UNC0638

Protein target name:

EHMT2, EHMT1

Mechanism of action:

protein substrate competitive inhibitorSAM noncompetitive inhibitor

Probe Availability

MedChem Express (1255580-76-7)

Tocris (1255580-76-7)

Abcam (1255580-76-7)

MilliporeSigma (UNC0638 hydrate)

Cayman (1255580-76-7)

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Protein methyltransferase

HUGO ID:

HGNC:24650 HGNC:14129

Entrez gene ID:

79813 10919

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:

CPP56

Probe alias:

UNC 0638, 1255580-76-7, UNC-0638

PubChem CID:

46224516

ChEMBL ID:

CHEMBL1231795

SMILES string:

CC(C)N1CCC(CC1)NC2=NC(=NC3=CC(=C(C=C32)OC)OCCCN4CCCC4)C5CCCCC5

InChi Key:

QOECJCJVIMVJGX-UHFFFAOYSA-N

Control compound:

UNC0737

Orthogonal probe:

A-366BIX-01294UNC0642

PAINs evaluation:

PAINs free

Physico-chemical properties

NMR:

1H NMR (400 MHz, CDCl3) 7.15 (s, 1H), 6.83 (s, 1H), 5.22 (d, J = 7.2 Hz, 1H), 4.29–4.19 (m, 1H), 4.17 (t, J = 6.7 Hz, 2H), 3.93 (s, 3H), 2.96 (d, J = 11.8 Hz, 2H), 2.83 (sep, J = 6.6 Hz, 1H), 2.70 (tt, J = 11.6, 3.4 Hz, 1H), 2.62 (t, J = 7.4 Hz, 2H), 2.57–2.48 (m, 4H), 2.47–2.37 (m, 2H), 2.22 (d, J = 10.4 Hz, 2H), 2.09 (p, J = 7.0 Hz, 2H), 1.97 (d, J = 11.7 Hz, 2H), 1.86–1.58 (m, 11H), 1.45–1.27 (m, 3H), 1.11 (d, J = 6.6 Hz, 6H).

13C NMR (100 MHz, CDCl3) δ 169.32, 157.83, 153.52, 148.26, 147.27, 108.40, 106.65, 100.02, 67.46, 56.28, 54.45, 54.11, 52.81, 48.55, 47.84, 47.74, 32.61, 31.89, 28.39, 26.38, 26.21, 23.44, 18.47.

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

27 nM (EHMT2)

Potency assay:

SPR; also EHMT1 IC50 = 19 nM and EHMT2 IC50 < 15 nM in fluorescence-based SAHH coupled assay.

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

3RJW

Off target activity

Potency assay (off target):

UNC0638 was inactive against other H3K9 (SUV39H1 and SUV39H2), H3K27 (EZH2), H3K4 (SETD7, MLL and SMYD3), H3K79 (DOT1L) and H4K20 (SETD8) methyltransferases, as well as PRDM1, PRDM10 and PRDM12. In addition, UNC0638 was inactive against protein arginine methyltransferases PRMT1 and PRMT3, and HTATIP, a histone acetyltransferase. UNC0638 had weak but measurable activity against JMJD2E (IC50 = 4,500 ± 1,100 nM (n = 3)), a Jumonji protein demethylase and DNA methyltransferase DNMT1 (IC50 = 107,000 ± 6,000 nM (n = 2)). Nevertheless, the selectivity of UNC0638 for EHMT2 and EHMT1 over JMJD2E was >200-fold, and selectivity for EHMT2 and EHMT1 over DNMT1 was >5,000-fold.

Probe Selectivity in Vitro:

Selectivity of UNC0638 was evaluated across non-epigenetic targets, including GPCRs, ion channels, transporters and kinases. UNC0638 was clean (<30% inhibition at 1 μM) against 26 out of 29 targets in the Ricerca Selectivity Panel. At 1 μM concentration, UNC0638 showed 64%, 90% and 69% inhibition of muscarinic M2, adrenergic α1A and adrenergic α1B receptors, respectively. Because UNC0638 hit three GPCRs in the Ricerca Selectivity Panel, we further assessed its selectivity against GPCRs by testing UNC0638 in the NIMH's Psychoactive Drug Screen Program Selectivity Panel, which consists of a total of 45 targets, including 36 GPCRs. UNC0638 had <50% inhibition at 1 μM against 39 targets in the panel, and >50% inhibition at 1 μM against six targets in the panel. Ki in the radioligand binding assay for each of the six interacting GPCRs was subsequently determined. The Ki measurements showed UNC0638 was at least 100-fold selective for EHMT2 over these six GPCRs. In M1, M2 and M4 functional assays, UNC0638 had no agonist activity, low antagonist potency against M1 and M4 (IC50 > 10,000 nM (n = 2)), and modest antagonist potency against M2 (IC50 = 480 ± 10 nM (n = 2)). Furthermore, UNC0638 was tested against a panel of 24 kinases and showed <10% inhibition at 1 μM against these kinases.

In cell validation

On target activity in cells

Potency in cells:

IC50

81 nM

Potency assay (cells):

In-cell western of protein substrate methylation assay

Target engagement assay (cells):

Indirect, in cell protein methylation assay

Toxicity

Cytoxicity assay:

Yes

Notes on cytotoxicity:

EC50 = 11,000 nM in standard resazurin reduction assay (Alamar blue)

In vivo validation

Organism:

Swiss albino mice

Dose:

1 mg/mL

Route of delivery:

Intravenous

Plasma half life:

8.8 h

Systemic clearance:

306 mL/min/kg

Cmax:

75.1 ng/mL

Primary Reference

Reference (doi):

10.1021/jm401480r 10.1038/nchembio.599

Reference (PMID):

24900801 21743462

External Resources

Available data:

GDSC1 GDSC1

UNC0638

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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UNC0638

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?