the antitumor activity of Ceralasertib in vivo, results showed significant tumor growth inhibition at 50 mg/kg dose
Cancer Res. 2022 Mar 15;82(6):1140-1152.
Mice
Subcutaneous and orthotopic LL2-luc tumour models
Intraperitoneal injection
15 mg/kg
Daily for 6 days
To evaluate the antitumor efficacy of AZD6738 in combination with radiotherapy in subcutaneous and orthotopic LL2-luc tumour models, results showed that AZD6738 enhanced the antitumor efficacy of radiotherapy, but the effect was limited in orthotopic tumour models.
Br J Cancer. 2020 Sep;123(5):762-771.
Mice
MIA PaCa-2 subcutaneous xenograft model
Oral
50 mg/kg
Once daily for 5 consecutive days a week for 3 weeks
To evaluate the efficacy of ATR inhibitor AZD6738 monotherapy in ATM-deficient xenografts, results showed significant growth delay in ATM-deficient tumors.
Br J Cancer. 2020 Oct;123(9):1424-1436.
Mice
K8484 subcutaneous allograft model
Oral
25 mg/kg
5 days/week for 3 weeks
AZD6738 in combination with gemcitabine significantly inhibited tumor growth and extended survival in mice
搜索
