EPZ011989

Reagent Authentication Certificate:

Chemical Probes Portal EPZ011989.pdf

Protein target name:

EZH1, EZH2

Mechanism of action:

Inhibitor

Recommended Concentration for use in cells:

100-600 nM

Probe Availability

MedChem Express (1598383-40-4)

Cayman (1598383-40-4)

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Protein methyltransferase

HUGO ID:

HGNC:3526 HGNC:3527

Entrez gene ID:

2145 2146

UNIPROT ID:

Q15910 Q92800

Target alias:

WVS, ENX1, EZH1, KMT6, WVS2, ENX-1, EZH2b, KMT6A

Probe Details

Chemical Probes Portal ID:

CPP50

PubChem CID:

73670548

ChEMBL ID:

CHEMBL3672841

SMILES string:

O=C(NCC1=C(C)C=C(C)NC1=O)C2=CC(C#CCN3CCOCC3)=CC(N(CC)[C@H]4CC[C@H](N(CCOC)C)CC4)=C2C

InChi Key:

XQFINGFCBFHOPE-UHFFFAOYSA-N

Orthogonal probe:

UNC1999GSK343CPI-169

PAINs evaluation:

PAINs free

Physico-chemical properties

NMR:

1H NMR (MeOD, 400 MHz) δ 7.23 (s, 1H), 7.09 (s, 1H), 6.11 (s, 1H), 4.46 (s, 2H), 3.74-3.72 (m, 4H), 3.51 (s, 2H), 3.47 (t, J=5.6 Hz, 2H), 3.32 (s, 3H), 3.07 (q, J=7.2 Hz, 2H), 2.64-2.63 (m, 7H), 2.38 (m,1H), 2.37 (s, 3H), 2.27 (s, 3H), 2.26 (s, 3H), 2.25 (s, 3H), 1.89-1.86 (m, 4H), 1.50-1.30 (m, 4H), 0.83 (t, J=7.2 Hz, 3H)

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

< 3 nM

Potency assay:

IC50=6 nM in ELISA-based protein substrate methylation assay.

Structure-activity relationship:

Yes.

Off target activity

Off target protein and potency:

EZH1 IC50 = 103 nM

Potency assay (off target):

15-fold selective for EZH2 > EZH1 in protein substrate methylation assay. 3000-fold selective for EZH2 > 20 other protein methyltransferases (CARM1, DOT1L, EHMT1, EHMT2, PRMT1, PRMT3, PRMT5, PRMT6, PRMT8, SETD2, SETD7, SUV39H1, SMYD2, SMYD3, WHSC1, WHSC1L1).

In cell validation

On target activity in cells

Potency in cells:

IC50

< 100 nM

Potency assay (cells):

Protein substrate methylation in WSU-DLCL2 cells

Target engagement assay (cells):

Indirect, protein substrate methylation assay

In vivo validation

Organism:

Rat (and SCID mouse)

Dose:

30 mg/kg - 300 mg/kg (rat)125 - 1000 mg/kg (mice)

Route of delivery:

Oral

Plasma half life:

3.7 - 4.7 h (rat)

Fraction of probe bound to plasma protein:

0.09 (rat), 0.2 (mouse)

Cmax:

240-2900 ng/mL (rat)

Tmax:

2 - 2.7 h (rat)

Organ of Interest

Organ (O):

bone marrowKARPAS-422 DLBCL tumor cell xenograft

Target engagement assay:

Indirect, protein substrate methylation in bone marrow and in tumor (xenograft).

Primary Reference

Reference (doi):

10.1021/acsmedchemlett.5b00037

Reference (PMID):

26005520

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 small molecule represents an additional probe for testing the effects of EZH2 inhibition in cells and in vivo. Oral bioavailability and improved metabolic stability are key features that add to its value for use as a probe. The concentration range in mouse models seems somewhat high, but given its clearance from plasma, this is required to maintain predicted efficacious levels in an animal. The lack of a negative control compound for direct comparison makes this compound less ideal than other EZH2 inhibitors currently available (especially UNC-1999).

Jun 12 2016 - 4:00am

Users should note the salt and vehicle preparation for any in vivo use (0.5% methyl cellulose and 0.1% Tween-80 was used in original manuscript).

Jun 30 2016 - 3:58am

This is a good tool compound for a couple of interesting targets. I'm not concerned by lack of a negative control compounds, since such compounds often offer a false confidence (if they lack the same off targets as their positive compounds).

Aug 17 2016 - 9:07pm

EPZ011989

Probe Availability

Probe Information

Target Details

WVS, ENX1, EZH1, KMT6, WVS2, ENX-1, EZH2b, KMT6A

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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EPZ011989

Probe Availability

Probe Information

Target Details

WVS, ENX1, EZH1, KMT6, WVS2, ENX-1, EZH2b, KMT6A

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?