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Purity: ≥98%
Nutlin-3, a tetra-substituted imidazoline, is a novel potent and selective small-molecule antagonist/inhibitor of murine double minute 2 (MDM2) with potential antitumor activities. In essence, Nutlin-3 is a protein-protein interaction (PPI) inhibitor. It works by occupying the p53 binding site in MDM2 and preventing MDM2 from binding to p53, which breaks down the autoregulator feedback loop and strengthens the p53 tumor suppressor network. Additionally, it interacts with murine double minute 4 (MDM4), a different element of the p35 tumor surveillance pathway. By attaching to MDM2 in the TP53-binding pocket, Nutlin-3 prevents MDM2-directed TP53 degradation. disrupts its interaction with p53, which causes p53 to become stabilized and active.
| Targets |
MDM2 (IC50 = 180 nM)
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| ln Vitro |
Nutlin-3 potently inhibits the MDM2-p53 interaction, leading to the activation of the p53 pathway. 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, does nutlin-3 treatment induce the expression of MDM2 and p21 and exhibit strong antiproliferative activity with an IC50 of 1.5 μM. A 48-hour treatment with 10 μM of Nutlin-3 significantly increases the amount of caspase-dependent cell apoptosis in SJSA-1 cells by about ~45%. Although Nutlin-3 also inhibits the growth and viability of human skin (1043SK) and mouse embryo (NIH/3T3) with IC50 values of 2.2 M and 1.3 M, respectively, cells remain viable one week after treatment even at 10 M of Nutlin-3, in contrast to the SJSA-1 cells, whose viability was lost at a concentration of 3 M of Nutlin-3 treatment. [1] When compared to phosphorylated p53 produced by the genotoxic drugs doxorubicin and etoposide, nutlin-3 does not cause the phosphorylation of p53 on critical serine residues. It also shows no difference in their capacity to transactivate p53 target genes and sequence-specific DNA binding. These findings show that phosphorylation of p53 on critical serines is not necessary for transcriptional activation and apoptosis.[2] Nutlin-3 has a lower affinity for MDMX than it does for MDM2, but it still has the ability to block the MDMX-p53 interaction and activate the p53 pathway in retinoblastoma cells (Weri1) with an IC50 of 0.7 μM. [3] Apoptosis is induced and cell growth is inhibited in a dose-dependent manner in cells lacking wild-type p53 due to nutlin-3 at a concentration of 30 M, which also disrupts the interaction between endogenous p73 and HDM2 and increases the stability and proapoptotic activities of p73. [4]
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| ln Vivo |
Nutlin-3 200 mg/kg orally twice daily for three weeks significantly slows the growth of SJAS-1 xenograft tumors by 90%, which is comparable to the effect of doxorubicin treatment, which inhibits tumor growth by 81%. [1]
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| Enzyme Assay |
Competition assay is performed on a Biacore S51. A PentaHis antibody is immobilized on a Series S Sensor chip CM5 in order to capture p53 that has been His-tagged. The level of capture is 200 response units (one response unit equals one pg of protein per mm2). MDM2 protein is maintained at a constant concentration of 300 nM. In order to create a concentration series of Nutlin-3 in each MDM2 test sample, Nutlin-3 is first dissolved in DMSO at a concentration of 10 mM and then further diluted. The assay is carried out 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 calculated.
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| Cell Assay |
Nutlin-3 is present in cells for 8, 24, and 48 hours at various concentrations. Real-time PCR and western blotting are used to assess the protein levels of the p21 and MDM2 genes' transcriptional activity. Using the MTT assay, cell viability is evaluated. Deoxyuridine triphosphate nick end labeling (TUNEL) staining, which is carried out by terminal deoxynucleotidyl transferase and used in flow cytometry and fluorescence microscopy, is used to identify cell apoptosis.
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| Animal Protocol |
Athymic female nude mice (Nu/Nu-nuBR) injected subcutaneously with SJSA-1 cells
200 mg/kg Orally, twice a day |
| References | |
| Additional Infomation |
4-[[4,5-bis(4-chlorophenyl)-2-(4-methoxy-2-propan-2-yloxyphenyl)-4,5-dihydroimidazol-1-yl]-oxomethyl]-2-piperazinone is a stilbenoid.
Nutlin-3 is a small molecule inhibitor that targets p53-Mdm2 interaction. |
| Molecular Formula |
C30H30CL2N4O4
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|---|---|
| Molecular Weight |
581.5
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| Exact Mass |
580.164
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| CAS # |
890090-75-2
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| Related CAS # |
Nutlin-3a;675576-98-4;Nutlin-3;548472-68-0
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| PubChem CID |
216345
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| Appearance |
White to off-white solid powder
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| Density |
1.4±0.1 g/cm3
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| Index of Refraction |
1.648
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| LogP |
2.77
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
40
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| Complexity |
919
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
BDUHCSBCVGXTJM-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C30H30Cl2N4O4/c1-18(2)40-25-16-23(39-3)12-13-24(25)29-34-27(19-4-8-21(31)9-5-19)28(20-6-10-22(32)11-7-20)36(29)30(38)35-15-14-33-26(37)17-35/h4-13,16,18,27-28H,14-15,17H2,1-3H3,(H,33,37)
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| Chemical Name |
4-[4,5-bis(4-chlorophenyl)-2-(4-methoxy-2-propan-2-yloxyphenyl)-4,5-dihydroimidazole-1-carbonyl]piperazin-2-one
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| Synonyms |
Nutlin-3
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
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| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7197 mL | 8.5985 mL | 17.1969 mL | |
| 5 mM | 0.3439 mL | 1.7197 mL | 3.4394 mL | |
| 10 mM | 0.1720 mL | 0.8598 mL | 1.7197 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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