MI-2 (MI2, Menin-MLL Inhibitor) is a novel, potent and selective inhibitor of menin-MLL interaction with antineoplastic activity. It inhibits menin-MLL interaction with an IC50 of 446 nM. MI-2 acts by binding to the wild-type menin but doesn't bind M278K and Y323K menin mutants. In HEK293 cells transfected with Flag-MLL-AF9, MI-2 effectively inhibited the menin-MLL-AF9 interaction without affecting the expression level of menin and MLL-AF9.

Physicochemical Properties

Molecular Formula	C18H25N5S2
Molecular Weight	375.55
Exact Mass	375.155
CAS #	1271738-62-5
Related CAS #	1271738-62-5
PubChem CID	54765302
Appearance	Light yellow to yellow solid powder
Density	1.4±0.1 g/cm3
Boiling Point	538.0±60.0 °C at 760 mmHg
Flash Point	279.2±32.9 °C
Vapour Pressure	0.0±1.4 mmHg at 25°C
Index of Refraction	1.712
LogP	3.59
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	4
Heavy Atom Count	25
Complexity	504
Defined Atom Stereocenter Count	0
InChi Key	SRQYLNYQAPCPIR-UHFFFAOYSA-N
InChi Code	InChI=1S/C18H25N5S2/c1-4-5-13-10-14-15(20-12-21-16(14)24-13)22-6-8-23(9-7-22)17-19-11-18(2,3)25-17/h10,12H,4-9,11H2,1-3H3
Chemical Name	4-[4-(5,5-dimethyl-4H-1,3-thiazol-2-yl)piperazin-1-yl]-6-propylthieno[2,3-d]pyrimidine
Synonyms	MI2 ; Menin-MLL inhibitor 2;MI 2 ; Menin MLL inhibitor 2;MI-2 ; Menin-MLL inhibitor-2;
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	The menin-MLL inhibitor MI-2, with GI50 values of approximately 5 μM, very efficiently inhibits the proliferation of MLL-AF9 and MLL-ENL transduced BMC. Several compounds with IC50 values in the nanomolar range were developed as a result of the assessment of various hydrophobic groups at R1. These compounds include MI-2 (IC50= 446±28 nM) and MI-3 (IC50= 648±25 nM). The dissociation constants (Kd=158 nM for MI-2) for the menin-MLL inhibitors have been measured at the nanomolar range. Inhibiting the menin-MLL-AF9 connection in human cells is a highly effective way that MI-2 can access the protein target. Moreover, MI-2 exhibits minimal impact on the proliferation of BMC transduced with E2A-HLF (GI50>50 μM). This could potentially be attributed to the inhibition of menin interaction with wild-type MLL. After MI-2 treatment, GI50 values for MV4;11 (harboring MLL-AF4; GI50=9.5 μM), KOPN-8 (MLL-ENL; GI50=7.2 μM), ML-2 (MLL-AF6; GI50=8.7 μM), and MonoMac6 (MLL-AF9; GI50=18 μM) are less than 10 μM[1].
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.
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.
Additional Infomation	4-[4-(5,5-dimethyl-4H-thiazol-2-yl)-1-piperazinyl]-6-propylthieno[2,3-d]pyrimidine is a N-arylpiperazine and a thienopyrimidine.

Solubility Data

Solubility (In Vitro)

DMSO: 75 mg/mL (199.7 mM) Water:<1 mg/mL Ethanol: NA

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.66 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 (6.66 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 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 (6.66 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	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.