JQEZ5

Protein target name:

EZH2

Mechanism of action:

SAM competitive inhibitor

Recommended Concentration for use in cells:

1-10 uM

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Protein methyltransferase

HUGO ID:

HGNC:3527

Entrez gene ID:

2146

UNIPROT ID:

Q15910

Target alias:

ENX-1, ENX1, EZH1b, KMT6, KMT6A, WVS, WVS2, EZH2

Probe Details

Chemical Probes Portal ID:

CPP575

PubChem CID:

121322599

SMILES string:

CC(C)N1C2=NC(C3=CC=C(N4CCN(C)CC4)N=C3)=CC(C(NCC5=C(CCC)C=C(C)NC5=O)=O)=C2C=N1

InChi Key:

LQTWDAYNGMMHLV-UHFFFAOYSA-N

Control compound:

JQEZ23

Orthogonal probe:

GSK343CPI-360EPZ-6438

Physico-chemical properties

Solubility - preferred solvent:

DMSO

Solubility - concentration:

10 mM

NMR:

1H NMR (500 MHz, CDCl3) δ 9.00 (d, J = 2.5 Hz, 1H), 8.33 (s, 1H), 8.31 (dd, J = 9.0 and 2.5 Hz, 1H), 8.14 (t, J = 6.0 Hz, 1H), 7.87 (s, 1H), 6.75 (d, J = 9.0 Hz, 1H), 6.02 (s, 1H), 5.38 (m, 1H), 4.69 (d, J = 6.0 Hz, 2H), 3.88 (m, 4H), 2.92 (m, 4H), 2.75 (m, 2H), 2.62 (s, 3H), 2.27 (s, 3H), 1.67 (m, 2H), 1.62 (d, J = 6.5 Hz, 6H), 1.05 (t, J = 7.0 Hz, 3H) ppm.

Storage Recommendations

Storage (dry powder):

4 oC

Storage (solution):

-20 oC

Storage sensitivities:

Stable for 3 months in DMSO solution

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

EZH2

11 nM

Potency assay:

Radiometric Scintillation Proximity Assay (SPA), Biocore SPR (87 nM)

Off target activity

Potency assay (off target):

Limited activity against 22 Protein methyltransferases

Probe Selectivity in Vitro:

Not available

In cell validation

On target activity in cells

Potency in cells:

IC50

2.5 uM

Potency assay (cells):

Antiproliferation against lung cancer cells, and western blot assessing H3K27Me3 level with compound treatment

Target engagement assay (cells):

Not available

In vivo validation

Organism:

CD1 mouse

Dose:

5 mg/kg or 75 mg/kg

Route of delivery:

IntravenousIntraperitoneal

Plasma half life:

3.9 hr (IP)

Systemic clearance:

2.4 L/h*kg (5 mg/kg IV)

Cmax:

15 uM (75 mg/kg IP)

Primary Reference

Reference (doi):

10.1158/2159-8290.CD-16-0164

Reference (PMID):

27312177 27135738

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

An evaluation of the selectivity of this compound for other targets is missing. Due to the close structural analogy with other known EZH2 inhibitors (UNC1999 and GSK343) that showed no off-target activity with other receptors/enzymes, we could expect the compound to be as well selective for its target. However, I would recommend additional screening to rule out any off-target effects.

Aug 5 2016 - 3:04pm

SAR data are not presented but could presumably have been as other analogs were made. The level of in vitro cellular activity (viability) is relatively low (50% reduction, Fig 6D) given that the compound is dosed 57-fold above the Ki. It would be straightforward to measure cellular uptake and metabolic stability of the compound. The xenograft data are compelling, but off-target effects are worrisome, especially in the absence of in vivo metabolic stability data and additional panel screening (e.g., CEREP). Little data is presented to show that suppression of EZH2 activity in lung tissue does not have a deleterious effect. It would also help to have broader testing in representative, high-expressing EZH2 NSCLC lines.

Sep 11 2016 - 4:08am

JQEZ5

Probe Information

Target Details

ENX-1, ENX1, EZH1b, KMT6, KMT6A, WVS, WVS2, EZH2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?

Search form

JQEZ5

Probe Information

Target Details

ENX-1, ENX1, EZH1b, KMT6, KMT6A, WVS, WVS2, EZH2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?