Home Apoptosis MDM2/MDMX antagonist - E3 Ligase antagonist Nutlin-3 For research use only.

Nutlin-3

Nutlin-3 is a potent and selective Mdm2 (RING finger-dependent ubiquitin protein ligase for itself and p53) antagonist with IC50 of 90 nM in a cell-free assay; stabilizes p73 in p53-deficient cells.

Nutlin-3 Chemical Structure

Nutlin-3 Chemical Structure

CAS No. 890090-75-2

Purity & Quality Control

Products often used together with Nutlin-3

Tozasertib (VX-680)

Nutlin-3 and Tozasertib combined therapy selectively kills p53-compromised cells and inhibits the proliferation of cells expressing wild-type p53.

Cheok CF, et al. Cell Death Differ. 2010 Sep;17(9):1486-500.

Nutlin-3 Related Products

Signaling Pathway

MDM2/MDMX Signaling Pathways

MDM2/MDMX Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
SMMC-7721  Function Assay 10 μM 36 h DMSO down-regulates the protein expression level of phospho-Ser392-p53 24286312
HuH-7 Apoptosis Assay 20 μM 48 h DMSO induces apoptosis 24286312
SMMC-7721  Apoptosis Assay 20 μM 48 h DMSO induces apoptosis 24286312
HuH-7 Cell Viability Assay 1.25-20 μM 24/48/72 h DMSO inhibits cell proliferation dose and time dependently 24286312
SMMC-7721  Cell Viability Assay 1.25-20 μM 24/48/72 h DMSO inhibits cell proliferation dose and time dependently 24286312
RKO Function Assay 20 μM 24 h induces HO-1 expression at the level of transcription 24366007
U2OS  Function Assay 20 μM 24 h induces HO-1 expression at the level of transcription 24366007
RKO Function Assay 20 μM 24 h increases the levels of the HO-1 protein as well as the p53 protein 24366007
U2OS  Function Assay 20 μM 24 h increases the levels of the HO-1 protein as well as the p53 protein 24366007
MOLM Function Assay 10 μM 24 h upregulates the SOCS-1 expression 24473562
OCI Function Assay 10 μM 24 h upregulates the SOCS-1 expression 24473562
BeWo Apoptosis Assay 30 µM 24 h increases apoptosis 24498154
BeWo Function Assay 30 µM 24 h increases p53, Mdm2, p21 and Puma at the protein level 24498154
MOML13 Function Assay 10μM 2/4 h increases the level of p53 24659749
AML3 Function Assay 10μM 2/4 h increases the level of p53 24659749
AML2 Function Assay 10μM 2/4 h increases the level of p53 24659749
MOML13 Apoptosis Assay 2/10 μM 24/48 h induces apoptosis 24659749
AML2 Apoptosis Assay 2/10 μM 24/48 h induces apoptosis 24659749
U2OS  Function Assay 20 μM 24 h increases the mRNA levels of BCL2A1, BCLXL andBCLW 24867259
Hep3B Apoptosis Assay induces apoptosis 24884809
Huh-7 Apoptosis Assay induces apoptosis 24884809
SMMC7721 Apoptosis Assay induces apoptosis 24884809
HepG2 Apoptosis Assay induces apoptosis 24884809
Hep3B Growth Inhibition Assay 72 h DMSO IC50=20.18 ± 1.84 μM 24884809
Huh-7 Growth Inhibition Assay 72 h DMSO IC50=33.96 ± 3.9 μM 24884809
SMMC7721/Ac Growth Inhibition Assay 72 h DMSO IC50=55.21 ± 5.03 μM 24884809
SMMC7721 Growth Inhibition Assay 72 h DMSO IC50=31.28 ± 4.2 μM 24884809
HepG2/As Growth Inhibition Assay 72 h DMSO IC50=68.13 ± 9.6 μM 24884809
HepG2 Growth Inhibition Assay 72 h DMSO IC50=35.86 ± 2.9 μM 24884809
MOLM-13 Function Assay 6 μM 6 h DMSO enhances the acetylation of histone H2B and heat shock proteins Hsp27 and Hsp90 24885082
MOLM-13 Function Assay 6 μM 0-8 h DMSO increases the levels of p53, MDM2, p21 and acetylated p53 24885082
115 Function Assay 5 μM 48 h DMSO induces p53-dependent senescence  25067787
A498 Function Assay 5 μM 48 h DMSO induces p53-dependent senescence  25067787
Caki-2 Function Assay 5 μM 48 h DMSO induces p53-dependent senescence  25067787
ACHN Function Assay 5 μM 48 h DMSO induces p53-dependent senescence  25067787
115 Growth Inhibition Assay 5 μM 48 h DMSO induces cell cycle arrest  25067787
A498 Growth Inhibition Assay 5 μM 48 h DMSO induces cell cycle arrest  25067787
Caki-2 Growth Inhibition Assay 5 μM 48 h DMSO induces cell cycle arrest  25067787
ACHN Growth Inhibition Assay 5 μM 48 h DMSO induces cell cycle arrest  25067787
115 Function Assay 0.5/1/5 μM 48 h DMSO leads to increased expression of p53 and some p53 target genes: MDM2, and p21 25067787
A498 Function Assay 0.5/1/5 μM 48 h DMSO leads to increased expression of p53 and some p53 target genes: MDM2, and p21 25067787
Caki-2 Function Assay 0.5/1/5 μM 48 h DMSO leads to increased expression of p53 and some p53 target genes: MDM2, and p21 25067787
ACHN Function Assay 0.5/1/5 μM 48 h DMSO leads to increased expression of p53 and some p53 target genes: MDM2, and p21 25067787
117 Cell Viability Assay 0.5-10 μM 0-6 d DMSO inhibits proliferation in a dose-dependent manner 25067787
115 Cell Viability Assay 0.5-10 μM 0-6 d DMSO inhibits proliferation in a dose-dependent manner 25067787
A498 Cell Viability Assay 0.5-10 μM 0-6 d DMSO inhibits proliferation in a dose-dependent manner 25067787
Caki-2 Cell Viability Assay 0.5-10 μM 0-6 d DMSO inhibits proliferation in a dose-dependent manner 25067787
ACHN Cell Viability Assay 0.5-10 μM 0-6 d DMSO inhibits proliferation in a dose-dependent manner 25067787
MCF7  Function Assay 2.5 µM 48 h DMSO decreases the homologous DSB repair frequencies 25085902
MCF7  Cell Viability Assay 2.5 µM 5 d DMSO sensitizes MCF7 to PARP inhibition 25085902
RAW 264.7 Function Assay 10 μM 30 min inhibits LPS-induced NO production  25172547
RAW 264.7 Function Assay 10 μM 30 min reduces the LPS-augmented the NF-κB luciferase reporter gene activity 25172547
RAW 264.7 Function Assay 10 μM 30 min prevents the p53 reduction in response to LPS 25172547
SUM102PT Function Assay 10 μM 24 h DMSO inhibits TGF-β3-induced EPHB2 mRNA and protein expression 25257729
SK-BR-7 Function Assay 10 μM 24 h DMSO inhibits TGF-β3-induced EPHB2 mRNA and protein expression 25257729
MCF-10CA1a Function Assay 10 μM 24 h DMSO inhibits TGF-β3-induced EPHB2 mRNA and protein expression 25257729
MCF-10CA1a Function Assay 10 μM 24 h DMSO decreases the TGF-β3-induced mRNA levels ofMMP2, MMP9, and integrin β 3 25257729
MCF-10A1  Function Assay 10 μM 24/48 h DMSO inhibits migration of normal breast epithelial  25257729
MCF-10CA1a Function Assay 10 μM 48 h DMSO inhibits basal invasion and reduced TGF-β3-induced invasion to basal levels 25257729
HCT116  Function Assay 10 µM 24 h causes a p53-dependent tetraploid G1-arrest in diploid HCT116 clones D3 and D8 25380055
A2780 Function Assay 10 μM 21h  DMSO decreases the FoxM1 levels 25426548
NCI-H23 Function Assay 10 μM 21h  DMSO decreases the FoxM1 levels 25426548
A2780 Function Assay 10 μM 21h  DMSO increases p53 protein levels 25426548
NCI-H23 Function Assay 10 μM 21h  DMSO increases p53 protein levels 25426548
OVCAR10 Function Assay 10 μM 21h  DMSO increases p53 protein levels 25426548
MCF-7 Function Assay 10 μM 0-24 h induces p53 and p21/Cip1 25482373
SMMC-7721 Function Assay 10 μM 36 h DMSO causes the ectopic expression of IFI16 25544361
SMMC-7721 Function Assay 10 μM 48 h DMSO increases the expression level of IFNB1 mRNA 25544361
SMMC-7721 Function Assay 10 μM 48 h DMSO induces the chromatin-bound protein IFI16 to partially localize in the cytoplasm  25544361
SMMC-7721 Function Assay 10 μM 48 h DMSO causes DNA DSB damage 25544361
DU4475  Function Assay 5/10/20 μM 24 h downregulates Toca-1 dose dependently 25547174
MCF-7  Function Assay 10 μM DMSO inhibits cyclin D1 and Dicer  25702703
C2C12 Growth Inhibition Assay 10 μM  24/48/72 h DMSO inhibits cells proliferation and differentiation 25871794
L6 Growth Inhibition Assay 10 μM  24/48/72 h DMSO inhibits cells proliferation and differentiation 25871794
CRL-5908 Growth Inhibition Assay 24 h IC50=38.71 ± 2.43 μM 26125230
A549-920 Growth Inhibition Assay 24 h IC50=33.85 ± 4.84 μM 26125230
A549-NTC Growth Inhibition Assay 24 h IC50=19.42 ± 1.96 μM 26125230
A549 Growth Inhibition Assay 24 h IC50=17.68 ± 4.52 μM 26125230
C666-1 Apoptosis Assay 10 µM 48/72 h DMSO sensitizes C666-1 cells to cisplatin-induced apoptosis 26252575
C666-1  Function Assay 10 µM 24 h DMSO activates the p53 pathway, upregulating p53, p21 and Mdm2 26252575
C666-1 Cell Viability Assay 10 µM 48 h DMSO sensitizes C666-1 cells to the cytotoxic effect of cisplatin 26252575
C666-1 Growth Inhibition Assay IC50=19.95±8.93 μM 26252575
NP460 Growth Inhibition Assay IC50=22.85±1.18 μM 26252575
NP69 Growth Inhibition Assay IC50=31.69±2.54 μM 26252575
MCF7 Growth Inhibition Assay 5 μM 48 h blocks 27-OHC-induced cell proliferation comparable to that of basal levels 26350565
HuH-7 Function Assay 10 μM 36 h DMSO down-regulates the protein expression level of phospho-Ser392-p53 24286312
AT2 Function Assay 5/10 μM leads to a substantial accumulation of the p53 protein as well as the expression of its direct targets p21, MDM2 and the pro-apoptotic BAX and PUMA proteins  24240203
REH Function Assay 5/10 μM leads to a substantial accumulation of the p53 protein as well as the expression of its direct targets p21, MDM2 and the pro-apoptotic BAX and PUMA proteins  24240203
UoCB6 Function Assay 5/10 μM leads to a substantial accumulation of the p53 protein as well as the expression of its direct targets p21, MDM2 and the pro-apoptotic BAX and PUMA proteins  24240203
AT2 Cell Viability Assay 0-25 μM inhibits cell viability dose dependently 24240203
REH Cell Viability Assay 0-25 μM inhibits cell viability dose dependently 24240203
UoCB6 Cell Viability Assay 0-25 μM inhibits cell viability dose dependently 24240203
A2780 Function Assay 5/10/20 μM 24 h upregulates p53, MDM2, p21 and DR5 protein levels dose dependently  24136147
H460 Function Assay 5/10/20 μM 24 h upregulates p53, MDM2, p21 and DR5 protein levels dose dependently  24136147
Lovo  Function Assay 5/10/20 μM 24 h upregulates p53, MDM2, p21 and DR5 protein levels dose dependently  24136147
A2780 Apoptosis Assay 5/10/20 μM 24 h enhances apoptosis induction by D269H/E195R over rhTRAIL 24136147
H460 Apoptosis Assay 5/10/20 μM 24 h enhances apoptosis induction by D269H/E195R over rhTRAIL 24136147
Lovo  Apoptosis Assay 5/10/20 μM 24 h enhances apoptosis induction by D269H/E195R over rhTRAIL 24136147
U87MG Function assay 10 mins Binding affinity to MDM2 in human U87MG cells assessed as inhibition of MDM2/p53 protein interaction after 10 mins by quantitative sandwich immuno assay, IC50 = 0.1045 μM. 27050782
U87MG Antiproliferative assay 48 hrs Antiproliferative activity against human U87MG cells expressing wild type p53 after 48 hrs by MTS assay, IC50 = 6.5 μM. 27050782
U343MG Antiproliferative assay 48 hrs Antiproliferative activity against human U343MG cells expressing wild type p53 after 48 hrs by MTS assay, IC50 = 12.6 μM. 27050782
SW620 Cytotoxicity assay 72 hrs Cytotoxicity against human SW620 cells after 72 hrs by SRB assay, IC50 = 0.38 μM. ChEMBL
A549 Cytotoxicity assay 72 hrs Cytotoxicity against human A549 cells after 72 hrs by SRB assay, IC50 = 0.41 μM. ChEMBL
SJSA1 Cytotoxicity assay 72 hrs Cytotoxicity against human SJSA1 cells after 72 hrs by SRB assay, IC50 = 1.81 μM. ChEMBL
HCT116 Cytotoxicity assay 72 hrs Cytotoxicity against human HCT116 cells after 72 hrs by SRB assay, IC50 = 4.63 μM. ChEMBL
PC3 Cytotoxicity assay 72 hrs Cytotoxicity against human PC3 cells after 72 hrs by SRB assay, IC50 = 6.37 μM. ChEMBL
Click to View More Cell Line Experimental Data

Biological Activity

Description Nutlin-3 is a potent and selective Mdm2 (RING finger-dependent ubiquitin protein ligase for itself and p53) antagonist with IC50 of 90 nM in a cell-free assay; stabilizes p73 in p53-deficient cells.
Targets
MDM2 [5]
(Cell-free assay)
180 nM
In vitro
In vitro

Nutlin-3 potently inhibits the MDM2-p53 interaction, leading to the activation of the p53 pathway. Nutlin-3 treatment induces the expression of MDM2 and p21, and displays potent antiproliferative activity with IC50 of ~1.5 μM, only in cells with wild-type p53 such as HCT116, RKO and SJSA-1, but not in the mutant p53 cell lines SW480 and MDA-MB-435. In SJSA-1 cells, Nutlin-3 treatment at 10 μM for 48 hours significantly induces caspase-dependent cell apoptosis by ~45%. Although Nutlin-3 also inhibits the growth and viability of human skin (1043SK) and mouse embryo (NIH/3T3) with IC50 of 2.2 μM and 1.3 μM, respectively, cells remain viable 1 week post-treatment even at 10 μM of Nutlin-3, in contrast with the SJSA-1 cells with viability lost at 3 μM of Nutlin-3 treatment. [1] Nutlin-3 does not induce the phosphorylation of p53 on key serine residues and reveals no difference in their sequence-specific DNA binding and ability to transactivate p53 target genes compared with phosphorylated p53 induced by the genotoxic drugs doxorubicin, demonstrating that phosphorylation of p53 on key serines is dispensable for transcriptional activation and apoptosis. [2] Although binding less efficiently to MDMX than to MDM2, Nutlin-3 can block the MDMX–p53 interaction and induce the p53 pathway in retinoblastoma cells (Weri1) with IC50 of 0.7 μM. [3] Nutlin-3 at 30 μM also disrupts endogenous p73-HDM2 interaction and enhances the stability and proapoptotic activities of p73, leading to the dose-dependent cell growth inhibition and apoptosis induction in cells without wild-type p53. [4]
Kinase Assay Biacore study
Competition assay is performed on a Biacore S51. A Series S Sensor chip CM5 is utilized for the immobilization of a PentaHis antibody for capture of the His-tagged p53. The level of capture is ~200 response units (1 response unit corresponds to 1 pg of protein per mm2). The concentration of MDM2 protein is kept constant at 300 nM. Nutlin-3 is dissolved in DMSO at 10 mM and further diluted to make a concentration series of Nutlin-3 in each MDM2 test sample. The assay is run at 25 °C in running buffer (10 mM Hepes, 0.15 M NaCl, 2% DMSO). MDM2-p53 binding in the presence of Nutlin-3 is calculated as a percentage of binding in the absence of Nutlin-3 and IC50 is calculate
Cell Research Cell lines HCT116, RKO, SJSA-1, SW480, and MDA-MB-435
Concentrations Dissolved in DMSO, final concentrations ~ 30 μM
Incubation Time 8, 24, and 48 hours
Method

Cells are exposed to various concentrations of Nutlin-3 for 8, 24 and 48 hours. The transcriptional levels of p21 and MDM2 genes are analyzed by real-time PCR, and protein levels by western blotting. Cell viability is measured by the MTT assay. Cell apoptosis is determined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) staining with flow cytometry and fluorescence microscopy.
Experimental Result Images Methods Biomarkers Images PMID
Western blot MDM2 / p53 / ALKBH2 / p21 / PUMA RRM1 / RRM2 / p53R2 / p21 / p53 / pS6 / S6 23258843
Immunofluorescence Lamin A / Lamin C / p16 / H3K9me3 Merlin / cyclin D1 / p53 / MDM2 p53 30728349
Growth inhibition assay Cell viability 29286113
In Vivo
In vivo

Oral administration of Nutlin-3 at 200 mg/kg twice daily for 3 weeks significantly inhibits the tumor growth of SJAS-1 xenografts by 90%, comparable with the effect of doxorubicin treatment with 81% inhibition of tumor growth. [1]
Animal Research Animal Models Athymic female nude mice (Nu/Nu-nuBR) injected subcutaneously with SJSA-1 cells
Dosages 200 mg/kg
Administration Orally, twice a day

Chemical Information & Solubility

Molecular Weight 581.5 Formula

C30H30Cl2N4O4
CAS No. 890090-75-2 SDF Download Nutlin-3 SDF
Smiles CC(C)OC1=C(C=CC(=C1)OC)C2=NC(C(N2C(=O)N3CCNC(=O)C3)C4=CC=C(C=C4)Cl)C5=CC=C(C=C5)Cl
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 100 mg/mL ( (171.96 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Ethanol : 100 mg/mL

Water : Insoluble


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In vivo
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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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Frequently Asked Questions

Question 1:
Is this a racemic mixture of Nutlin-3a and Nutlin-3b or just the Nutlin-3a enantiomer?

Answer:
It is a racemate.

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