Nutlin-3

製品コードS1061 バッチS106108

印刷

化学情報

Chemical Structure Synonyms N/A Storage
(From the date of receipt)		 3 years -20°C powder
1 years -80°C in solvent
化学式

C30H30Cl2N4O4
分子量 581.5 CAS No. 890090-75-2
Solubility (25°C)* 体外 DMSO 100 mg/mL (171.96 mM)
Ethanol 100 mg/mL (171.96 mM)
Water Insoluble
体内 （毎回新しく調製した物を用意してください）
Homogeneous suspension
CMC-NA
≥5mg/ml Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution
5%DMSO 40%PEG300 5%Tween80 50%ddH2O

この製剤はselleckのラボで検証済みです。上記の溶解方法がご要望を満たさない場合、selleckの営業担当までお問い合わせ頂ければ、個別の試験を行います。

 5.000mg/ml (8.60mM) Taking the 1 mL working solution as an example, add 50 μL of 100 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to make it clear; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results. 
* <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.

溶剤液（一定の濃度）を調合する

生物活性

製品説明 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.
in vitro

Nutlin-3 potently inhibits the MDM2-p53 interaction, leading to the activation of the p53 pathway. This compound 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, this compound treatment at 10 μM for 48 hours significantly induces caspase-dependent cell apoptosis by ~45%. Although it 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 this chemical, in contrast with the SJSA-1 cells with viability lost at 3 μM of this chemical treatment. [1] It 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, this compound can block the MDMX–p53 interaction and induce the p53 pathway in retinoblastoma cells (Weri1) with IC50 of 0.7 μM. [3] This chemical 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]

プロトコル(参考用のみ)

キナーゼアッセイ 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 this compound 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 this chemical is calculated as a percentage of binding in the absence of it and IC50 is calculate
細胞アッセイ 細胞株 HCT116, RKO, SJSA-1, SW480, and MDA-MB-435
濃度 Dissolved in DMSO, final concentrations ~ 30 μM
反応時間 8, 24, and 48 hours
実験の流れ

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.
動物実験 動物モデル Athymic female nude mice (Nu/Nu-nuBR) injected subcutaneously with SJSA-1 cells
投薬量 200 mg/kg
投与方法 Orally, twice a day

参考

  • https://pubmed.ncbi.nlm.nih.gov/14704432/
  • https://pubmed.ncbi.nlm.nih.gov/15471885/
  • https://pubmed.ncbi.nlm.nih.gov/17080083/
  • https://pubmed.ncbi.nlm.nih.gov/17700533/
  • http://eprints.lib.hokudai.ac.jp/dspace/bitstream/2115/54893/1/Masaki_Miyazaki.pdf

カスタマーフィードバック

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]

Selleckの高級品が、幾つかの出版された研究調査結果（以下を含む）で使われた：

Disparate Pathways for Extrachromosomal DNA Biogenesis and Genomic DNA Repair [ Cancer Discov, 2025, 15(1):69-82] PubMed: 39109936
MDM2 inhibitors induce apoptosis by suppressing MDM2 and enhancing p53, Bax, Puma and Noxa expression levels in imatinib‑resistant chronic myeloid leukemia cells [ Biomed Rep, 2025, 22(4):65] PubMed: 39991005
Alanyl-tRNA synthetase, AARS1, is a lactate sensor and lactyltransferase that lactylates p53 and contributes to tumorigenesis [ Cell, 2024, 187(10):2375-2392.e33] PubMed: 38653238
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
Decreased plasma gelsolin fosters a fibrotic tumor microenvironment and promotes chemoradiotherapy resistance in esophageal squamous cell carcinoma [ J Biomed Sci, 2024, 31(1):90] PubMed: 39261905
Multiomics analyses reveal adipose-derived stem cells inhibit the inflammatory response of M1-like macrophages through secreting lactate [ Stem Cell Res Ther, 2024, 15(1):485] PubMed: 39696485
A CRISPR/Cas9 screen in embryonic stem cells reveals that Mdm2 regulates totipotency exit [ Commun Biol, 2024, 7(1):809] PubMed: 38961268
NEK2 promotes TP53 ubiquitination to enhance the proliferation and migration of TP53 wild-type glioblastoma cells [ Neoplasma, 2024, 71(3):255-265] PubMed: 38764296
MYC and p53 alterations cooperate through VEGF signaling to repress cytotoxic T cell and immunotherapy responses in prostate cancer [ bioRxiv, 2024, 2024.07.24.604943] PubMed: 39091883

長期の保管のために-20°Cの下で製品を保ってください。

人間や獣医の診断であるか治療的な使用のためにでない。

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