MI-3 (Menin-MLL Inhibitor 3) is a novel and potent inhibitor of menin-MLL interaction with potential antitumor activity. It inhibits menin-MLL interaction with an IC50 of 648 nM. It exhibits excellent antiproliferative activity against various cancer cells.

Physicochemical Properties

Molecular Formula	C18H25N5S2
Molecular Weight	375.55
Exact Mass	375.155
CAS #	1271738-59-0
Related CAS #	1271738-59-0
PubChem CID	51001299
Appearance	Light yellow to yellow solid powder
Density	1.4±0.1 g/cm3
Boiling Point	527.9±60.0 °C at 760 mmHg
Flash Point	273.1±32.9 °C
Vapour Pressure	0.0±1.4 mmHg at 25°C
Index of Refraction	1.713
LogP	3.23
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	3
Heavy Atom Count	25
Complexity	516
Defined Atom Stereocenter Count	0
InChi Key	FUGQNAUKABUDQI-UHFFFAOYSA-N
InChi Code	InChI=1S/C18H25N5S2/c1-12(2)14-9-13-15(20-11-21-16(13)24-14)22-5-7-23(8-6-22)17-19-10-18(3,4)25-17/h9,11-12H,5-8,10H2,1-4H3
Chemical Name	4-[4-(5,5-dimethyl-4H-1,3-thiazol-2-yl)piperazin-1-yl]-6-propan-2-ylthieno[2,3-d]pyrimidine
Synonyms	MI 3; MI-3; MI3
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	In human cells, treatment with MI-3 (12.5-50 μM; HEK293 cells) efficiently suppresses the menin-MLL-AF9 interaction[1]. In KOPN-8, MV4, and ME-1 cells, MI-3 (0-1.6 μM; 72 hours; 11 cells) treatment demonstrates an efficient and dose-dependent growth inhibition[1]. The administration of MI-3 (12.5-50 μM; 48 hours; MV4; 11 cells) causes a significant, dose-dependent rise in the number of Annexin V and Annexin V/propidium iodide (PI) cells, indicating an increase in apoptosis[1]. The treatment of THP-1 cells with MI-3 (6.25-25 μM; 6 days) significantly lowers the expression of MEIS1 and HOXA9.
ln Vivo	MLL-AF9 transformed BMC that remained viable after 7 days of treatment with MI-2 and MI-3 showed substantial changes in morphology, indicative of monocytic differentiation, as evidenced by increased cell size, lower nuclear to cytoplasmic ratio and highly vacuolated cytoplasm. Consistent with the change in cell morphology, the expression of CD11b was substantially increased on MLL-AF9 transformed BMC after 7 days of treatment with MI-2 and MI-3.
Cell Assay	Western Blot Analysis[1] Cell Types: HEK293 cells Tested Concentrations: 12.5 μM, 25 μM, 50 μM Incubation Duration: Experimental Results: Very effectively inhibited the menin-MLL-AF9 interaction in human cells. Cell Viability Assay[1] Cell Types: KOPN-8 and MV4;11 cells Tested Concentrations: 0 μM, 0.4 μM, 0.8 μM, 1.2 μM, 1.6 μM Incubation Duration: 72 hrs (hours) Experimental Results: demonstrated an effective and dose-dependent growth inhibition in KOPN-8 and MV4;11 cells.
Animal Protocol	NA NA
References	[1]. Menin-MLL inhibitors reverse oncogenic activity of MLL fusion proteins in leukemia. Nature Chemical Biology (2012), 8(3), 277-284.

Solubility Data

Solubility (In Vitro)

DMSO: 19 mg/mL (50.6 mM) Water:<1 mg/mL Ethanol:19 mg/mL (50.6 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 0.83 mg/mL (2.21 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 8.3 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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: ≥ 0.83 mg/mL (2.21 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 8.3 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: ≥ 0.83 mg/mL (2.21 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 8.3 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	2.6628 mL	13.3138 mL	26.6276 mL
	5 mM	0.5326 mL	2.6628 mL	5.3255 mL
	10 mM	0.2663 mL	1.3314 mL	2.6628 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.