CPI-0610

Reagent Authentication Certificate:

Chemical Probes Portal CPI0610.pdf

Protein target name:

BRD2, BRD3, BRD4, BRDT

Mechanism of action:

AntagonistReversible, competitive inhibitor

Recommended Concentration for use in cells:

10-500 nM

Probe Availability

Cayman (1380087-89-7)

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Bromodomain

HUGO ID:

HGNC:13575 HGNC:1103 HGNC:1104 HGNC:1105

Entrez gene ID:

6046 8019 23476 676

UNIPROT ID:

P25440 O60885 Q15059 Q58F21

Target alias:

D6S113E, FSH, FSRG1, NAT, O27.1.1, RING3, RNF3, ORFX, RING3L, CAP, HUNK1, HUNKI, MCAP, BRD6, CT9

Probe Details

Chemical Probes Portal ID:

CPP1312

Probe alias:

Cpi-0610

PubChem CID:

57389999

SMILES string:

CC1=NOC2=C1C3=CC=CC=C3C(=N[C@H]2CC(=O)N)C4=CC=C(C=C4)Cl

InChi Key:

GCWIQUVXWZWCLE-INIZCTEOSA-N

Orthogonal probe:

JQ1MT1PFI-1

Physico-chemical properties

NMR:

1H NMR (400 MHz, Acetone-D6) δ 7.85 (d, J = 7.60 Hz, 1H), 7.73─7.68 (m, 1H), 7.47─7.41 (m, 6H), 7.20 (br. s., 1H), 6.41 (br. s., 1H), 4.50 (br. s.,1H), 3.42─3.37 (m, 2H), 2.55 (s, 3H); 13C NMR (500 MHz, d6-DMSO) d 171.3, 168.6, 167.4, 157.3, 139.3, 134.7, 133.4, 131.4, 131.1, 130.7, 130, 28, 126.8, 125.8, 112.5, 53.5, 36.4; 10.6

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

BRD4: 39 nM

Potency assay:

Time-resolved FRET assay. IC50 values in Alphalisa assays were also determined: BRD4 BD1=0.12 uM, BD2-0.018 uM; BRD2 BD1=0.16 uM, BD2=0.025 uM; BRD3 BD1=0.17 uM, BD2=0.021 uM; BRDT BD1=0.22 uM, BD2=0.13 uM.

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

5HLS

Structure-activity relationship:

Yes, see J Med Chem paper

Off target activity

Potency assay (off target):

CPI-0610 had IC50s>15 uM against other bromodomain proteins in Alphalisa assays (CBP, BRD9, BRPF1, PCAF, BRG1, ATAD2, TRIM24, and BRD8.

Probe Selectivity in Vitro:

CPI-0610 had no significant activity in Cerep panel of 50 GPCRs, ion channels and transporters or against common P450 enzyme isoforms at 10 uM.

In cell validation

On target activity in cells

Potency in cells:

EC50

180 nM

Potency assay (cells):

Quantigene MYC cellular assay: MV-4-11 cells were plated at 10,000 cells/well in 96-well plates containing compounds in 100 μL RPMI medium supplemented with 10% FBS. Cells were incubated with compound for 4 hours at 37◦C and 5% CO2 prior to analysis of MYC transcript levels using the QuantiGene 2.0 Reagent system (Affymetrix) according to the manufacturer’s instructions. Inhibition was calculated relative to DMSO treated MYC levels.

In vivo validation

Organism:

Balb/c nude mouse

Dose:

30 mg/kg (oral), 10 mg/kg (SC)

Route of delivery:

OralSubcutaneous

Cmax:

24 uM (oral)8.4 uM (SC)

Tmax:

0.4 h (oral)1.3 h (SC)

Organ of Interest

Organ (O):

MV4-11 xenograft

Target engagement assay:

Indirect, Myc expression assay

Primary Reference

Reference (doi):

10.1021/acs.jmedchem.5b01882

Reference (PMID):

26815195

CPI-0610

Probe Availability

Probe Information

Target Details

D6S113E, FSH, FSRG1, NAT, O27.1.1, RING3, RNF3, ORFX, RING3L, CAP, HUNK1, HUNKI, MCAP, BRD6, CT9

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?

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CPI-0610

Probe Availability

Probe Information

Target Details

D6S113E, FSH, FSRG1, NAT, O27.1.1, RING3, RNF3, ORFX, RING3L, CAP, HUNK1, HUNKI, MCAP, BRD6, CT9

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?