EPZ-6438

Reagent Authentication Certificate:

Chemical Probes Portal EPZ-6438.pdf

Protein target name:

EZH2

Mechanism of action:

SAM competitive inhibitor

Recommended Concentration for use in cells:

30-1,000 nM

Probe Availability

MedChem Express (1403254-99-8)

Cayman (1403254-99-8)

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:

WVS; ENX1; EZH1; KMT6; WVS2; ENX-1; EZH2b; KMT6A, enhancer of zeste 2 polycomb repressive complex 2 subunit

Probe Details

Chemical Probes Portal ID:

CPP627

Probe alias:

1403254-99-8; EPZ6438; Tazemetostat

PubChem CID:

66558664

ChEMBL ID:

CHEMBL3414621

SMILES string:

CCN(C1CCOCC1)C2=CC(=CC(=C2C)C(=O)NCC3=C(C=C(NC3=O)C)C)C4=CC=C(C=C4)CN5CCOCC5

InChi Key:

NSQSAUGJQHDYNO-UHFFFAOYSA-N

Orthogonal probe:

GSK343CPI-169CPI-360

Physico-chemical properties

NMR:

Not available

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

EZH2 2.5 nM

Potency assay:

PRC2-containing enzyme assay. The Ki was comparable for wild-type enzyme and for the following mutants: A677G, A687V, Y641F, Y641H, Y641N, and Y641S.

Off target activity

Potency assay (off target):

EPZ-6438 is 35-fold selective for EZH2 over EZH1 and 4,500-fold selective over all other protein methyltransferases tested (CARM1,DOT1L, EHMT1, EHMT2, PRMT1, PRMT3, PRMT5, PRMT6, PRMT8, SETD7, SMYD2, SMYD3, WHSC1 WHSC1L1).

Probe Selectivity in Vitro:

Not available

In cell validation

On target activity in cells

Potency in cells:

IC50

EZH2 1.4-5.6 nM

Potency assay (cells):

Dose-dependent decrease in substrate (H3K27) methylation in multiple cell types and against multiple EZH2 mutants.

Target engagement assay (cells):

Indirect, inhibition of substrate methylation

Off target activity in cells

Potency assay, off target (cells):

No activity was detected by assessing other histone substrates (H3K9, H3K4, H3K36 and H3K79)

In vivo validation

Organism:

SCID mice

Dose:

125-500 mg/kg

Route of delivery:

Oral

Organ of Interest

Organ (O):

G401 xenograft

Target engagement assay:

Indirect, inhibition of substrate methylation in tumor

Primary Reference

Reference (doi):

10.1073/pnas.1303800110

Reference (PMID):

23620515

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

The compound shows good potency in the inhibition of H3K27 trimethylation in multiple cell lines, making it a potentially useful tool. Note that 4 days of dosing is required. Note also that the compound shows EZH1 (off-target) activity with ~35 fold selectivity. Data on broad selectivity would be useful; although the excellent potency/low concentration required in cells somewhat mitigates against concerns. In vivo experiments require high doses over an extended period (3-4 weeks), despite the stated "good oral bioavailability" of this compound. Therefore, I cannnot be entirely certain that the effects observed in the in vivo experiments are due solely to the compound's EZH2 activity. At the high concentrations used, activity against EZH1 or other unknown targets could be involved. No detailed pharmacokinetic information is provided (only concentrations at 2 time points after 21 days).

Aug 15 2016 - 10:15pm

This compound shows high in vitro potency for wild-type EZH2 (Ki = 2.5 nM) and for EZH2 mutations (Y641F, C, H, N, S and A677G); SAM competitive and non-competitive with peptide substrate; ca. 35-fold selective for EZH2 over EZH1 and more than 4,500-fold selective relative to other PMTs, including G9a, GLP, SETD7, SMYD2, SMYD3, MMSET, WHSC1L1, PRMTs, DOT1L. The compound is potent in cellular antiproliferation assays, displayed efficacy in a xenograft model (dosed for 28 days) and was well-tolerated, making it an interesting tool compound.

Sep 10 2016 - 3:29pm

EPZ-6438 is a potent, SAM-competitive inhibitor of wild-type and all known mutants of EZH2 with biochemical selectivity over EZH1 and other histone methyl transferases. Potent target engagement in cells (<5 nM) has been demonstrated by inhibition of the H3K27Me3 methylation mark, but this requires extended incubation (4 days). Selectivity in cells is shown by the lack of effect on other H3 histone methylation marks. No other off-target pharmacology data is available, and I cannot exclude off-target effects in cells, although this is mitigated by the selective apoptotic effects demonstrated in SMARCB1-deficient, EZH2-dependent cells (antiproliferation IC50s ca. 100 nM) compared to SMARCB1-proficient cells. I suggest the use of 30-1,000 nM concentrations in cell-based experiments, and the use of an orthogonal, EZH2-selective probe in parallel (e.g., GSK343). Dose-dependent anti-tumour efficacy and inhibition of H3K27Me3 in human tumour xenografts by EPZ-6438 has been shown following high oral doses for extended periods. Detailed in vivo pharmacokinetic parameters are not available but would be useful to interpret the in vivo biology. I cannot exclude the possibility of off-target effects at the high doses.

Dec 26 2016 - 12:44pm

EPZ-6438

Probe Availability

Probe Information

Target Details

WVS; ENX1; EZH1; KMT6; WVS2; ENX-1; EZH2b; KMT6A, enhancer of zeste 2 polycomb repressive complex 2 subunit

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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EPZ-6438

Probe Availability

Probe Information

Target Details

WVS; ENX1; EZH1; KMT6; WVS2; ENX-1; EZH2b; KMT6A, enhancer of zeste 2 polycomb repressive complex 2 subunit

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?