MMRi62, a ferroptosis inducer targeting MDM2-MDM4 (negative regulators of tumor suppressor p53). MMRi62 exhibits pro-apoptotic activity and induces autophagy in pancreatic ductal adenocarcinoma (PDAC) cells without the need for P53. As a result of MMRi62's induction of ferroptosis, the ferritin heavy chain (FTH1) is degraded in the lysosomes and reactive oxygen species are increased. In addition to inhibiting the orthotopic xenograft PDAC mouse model in vivo with high frequency mutation characteristics of KRAS and TP53, MMRi62 also causes the proteasomal degradation of mutant p53.12[1][2].

Physicochemical Properties

Exact Mass	395.0592
Elemental Analysis	C, 63.65; H, 3.82; Cl, 17.89; N, 10.60; O, 4.04
CAS #	352693-80-2
Related CAS #	352693-80-2
Appearance	White to off-white solid
InChi Key	FMUBUUSODRAEMS-UHFFFAOYSA-N
InChi Code	InChI=1S/C21H15Cl2N3O/c22-16-7-3-6-14(18(16)23)20(26-17-8-1-2-11-24-17)15-10-9-13-5-4-12-25-19(13)21(15)27/h1-12,20,27H,(H,24,26)
Chemical Name	7-[(2,3-dichlorophenyl)-(pyridin-2-ylamino)methyl]quinolin-8-ol
Synonyms	MMRi 62; MMRi62; MMRi-62
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 Note: This product requires protection from light (avoid light exposure) during transportation and storage.
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

ln Vitro	By causing cell death, MMRi62 prevents pancreatic ductal glands (PDAC) from proliferating, cloning, and growing generatively [1]. MMRi62 attaches to the cyclic heterodimer of MDM2 and MDM4 (3 nM-100 μM; 4 h). Albino cells are activated and inhibited by MMRi62 (10 nM-1 μM; 72 h), with IC50 values of 0.34 μM (HL60) and 0.22 μM (HL60VR)[2]. In a dose-dependent manner, 5 μM, 10μM; 24 hours) decreases MDM2B autoubiquitination and enhances MDM4 ubiquitination [2]. An E3 ligase modification called MMRi62 causes the substrate preference to change from MDM2 to MDM4 [2]. Cellular fluorescence is transmitted by MMRi62 (5 μM; 24, 72 h) without the aid of p53[2].
ln Vivo	MMRi62 showed anti-activity in an orthotopic xenograft PDAC tumor model, decreasing tumor development in mice by suppressing NCOA4 and mutant p53 transcription [1]. MMRi62 also totally prevents orthotopic tumor development [1].
Cell Assay	Western Blot Analysis[2] Cell Types: WT-p53 with MV4-11 cells; 293 cells transfected with MDM2B and MDM4 Tested Concentrations: 2, 2.5, 5, 10, 40, 80, 160 μM Incubation Duration: 24 hrs (hours) Experimental Results: 2 μM Levels of cleaved PARP protein and activated caspase 3 increased in wt-p53 MV4-11 cells for 24 hrs (hours). At 5 μM and 10 μM for 24 hrs (hours), MDM2B autoubiquitination diminished and MDM4 ubiquitination increased. In NALM6 cells, 5 μM induces MDM2-dependent degradation of MDM4 protein. Cell proliferation assay[2] Cell Types: primary AML patient cells, NALM6 cells and NALM6shp53 cells Tested Concentrations: 1, 10, 25 and 50 µM Incubation Duration: 24 hrs (hours) and 72 hrs (hours) Experimental Results: Primary cells induced in 24 hands NALM6 cells induced apoptosis for 72 hrs (hours) in AML patients.
References	[1]. Small-Molecule MMRi62 Induces Ferroptosis and Inhibits Metastasis in Pancreatic Cancer via Degradation of Ferritin Heavy Chain and Mutant p53. Mol Cancer Ther. 2022 Apr 1;21(4):535-545. [2]. Small molecule MMRi62 targets MDM4 for degradation and induces leukemic cell apoptosis regardless of p53 status. Front Oncol. 2022 Aug 5;12:933446.

Solubility Data

Solubility (In Vitro)

DMSO: ~62.5 mg/mL (~157.7 mM)

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.)