MDMX/MDM2-IN-2 is a potent dual p53-MDM2/MDMX inhibitor (antagonist) with Ki of 0.23 μM and 2.45 μM for MDM2 and MDMX, respectively. MDMX/MDM2-IN-2 inhibits/disrupts the binding of p53 and MDM2 proteins. MDMX/MDM2-IN-2 restores p53 function and induces cell cycle arrest and apoptosis. MDMX/MDM2-IN-2 inhibits cell migration and invasion and has anti-tumor activity.

Physicochemical Properties

Molecular Formula	C28H25CL3FN3O3
Molecular Weight	576.87
Appearance	Typically exists as solid at room temperature
HS Tariff Code	2934.99.9001
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)

Biological Activity

Targets	Ki: 0.23 μM (MDM2) and 2.45 μM (MDMX)[1]
ln Vitro	On HCT116 and SH-SY5Y cells, MDMX/MDM2-IN-2 had a moderate antiproliferative effect (IC50 values of 0.68 μM and 0.54 μM, correspondingly). When it comes to normal human lung epithelial BEAS-2B cells and LO2 hepatocytes, MDMX/MDM2-IN-2 exhibits minimal cytotoxicity (IC50 values of 17.96 μM and 15.93 μM, respectively)[1]. In HCT116 and SH-SY5Y cells, MDMX/MDM2-IN-2 (0.6-2.4 μM; 48 hours) causes apoptosis [1]. In the G1 phase, MDMX/MDM2-IN-2 (0.6-2.4 μM; 48 hours) stops the cell cycle [1]. MDMX/MDM2-IN-2 (0.6-2.4 μM; 48 hours) raises p53 levels and its targets downstream, which include cleaved-caspase3, MDM2, MDMX, and p21 [1]. HCT116 and SH-SY5Y cell colony formation, migration, and invasion are strongly inhibited by MDMX/MDM2-IN-2 (0.4-0.8 μM) [1].
Cell Assay	Apoptosis Analysis[1] Cell Types: HCT116 and SH-SY5Y cells Tested Concentrations: 0.6, 1.2, 2.4 μM Incubation Duration: 48 h Experimental Results: The percentages of apoptotic HCT116 and SH-SY5Y cells were 13.63% and 15.69% with 0.6 μM. The percentage of apoptotic cells correspondingly increased to 37.6% and 40.8% with 2.4 μM. Cell Cycle Analysis[1] Cell Types: HCT116 and SH-SY5Y cells Tested Concentrations: 0.6, 1.2, 2.4 μM Incubation Duration: 48 h Experimental Results: There was an increase in the percentage of cancer cells at the G1 phase. Meanwhile, the percentage of G2 phase cells was relatively diminished. Western Blot Analysis[1] Cell Types: HCT116 and SH-SY5Y cells Tested Concentrations: 0.6, 1.2, 2.4 μM Incubation Duration: 48 h Experimental Results: Increased the levels of p53 and its downstream targets, MDM2, MDMX, p21 and cleaved-caspase3. Cell Migration Assay [1] Cell Types: HCT116 and SH-SY5Y cells Tested Concentrations: 0.4, 0.6, 0.8 μM Incubation Duration: 48 h Experimental Results: Dramatically inhibited the migration and invasion in a dose-dependent manner.
References	[1]. Structure-based discovery of novel α-aminoketone derivatives as dual p53-MDM2/MDMX inhibitors for the treatment of cancer. Eur J Med Chem. 2023 Apr 5;252:115282.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.7335 mL	8.6675 mL	17.3349 mL
	5 mM	0.3467 mL	1.7335 mL	3.4670 mL
	10 mM	0.1733 mL	0.8667 mL	1.7335 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.