Nutlin-3

Catalog No.S1061 Batch:S106107

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Technical Data

Formula

C30H30Cl2N4O4
Molecular Weight 581.5 CAS No. 890090-75-2
Solubility (25°C)* In vitro DMSO 100 mg/mL (171.96 mM)
Ethanol 100 mg/mL (171.96 mM)
Water Insoluble
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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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 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 and etoposide, 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]
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]

Protocol (from reference)

Kinase Assay:

[1]
  • 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 Assay:

[1]
  • 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.
Animal Study:

[1]
  • Animal Models

    Athymic female nude mice (Nu/Nu-nuBR) injected subcutaneously with SJSA-1 cells

  • Dosages

    200 mg/kg

  • Administration

    Orally, twice a day

Customer Product Validation

Data from [Data independently produced by Int J Oncol, 2014, 44, 761-8]

Data from [Cell Death Dis, 2012, 3, e294]

Data from [Cell Death Dis, 2011, 2, e243]

Data from [Data independently produced by , , J Cell Physiol, 2018, 233(9):7424-7434]

Selleck's Nutlin-3 has been cited by 100 publications

High-throughput evaluation of genetic variants with prime editing sensor libraries [ Nat Biotechnol, 2024, 10.1038/s41587-024-02172-9] PubMed: 38472508
A homoeostatic switch causing glycerol-3-phosphate and phosphoethanolamine accumulation triggers senescence by rewiring lipid metabolism [ Nat Metab, 2024, 6(2):323-342] PubMed: 38409325
Senescence Rewires Microenvironment Sensing to Facilitate Antitumor Immunity [ Cancer Discov, 2023, 13(2):432-453] PubMed: 36302222
Senescence Rewires Microenvironment Sensing to Facilitate Antitumor Immunity [ Cancer Discov, 2023, 13(2):432-453] PubMed: 36302222
Comprehensive analysis of pyroptosis-related gene signatures for glioblastoma immune microenvironment and target therapy [ Cell Prolif, 2023, e13376.] PubMed: 36681858
Tumor suppressor p53 mediates interleukin-6 expression to enable cancer cell evasion of genotoxic stress [ Cell Death Discov, 2023, 9(1):340] PubMed: 37696858
Tumor suppressor p53 mediates interleukin-6 expression to enable cancer cell evasion of genotoxic stress [ Cell Death Discov, 2023, 9(1):340] PubMed: 37696858
Carnitine o-octanoyltransferase is a p53 target that promotes oxidative metabolism and cell survival following nutrient starvation [ J Biol Chem, 2023, 299(7):104908] PubMed: 37307919
Carnitine o-octanoyltransferase is a p53 target that promotes oxidative metabolism and cell survival following nutrient starvation [ J Biol Chem, 2023, 299(7):104908] PubMed: 37307919
p53 controls choice between apoptotic and non-apoptotic death following DNA damage [ bioRxiv, 2023, 2023.01.17.524444] PubMed: 36712034

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.