UNC0642

Reagent Authentication Certificate:

Chemical Probes Portal UNC0642.pdf

Protein target name:

EHMT2, EHMT1

Mechanism of action:

Peptide substrate competitive inhibitorSAM noncompetitive inhibitor

Recommended Concentration for use in cells:

100 nM - 1 uM

Probe Availability

MedChem Express (1481677-78-4)

Tocris (1481677-78-4)

Cayman (1481677-78-4)

MilliporeSigma (UNC0642)

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Protein methyltransferase

HUGO ID:

HGNC:14129 HGNC:24650

Entrez gene ID:

10919 79813

UNIPROT ID:

Q96KQ7 Q9H9B1

Target alias:

G9A, G9a, G9a-GLP, G9a/GLP, BAT8, GAT8, NG36, KMT1C, euchromatic histone-lysine N-methyltransferase 2, GLP, GLP1, KMT1D, FP13812, EUHMTASE1, Eu-HMTase1, bA188C12.1, euchromatic histone-lysine N-methyltransferase 1

Probe Details

Chemical Probes Portal ID:

CPP647

Probe alias:

UNC 0642; 1481677-78-4; GTPL7017

PubChem CID:

53315878

ChEMBL ID:

CHEMBL2441082

SMILES string:

CC(C)N1CCC(CC1)NC(=NC1N2CCC(CC2)(F)F)C(C(N=1)=CC(=C1OC)OCCCN2CCCC2)=C1

InChi Key:

RNAMYOYQYRYFQY-UHFFFAOYSA-N

Orthogonal probe:

UNC0638BIX-01294A-366

Physico-chemical properties

NMR:

1H NMR (400 MHz, CDCl3) δ 7.12 (s, 1H), 6.78 (s, 1H), 5.36 (d, J = 7.7 Hz, 1H), 4.31 – 4.20 (m, 1H), 4.17 (t, J = 6.7 Hz, 2H), 3.96 (s, 3H), 2.90 (d, J = 11.9 Hz, 2H), 2.84 – 2.73 (m,
1H), 2.67 – 2.59 (m, 2H), 2.57 – 2.46 (m, 4H), 2.45 – 2.34 (m, 2H), 2.22 – 2.05 (m, 4H), 1.84 – 1.74 (m, 4H), 1.07 (d, J = 6.6 Hz, 6H).

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

EHMT2 3.7 nM

Potency assay:

EHMT1 and EHMT2 IC50 < 2.5 nM in enzyme activity assays.

Structure-activity relationship:

Yes

Off target activity

Potency assay (off target):

UNC0642 was 2,000-fold selective for the targets over EZH2/PRC2 and 20,000-fold over 15 other protein methyltransferases

Probe Selectivity in Vitro:

No activity was detected against 50 kinases. <50% inhibitory activity was detected against a panel of 44 GPCRs, ion channels, and transporters at 1 uM, except histamine H3 receptor (Ki = 45 nM).

In cell validation

On target activity in cells

Potency in cells:

IC50

110 nM

Potency assay (cells):

Inhibition of protein substrate (H3K9) methylation assessed by in cell western in MDA-MB-231 cells. Comparable results were also obtained in U20S, PC3 and PANC-1 cells.

Target engagement assay (cells):

Indirect, inhibition of protein substrate methylation

Toxicity

Cytoxicity assay:

Yes

Notes on cytotoxicity:

Cytotoxicity was assessed by Alamar blue assay, yielding desirable efficacy: toxicity ratios in multiple cell types.

In vivo validation

Organism:

Swiss albino mice

Dose:

5 mg/kg

Route of delivery:

Intraperitoneal

Ratio of probe in organ to plasma (O:P):

0.33

Cmax:

947 ng/mL

Organ of Interest

Organ (O):

brain

Primary Reference

Reference (doi):

10.1021/jm401480r

Reference (PMID):

24102134

External Resources

Available data:

GDSC1

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

UNC0642 potently and specifically inhibits G9a and GLP compared with a wide range of pharmacologically important proteins (except for the histamine H3 receptor) in cell-free assays. Importantly, the compound shows clear concentration differences between the cellular functional activity (on-target activity) and cytotoxic activity (approximately three orders of magnitude). Therefore, chemical scaffold-induced, non-specific cellular effects would be avoided when the compound is used at the concentrations ranging from 100 nM to 1 microM. The roughly obtained PK values seem acceptable for in vivo mouse studies; however, whether i.p. administration of UNC0642 modulates the pharmacodynamics of the target (assayed by looking at levels of H3H9me2 in major tissues) would be worthwhile to examine before in vivo applications are pursued.

Sep 7 2016 - 8:01am

The in vitro potency and selectivity against other family members is good. The margin between concentrations that yield a functional effect and cellular toxicity was determined and is at least 80-fold. Limited PK data are provided in the original paper; however, dosing is limited to IP administration, and only Cmax and AUC(0-24hr) are reported. Plasma Cmax at this dose is 1.7 uM. In the absence of protein-binding data at this concentration, relative in vitro potency cannot be estimated, and direct pharmacodynamic data for in vivo target modulation are absent. A second publication (doi:10.1038/nm.4257) reports the in vivo use of this compound using the same dosing regimen, and it shows modulation of maternally repressed gene expression consistent with the in vitro cellular data. Direct demonstration of histone methylation modulation in vivo would increase my confidence in the mechanism of action of this probe.

Jan 23 2017 - 1:25pm

This is a well characterised probe with dual activity against the related targets GLP and G9a. The probe has been evaluated for selectivity against a variety of off-targets including kinases, GPCRs, etc. and appears to be suitably selective. Moreover, selectivity against other related epigenetic targets has also been undertaken and appears satisfactory. Reversibility, cellular activity, etc. are well described and some in vivo data support the use of the probe in model organisms. However, it is worth noting that the probe is recommended for IP dosing only.

Jun 19 2017 - 12:54pm

UNC0642

Probe Availability

Probe Information

Target Details

G9A, G9a, G9a-GLP, G9a/GLP, BAT8, GAT8, NG36, KMT1C, euchromatic histone-lysine N-methyltransferase 2, GLP, GLP1, KMT1D, FP13812, EUHMTASE1, Eu-HMTase1, bA188C12.1, euchromatic histone-lysine N-methyltransferase 1

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Toxicity

In vivo validation

Primary Reference

External Resources

Supporting Organizations

Interested in supporting the portal?

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UNC0642

Probe Availability

Probe Information

Target Details

G9A, G9a, G9a-GLP, G9a/GLP, BAT8, GAT8, NG36, KMT1C, euchromatic histone-lysine N-methyltransferase 2, GLP, GLP1, KMT1D, FP13812, EUHMTASE1, Eu-HMTase1, bA188C12.1, euchromatic histone-lysine N-methyltransferase 1

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Toxicity

In vivo validation

Primary Reference

External Resources

Supporting Organizations

Interested in supporting the portal?