EED226

Reagent Authentication Certificate:

Chemical Probes Portal EED226.pdf

Protein target name:

EED

Mechanism of action:

AllostericNoncompetitive with SAM and peptide substrate

Recommended Concentration for use in cells:

100 nM - 1 uM

Probe Availability

MedChem Express (2083627-02-3)

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Polycomb family

HUGO ID:

HGNC:3188

Entrez gene ID:

8726

UNIPROT ID:

O75530

Target alias:

Embryonic ectoderm development, HEED, WAIT-1, WD protein associating with integrin cytoplasmic tails 1

Probe Details

Chemical Probes Portal ID:

CPP1283

PubChem CID:

123132228

SMILES string:

CS(=O)(=O)C1=CC=C(C=C1)C2=CN=C(N3C2=NN=C3)NCC4=CC=CO4

InChi Key:

DYIRSNMPIZZNBK-UHFFFAOYSA-N

Orthogonal probe:

A-395

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

82 nM

Potency assay:

Isothermal calorimetry. In in vitro enzyme assays, EED226 had an IC50=23.4 nM for PRC2

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

5GSA

Structure-activity relationship:

Not available

Off target activity

Potency assay (off target):

EED226 had limited activity against 21 protein methyltransferases (IC50 >100 uM). EED226 inhibited EZH1-PRC2 complex.

Probe Selectivity in Vitro:

EED226 had limited activity against 23 kinases (IC50 >10 uM), and 22 GPCRs, ion channels, nuclear receptors (IC50 >30 uM).

In cell validation

On target activity in cells

Potency assay (cells):

In G401 cells, EED226 dose-dependently decreased global H3K27me3 and H3K27me2 levels.

Target engagement assay (cells):

In chemoproteomic experiments with biotinylated EE226 in Karpas442 cells, EZH2, SUZ12 and EED (PRC2 components) and MTF2 (a PRC2 partner) were the only proteins pulled down.

In vivo validation

Organism:

Athymic BALB/c nude mice

Dose:

4 and 40 mg/kg

Route of delivery:

Oral

Cmax:

~10 uM (4 mg/kg)

Tmax:

~1 h (4 mg/kg)

Organ of Interest

Organ (O):

Karpas422 xenograft

Target engagement assay:

EED226 led to dose-dependent decrease in global H3K27me3 levels.

Primary Reference

Reference (doi):

10.1038/nchembio.2304

Reference (PMID):

28135235

Endorsed probe

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

SAB Rating

Rating for use in Cells

4 review(s)

Rating for use in Model Organisms

4 review(s)

SAB Members

SAB Comments

The fact that this compound is non-competitive with respect to SAM and peptide substrate is supported by an X-ray structure and enzymatic competition assays. The furane ring with electron-donating group is supposed to be unstable, but the structure was confirmed to be stable under even an acidic condition reported in the supplementary note. I think the chemical probe could prove to be very interesting and useful.

Jun 12 2017 - 11:04am

This allosteric inhibitor of PRC2 competes with the H3K27me3 peptide for binding to EED. So far, it seems to be a very clean inhibitor. The only data on this compound has been published by a single group, so validation by other groups will increase enthusiasm for its use as an orthogonal inhibitor to SAM-competitive EZH2 inhibitors now in clinical trials.

Jun 15 2017 - 12:28pm

EED226

Probe Availability

Probe Information

Target Details

Embryonic ectoderm development, HEED, WAIT-1, WD protein associating with integrin cytoplasmic tails 1

Probe Details

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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EED226

Probe Availability

Probe Information

Target Details

Embryonic ectoderm development, HEED, WAIT-1, WD protein associating with integrin cytoplasmic tails 1

Probe Details

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?