JNK-IN-8

Reagent Authentication Certificate:

Chemical Probes Portal JNK-IN-8.pdf

Protein target name:

MAPK8, MAPK9, MAPK10

Mechanism of action:

Covalent inhibitor

Recommended Concentration for use in cells:

1 nM - 1 uM

Probe Availability

MedChem Express (1410880-22-6)

MilliporeSigma (JNKIN8)

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

CMGC

HUGO ID:

HGNC:6881 HGNC:6886 HGNC:6872

Entrez gene ID:

5599 5601 5602

UNIPROT ID:

P45983 P45984 P53779

Target alias:

JNK1, JNK2, JNK3

Probe Details

Chemical Probes Portal ID:

CPP97

Probe alias:

GTPL8058

PubChem CID:

57340686

ChEMBL ID:

CHEMBL2216824

SMILES string:

CC1=C(C=CC(=C1)NC(=O)C2=CC(=CC=C2)NC(=O)/C=C/CN(C)C)NC3=NC=CC(=N3)C4=CN=CC=C4

InChi Key:

GJFCSAPFHAXMSF-UXBLZVDNSA-N

PAINs evaluation:

PAINs free

Physico-chemical properties

NMR:

1H NMR (600 MHz, DMSO-d6) 10.54 (s, 1 H), 10.20 (s, 1 H), 9.24 (s, 1 H), 8.95 (s, 1 H), 8.70 (d, J = 3.0 Hz, 1 H), 8.47 (d, J = 4.8 Hz, 1 H), 8.44 (d, J = 7.8 Hz, 1 H), 8.17 (s, 1 H), 7.89 (d, J = 7.8 Hz, 1 H), 7.77 (d, J = 7.8 Hz, 1 H), 7.63 (d, J = 1.8 Hz, 1 H), 7.49-7.44 (m, 2 H), 7.40 (d, J = 5.4 Hz, 1 H), 6.76 (m, 1 H), 6.46 (d, J = 15.6 Hz, 1 H), 3.94 (d, J = 6.0 Hz, 2 H), 2.79 (s, 6 H), 2.23 (s, 3 H).

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

MAPK8 14.7 nM, MAPK9 18.7 nM, MAPK10 1 nM

Potency assay:

In vitro kinase assay

Off target activity

Potency assay (off target):

442 kinases were tested in DiscoveRx panel, and 121 kinases were tested in radiometric assays. With cellular data, JNK-IN-8 did not inhibit any off-target kinases with an IC50< 1 uM.

In cell validation

On target activity in cells

Potency in cells:

IC50

338-486 nM

Potency assay (cells):

In HeLa and A375 cells, JNK-IN-8 inhibited phosphorylation of protein substrates

Target engagement assay (cells):

Indirect: inhibition of protein substrate phosphorylation Direct: pull down of JNK proteins with a biotinylated-derivative of the probe, and expression of JNK C116S resistant mutant raises the cellular IC50 100-fold.

Off target activity in cells

Potency assay, off target (cells):

200 kinases were assays in A375 cells using KiNativ. With in vitro data, JNK-IN-8 did not inhibit any off-target kinase with an IC50< 1 uM.

Primary Reference

Reference (doi):

10.1016/j.chembiol.2011.11.010

Reference (PMID):

22284361

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

This is a potent JNK1/2/3 inhibitor with some off target activity (MNK2, FMS at 200-500 nM).

Jun 29 2016 - 7:53pm

JNK-IN-8 is a covalent and highly selective JNK1/2/3 inhibitor with well validated activity in cells. The JNK family of kinases (also known as the stress-activated protein kinases, SAPK) play a significant role in the cellular response to cytokines and environmental stress. The literature surrounding these targets is vast, with JNK inhibition possibly being of utility in cancer, diabetes, Alzheimer’s disease and endometriosis. Unfortunately, the literature is also sometimes confusing and contradictory. Good small molecule tools like JNK-IN-8 are essential to the study of these important targets. JNK-IN-8 should prove a useful tool when used alongside other validated JNK inhibitors including bentamapimod, XG-102 and tanzisertib.

Sep 6 2016 - 8:05pm

JNK-IN-8 is a potent pan-JNK inhibitor (biochemical IC50 values: 4.7 nM (JNK1), 18.7 nM (JNK2), 1 nM (JNK3)) with good selectivity against the broader kinome. It does inhibit other kinases such as MNK2 and FMS in the range of 200-300 nM. Its binding mode is well understood; it is a covalent irreversible inhibitor. Inhibition of phosphorylation of c-Jun, a direct substrate of JNK, indicates target engagement in cells. This compound is a useful tool for cellular assays.

Sep 10 2016 - 4:07pm

JNK-IN-8 is an imatinib-derived pan-JNK inhibitor. Its covalent-irreversible binding mode is reasonable validated by X-ray structures and protein mass spectroscopy of close analogues. While showing high potency in biochemical assays on the isolated JNKs, the compound suffers from a surprisingly low inhibitory activity in a cellular environment (IC50 300-500 nM). Kinome-wide screening revealed several highly potent off target hits, but most of them could not be validated in subsequent IC50 or Kd determinations (e.g., FMS and MNK2, ~200-300 nM). JNK-IN-8 may serve as a chemical probe in cells to investigate unselective JNK inhibition. It should be used with caution along with another positive control compound.

May 30 2017 - 12:42pm

JNK-IN-8

Probe Availability

Probe Information

Target Details

JNK1, JNK2, JNK3

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

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JNK-IN-8

Probe Availability

Probe Information

Target Details

JNK1, JNK2, JNK3

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?