SGC-CBP30

Reagent Authentication Certificate:

Chemical Probes Portal SGC-CBP30.pdf

Protein target name:

CREBBP, EP300

Mechanism of action:

Antagonist

Recommended Concentration for use in cells:

100 nM - 1 uM

Probe Availability

MedChem Express (1613695-14-9)

Tocris (SGCCBP30)

MilliporeSigma (SGCCBP30)

Cayman (1613695-14-9)

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Bromodomain

HUGO ID:

HGNC:2348 HGNC:3373

Entrez gene ID:

1387 2033

UNIPROT ID:

Q09472 Q92793

Target alias:

CBP, KAT3A, RSTS, KAT3B, RSTS2, p300

Probe Details

Chemical Probes Portal ID:

CPP70

PubChem CID:

72201027

ChEMBL ID:

CHEMBL3622373

SMILES string:

CC1=C(C(=NO1)C)C2=CC3=C(C=C2)N(C(=N3)CCC4=CC(=C(C=C4)OC)Cl)C[C@H](C)N5CCOCC5

InChi Key:

GEPYBHCJBORHCE-SFHVURJKSA-N

Orthogonal probe:

GNE-049CPI-1612I-CBP112

PAINs evaluation:

PAINs free

Physico-chemical properties

Solubility - preferred solvent:

DMSO

Solubility - concentration:

100 mM

Storage Recommendations

Storage (dry powder):

2 years -20 oC

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

38 nM (CREBBP), 47 nM (EP300)

Potency assay:

Biolayer interference.

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

4NR7

Off target activity

Potency assay (off target):

In DSF binding assays with 45 bromodomains, SGC-CBP30 had no significant activity except against BRD2, BRD3, and BRD4.

Probe Selectivity in Vitro:

SGC-CPB30 had minimal activity against 136 GPCRs, ion channels, enzymes, and kinases with IC50s >1 uM, except Adrenergic receptor alpha 2C (0.11 uM), alpha 2A (0.57 uM), PDE5 (0.15 uM), and PAF (0.51 uM).

In cell validation

On target activity in cells

Potency assay (cells):

In HeLa cells, SGC-CBP30 displaced CREBBP from chromatin in a FRAP assay.

Target engagement assay (cells):

FRAP chromatin-displacement assay

Toxicity

Cytoxicity assay:

Yes

Notes on cytotoxicity:

SGC-CBP30 had modest cytotoxicity in U2OS cells; CC50 = 80 uM in MTT assay

Primary Reference

Reference (doi):

10.1021/ja412434f 10.1073/pnas.1501956112

Reference (PMID):

24946055 26261308

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

no rating

SAB Members

SAB Comments

SGC-CBP30 has BRD4(1) Kd of 850 nM; at 40-fold, this was the largest selectivity window over BET bromodomains reported until recently. BET-family bromodomain inhibitors have strong effects on cell proliferation and gene expression with IC50s similar to BET-binding affinity, making selectivity over this subfamily especially important for clean interpretation of biological results. Recently, an alternative CBP inhibitor, CPI-637 (http://dx.doi.org/10.1021/acsmedchemlett.6b00075), has stretched this window to ~700x BET selectivity, so it should be more suitable for distinguishing CBP-driven effects from BET, and it also has a negative enantiomer control. CPI-637 is not completely selective either, having 730 nM BRD9 affinity, but potent BRD9 chemical probes such as BI-9564 (http://dx.doi.org/10.1021/acs.jmedchem.5b01865) are available as controls, and their reported effects including on cell proliferation appear to be relatively modest compared to BET inhibitors of comparable affinity.

Jun 16 2016 - 5:34pm

While the existing data suggest this is a high-quality probe, users should realize it was only screened versus 10 representatives within each BRD family, not the entire target class. Also, there is some activity versus GPCRs and PDEs: When tested against 136 GPCR, ion channel, enzyme, and kinase targets, compound 59 showed an IC50 < 1 μM only for the adrenergic receptors α2C (0.11 μM) and α2A (0.57 μM), phosphodiesterase-5 (PDE5, 0.15 μM), and platelet-activating factor (PAF) (0.54 μM). In a human liver microsome (HLM) stability assay, no compound was detected after 60 min, implying that the metabolism of compound 59 may be too rapid for it to be useful as an oral in vivo probe.

Jun 29 2016 - 7:39pm

Users of this probe are advised to carefully consider the concentration range appropriate for their experiment. Although the recommendation states a 100-1000nM range in cells, pIC50 in a reporter assay is 1540nM (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4183655/), and 2uM is used in a later publication (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4553799/pdf/pnas.201501956.pdf); however profiles of activity appear to suggest that the weaker BRD activity may become significant at concentrations of 3.3 and 10 micromolar in cells (ibid), leaving a narrow window between on-target and off-target effects. Use of alternative probes for this target in parallel is recommended to aid deconvolution of possible BRD effects - use caution in interpreting results.

Jun 30 2016 - 4:21am

SGC-CBP30

Probe Availability

Probe Information

Target Details

CBP, KAT3A, RSTS, KAT3B, RSTS2, p300

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Toxicity

Primary Reference

Supporting Organizations

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SGC-CBP30

Probe Availability

Probe Information

Target Details

CBP, KAT3A, RSTS, KAT3B, RSTS2, p300

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Toxicity

Primary Reference

Supporting Organizations

Interested in supporting the portal?