EPZ-5676

Reagent Authentication Certificate:

Chemical Probes Portal EPZ-5676.pdf

Protein target name:

DOT1L

Mechanism of action:

Inhibitor

Recommended Concentration for use in cells:

Up to 100 nM

Probe Availability

MedChem Express (1380288-87-8)

Cayman (1380288-87-8)

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

Probe Details

Chemical Probes Portal ID:

CPP54

Probe alias:

Pinometostat

PubChem CID:

57345410

ChEMBL ID:

CHEMBL3087499

SMILES string:

CC(C)N(C[C@@H]1[C@H]([C@H]([C@@H](O1)N2C=NC3=C2N=CN=C3N)O)O)C4CC(C4)CCC5=NC6=C(N5)C=C(C=C6)C(C)(C)C

InChi Key:

LXFOLMYKSYSZQS-XKHGBIBOSA-N

Orthogonal probe:

SGC0946EPZ004777

PAINs evaluation:

PAINs free

Physico-chemical properties

Storage Recommendations

Storage (solution):

-20 oC 2 years

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

0.08 nM

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

4HRA

Off target activity

Potency assay (off target):

Minimal off-target activity against 16 protein methyltransferases up to 10 uM EPZ5676. At 10 uM, PRMT5 activity was inhibited by 40%.

In cell validation

On target activity in cells

Potency in cells:

IC50

3 nM

Potency assay (cells):

ELISA to detect protein substrate methylation

Target engagement assay (cells):

Indirect, via protein substrate methylation assay

In vivo validation

Organism:

NCr nu/nu mouse and Sprague-Dawley rat

Dose:

20 mg/kg (mice)1 mg/kg (rat)

Route of delivery:

Intraperitoneal

Other route of delivery:

IV for rat

Plasma half life:

5.22 h (mouse), 3.73 h (rat)

Systemic clearance:

4.07 L/h/kg (rat)

Fraction of probe bound to plasma protein:

0.47 (mouse)

Cmax:

4.93 uM (mouse), 2.04 uM (rat)

Tmax:

0.03 h (mouse), 0.05 h (rat)

Organ of Interest

Organ (O):

xenograft tumor

Target engagement assay:

Indirect, protein substrate methylation assay with inhibitory activity detected in the tumor and surrogate tissues, bone marrow and PBMCs

Primary Reference

Reference (doi):

10.1182/blood-2013-04-497644

Reference (PMID):

23801631

Endorsed probe

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

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

EPZ-5676 (pinometostat) is a first-in-class inhibitor of DOT1L activity. EPZ-5676 is a potent and selective inhibitor of DOT1L with activity in both cellular and rodent MLL-models. Nonclinical in vivo pharmacokinetics of EPZ-5676 in mouse, rat and dog showed moderate to high clearance and low oral bioavailability, so sustained plasma exposures in rat require IV fusion. For cellular experiments, I recommend using EPZ-5676 in parallel with a second chemical probe SGC0946 to seek a consistent outcome, as both are potent inhibitors of DOT1L activity. The 'inactive control' SGC-0649 could also be used but only at relatively low concentrations because it still retains some DOT1L activity (IC50=390 nM). For in vivo PK-PD experiments, I recommend administering EPZ-5676 by continuous IV infusion to achieve sustained plasma levels above concentrations shown to be efficacious in cell models.

Jun 30 2016 - 12:34am

EPZ-5676 continuous IV infusion for 21 days in a xenograft model of MLL-rearranged leukemia leads to dose-dependent anti-tumor activity. At the highest dose of 70.5 mg/kg/day, complete tumor regression is achieved with no regrowth for up to 32 days after the cessation of treatment. No significant weight loss or obvious toxicity is observed in rats treated with EPZ-5676 during an efficacy study. The selectivity of this agent (>37,000-fold selectivity against all other PMTs tested) makes it a useful cellular tool that is appropriate for rodent models.

Jun 30 2016 - 4:19am

EPZ-5676

Probe Availability

Probe Information

Target Details

KMT4

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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EPZ-5676

Probe Availability

Probe Information

Target Details

KMT4

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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