BAZ2-ICR

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Chemical Probes Portal BAZ2-ICR.pdf

Protein target name:

BAZ2A, BAZ2B

Mechanism of action:

AntagonistMimics acetylated lysine

Recommended Concentration for use in cells:

500 nM - 1 uM

Probe Availability

Tocris (BAZ2-ICR)

Cayman (1665195-94-7)

MilliporeSigma (BAZ2-ICR)

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Bromodomain

HUGO ID:

HGNC:962 HGNC:963

Entrez gene ID:

11176 29994

UNIPROT ID:

Q9UIF9 Q9UIF8

Target alias:

TIP5, WALp3, WALp4

Probe Details

Chemical Probes Portal ID:

CPP66

Probe alias:

GTPL8571, AKOS025142070, ZINC226098129, Inhibitor 13

PubChem CID:

91654625

SMILES string:

CN1C=C(C=N1)CCN2C=NC(=C2C3=CC=C(C=C3)C#N)C4=CN(N=C4)C

InChi Key:

RRZVGDGTWNQAPW-UHFFFAOYSA-N

Orthogonal probe:

GSK2801

Physico-chemical properties

NMR:

1H NMR (500 MHz, MeODd4)
 8.09 (1H, brs, protonated), 8.07 (1H, s, ArH), 7.88 (2H, dat, J = 1.5, 1.5, 8.5 Hz, ArH),
7.0 (2H, dat, J = 1.5, 1.5, 8.5 Hz, ArH), 7.50 (1H, brs, ArH), 7.32 (1H, d, J = 0.5 Hz, ArH),
7.19 (1H, brs, ArH), 7.03 (1H, brs, ArH), 4.15 (2H, t, J = 6.5 Hz, CH2), 3.81 (3H, s, CH3), 3.78
(3H, s, CH3), 2.71 (2H, t, J = 6.5 Hz, CH2) ppm; 13C NMR (126 MHz, MeOD-d4)  139.6,
138.7, 137.9, 135.1, 134.1, 132.8, 131.3, 131.1, 129.8, 127.5, 119.3, 117.8, 115.2, 114.1,
48.3, 38.9, 38.7, 26.4 ppm

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

BAZ2A 109 nM, BAZ2B 170 nM

Potency assay:

Isothermal calorimetry. Activity was also assessed in an Alphascreen with IC50s 130 nM (BAZ2A) and 180 nM (BAZ2B).

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

4XUA, 4XUB

Structure-activity relationship:

Yes

Off target activity

Potency (off target):

Off target protein and potency:

CECR2 1.55 uM

Potency assay (off target):

In thermal shift assay of 47 human bromodomains, BAZ2-ICR was active against one off-target protein, CECR2, but 15-fold selective for the targets.

Probe Selectivity in Vitro:

In Cerep screen of 55 receptors, BAZ2-ICR had a clean off-target profile.

In cell validation

On target activity in cells

Potency assay (cells):

In FRAP assay in U2OS cells, BAZ2-ICR displaced the targets from chromatin.

Target engagement assay (cells):

Indirect, FRAP assay

Off target activity in cells

Potency assay, off target (cells):

In FRAP assay, BAZ2-ICR did not displace BRD4 from chromatin.

Probe Selectivity in Cell:

Not available

In vivo validation

Organism:

Mouse

Dose:

5 mg/kg

Route of delivery:

OralIntravenous

Plasma half life:

0.93 h (IV), 0.90 h (oral)

Systemic clearance:

36.2 mL/min/kg (IV), 36.0 mL/min/kg (oral)

Cmax:

11,272.7 nmol/L (IV), 5,824.3 nmol/L (oral)

Tmax:

0.083 h (IV and oral)

Primary Reference

Reference (doi):

10.1021/jm501963e

Reference (PMID):

25719566

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

BAZ2A/B (bromodomain adjacent to zinc finger domain, 2A/B) is a member of the nuclear remodeling complex (NoRC), but little is known about the function. Elevated levels of BAZ2A expression are observed in prostate cancer. BAZ2-ICR is a dual BAZ2A and BAZ2B bromodomain small-molecule inhibitor. It displays moderate potency (BAZ2A, ITC Kd = 109 nM; IC50 = 130 nM; BAZ2B, ITC Kd = 170 nM; IC50 = 180 nM) and good selectivity against a broad panel of 47 bromodomains: 15-fold selectivity for binding BAZ2A/B over CECR2 and >100-fold selectivity over all other bromodomains. BAZ2-ICR displaces BAZ2 bromodomains in living cells as demonstrated by accelerated FRAP recovery at 1 µM in the BAZ2A-FRAP assay. (PK data: LogD = 1.05, solubility 25 mM (D2O), F = 70%.) In comparison with another BAZA/B small-molecule inhibitor (GSK2801), it displays a better selectivity. To assess the compound's activity in a model organism, further experiments are required.

Aug 15 2016 - 10:13pm

In cells, the probe shows good selectivity, but the compound has only a moderate binding affinity ~100 nM and cellular activity (1 µM in FRAP assay). BAZ2-ICR has been shown by ITC to bind to BAZ2A with a KD of 109 nM and to BAZ2B with a KD of 170 nM. The compound has good selectivity; it only binds one off-target bromodomain: CECR2 (Kd= 1.55 μM). The compound (10 µM) displayed no activity against a panel of 55 receptors and ion channels. BAZ2-ICR shows good solubility, high stability in mouse microsomes and is cell permeable. BAZ2-ICR was tested in mouse model using iv and oral routes of administration (5 mg/kg); the compound showed good bioavailability (70%) and moderate clearance (∼50% of mouse liver blood flow). The maximum tolerated dose (MTD) was not determined. Considering the compound's cellular activity, a higher in vivo dose should be tested to see if concentration > 1µM could be maintained for longer time .

Oct 27 2016 - 1:38am

This probe has modest affinity to BZA2A (130 nM) and BZA2B (180 nM). It has good selectivity against 45 other bromodomains with the exception of CERC2 against which BAZ2-ICR is only 10-15 fold selective. In a cellular FRAP assay, activity was observed at a concentration of 1 uM; however, only a single concentration was reported. In the absence of dose-response data, absolute potency cannot be established.

Jun 15 2017 - 3:11am

BAZ2-ICR

Probe Availability

Probe Information

Target Details

TIP5, WALp3, WALp4

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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BAZ2-ICR

Probe Availability

Probe Information

Target Details

TIP5, WALp3, WALp4

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?