GSKJ1 (GSK-J1; GSK-J 1; GSK-J-1) is a novel, highly selective and potent inhibitor of histone demethylase (H3K27me3/me2-demethylases JMJD3/KDM6B and UTX/KDM6A) with potential antineoplastic activity. It that has significant activity (IC50 60 nM for human JmjD3) in vitro and in cell assays using an ester prodrug derivative (GSK-J4: 1 µM < IC50 < 10 µM; e.g. 9 µM in primary human macrophages). The methyl groups from tri- and dimethylated lysine 4 of histone H3 are removed by the KDM5/JARID1 family of Fe(II)- and α-ketoglutarate-dependent demethylases. KDM5A (JARID1A/RBP2) and KDM5B (JARID1B/PLU1) may play a role as oncogenic drivers, according to mounting evidence from primary tumors and model systems. As an ester derivative (GSK-J5) in cells and a control for target effects in vitro, the pyridine regioisomer GSK-J2 exhibits significantly less on-target activity (IC50 > 100 µM for human JmjD3). Recent data against H3K4me3/2/1 demethylases indicates that GSK-J1 also exhibits some activity (IC50 950 nM for Jarid1b, IC50 1.76 uM for Jarid1c).

Physicochemical Properties

Molecular Formula	C22H23N5O2
Molecular Weight	389.45
Exact Mass	389.185
Elemental Analysis	C, 67.85; H, 5.95; N, 17.98; O, 8.22
CAS #	1373422-53-7
Related CAS #	1373422-53-7
PubChem CID	56963315
Appearance	White to yellow solid powder
Density	1.3±0.1 g/cm3
Boiling Point	608.9±55.0 °C at 760 mmHg
Flash Point	322.0±31.5 °C
Vapour Pressure	0.0±1.8 mmHg at 25°C
Index of Refraction	1.653
LogP	2.75
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	6
Heavy Atom Count	29
Complexity	517
Defined Atom Stereocenter Count	0
SMILES	O([H])C(C([H])([H])C([H])([H])N([H])C1=C([H])C(=NC(C2=C([H])C([H])=C([H])C([H])=N2)=N1)N1C([H])([H])C([H])([H])C2=C([H])C([H])=C([H])C([H])=C2C([H])([H])C1([H])[H])=O
InChi Key	AVZCPICCWKMZDT-UHFFFAOYSA-N
InChi Code	InChI=1S/C22H23N5O2/c28-21(29)8-12-24-19-15-20(26-22(25-19)18-7-3-4-11-23-18)27-13-9-16-5-1-2-6-17(16)10-14-27/h1-7,11,15H,8-10,12-14H2,(H,28,29)(H,24,25,26)
Chemical Name	3-[[2-pyridin-2-yl-6-(1,2,4,5-tetrahydro-3-benzazepin-3-yl)pyrimidin-4-yl]amino]propanoic acid
Synonyms	GSKJ1; GSKJ 1; GSKJ-1
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

Targets	JMJD3 ( IC50 = 60 nM )
ln Vitro	GSK-J1 blocks the functions of JMJD3 and UTX that are transiently transfected in HEK-293 cells. GSK-J1 also raises total nuclear H3K27me3 levels, which prevents human primary macrophages from producing TNF-α.[1] GSK-J1 raises global levels of H3K27me3 and decreases Runx2 and Osterix expressions as well as ALP activity in MC3T3-E1 cells.[2]
ln Vivo	GSK-J1, a JMJD3 small molecule inhibitor, directly disrupted the transcription of genes linked to inflammation by modifying their promoters with histone modification H3K27me3.
Enzyme Assay	Purified JmjD3 (1 μM) and UTX (3 μM) are incubated with a 10 μM peptide [Biotin-KAPRKQLATKAARK(me3)SAPATGG]. in 50 mM HEPES pH 7.5, 150 mM KCl, 50μM (NH4)2SO4·FeSO4·H2O, 1 mM 2-oxoglutarate, and 2 mM ascorbate (JmjD3, 3 minutes at 25°C; UTX, 20 minutes at 25°C) had different inhibitor concentrations (0, 0.005, 0.01, 0.02, 0.05, 0.1 μM). To halt the reaction, add 10 mM EDTA. Utilizing a zip tip to desalt the reaction, the reaction is then spotted using an α-cyano-4-hydroxycinnamic acid MALDI matrix on a MALDI plate. Via the MALDI-TOF R system, samples are analyzed.
Animal Protocol	100 mg/kg/day; i.p.; for 10 days Mice harboring subcutaneous SF8628 K27M xenografts
References	[1]. Nature . 2012 Aug 16;488(7411):404-8. [2]. J Cell Biochem . 2015 Nov;116(11):2628-36. [3]. J Biol Chem . 2022 Jun;298(6):102017.
Additional Infomation	3-[[2-(2-pyridinyl)-6-(1,2,4,5-tetrahydro-3-benzazepin-3-yl)-4-pyrimidinyl]amino]propanoic acid is an organonitrogen heterocyclic compound.

Solubility Data

Solubility (In Vitro)

DMSO: 50~77 mg/mL (128.4~197.7 mM) Water: <1 mg/mL Ethanol: <1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.42 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 (6.42 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.5677 mL	12.8386 mL	25.6772 mL
	5 mM	0.5135 mL	2.5677 mL	5.1354 mL
	10 mM	0.2568 mL	1.2839 mL	2.5677 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.