JIB-04 (also known as NSC693627) is a novel, potent and pan-selective inhibitor of Jumonji histone demethylase (JMJD) inhibitor with antineoplastic activity. As an epigenetic modifier, it inhibits JARID1A, JMJD2E, JMJD3, JMJD2A, JMJD2B, JMJD2C, and JMJD2D with IC50 values of 230, 340, 855, 445, 435, 1100, and 290 nM, respectively. JIB-04 inhibits cancer cell growth in vitro selectively and reduces in vivo tumor growth in H358 and A549 mouse xenograft models. It also prolongs the survival of mice with breast cancer.

Physicochemical Properties

Molecular Formula	C17H13CLN4
Molecular Weight	308.76
Exact Mass	308.082
CAS #	199596-05-9
Related CAS #	(Z)-JIB-04;199596-24-2
PubChem CID	6519698
Appearance	Off-white to yellow solid powder
Density	1.2±0.1 g/cm3
Boiling Point	472.9±55.0 °C at 760 mmHg
Flash Point	239.8±31.5 °C
Vapour Pressure	0.0±1.2 mmHg at 25°C
Index of Refraction	1.644
LogP	3.92
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	4
Heavy Atom Count	22
Complexity	368
Defined Atom Stereocenter Count	0
SMILES	C1=CC=C(C=C1)/C(=N\NC2=NC=C(C=C2)Cl)/C3=CC=CC=N3
InChi Key	YHHFKWKMXWRVTJ-OQKWZONESA-N
InChi Code	InChI=1S/C17H13ClN4/c18-14-9-10-16(20-12-14)21-22-17(13-6-2-1-3-7-13)15-8-4-5-11-19-15/h1-12H,(H,20,21)/b22-17+
Chemical Name	(E)-5-chloro-2-(2-(phenyl(pyridin-2-yl)methylene)hydrazinyl)pyridine
Synonyms	NSC-693627; NSC 693627; NSC693627; JIB-04; JIB 04; JIB04;
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	Compared to HBEC and PrSC/PrEC, JIB-04 was consistently more selective for cancer and normal cells, as demonstrated by its increased sensitivity in lung and prostate cancer cell lines (IC50 as low as 10nM). Jumonji demethylase activity in cells is inhibited by JIB-04, and JIB-04's function in cells is influenced by Jumonji levels [1]. The proliferation of GB cell lines and stem cell-enriched cultures is markedly inhibited by JIB-04. JIB-04 causes H3K4 hypermethylation, controls the expression of genes involved in limiting the proliferation of cancer cells, and has the strongest inhibitory effect on KDM5A. Furthermore, JIB-04 (2500 nM) inactivates PI3K and activates the pathways involved de autophagy and apoptosis. JIB-04 and TMZ work together to destroy GB cells as well[2].
ln Vivo	JIB-04 significantly lowers the rates of cancer-related mortality in mice, extending their lifespan[1]. The mice's brains contain bioactive concentrations of JIB-04 (60, 40, and 20 mg/kg, ip). With a Hazard Ratio of 0.5, the orthotopic GB xenograft model indicates a tendency toward prolonged survival in mice treated with JIB-04[2].
Animal Protocol	Dissolved in 12.5% Cremophor EL, 12.5% DMSO, 10% DMSO 90% sesame oil (H358, i.p.); 110 mg/kg (H358, i.p.), 55 mg/kg (A549, Oral gavage); i.p. or p.o. Mice harboring H358 xenografts or A549 xenografts
References	[1]. A small molecule modulates Jumonji histone demethylase activity and selectively inhibits cancer growth. Nat Commun, 2013. 4: p. 2035. [2]. Small molecules targeting histone demethylase genes (KDMs) inhibit growth of temozolomide-resistant glioblastoma cells. Oncotarget. 2017 Apr 4.

Solubility Data

Solubility (In Vitro)

DMSO: 25 mg/mL (80.9 mM) Water:<1 mg/mL Ethanol:<1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (6.74 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 20.8 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: ≥ 1.25 mg/mL (4.05 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 12.5 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 3: 10 mg/mL (32.39 mM) in 0.5% CMC-Na/saline water (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. 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	3.2388 mL	16.1938 mL	32.3876 mL
	5 mM	0.6478 mL	3.2388 mL	6.4775 mL
	10 mM	0.3239 mL	1.6194 mL	3.2388 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.