EPZ004777

Reagent Authentication Certificate:

Chemical Probes Portal EPZ004777.pdf

Protein target name:

DOT1L

Mechanism of action:

SAM competitive inhibitor

Recommended Concentration for use in cells:

1 uM

Probe Availability

MedChem Express (1338466-77-5)

Tocris (1338466-77-5)

Cayman (EPZ004777 formic acid salt)

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Protein methyltransferase

HUGO ID:

HGNC:24948

Entrez gene ID:

84444

UNIPROT ID:

Q8TEK3

Target alias:

KMT4, DOT1 like histone H3K79 methyltransferase

Probe Details

Chemical Probes Portal ID:

CPP625

Probe alias:

1338466-77-5; EPZ 004777; EPZ-004777

PubChem CID:

56962336

ChEMBL ID:

CHEMBL2169919

SMILES string:

CC(C)N(CCCNC(=O)NC1=CC=C(C=C1)C(C)(C)C)C[C@@H]2[C@H]([C@H]([C@@H](O2)N3C=CC4=C3N=CN=C4N)O)O

InChi Key:

WXRGFPHDRFQODR-ICLZECGLSA-N

Orthogonal probe:

SGC0946EPZ-4676

Physico-chemical properties

Solubility - preferred solvent:

DMSO

Solubility - concentration:

50 - 100 mM

NMR:

Not available

Storage Recommendations

Storage (solution):

-20 oC

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

400 pM

Potency assay:

DOT1L enzyme activity assay with PRC2

Structure-activity relationship:

Not available

Off target activity

Potency assay (off target):

EPZ004777 is >1,000-fold selective for DOT1L over other PMTs tested (CARM1, EHMT2, EZH1, EZH2, PRMT1, PRMT5, PRMT8, SETD7, WHSC1).

Probe Selectivity in Vitro:

Not available

In cell validation

On target activity in cells

Potency in cells:

IC50

700 nM

Potency assay (cells):

Dose-dependent decrease in the expression of HOXA9 and MEIS1 transcripts. In MOLM-13, MLL-AF9, MV4-11, MLL-AF4, and Jurkat cells, EPZ004777 dose-dependently decreased DOT1L substrate methylation (H3K79) over 4-5 days as assessed by immunoblot.

Target engagement assay (cells):

Indirect, inhibition of substrate methylation in cells

Off target activity in cells

Potency assay, off target (cells):

PRMT5 (H4R3), PRDMs ((H3K9) and SMYDs (H3K4) protein substrates were not impacted in cells.

Probe Selectivity in Cell:

Not available

In vivo validation

Organism:

Mice

Dose:

50 mg/mL continuous infusion

Route of delivery:

Subcutaneous

Organ of Interest

Organ (O):

MV4-11 xenograft

Target engagement assay:

Indirect, decrease in substrate methylation in the tumor. Authors noted that EPZ004777 had a poor PK profile, which necessitated delivery by continuous infusion.

Primary Reference

Reference (doi):

10.1016/j.ccr.2011.06.009

Reference (PMID):

21741596

External Resources

Available data:

GDSC2

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 DOT1L inhibitor was the first potent DOT1L ligand disclosed (by Epizyme) in 2011. EPZ004777 is an extremely potent and selective DOT1L inhibitor, making this an ideal probe for cellular use. The original publication shows that EPZ004777 effectively inhibits the proliferation of MLL-rearranged cell lines; however, due to restrictions of the pharmacokinetic properties of this molecule, whole animal studies needed to use subcutaneous osmotic pumps to attain useful compound exposure. Significant data on the non-epigenetic off-target profiling could not be found, and no closely related control compound is available.

Aug 17 2016 - 8:03pm

EPZ004777 is an excellent probe for cellular experiments. It specifically affected the MLL-translocated leukemia lines that are dependent on DOT1L recruitment and enzymatic function. Due to the PK limitationsof EPZ004777, EPZ-5676 would be the best choice of DOT1L in vivo probe.

Jan 23 2017 - 1:16pm

EPZ-5676, a derivative with a better pharmacokinetic profile than this probe, would be the better choice for a DOT1L chemical probe. EPZ-5676 is available commercially and is listed in the Portal.

Mar 28 2017 - 12:56pm

There are other derivatives which are better suited to in vivo model use.

Apr 27 2017 - 9:33pm

EPZ004777

Probe Availability

Probe Information

Target Details

KMT4, DOT1 like histone H3K79 methyltransferase

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

External Resources

Supporting Organizations

Interested in supporting the portal?

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EPZ004777

Probe Availability

Probe Information

Target Details

KMT4, DOT1 like histone H3K79 methyltransferase

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

External Resources

Supporting Organizations

Interested in supporting the portal?