p53 is a tumor suppressor protein. Under stressful conditions, p53 tightly regulates cell growth by promoting apoptosis and DNA repair. When p53 becomes mutated, it loses its function, resulting in abnormal cell proliferation and tumor progression[3]. Eprenetapopt (APR-246) is a first-in-class, small molecule that restores wild-type p53 functions in TP53-mutant cells. Eprenetapopt triggers apoptosis in tumor cells. Eprenetapopt also targets the selenoprotein thioredoxin reductase 1 (TrxR1), a key regulator of cellular redox balance[4]. Eprenetapopt inhibits both recombinant TrxR1 in vitro and TrxR1 in cells. Cellular TrxR1 activity is inhibited by Eprenetapopt irrespective of p53 status. Eprenetapopt can directly affect cellular redox status via targeting of TrxR1. Several small molecules have been shown to restore wild-type activity to mutant p53, including CP-31398, PRIMA-1 and Eprenetapopt, MIRA, STIMA, PhiKan-083 and NSC319726. PRIMA-1 and its methylated analog Eprenetapopt promote correct folding of mutant p53, induce cell death by apoptosis, and inhibit tumor growth in mice. Eprenetapopt has also been shown to reactivate mutant forms of the p63 and p73 proteins that share high structural homology with p53[5]. Eprenetapopt is a powerful apoptosis-inducing agent. Eprenetapopt can enhance apoptosis in mutant p53 carrying cells, compared to the p53 null parental cells. Most p53 mutants are in complex with Hsp70 proteins. Eprenetapopt treatment increases Hsp70 expression and nucleolar translocation, in parallel with the induction of nucleolar accumulation of mutant p53. Several lines of evidence suggest that Eprenetapopt can also act independently of the p53 status of the cell. It can radiosensitize prostate carcinoma cell lines with mutant or wild type p53 and p53-/- cells as well. Introduction of mutant p53 (p53ser249 or p53gln248) into p53-/- hepatocarcinoma cells increases sensitivity to Eprenetapopt without the induction of p53 target genes.Eprenetapopt regularly induces apoptosis in mutant p53 expressing cells[6].
Eprenetapopt 细胞实验
Cell Line
Concentration
Treated Time
Description
References
BT-20
100 µM
12 hours
APR-246 enhanced the expression of IFN signalling genes
Br J Cancer. 2022 Nov;127(11):2060-2071.
BT-549
40 µM
12 hours
APR-246 enhanced the expression of IFN signalling genes
Br J Cancer. 2022 Nov;127(11):2060-2071.
DLD-1
90 µM
24 hours
APR-246 enhanced the expression of IFN signalling genes
Br J Cancer. 2022 Nov;127(11):2060-2071.
Recombinant rat TrxR1
50 µM
10 minutes
APR-246 inhibits TrxR1 activity, with MQ and preheated APR-246 being more potent inhibitors.
Cell Death Dis. 2013 Oct 24;4(10):e881.
SKBR-3
40 µM
12 hours
APR-246 repressed the expression of immune checkpoint molecules
Br J Cancer. 2022 Nov;127(11):2060-2071.
MDAMB-231
200 µM
12 hours
APR-246 repressed the expression of immune checkpoint molecules
Br J Cancer. 2022 Nov;127(11):2060-2071.
MDAMB-468
60 µM
12 hours
APR-246 repressed the expression of immune checkpoint molecules
Br J Cancer. 2022 Nov;127(11):2060-2071.
OACM5.1 cells
50 µM
12 hours
To study the effect of Eprenetapopt on cellular metabolism, results showed that Eprenetapopt decreased reduced GSH levels without increasing oxidized GSH (GSSG) levels, indicating total GSH depletion.
Sci Adv. 2022 Sep 16;8(37):eabm9427.
MOLM14
60 µM
16 hours
To investigate the mechanisms underlying APR-246-induced cell death, results showed that iron chelators and lipophilic antioxidants almost completely rescued cell viability, indicating that cell death is both iron and ROS-dependent.
Haematologica. 2022 Feb 1;107(2):403-416.
OVCAR-3
0.0025, 0.01, 0.05, 0.2, 0.8, 3.1, 12.5 µM
2 hours
MS-CETSA data indicated that ASNS is a direct or indirect target of APR-246 via the active product MQ. MQ treatment resulted in increased thermal stability of ASNS, suggesting that MQ may directly or indirectly affect ASNS activity.
Cell Death Dis. 2021 Jul 15;12(7):709.
CCRF-CEM
10, 15 µM
2 hours
WB-CETSA results showed that MQ treatment induced a temperature-dependent stabilization of ASNS at 57 and 59 °C, indicating that MQ directly or indirectly modulates ASNS.
Cell Death Dis. 2021 Jul 15;12(7):709.
Neuroblastoma cell lines
16.1 µM
24 hours
To investigate the ability of PRIMA-1MET to induce NB cell death, results showed that PRIMA-1MET induces cell death at low concentrations
J Exp Clin Cancer Res. 2019 Feb 12;38(1):69.
Keratinocytes
65.8 µM (IC50)
24 hours
To compare the specificity of PRIMA-1MET for tumor cells versus normal cells, results showed that normal cells are less sensitive to PRIMA-1MET
J Exp Clin Cancer Res. 2019 Feb 12;38(1):69.
Fibroblasts
78.3 µM (IC50)
24 hours
To compare the specificity of PRIMA-1MET for tumor cells versus normal cells, results showed that normal cells are less sensitive to PRIMA-1MET
J Exp Clin Cancer Res. 2019 Feb 12;38(1):69.
H1299 cells
50 µM
24 hours
To investigate whether Eprenetapopt-induced cell death could be reversed by ferroptosis inhibitors, results showed that ferroptosis inhibitors could reverse Eprenetapopt-induced cell death.
Sci Adv. 2022 Sep 16;8(37):eabm9427.
HL60
11 µM to >200 µM
24 hours
To evaluate the activity of APR-246 in AML cells, results showed that most AML cell lines and primary AML cells were sensitive to cell death induction by APR-246.
Haematologica. 2022 Feb 1;107(2):403-416.
TE1
10.5 µM
48 hours
APR-246 induced significant apoptosis by upregulating p73 and Noxa via ROS induction in ESCC cell lines harbouring p53 missense mutations.
Br J Cancer. 2021 Nov;125(11):1523-1532.
KYSE410
31.6 µM
48 hours
APR-246 showed minimal effectiveness in ESCC with wild-type p53, and did not significantly induce apoptosis.
Br J Cancer. 2021 Nov;125(11):1523-1532.
FLO-1
25 µM(GI90)
6 hours
To detect intracellular ROS levels after APR-246 treatment, results showed that APR-246 significantly increased ROS levels in FLO-1 cells.
Nat Commun. 2017 Mar 28;8:14844.
JH-EsoAd1
40 µM(GI90)
6 hours
To detect intracellular ROS levels after APR-246 treatment, results showed that APR-246 significantly increased ROS levels in JH-EsoAd1 cells.
Nat Commun. 2017 Mar 28;8:14844.
Keratinocytes from EEC syndrome patients
10–30 µM
7 days
APR-246 partially restored differentiation defects in p63 mutant keratinocytes
Proc Natl Acad Sci U S A. 2013 Feb 5;110(6):2157-62.
UMSCC10A
0-50 µM
72 hours
To evaluate the effect of APR-246 combined with PL on HNSCC cells, the results showed that the combination significantly reduced cell viability and increased cell death.
Oncogene. 2018 Jun;37(25):3384-3398.
FaDu
0-50 µM
72 hours
To evaluate the effect of APR-246 combined with PL on HNSCC cells, the results showed that the combination significantly reduced cell viability and increased cell death.
Oncogene. 2018 Jun;37(25):3384-3398.
A2780-CP20
8 µM, 20 µM, 30 µM, 40 µM, 60 µM
72 hours
APR-246 completely restored the sensitivity of ovarian cancer cells to cisplatin at clinically relevant concentrations.
Cell Death Dis. 2015 Jun 18;6(6):e1794.
OVCAR-3
2 µM, 20 µM, 30 µM
72 hours
APR-246 resensitized OVCAR-3 cells to cisplatin and increased the efficacy of cisplatin.
Cell Death Dis. 2015 Jun 18;6(6):e1794.
Neuroblastoma cells
3.8 µM
96 hours
To assess the cytotoxicity of APR-246 on ALT cells, results showed ALT cells were significantly more sensitive to APR-246
Cancer Res. 2022 Sep 16;82(18):3345-3358.
Rhabdomyosarcoma cells
3.8 µM
96 hours
To assess the cytotoxicity of APR-246 on ALT cells, results showed ALT cells were significantly more sensitive to APR-246
Cancer Res. 2022 Sep 16;82(18):3345-3358.
Carcinoma cells
3.8 µM
96 hours
To assess the cytotoxicity of APR-246 on ALT cells, results showed ALT cells were significantly more sensitive to APR-246
Cancer Res. 2022 Sep 16;82(18):3345-3358.
FLO-1 cells
50 μM
24 hours
Evaluate eprenetapopt-induced cell death, results showed ferroptosis inhibitors could reverse cell death
Sci Adv. 2022 Sep 16;8(37):eabm9427.
H1299 cells
50 μM
24 hours
Evaluate eprenetapopt-induced cell death, results showed ferroptosis inhibitors could reverse cell death
Sci Adv. 2022 Sep 16;8(37):eabm9427.
OACM5.1 cells
50 μM
12 hours
Evaluate the effect of eprenetapopt on GSH levels, results showed decreased GSH levels
Sci Adv. 2022 Sep 16;8(37):eabm9427.
OVCAR-3 ovarian cancer cells
2-30 μM
72 hours
APR-246 increased the sensitivity of OVCAR-3 cells to cisplatin and reduced the survival index plateau at higher concentrations.
Cell Death Dis. 2015 Jun 18;6(6):e1794.
A2780-CP20 ovarian cancer cells
8-60 μM
72 hours
APR-246 completely restored the sensitivity of ovarian cancer cells to cisplatin at clinically relevant concentrations.
Cell Death Dis. 2015 Jun 18;6(6):e1794.
Eprenetapopt 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Mice
FLO-1 xenograft model
Intraperitoneal injection
100 mg/kg
Once daily for 3 weeks
To evaluate the anti-tumour activity of APR-246 in the FLO-1 xenograft model, results showed that APR-246 significantly inhibited tumour growth.
Nat Commun. 2017 Mar 28;8:14844.
BALB/C mice
MCO4 fibrosarcoma model
Intraperitoneal injection
100 mg/kg
Daily for 4 days
APR-246 promoted CD4+ T cells infiltration and decreased the exhaustion of CD8+ T cells in vivo
Br J Cancer. 2022 Nov;127(11):2060-2071.
NSG mice
FLO-1 LM tumor model
Intraperitoneal injection
100 mg/kg
Daily for 35 days
To study the effect of Eprenetapopt combined with serine and glycine dietary restriction on tumor growth, results showed that the combination significantly inhibited primary tumor growth and delayed the onset of metastatic disease.
Sci Adv. 2022 Sep 16;8(37):eabm9427.
NOD/SCID IL-2 receptor γ-chain-null mice
MOLM14 xenograft model
Intraperitoneal injection
100 mg/kg
Once daily for 4 days
To evaluate the synergistic anti-leukemic activity of APR-246 combined with SLC7A11 inhibition, results showed that SLC7A11 knockdown significantly reduced tumor cell burden in the BM, and this effect was enhanced when combined with APR-246 treatment.
Haematologica. 2022 Feb 1;107(2):403-416.
C57BL/6J mice
B16 melanoma model
Intraperitoneal injection
100 mg/kg
Daily for 6 days
To study the effects of APR-246 on p53 expression in the tumor microenvironment and its combination with immune checkpoint blockade
J Clin Invest. 2022 Sep 15;132(18):e148141
SCID mice
UMSCC10A xenograft model
Intraperitoneal injection
100 mg/kg/day
Daily for 24 days
To evaluate the effect of APR-246 combined with PL on HNSCC xenograft tumors, the results showed that the combination significantly suppressed tumor growth.
Oncogene. 2018 Jun;37(25):3384-3398.
Mice
Xenograft model
Intraperitoneal injection
25 mg/kg/day
Once daily for 21 days
Combination of APR-246 and 5-FU showed synergistic antitumour effects in p53 missense mutant ESCC xenograft models through activation of ROS and the p73-Noxa pathway.
Br J Cancer. 2021 Nov;125(11):1523-1532.
Athymic nu/nu mice
Xenograft model
Intraperitoneal injection
250 mg/kg
Twice daily for 7 days, 21-day cycle, total of 3 cycles
To evaluate the efficacy of APR-246 combined with irinotecan in ALT xenograft models, results showed complete responses in ALT models
Cancer Res. 2022 Sep 16;82(18):3345-3358.
Mice
A2780-CP20 tumor xenograft model
Intravenous injection
400 mg/kg/day
APR-246: once daily for 7 days; cisplatin: once on day 2 and day 6
APR-246 in combination with cisplatin significantly inhibited tumor growth, indicating at least an additive effect.
Cell Death Dis. 2015 Jun 18;6(6):e1794.
Mice
Eso26 esophageal cancer cell xenograft model
Intraperitoneal injection
50 mg/kg APR-246, 50 mg/kg MK-571
APR-246 daily for 16 days; MK-571 daily except on days 6–7 and 13–14
Combination treatment with APR-246 and MK-571 significantly suppressed the growth of Eso26 xenograft tumors and prolonged the survival of mice.
EMBO Mol Med. 2021 Feb 5;13(2):e10852
NSG mice
FLO-1 LM xenograft model
Intraperitoneal injection
100 mg/kg
Daily for 35 days
Evaluate the effect of combining eprenetapopt with serine and glycine dietary restriction, results showed combination therapy significantly inhibited tumor growth
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