BAY-299

Protein target name:

BRD1, TAF1, TAF1L

Mechanism of action:

Antagonist

Recommended Concentration for use in cells:

2-500 nM

Probe Availability

Tocris (2080306-23-4)

Cayman (2080306-23-4)

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Bromodomain

HUGO ID:

HGNC:1102 HGNC:11535 HGNC:18056

Entrez gene ID:

23774 6872 138474

UNIPROT ID:

O95696 P21675 Q8IZX4

Target alias:

BRL, BRPF1, BRPF2, BA2R, CCG1, CCGS, DYT3, DYT3/TAF1, KAT4, MRXS33, N-TAF1, NSCL2, OF, P250, TAF(II)250, TAF2A, TAFII-250, TAFII250, XDP, TAF(II)210, TAF2A2

Probe Details

Chemical Probes Portal ID:

CPP1557

PubChem CID:

122705990

SMILES string:

OCCCC1=CC=C(C(N2C3=C(C)C=C(N(C)C(N4C)=O)C4=C3)=O)C5=C1C=CC=C5C2=O

InChi Key:

OFWWWKWUCDUISA-UHFFFAOYSA-N

Control compound:

BAY-364

Orthogonal probe:

NI-57 (inhibits BRPF family)OF-1 (inhibits BRPF family)

Physico-chemical properties

Solubility - preferred solvent:

DMSO

Solubility - concentration:

6.5 mg/L

NMR:

1H NMR (400 MHz, [D]6DMSO): δ = 1.88 (mc, 2 H), 2.08, 3.29 (2 s, 3 H each), 3.30 (mc, 2 H), 3.38 (s, 3 H), 3.54 (mc, 2 H), 4.67 (t, J = 5.2 Hz, 1 H), 7.17 (mc, 2 H), 7.78 (d, J = 7.6 Hz, 1 H), 7.94 (dd, J = 7.4, 8.6 Hz, 1 H), 8.46 (d, J = 7.6 Hz, 1 H), 8.55 (dd, J = 1.0, 7.4 Hz, 1 H), 8.69 (dd, J = 1.0, 8.4 Hz, 1 H) ppm

Storage Recommendations

Storage sensitivities:

Storage: -20°C; Shipping: Wet ice in the continental US, may vary elsewhere; Stability: ≥2 years

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

BRD1: BD1=67 nM; TAF1: BD2=8 nM; TAF1L: BD2=106 nM

Potency assay:

Time-resolved fluorescence resonance energy transfer (TR-FRET) assay

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

5N49, 5MG2

Structure-activity relationship:

The primary publication lists SAR of position 6, 5', and 6' of the 1,3-dimethyl-benzimidazolone core and naphthalimide moiety, respectively. Position 6 SAR was done to optimize for BRPF2 BD, BRPF1 BD, TAF1 BD2 and BRD4 BD1 activity while position 5' and 6' were used to optimize BRPF2 BD, BRPF1 BD, TAF1 BD2 and BRD4 BD1 activity. Another SAR was done to improve solubility. TF-FRET was used to generate all SAR.

Off target activity

Potency (off target):

Off target protein and potency:

CREBBP: 1,390 nM

Potency assay (off target):

Thermal Shift Assay on a panel of 48 bromodomains and BROMOscan panel.

Probe Selectivity in Vitro:

LeadProfilingScreen (Cerep) containing 68 potential protein targets at 10 uM, with no inhibition or stimulation >25% observed. At 10 uM, BAY-299 showed <50% inhibitory activity in a panel of 300 kinases for all proteins.

In cell validation

On target activity in cells

Potency in cells:

IC50

BRD1: BD H4=575 nM, BD H3.3=825 nMTAF1: BD2 H4=970 nM, BD2 H3.3=1,400 nM

Potency assay (cells):

NanoBRET assay in HCT116 cells

Target engagement assay (cells):

NanoBRET: The NanoLuc luciferase was C-terminally fused to BRPF2 BD (a.a. 560-666), BRPF1 BD (a.a. 626-732) or TAF1 BD2 (a.a. 1528-1640). A BRD4-NanoLuc fusion was used as control. The HaloTag was C-terminally fused to histone H3.3 or H4. The resulting plasmids were transfected into HCT116 cells using standard procedures. Following addition of the HaloTag ligand, the cells were treated with compound (10 nM-10 μM) for 4 or 24 h, and the IC50 values were calculated.

Off target activity in cells

Potency assay, off target (cells):

BAY-299 showed no activity against BRD4-H4 or BRPF1-H4 in NanoBRET assays up to 10 uM.

In vivo validation

Organism:

Male Wistar rats

Route of delivery:

IntravenousOther

Other route of delivery:

Intragastrally

Plasma half life:

10 h (IV)

Systemic clearance:

0.73 L/h/kg (IV)

Primary Reference

Reference (doi):

10.1021/acs.jmedchem.7b00306

Reference (PMID):

28402630

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

Cellular data show activity of BAY-299 on BRPF2 BD/H3.3, BRPF2 BD/H4, TAF1 BD2/H3.3, and TAF1 BD2/H4. However, little or no anti-proliferative affect was detected.

Jul 5 2017 - 2:39pm

Because the compound showed similar binding affinity for two types of Bromodomains (BDs) from two proteins with very different roles in transcriptional regulation, it is not amenable for use as a chemical probe to investigate the functions of these types of BDs.

Aug 3 2017 - 3:33pm

As the compound showed similar binding affinity between two type of Bromo domains (BDs) from two proteins with very different roles in transcriptional regulation means that it is not amenable for use as a chemical probe to investigate the functions of these types of BDs.

Jul 9 2020 - 10:19am

BAY-299

Probe Availability

Probe Information

Target Details

BRL, BRPF1, BRPF2, BA2R, CCG1, CCGS, DYT3, DYT3/TAF1, KAT4, MRXS33, N-TAF1, NSCL2, OF, P250, TAF(II)250, TAF2A, TAFII-250, TAFII250, XDP, TAF(II)210, TAF2A2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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BAY-299

Probe Availability

Probe Information

Target Details

BRL, BRPF1, BRPF2, BA2R, CCG1, CCGS, DYT3, DYT3/TAF1, KAT4, MRXS33, N-TAF1, NSCL2, OF, P250, TAF(II)250, TAF2A, TAFII-250, TAFII250, XDP, TAF(II)210, TAF2A2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?