JQ1

Reagent Authentication Certificate:

Chemical Probes Portal JQ1.pdf

Protein target name:

BRD2, BRD3, BRD4, BRDT

Mechanism of action:

Antagonist

Recommended Concentration for use in cells:

10 nM - 1 uM

Probe Availability

Tocris (1268524-70-4)

MedChem Express (1268524-70-4)

Abcam (1268524-70-4)

MilliporeSigma (1268524-69-1)

Cayman (1268524-70-4)

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Bromodomain

HUGO ID:

HGNC:1103 HGNC:1104 HGNC:13575 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:

CPP1

Probe alias:

(+)-JQ-1,

PubChem CID:

46907787

ChEMBL ID:

CHEMBL1957266

SMILES string:

CC1=C(SC2=C1C(=N[C@H](C3=NN=C(N32)C)CC(=O)OC(C)(C)C)C4=CC=C(C=C4)Cl)C

InChi Key:

DNVXATUJJDPFDM-KRWDZBQOSA-N

Control compound:

(-)-JQ1

PAINs evaluation:

PAINs free

Physico-chemical properties

Solubility - preferred solvent:

DMSO

Solubility - concentration:

50 mM

Storage Recommendations

Storage (dry powder):

-20 oC 2 years

Storage sensitivities:

Avoid freeze-thaw cycles

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

BRD4 (N): 49.0 nM

Potency assay:

KD: BRD2 128 nM, BRD3 (N) 59.1 nM, BRD3 (C) 82.0 nM, BRD4 (C) 90.1 nM; BRDT (N): 190.1 nM, ITC assay IC50: BRD2 17.7 nM, BRD4 (N) 76.9 nM, BRD4 (C) 32.6 nM, alpha screen

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

2OSS, 3MXF, 3ONI

Off target activity

Off target protein and potency:

ATAD2: 0.18CREBBP: 1.04EP300: 0.07KIAA1240: 0.05PB1/3: 0.79SMARCA2: 0.93SMARCA4: 0.15TAF1/2: 0.28TAF1/3: 0.50TAF1L/2: 0.04

Potency assay (off target):

Differential scanning fluorimetry; for comparison BRD2/1: 6.47, BRD2/2: 7.97, BRD3/1: 8.27, BRD3/2: 8.39, BRD4/1: 9.35, BRD4/2: 7.44, BRDT/1: 3.93. No activity was detected against: ASH1L, BAZ2B, BRD1, BRD9, BRPF1, CECR2, FALZ, GCN5L2, LOC93349, PB1/1, PB1/2, PB1/5, PB1/6, PCAF, PHIP/2, SP140, TAF1L/3, TIF1, WDR9

Probe Selectivity in Vitro:

In ExpressProfile, negligible activity was detected: H2 [cimetindine]: -27, Neurokinin NK3 [SB 222200]: -23, Somatostatin SST [somatostatin-14]: -22
Benzodiazepine [diazepam]: -20, Prostanoid TP (TXA2/PGH2)[U 44069]: -19, Serotonin 5-HT transporter [imipramine]: -19, Serotonin 5-HT1A [8-OH-DPAT]: : -14, Muscarinic M1 [pirenzepine]: -14, GABA [GABA]: -12, Serotonin 5-HT7 [serotonin]: -12, Norepinephrine transporter [protriptyline]: -10, Adenosine A1 [DPCPX]: -10, Adenosine A2A [NECA]: -9, VPAC1 [VIP]: -9, Dopamine transporter [BTCP]: -7, Alpha-2 Adrenergic (nonselective) [yohimbine]:-7, Serotonin 5-HT5a [serotonin]: -7, Opioid Delta 2 (DOP) [DPDPE]: -6, Serotonin 5-HT3 [MDL 7222]: -6, Chemokine CXCR2 (IL-8B) [IL-8]: -5
Alpha-1 Adrenergic (nonselective) [prazosin]: -5, Histamine H1 [pyrilamine]: -5, Vasopressin V1a [[d(CH2)51Tyr(Me)2]-AVP]: -5, Ca2+ channel [D 600]: -5
Chemokine CCR1 [MIP-1alpha]: -4, Opioid Mu (MOP) [DAMGO]: -4, K+ (KV) channel [alpha-dendrotoxin]: -4, Serotonin 5-HT1B [serotonin]: -4,, Muscarinic M3 [4-DAMP]: -4, Neurotensin NTS1 (NT1) [neurotensin]: -4, Bradykinin B2 [NPC 567]: -3, Opioid-like NOP (ORL1) [nociception]: -2, Dopamine D1 [SCH 23390]: -2, Dopamine D2S [(+)-butaclamol]:-2, Neuropeptide Y1 [NPY]: -2, Serotonin 5-HT2B [DOI]: -2, Angiotensin-II AT1 [saralasin]: 0, Cannabinoid CB1 [CP 55940]: 0, Melanocortin MC4 [NDP-alpha-MSH]: 0, Beta-1 Adrenergic [atenolol]: +1, Muscarinic M2 [methoctramine]: +2, Serotonin 5-HT6 [serotonin]: +2, Cl- channel (GABA gated) [picrotoxinin]: +3, K+ Channel SKCa [apamin]: +3, Opioid Kappa (KOP) [U 50488]: +3
Beta-2 Adrenergic [ICI 118551]: +4, Galanin GAL2 [galanin]: +5, Cholecystokinin CCK1 (CCKA) [CCK-8s]: +9, Neuropeptide Y2 [NPY]: +10, Endothelin ETA [endothelin-1]: +16, Na+ channel [veratridine]: +18, Serotonin 5-HT2A [ketanserin]: +24, Melatonin MT1( ML1A) [melatonin]: +29, Neurokinin NK2 [[Nleu10]-NKA (4010)]: +56, Adenosine A3 [IB-MECA]: +61

In cell validation

On target activity in cells

Potency assay (cells):

Human osteosarcoma cells (U2OS) transfected with GFP–BRD4 show a time-dependent recovery of fluorescence intensity in Fluorescence recovery after photobleaching (FRAP) assay. In the presence of JQ1 (500 nM), the observed recovery is immediate, indicating displaced and freely diffusing nuclear BRD4. Cellular FRAP studies confirmed that the effects on the mobile fraction of BRD4 are limited to the biochemically active (1)-JQ1 stereoisomer.

Target engagement assay (cells):

Indirect: FRAP assay

Toxicity

Cytoxicity assay:

Yes

Notes on cytotoxicity:

At 500 nM, JQ1 leads to on target, BRD4-dependent apoptosis.

In vivo validation

Organism:

Mouse

Dose:

50 mg/kg

Route of delivery:

Oral

Other route of delivery:

Intravenous

Plasma half life:

0.897 hr (IV), 1.39 hr (oral)

Systemic clearance:

2.35 L/hr/kg (IV)

Cmax:

1,180 ng/mL (oral)

Tmax:

0.250 hr (oral)

Organ of Interest

Organ (O):

xenograft

Target engagement assay:

Indirect: effacement of NUT nuclear speckles, consistent with competitive binding to nuclear chromatin

Primary Reference

Reference (doi):

10.1038/nature09504

Reference (PMID):

20871596

External Resources

Available data:

GDSC1 GDSC1 GDSC2

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

This is an excellent selective cellular probe for the BET family of bromodomains, binding potently to all eight (BD1 and BD2 of BRD2/3/4/T). It has been extensively used in cells as a positive control. Its drawback is its suboptimal in vivo pharmacokinetics (IV t1/2 0.9 hours: source http://dx.doi.org/10.1038%2Fnature09504), so for in vivo studies, I would recommend using other BET-family bromodomain inhibitors.

Jun 12 2016 - 1:45pm

JQ1 is very potent probe for the BET family of proteins that can be used in cells.

Jun 12 2016 - 1:46pm

JQ1

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

Toxicity

In vivo validation

Primary Reference

External Resources

Supporting Organizations

Interested in supporting the portal?

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JQ1

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

Toxicity

In vivo validation

Primary Reference

External Resources

Supporting Organizations

Interested in supporting the portal?