JQ-EZ-05 (also abbreviated as JQEZ5) is a novel, potent, specific and reversible inhibitor of EZH2 (overexpressed in tumor cells such as lung cancer). Epigenetic targets are exciting new avenues for the discovery of targeted anticancer drugs. Zhang etc. have designed the open-source EZH2 inhibitor JQEZ5 which showed potent antitumor efficacy in vitro and in vivo in preclinical studies in murine and human lung adenocarcinoma models expressing high levels of EZH2. EZH2 is a lysine methyltransferase and acts as master regulator of chromatin function, it orchestrates transcriptional silencing of developmental gene networks. Overexpression of EZH2 is commonly observed in human epithelial cancers, such as non-small cell lung carcinoma (NSCLC), yet definitive demonstration of malignant transformation by deregulated EZH2 remains elusive.

Physicochemical Properties

Molecular Formula	C30H38N8O2
Molecular Weight	542.675125598907
Exact Mass	542.311
CAS #	1913252-04-6
Related CAS #	1913252-04-6
PubChem CID	121322599
Appearance	Light yellow to yellow solid powder
LogP	3.3
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	8
Heavy Atom Count	40
Complexity	988
Defined Atom Stereocenter Count	0
SMILES	O=C(C1C=C(C2=CN=C(C=C2)N2CCN(C)CC2)N=C2C=1C=NN2C(C)C)NCC1C(NC(C)=CC=1CCC)=O
InChi Key	LQTWDAYNGMMHLV-UHFFFAOYSA-N
InChi Code	InChI=1S/C30H38N8O2/c1-6-7-21-14-20(4)34-30(40)24(21)17-32-29(39)23-15-26(35-28-25(23)18-33-38(28)19(2)3)22-8-9-27(31-16-22)37-12-10-36(5)11-13-37/h8-9,14-16,18-19H,6-7,10-13,17H2,1-5H3,(H,32,39)(H,34,40)
Chemical Name	N-[(6-methyl-2-oxo-4-propyl-1H-pyridin-3-yl)methyl]-6-[6-(4-methylpiperazin-1-yl)pyridin-3-yl]-1-propan-2-ylpyrazolo[3,4-b]pyridine-4-carboxamide
Synonyms	JQEZ5
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	JQEZ5 inhibits PRC2's enzymatic functioning with a biochemical IC50 of 80nM. H661 cells treated with increasing dosages of JQEZ5 show markedly lower levels of H3K27me3 without changing H3K27 mono- or di-methylation. After 4 days of treatment, JQEZ5 inhibits the proliferation of EZH2-overexpressing H661 and H522 cells while having no effect on the proliferation of cell lines that were previously thought to be resistant to EZH2 knockdown [1].
ln Vivo	During the course of the three-week treatment, there is a noticeable and swift reduction in tumor size with JQEZ5 (75 mg/kg; intraperitoneal injection; daily). Additionally, the treatment significantly lowers H3K27me3, supporting JQEZ5's on-target effect in mice[1].
Animal Protocol	JQEZ5 was dissolved in DMSO and then diluted 1:10 in 10% (2-Hydroxypropyl)-β-cyclodextrin; 75 mg/kg/d, i.p. H661 Tumor-bearing GEMMs mice
References	[1]. Oncogenic Deregulation of EZH2 as an Opportunity for Targeted Therapy in Lung Cancer. Cancer Discov. 2016 Sep;6(9):1006-21.

Solubility Data

Solubility (In Vitro)

DMSO: Water: Ethanol:

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.61 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.8427 mL	9.2135 mL	18.4271 mL
	5 mM	0.3685 mL	1.8427 mL	3.6854 mL
	10 mM	0.1843 mL	0.9214 mL	1.8427 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.