MI-773 (2'S,3R isomer, SAR405838) is a novel, potent, specific and orally bioavailable small molecule antagonist of MDM2/piro-oxindole HDM2 (Murine double minute 2/human double minute 2) with a Ki value of 0.88 nM. There may be some anticancer properties. Wild-type p53, a tumor suppressor, is primarily negatively regulated by MDM2, a protein. The p53 trans-activation domain (TAD) is blocked by MDM2, an E3 ubiquitin ligase that also promotes p53 degradation.

Physicochemical Properties

Molecular Formula	C29H34CL2FN3O3
Molecular Weight	562.50
Exact Mass	561.196
Elemental Analysis	C, 61.92; H, 6.09; Cl, 12.60; F, 3.38; N, 7.47; O, 8.53
CAS #	1303607-60-4
Related CAS #	SAR405838-d10
PubChem CID	53476877
Appearance	White to off-white solid powder
Density	1.4±0.1 g/cm3
Boiling Point	732.1±60.0 °C at 760 mmHg
Flash Point	396.6±32.9 °C
Vapour Pressure	0.0±2.5 mmHg at 25°C
Index of Refraction	1.627
LogP	5.79
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	5
Heavy Atom Count	38
Complexity	895
Defined Atom Stereocenter Count	4
SMILES	O=C([C@H](N[C@H]1CC(C)(C)C)[C@H](C2=CC=CC(Cl)=C2F)[C@@]31C(NC4=C3C=CC(Cl)=C4)=O)N[C@@H]5CC[C@@H](O)CC5
InChi Key	IDKAKZRYYDCJDU-AEPXTFJPSA-N
InChi Code	InChI=1S/C29H34Cl2FN3O3/c1-28(2,3)14-22-29(19-12-7-15(30)13-21(19)34-27(29)38)23(18-5-4-6-20(31)24(18)32)25(35-22)26(37)33-16-8-10-17(36)11-9-16/h4-7,12-13,16-17,22-23,25,35-36H,8-11,14H2,1-3H3,(H,33,37)(H,34,38)/t16?,17?,22-,23-,25+,29+/m0/s1
Chemical Name	(2'R,3R,3'S,5'S)-6-chloro-3'-(3-chloro-2-fluorophenyl)-5'-(2,2-dimethylpropyl)-N-(4-hydroxycyclohexyl)-2-oxospiro[1H-indole-3,4'-pyrrolidine]-2'-carboxamide
Synonyms	MI773; MI-773; MI 773; MI 77301; MI77301; MI-77301; SAR-405838; SAR 405838; SAR405838
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: (1). This product is not stable in solution, please use freshly prepared working solution for optimal results.(2). 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 (Ki = 0.88 nM); MDM2 (Kd = 8.2 nM)
ln Vitro	SAR405838 (MI-77301) potently inhibits cell growth in cancer cell lines, including SJSA-1 (IC50, 0.092 μM), RS4;11 (IC50, 0.089 μM), LNCaP (IC50, 0.27 μM), and HCT-116 (IC50, 0.20 μM) cells, and displays high selectivity over cancer cell lines with mutated or deleted p53, including SAOS-2 (IC50, >10 μM), PC-3 (IC50, >10 μM), SW620 (IC50, >10 μM), and HCT-116 (p53-/-) (IC50, >20 μM) cells[1]. SAR405838 has a modestly reduced potency when compared to the control RS4;11 cell line, but it still effectively inhibits cell growth and induces dose-dependent apoptosis in the ABTR1 and ABTR2 sublines[2].
ln Vivo	SAR405838 inhibits the growth of tumors completely or permanently in mouse xenograft models of SJSA-1 osteosarcoma, RS4;11 acute leukemia, LNCaP prostate cancer, and HCT-116 colon cancer at dose schedules that are well tolerated. Surprisingly, SAR405838 only requires one oral dose to completely reverse tumor growth in the SJSA-1 model. MI-773 (p.o.) effectively inhibits tumor growth in a dose-dependent manner in the SJSA-1 osteosarcoma, acute lymphoblastic leukemia RS4;11, LNCaP prostate cancer, and HCT-116 colon cancer xenograft model (10 mg/kg, 30 mg/kg, 50 mg/kg, 100 mg/kg, and 200 mg/kg)[1].
Enzyme Assay	Using a Fluorescence-polarization (FP) binding assay, binding affinities of MDM2 inhibitors and p53 peptide to MDM2 protein are assessed. MI-773's binding affinities to Bcl-2, Bcl-xL, Mcl-1, and β-catenin are assessed using a competitive FP-based assay, and its affinity for MDMx is assessed using Biolayer Interferometry technology.
Cell Assay	In a water-soluble tetrazolium-based assay, cell growth inhibition activity is assessed. Trypan blue staining is used to measure cell death, and a kit for staining with Annexin V-FLUOS determines apoptosis.
Animal Protocol	10% PEG400: 3% Cremophor: 87% PBS, or 2% TPGS: 98% PEG200; 200 mg/kg; Oral SCID mice with SJSA-1 osteosarcoma (females), acute lymphoblastic leukemia RS4;11 (females), LNCaP prostate cancer (males), or HCT-116 colon cancer (females) xenograft model
References	[1]. SAR405838: an optimized inhibitor of MDM2-p53 interaction that induces complete and durable tumor regression. Cancer Res. 2014 Oct 15;74(20):5855-5865. [2]. Elucidation of Acquired Resistance to Bcl-2 and MDM2 Inhibitors in Acute Leukemia In Vitro and In Vivo. Clin Cancer Res. 2015 Jun 1;21(11):2558-2568.
Additional Infomation	SAR405838 has been used in trials studying the treatment of Neoplasm Malignant. p53-HDM2 Interaction Inhibitor MI-773 is an orally available spiro-oxindole HDM2 (human double minute 2) antagonist with potential antineoplastic activity. Upon oral administration, the p53-HDM2 protein-protein interaction inhibitor MI-773 binds to HDM2, preventing the binding of the HDM2 protein to the transcriptional activation domain of the tumor suppressor protein p53. By preventing this HDM2-p53 interaction, the proteasome-mediated enzymatic degradation of p53 is inhibited and the transcriptional activity of p53 is restored, which may result in the restoration of p53 signaling and lead to the p53-mediated induction of tumor cell apoptosis. HDM2, a zinc finger protein and a negative regulator of the p53 pathway, is often overexpressed in cancer cells. It has been implicated in cancer cell proliferation and survival.

Solubility Data

Solubility (In Vitro)

DMSO: ~100 mg/mL (~177.8 mM) Water: <1 mg/mL (slightly soluble or insoluble) Ethanol: ~31 mg/mL warming (~55.1 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.44 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 25.0 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: 2.5 mg/mL (4.44 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (4.44 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 25.0 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	1.7778 mL	8.8889 mL	17.7778 mL
	5 mM	0.3556 mL	1.7778 mL	3.5556 mL
	10 mM	0.1778 mL	0.8889 mL	1.7778 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.