CPI-360

Protein target name:

EZH2

Mechanism of action:

SAM competitive inhibitor

Recommended Concentration for use in cells:

50 nM - 1 uM

Probe Availability

MedChem Express (1802175-06-9)

Cayman (1802175-06-9)

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:

CPP632

PubChem CID:

73442743

ChEMBL ID:

CHEMBL3605456

SMILES string:

CC1=CC(=C(C(=O)N1)CNC(=O)C2=C(N(C3=CC=CC=C32)[C@H](C)C4CCOCC4)C)OC

InChi Key:

PFPSFENQCNMITC-MRXNPFEDSA-N

Orthogonal probe:

GSK343CPI-1205EPZ-6438

Physico-chemical properties

NMR:

1H NMR (400MHz, DMSO-d6) δ 11.60 (s, 1H), 7.73-7.62 (m, 3H), 7.60 (d, 2H) 7.07-7.05 (m, 2H), 6.15 (s, 1H) 4.33 (s, 1H), 4.21- 4.11 (m, 1H), 3.92 (br.d., 1H), 3.65 (d, 1H), 3.34-3.32 (m, 1H), 3.02 (t, 1H), 2.61 (s, 3H),
2.48-2.44 (m, 1H), 2.20 (s, 3H), 1.84-1.81 (m, 1H), 1.54 (d, 3H), 1.40-1.38 (m, 1H), 1.25- 1.22 (m, 1H), 1.08-1.04 (m, 1H), 0.86 (br. s., 1H), 0.58 (br. d., 1H)

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

EZH2 wild-type 0.07 nM, EZH2 Y641N 2.5 nM

Potency assay:

PRC2 enzyme inhibition assay

Off target activity

Potency assay (off target):

CPI-360 is 100-fold less potent against EZH1 than EZH2 and showed minimal activity against 30 additional protein methyltransferases.

Probe Selectivity in Vitro:

Not available

In cell validation

On target activity in cells

Potency in cells:

EC50

0.056 - 0.06 uM

Potency assay (cells):

ELISA-based detection of protein substrate (H3K27) methylation, CETSA, and NanoBRET assay.

Target engagement assay (cells):

Indirect via inhibition of protein substrate methylation, and direct via NanoBRET assay.

Primary Reference

Reference (doi):

10.1016/j.chembiol.2014.09.017

Reference (PMID):

25457180

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 molelcule has excellent selectivity for H3K27Me3 (EZH2) over H3K27Me1 (EZH1) in cells, so this appears to be an excellent orthogonal molecule to existing probes.

Sep 12 2016 - 8:04am

CPI-360 appears to be a potent and selective inhibitor of EZH2 in both biochemical and cellular assays, with selectivity over EZH1 and several other methytransferases. As such, it offers a useful tool to complement compounds such as GSK343. The compound appears to show in vivo efficacy in xenograft models, but at very high dosing (200 mg/kg compared with a cellular potency of 50 nM), suggesting potential deficiencies in the PK profile. Disappointingly, no PK data are given to allow assessment of this aspect, and as such, any use in vivo should proceed with caution, ideally only after gaining an understanding of the full PK and dose/exposure relationships.

Jan 6 2017 - 11:58am

This probe has good selectivity and cell activity. However, the discoverers noted that in mice even at the very high subcutaneous dose of 200 mg/kg the "inadequate pharmacological properties of CPI-360 did not allow for complete target coverage." This appears to be in part due to poor microsomal stability, which is given as 166/138/106/110 μl/min/mg (for m/r/d/h) in Supplementary Figure S5. The closely related compound CPI-169 is more stable, shows better activity in a xenograft model, and appears to have a similarly favorable selectivity and potency profile, suggesting that CPI-169 may be a superior probe.

Jan 13 2017 - 1:14pm

The compound is very potent. The in vivo dose might need to adjusted when the in vivo exp need to be longer.

Nov 4 2020 - 11:41am

CPI-360

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

Primary Reference

Supporting Organizations

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CPI-360

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

Primary Reference

Supporting Organizations

Interested in supporting the portal?