UNC0224 (UNC-0224) is a novel, potent and selective G9a histone methyltransferase inhibitor with anticancer activity. It inhibits G9a with IC50 of 15 nM in in the G9a Thioglo assay. In an ITC experiment to measure the binding affnityaffinity, UNC0224 showed 5-fold higher binding affinity than BIX01294. Through the first X-ray crystal structure of the G9a-UNC0224 complex, it was found that the 7-dimethy -laminopropoxy side chain of UNC0224 indeed occupied the lysine binding channel of G9a nicely, thus validating previous SAR experimental conclusions. In summary, UNC0224 is a significantly more potent G9a inhibitor than BIX-01294. The discovery of UNC0224 and the emergence of the first X-ray crystal structure of G9a with UNC0224 paved the way for studying the mechanism of G9a protein in vivo, which will enable structure-based design of novel inhibitors.

Physicochemical Properties

Molecular Formula	C26H43N7O2
Molecular Weight	485.66532
Exact Mass	485.347
CAS #	1197196-48-7
PubChem CID	44251522
Appearance	White to yellow solid powder
Density	1.1±0.1 g/cm3
Boiling Point	641.0±65.0 °C at 760 mmHg
Flash Point	341.5±34.3 °C
Vapour Pressure	0.0±1.9 mmHg at 25°C
Index of Refraction	1.589
LogP	1.3
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	9
Heavy Atom Count	35
Complexity	619
Defined Atom Stereocenter Count	0
InChi Key	XIVUGRBSBIXXJE-UHFFFAOYSA-N
InChi Code	InChI=1S/C26H43N7O2/c1-30(2)10-7-17-35-24-19-22-21(18-23(24)34-5)25(27-20-8-13-32(4)14-9-20)29-26(28-22)33-12-6-11-31(3)15-16-33/h18-20H,6-17H2,1-5H3,(H,27,28,29)
Chemical Name	7-[3-(dimethylamino)propoxy]-2-(hexahydro-4-methyl-1H-1,4-diazepin-1-yl)-6-methoxy-N-(1-methyl-4-piperidinyl)-4-quinazolinamine
Synonyms	UNC-0224; UNC0224; UNC 0224
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 the G9a ECSD and CLOT tests, UNC0224 (compound 10) had IC50 values of 43 nM and 57 nM, respectively. In the GLP ECSD and CLOT tests, UNC0224's IC50 values were 50 nM and 58 nM, respectively. The Kd value of UNC0224 for the G9a protein in the ITC experiment was 23 nM[2].
References	[1]. Discovery of a 2,4-diamino-7-aminoalkoxyquinazoline as a potent and selective inhibitor of histone lysine methyltransferase G9a. J Med Chem. 2009 Dec 24;52(24):7950-3. [2]. Protein lysine methyltransferase G9a inhibitors: design, synthesis, and structure activity relationships of 2,4-diamino-7-aminoalkoxy-quinazolines. J Med Chem. 2010 Aug 12;53(15):5844-57.

Solubility Data

Solubility (In Vitro)

DMSO : ~16.67 mg/mL (~34.32 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 1.67 mg/mL (3.44 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 16.7 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: ≥ 1.67 mg/mL (3.44 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 16.7 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: ≥ 1.67 mg/mL (3.44 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 16.7 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	2.0590 mL	10.2951 mL	20.5901 mL
	5 mM	0.4118 mL	2.0590 mL	4.1180 mL
	10 mM	0.2059 mL	1.0295 mL	2.0590 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.