UNC0638 (UNC-0638) is a selective and cell-permeable inhibitor of G9a histone methyltransferase and GLP with IC50 of<15 nM and 19 nM, respectively. Protein lysine methyltransferases G9a and GLP modulate the transcriptional repression of a variety of genes via dimethylation of Lys9 on histone H3 (H3K9me2) as well as dimethylation of non-histone targets. UNC-0638 shows selectivity over a wide range of epigenetic and non-epigenetic targets. UNC0638 showed balanced physicochemical properties and potency aiding cell penetration in vitro, had high potency in cellular assays. It was much less toxic than BIX01294 to cells. In MDA-MB-231 cells, in a concentration-dependent manner, exposure to UNC0638 for 48 h reduced H3K9me2 levels with an IC50 value of 81 ± 9 nM (n= 3), which showed considerably higher potency than BIX01294 (IC50= 500 ± 43 nM (n= 3)). In reducing H3K9me2 levels, UNC0638 was of greater maximum effect than BIX01294. This effect is close, but not equal, to the effect on the double knockdown of G9a and GLP via shRNA.

Physicochemical Properties

Molecular Formula	C30H47N5O2
Molecular Weight	509.73
Exact Mass	509.372
CAS #	1255580-76-7
Related CAS #	1255580-76-7
PubChem CID	46224516
Appearance	White to yellow solid powder
Density	1.1±0.1 g/cm3
Boiling Point	563.9±50.0 °C at 760 mmHg
Melting Point	93-94 ºC
Flash Point	294.8±30.1 °C
Vapour Pressure	0.0±1.5 mmHg at 25°C
Index of Refraction	1.587
LogP	5.75
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	10
Heavy Atom Count	37
Complexity	660
Defined Atom Stereocenter Count	0
InChi Key	QOECJCJVIMVJGX-UHFFFAOYSA-N
InChi Code	InChI=1S/C30H47N5O2/c1-22(2)35-17-12-24(13-18-35)31-30-25-20-27(36-3)28(37-19-9-16-34-14-7-8-15-34)21-26(25)32-29(33-30)23-10-5-4-6-11-23/h20-24H,4-19H2,1-3H3,(H,31,32,33)
Chemical Name	2-cyclohexyl-6-methoxy-N-(1-propan-2-ylpiperidin-4-yl)-7-(3-pyrrolidin-1-ylpropoxy)quinazolin-4-amine
Synonyms	UNC-0638, UNC0638, UNC 0638;2-cyclohexyl-N-(1-isopropylpiperidin-4-yl)-6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)quinazolin-4-amine
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	UNC0638 is a highly selective and potent G9a and GLP inhibitor that works against a variety of both epigenetic and non-epigenetic targets. It is found that UNC0638's Ki is 3.0±0.05 nM (n=2). The Morrison Ki for UNC0638 is 3.7±0.2 nM (n=3), which is consistent with this. We assess UNC0638's selectivity over a broad spectrum of epigenetic targets. Notably, UNC0638 is not active against PRDM1, PRDM10, and PRDM12, nor against other methyltransferases of H3K9 (SUV39H1 and SUV39H2), H3K27 (EZH2), H3K4 (SETD7, MLL, and SMYD3), H3K79 (DOT1L), or H4K20 (SETD8). Furthermore, UNC0638 exhibits no activity against the histone acetyltransferase HTATIP as well as the protein arginine methyltransferases PRMT1 and PRMT3. It is noteworthy that UNC0638 exhibits modest but detectable activity against DNA methyltransferase DNMT1 (IC50=107,000±6,000 nM), a Jumonji protein demethylase, and JMJD2E (IC50=4,500±1,100 nM). Nevertheless, UNC0638 has >200-fold selectivity for G9a and GLP over JMJD2E and >5,000-fold selectivity for G9a and GLP over DNMT1[1]. A particular class of small chemical known as UNC0638 has the ability to precisely block the activity of the enzyme histone methyltransferase EHMT and lower cell levels of H3K9 dimethylation (H3K9me2)[2].
ln Vivo	In 6-week-old male athymic nude mice subcutaneously inoculated with BON cells, UNC0638 decreased H3K9me2 level. In organotypic cochlear cultures, rapid increase of H3K9me2 upon the damage of hair cells was observed. Both ex vivo and in vivo, UNC0638 effectively prevented aminoglycosides-induced hair cell damage.
Animal Protocol	DMPK studies in male Swiss albino mice (3 animals per data point) are conducted, following intravenous (IV, 1 mg/kg), oral (PO, 3 mg/kg), and intraperitoneal (IP, 2.5 mg/kg) administration of UNC0638. Male athymic nude mice
References	[1]. A chemical probe selectively inhibits G9a and GLP methyltransferase activity in cells. Nat Chem Biol. 2011 Jul 10;7(8):566-74. [2]. Effects of the Histone Methyltransferase Inhibitor UNC0638 on Histone H3K9 Dimethylation of Cultured Ovine Somatic Cells and Development of Resulting Early Cloned Embryos. Reprod Domest Anim. 2014 Apr;49(2):e21-5. [3]. Inhibition of EHMT2 Induces a Robust Antiviral Response Against Foot-and-Mouth Disease and Vesicular Stomatitis Virus Infections in Bovine Cells. J Interferon Cytokine Res. 2016 Jan;36(1):37-47.
Additional Infomation	UNC0638 is a member of the class of quinazolines that is quinazoline substituted by cyclohexyl, [1-(propan-2-yl)piperidin-4-yl]amino, methoxy, and 3-(pyrrolidin-1-yl)propoxy groups at positions 2, 4, 6 and 7, respectively. It inhibits G9a and GLP histone methyltransferase activity (IC50 = <15 nM and <20 nM, respectively). It has a role as an antineoplastic agent and an antiviral agent. It is a member of quinazolines, an aminopiperidine, an aromatic ether, a member of pyrrolidines, a tertiary amino compound and a secondary amino compound.

Solubility Data

Solubility (In Vitro)

DMSO:100 mg/mL (196.2 mM) Water:6 mg/mL (11.8 mM) Ethanol:100 mg/mL (196.2 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.90 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 (4.90 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 (4.90 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	1.9618 mL	9.8091 mL	19.6182 mL
	5 mM	0.3924 mL	1.9618 mL	3.9236 mL
	10 mM	0.1962 mL	0.9809 mL	1.9618 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.