MD-222 is a first-of-its-kind, highly efficient PROTAC-based MDM2 degrader. MD-222 consists of Cereblon and MDM2 ligand. MD-222 induces rapid degradation of MDM2 protein and activates wild-type p53 in cells. MD-222 has anti-neoplastic activities.

Physicochemical Properties

Molecular Formula	C48H47CL2FN6O6
Molecular Weight	893.83
Exact Mass	892.291
CAS #	2136246-72-3
PubChem CID	131988640
Appearance	White to off-white solid powder
LogP	6.4
Hydrogen Bond Donor Count	5
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	11
Heavy Atom Count	63
Complexity	1760
Defined Atom Stereocenter Count	3
SMILES	O=C1CCC(N2CC3C(CCCCCNC(C4=CC=C(NC(=O)[C@H]5[C@H](C6=C(C(Cl)=CC=C6)F)[C@@]6(C7C(=CC(Cl)=CC=7)NC6=O)C6(CCCCC6)N5)C=C4)=O)=CC=CC=3C2=O)C(=O)N1
InChi Key	RRSNDVCODIMOFX-MPKOGUQCSA-N
InChi Code	InChI=1S/C48H47Cl2FN6O6/c49-29-16-19-34-36(25-29)54-46(63)48(34)39(32-12-8-13-35(50)40(32)51)41(56-47(48)22-4-2-5-23-47)44(61)53-30-17-14-28(15-18-30)42(59)52-24-6-1-3-9-27-10-7-11-31-33(27)26-57(45(31)62)37-20-21-38(58)55-43(37)60/h7-8,10-19,25,37,39,41,56H,1-6,9,20-24,26H2,(H,52,59)(H,53,61)(H,54,63)(H,55,58,60)/t37?,39-,41+,48+/m0/s1
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: Please store this product in a sealed and protected environment, avoid exposure to moisture.
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	MDM2
ln Vitro	In RS4;11 and RS4;11/IRMI -2 cells, MD-222 (1-30 nM; 1-2 hours) treatment is particularly effective in lowering MDM2 protein levels and raising p53 levels in a time- and dose-dependent manner[1]. In RS4;11 cells, MD-222 (30-100 nM; 6 hours) is effective in upregulating the expression of multiple p53-targeted genes, including MDM2, CDKN1A, and PUMA, demonstrating robust p53 activation[2]. MD-222 (4 days) had no effect on RS4;11/IRMI-2, MDA-MB-231, or MDA-MB-468 cells, but has cell growth inhibitory activity in RS4;11 cells with an IC50 of 5.5 nM[1].
Cell Assay	Western Blot Analysis[1] Cell Types: RS4;11 and RS4;11/IRMI- 2 cells Tested Concentrations: 1 nM, 3 nM, 10 nM, 30 nM Incubation Duration: 1 hour, 2 hrs (hours) Experimental Results: decreased the level of MDM2 and increased the level of p53. RT-PCR[2] Cell Types: RS4;11 cells Tested Concentrations: 30 nM, 100 nM Incubation Duration: 6 hrs (hours) Experimental Results: Increased the expression of several p53-targeted genes, such as MDM2, CDKN1A, and PUMA.
References	[1]. Simple Structural Modifications Converting a Bona fide MDM2 PROTAC Degrader into a Molecular Glue Molecule: A Cautionary Tale in the Design of PROTAC Degraders. J Med Chem. 2019 Nov 14;62(21):9471-9487. [2]. Discovery of MD-224 as a First-in-Class, Highly Potent, and Efficacious Proteolysis TargetingChimera Murine Double Minute 2 Degrader Capable of Achieving Complete and Durable TumorRegression. J Med Chem. 2019 Jan 24;62(2):448-466.

Solubility Data

Solubility (In Vitro)

DMSO : 200 mg/mL (223.76 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 5 mg/mL (5.59 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 5 mg/mL (5.59 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.1188 mL	5.5939 mL	11.1878 mL
	5 mM	0.2238 mL	1.1188 mL	2.2376 mL
	10 mM	0.1119 mL	0.5594 mL	1.1188 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.