GSK-J4

Protein target name:

KDM6A, KDM6B

Mechanism of action:

pro-drug for GSK-J1competitive with alpha-ketoglutaratenoncompetitive with peptide substrate

Recommended Concentration for use in cells:

1 nM - 5 uM

Probe Availability

MedChem Express (1373423-53-0)

Tocris (1227911-45-6)

Abcam (1373423-53-0)

Cayman (1797983-09-5)

MilliporeSigma (1373423-53-0)

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Lysine demethylase

HUGO ID:

HGNC:12637 HGNC:29012

Entrez gene ID:

7403 23135

UNIPROT ID:

O15550 O15054

Target alias:

KABUK2, UTX, JMJD3

Probe Details

Chemical Probes Portal ID:

CPP107

Probe alias:

GSKJ4, GTPL8979

PubChem CID:

71729975

ChEMBL ID:

CHEMBL3183531

SMILES string:

CCOC(=O)CCNC1=NC(=NC(=C1)N2CCC3=CC=CC=C3CC2)C4=CC=CC=N4

InChi Key:

WBKCKEHGXNWYMO-UHFFFAOYSA-N

Control compound:

GSK-J5

PAINs evaluation:

PAINs free

Physico-chemical properties

Solubility - preferred solvent:

DMSO

Solubility - concentration:

20 mg/mL

Storage Recommendations

Storage (dry powder):

2 years -20 oC

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

60 nM

Potency assay:

Alpha screen

PDB ID for probe-target interaction (3D structure):

4ASK

Off target activity

Potency (off target):

IC50

Off target protein and potency:

KDM5B 950 nMKDM5C 1760 nM

Potency assay (off target):

Inactive against additional demethylases in thermal shift, MS and antibody-based assays

Probe Selectivity in Vitro:

GSK-J1 had limited activity at 30 uM against 100 protein kinases in competitive binding assay, and also against 60 additional proteins (chromatin-modifying enzymes and HDACs).

In cell validation

On target activity in cells

Potency in cells:

IC50

9 uM

Potency assay (cells):

Inhibition of TNF alpha release in primary human macrophages

Target engagement assay (cells):

Confirmed in probe affinity and quantitative MS assays, and demonstrated indirectly via inhibition of protein substrate demethylation

Primary Reference

Reference (doi):

10.1038/nature11262

Reference (PMID):

22842901

Endorsed probe

The Chemical Probes Portal endorses this compound as chemical probe for use in cells.

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

GSK-J4 is the ester prodrug of the active (acid) component GSK-J1. The ester is designed to facilitate entry into cells where it is rapidly hydrolysed to the active acid and studies from the authors demonstrate rapid hydrolysis in macrophages, the systems studied. They also note that in other cell systems, the degree and rate of conversion to acid should be assessed to ensure data generated is not misleading. It is a cell active inhibitor of KDM6 although more recently it has been demonstrated to exhibit inhibition of KDM5, albeit weaker. It can be recommended as a cell target probe with the caveats around moderate KDM selectivity, dependence on ester cleavage for activity, and also a not ideal level of target potency even when hydrolysed in cells.

Jun 30 2016 - 4:00am

I could not find a broad selectivity assessment of GSK-J4 or its active carboxylic acid form. A broader assessment of the ester prodrug and the carboxylate is necessary before using this compound as an in vivo tool; thus, this is not an ideal tool molecule. I recommend using it for cellular studies only.

Sep 10 2016 - 3:46pm

Tested in mice at 5mg/kg delivery by i.p. injection (https://doi.org/10.1093/hmg/ddu376) as well as at 0.5mg/kg by i.p injection ( 10.1186/s13075-018-1560-9) therefore recommend 0.5-5 mg/kg i.p. in mice.

Nov 16 2020 - 9:14pm

GSK-J4

Probe Availability

Probe Information

Target Details

KABUK2, UTX, JMJD3

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

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GSK-J4

Probe Availability

Probe Information

Target Details

KABUK2, UTX, JMJD3

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?