GSK4027

Reagent Authentication Certificate:

Chemical Probes Portal GSK4027.pdf

Protein target name:

PCAF, GCN5

Mechanism of action:

Antagonist

Recommended Concentration for use in cells:

50 nM - 1 uM

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Bromodomain

HUGO ID:

HGNC:8638 HGNC:4201

Entrez gene ID:

8850 2648

UNIPROT ID:

Q92831 Q92830

Target alias:

Lysine acetyltransferase 2B, CAF, P/CAF, PCAF, Lysine acetyltransferase 2A, GCN5, GCN5L2

Probe Details

Chemical Probes Portal ID:

CPP1290

PubChem CID:

126961735

SMILES string:

O=C1C(Br)=C(N[C@@H]2C[C@H](C3=CC=CC=C3)CN(C)C2)C=NN1C

InChi Key:

VZAFGXCWAWRULT-UONOGXRCSA-N

Control compound:

GSK4028

Orthogonal probe:

L-Moses

Physico-chemical properties

Solubility - preferred solvent:

DMSO

Solubility - concentration:

10 mM

NMR:

1H NMR (500 MHz, DMSO-d6) δ 7.87 (s, 1H), 7.33–7.20 (m, 5H), 5.81 (d, J = 9.1 Hz, 1H), 3.97–3.93 (m, 1H), 3.59 (s, 3H), 2.97–2.93 (m, 2H), 2.84 (dd, J = 10.8, 3.2 Hz, 1H), 2.24 (s, 3H), 2.02–1.87 (m, 3 H), 1.73 (q, J = 12.1 Hz, 1H); 13C NMR (125.8 MHz, DMSO-d6) δ 157.0, 145.5, 143.2, 128.4, 127.0, 126.4, 126.2, 97.3, 61.5, 59.8, 49.4, 45.5, 40.6, 39.6, 36.8

Storage Recommendations

Storage (dry powder):

Stable as solid at room temperature for >3 years

Storage (solution):

Stable at -20 oC in DMSO (10 mM) for > 3 years

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

40 nM

Potency assay:

TR-FRET binding competition assay using truncated PCAF bromodomain and a fluorescently tagged bromodomain ligand.

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

5MLJ

Structure-activity relationship:

SAR is available and was demonstrated using a TR-FRET binding competition assay and multiple small molecule compounds.

Off target activity

Potency (off target):

Off target protein and potency:

BRPF3: 100 nMBRD1: 110 nMFALZ: 130 nMBRPF1: 140 nM

Potency assay (off target):

BROMOscan assay run at DiscoverX. In the assay, PCAF and GCN5: Ki=1.4 nM, hence >70 fold window to other targets.

Probe Selectivity in Vitro:

GSK internal enhanced, cross-screening panel (eXP); a full curve against 53 biochemical and phenotypic assays did not reveal any off-target binding <3 µM.

In cell validation

On target activity in cells

Potency in cells:

IC50

60 nM

Potency assay (cells):

NanoBRET assay, measuring displacement of NanoLuc-tagged full length PCAF from Halo-tagged histone H3.3 in HEK293 cells (Promega).

Target engagement assay (cells):

NanoBRET assay (as described above).

Toxicity

Cytoxicity assay:

Yes

Notes on cytotoxicity:

Cellular health assay looking at mitochondrial integrity, nuclear size and membrane permeability found no changes up to 200 uM.

Primary Reference

Reference (doi):

10.1021/acs.jmedchem.6b01566

Reference (PMID):

28002667

Endorsed probe

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

SAB Rating

Rating for use in Cells

5 review(s)

Rating for use in Model Organisms

5 review(s)

SAB Members

SAB Comments

GSK4027 is a potent inhibitor of the bromodomain of PCAF and GCN5 with good selectivity over other BRD-containing proteins. GSK4027 has good aqueous solubility, cell permeability, and it shows cellular target engagement of PCAF as measured in a NanoBRET assay (IC50=60 nM). GSK4027 is non-cytotoxic up to 200 uM. GSK4027 (R,R-enantiomer) is partnered with the enantiomeric negative control GSK4028 (S,S-enantiomer). L-Moses represents an orthogonal chemical probe.

Mar 18 2017 - 8:16pm

GSK4027 represents an orthogonal method to inhibit binding of the bromodomains present in PCAF and GCN5 to their targets compared with L-Moses. This compound shows similar in vitro and cellular potency to L-Moses (~50 nM) but was developed on an entirely different structural scaffold. The compound shows good target engagement in a nanoBRET assay against full-length PCAF and excellent selectivity versus other bromodomains, including BRD4 (>10,000-fold) and BRPF2, BRPF3, and BPTF (>70-fold). Compared with L-Moses, this compound showed a stronger response in the cellular target-engagement assays, and its use should take priority over L-Moses. GSK4028 serves as a reliable negative control. As with L-Moses, the one caveat to GSK4027 is that it is difficult to distinguish between effects on PCAF and GCN5 with this inhibitor.

Apr 11 2017 - 11:59am

GSK4027

Probe Information

Target Details

Lysine acetyltransferase 2B, CAF, P/CAF, PCAF, Lysine acetyltransferase 2A, GCN5, GCN5L2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Toxicity

Primary Reference

Supporting Organizations

Interested in supporting the portal?

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GSK4027

Probe Information

Target Details

Lysine acetyltransferase 2B, CAF, P/CAF, PCAF, Lysine acetyltransferase 2A, GCN5, GCN5L2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Toxicity

Primary Reference

Supporting Organizations

Interested in supporting the portal?