MI-463 is a novel potent & orally bioactive inhibitor of Menin-MLL interaction with an IC50 value of 15.3 nM. MI-463 can reach the target protein in mammalian cells and effectively inhibit the menin-MLL-AF9 interaction at sub-micromolar concentrations. Treatment of murine bone marrow cells (BMC) transformed with the MLL-AF9 oncogene with MI-463 results in substantial growth inhibition, with GI50 of 0.23 μM. MI-463 achieves high level in peripheral blood following a single intravenous or oral dose, while also showing high oral bioavailability (45%).

Physicochemical Properties

Molecular Formula	C24H23F3N6S
Molecular Weight	484.54
Exact Mass	484.165
CAS #	1628317-18-9
Related CAS #	1628317-18-9
PubChem CID	90455046
Appearance	White to light yellow solid powder
Density	1.4±0.1 g/cm3
Boiling Point	663.2±55.0 °C at 760 mmHg
Flash Point	354.9±31.5 °C
Vapour Pressure	0.0±2.0 mmHg at 25°C
Index of Refraction	1.669
LogP	3.88
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	5
Heavy Atom Count	34
Complexity	751
Defined Atom Stereocenter Count	0
InChi Key	DZACSLYTXLZAAF-UHFFFAOYSA-N
InChi Code	InChI=1S/C24H23F3N6S/c1-14-15(2-3-21-19(14)8-17(11-28)31-21)12-33-6-4-16(5-7-33)32-22-20-9-18(10-24(25,26)27)34-23(20)30-13-29-22/h2-3,8-9,13,16,31H,4-7,10,12H2,1H3,(H,29,30,32)
Chemical Name	4-methyl-5-((4-((6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)methyl)-1H-indole-2-carbonitrile
Synonyms	MI-463; MI 463; MI463.
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

ln Vitro	At sub-micromolar concentrations, MI-463 can efficiently block the menin-mLL-AF9 interaction by binding to the target protein in mammalian cells. When MI-463 is applied to mouse bone marrow cells (BMC) that have been transformed with the mLL-AF9 oncogene, there is a significant growth inhibition observed, with a GI50 of 0.23 μM[1].
ln Vivo	After a single intravenous or oral dose, MI-463 exhibits a high level in peripheral blood and a high oral bioavailability of 45%. Through on-target activity, pharmacologic inhibition of the menin-mLL interaction significantly slows the progression of mLL leukemia in murine models without being toxic. MI-463 administered intraperitoneally (i.p.) once daily causes significant inhibition of tumor growth. After receiving MI-463 treatment, the expression of the genes MEIS1 and HOXA9 that are targets of mLL fusion proteins is greatly decreased. Twenty days of treatment with MI-463 in MV4;11 xenograft recipient mice also significantly delays the progression of leukemia, as evidenced by a significant decline in bioluminescence level, which is correlated with a significant reduction in the number of leukemic cells in bone marrow, spleen, and peripheral blood samples[1].
Animal Protocol	i.p. administration Mice
References	[1]. Pharmacologic inhibition of the Menin-MLL interaction blocks progression of MLL leukemia in vivo. Cancer Cell. 2015 Apr 13;27(4):589-602.

Solubility Data

Solubility (In Vitro)

DMSO:96 mg/mL (198.1 mM) Water:<1 mg/mL Ethanol:22 mg/mL (45.4 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.29 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 20.8 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: 2.08 mg/mL (4.29 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 20.8 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: ≥ 2.08 mg/mL (4.29 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 20.8 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	2.0638 mL	10.3191 mL	20.6381 mL
	5 mM	0.4128 mL	2.0638 mL	4.1276 mL
	10 mM	0.2064 mL	1.0319 mL	2.0638 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.