CPI-1205

Protein target name:

EZH2

Mechanism of action:

SAM-competitive inhibitor

Recommended Concentration for use in cells:

45 nM up to 1 uM

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:

Enhancer of zeste 2 polycomb repressive complex 2 subunit, ENX-1, ENX1, EZH1b, KMT6, KMT6A, WVS, WVS2, EZH2

Probe Details

Chemical Probes Portal ID:

CPP1537

Probe alias:

GTPL9115; SCHEMBL17329268; CPI1205; CPI 1205

PubChem CID:

78320408

SMILES string:

CC1=CC(=C(C(=O)N1)CNC(=O)C2=C(N(C3=CC=CC=C32)[C@H](C)C4CCN(CC4)CC(F)(F)F)C)OC

InChi Key:

HPODOLXTMDHLLC-QGZVFWFLSA-N

Orthogonal probe:

GSK343CPI-360EPZ-6438

Physico-chemical properties

NMR:

1H NMR (500 MHz, DMSO-d6) δ 11.59 (s, 1 H), 7.74 (d, J = 7.6 Hz, 1 H), 7.71−7.66 (m, 1 H), 7.61 (d, J = 7.8 Hz, 1 H), 7.13−7.01 (m, 2 H), 6.15 (s, 1 H), 4.32 (d, J = 4.9 Hz, 2 H), 4.22−4.12 (m, 1 H), 3.84 (s, 3 H), 3.15−2.95 (m, 3 H), 2.75−2.66 (m, 1 H), 2.60 (s, 3 H), 2.39−2.31 (m, 1 H), 2.20 (s, 3 H), 2.05−1.98 (m, 1 H), 1.92−1.84 (m, 1 H), 1.56−1.46 (m, 4 H), 1.42−1.32 (m, 1 H), 1.11−1.01 (m, 1 H), 0.69−0.62 (m, 1 H).

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

EZH2: 2 nM

Potency assay:

Scintillation proximity assay

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

5LS6

Structure-activity relationship:

Yes, see J Med Chem paper

Off target activity

Potency (off target):

IC50

Off target protein and potency:

EZH1: 52 nM

Potency assay (off target):

CPI-1205 was selective for EZH2 over 30 additional protein and DNA methyltransferases.

Probe Selectivity in Vitro:

CPI-1205 had limited activity against P450 enzymes as well as 54 transporters, receptors and ion channels at 10 uM. It had an IC50=21.3 uM for hERG in binding assays.

In cell validation

On target activity in cells

Potency in cells:

EC50

32 nM

Potency assay (cells):

In HeLa cells, CPI-1205 inhibited H3K27me3 levels.

Target engagement assay (cells):

Indirect, substrate methylation assay in HeLa cells

In vivo validation

Organism:

Male Balb/c mice

Dose:

1 mg/kg (IV), 100 mg/kg (oral)

Route of delivery:

OralIntravenous

Plasma half life:

0.402 h (IV)

Systemic clearance:

2.16 L/h/kg (IV)

Cmax:

67.36 uM (oral)

Tmax:

0.25 h (oral)

Organ of Interest

Organ (O):

Karpas-422 xenograft

Target engagement assay:

CPI-1205 decreased H3K27me3 levels in tumor tissue.

Primary Reference

Reference (doi):

10.1021/acs.jmedchem.6b01315

Reference (PMID):

27739677

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 tool molecule adds another EZH2 inhibitor with slightly different biochemical potency and selectivity over the closely related EZH1. As such, the suite of tools provides an increasingly robust mechanism for starting to interrogate the pathway within cells. The observation that the molecules are competitive with SAM gives a rationale for the decrease in potency within the cellular context and the slow turnover of the substrate in cells could explain the slow onset of effects in vivo. Key data that is missing is a listing of the ~50 enzymes and transporters used for the broad out-of-family selectivity profiling. In addition, the lack of detail around the cellular data, particularly the Hill slope, limits the utility in translating from a cellular context to in vivo. A detailed PK profile is presented that could have been used to project how much inhibition is required to see a sustainable effect over a period of time, but this cannot be extrapolated as only the cellular IC50 is presented. The in vivo efficacy data are less convincing than that for some of the other EZH2 inhibitors, but it is not clear whether this is due to different models or different profiles of the molecules, in terms of potency, cell penetration, or PK profile.

Jul 18 2017 - 11:41am

CPI-1205 is a Phase I/II agent designed to selectively target the EZH2 component of the polycomb repressor complex (PRC). It is a well designed, and well described pharmacological tool with well defined pharmacokinetic properties. The availability of several other probes, with differing chemotypes, against this target allows conformation of on-target pharmacology and CPI-1205 may be useful as a standalone agent, or for comparative pharmacology studies with probes such as EPZ-6438. It also offers a contrast to the EZH12 inhibitors such as GSK343 and EPZ011989. CPI-1205 inhibits methyltransferase activity through competition with SAM, and in doing so complements agents such as EED-226, which inhibits EZH2 activity by disrupting assembly of the polycomb repressor complex. Taken together, these two offer useful tools to dissect and understand some of the biology around the PRC, and the differential pharmacology between these complementary modes of action.

Dec 2 2020 - 9:56am

CPI-1205

Probe Information

Target Details

Enhancer of zeste 2 polycomb repressive complex 2 subunit, ENX-1, ENX1, EZH1b, KMT6, KMT6A, WVS, WVS2, EZH2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?

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CPI-1205

Probe Information

Target Details

Enhancer of zeste 2 polycomb repressive complex 2 subunit, ENX-1, ENX1, EZH1b, KMT6, KMT6A, WVS, WVS2, EZH2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?