OICR-9429

Reagent Authentication Certificate:

Chemical Probes Portal OICR-9429.pdf

Protein target name:

WDR5

Recommended Concentration for use in cells:

500 nM - 5 uM

Probe Availability

MedChem Express (1801787-56-3)

Tocris (OICR9429)

MilliporeSigma (OICR9429)

Cayman (1801787-56-3)

Probe Information

Target and Probe Information

Target Details

Target class:

Structural protein

Subclass:

WD40-repeat proteins

HUGO ID:

HGNC:12757

Entrez gene ID:

11091

UNIPROT ID:

P61964

Target alias:

BIG-3, CFAP89, SWD3

Probe Details

Chemical Probes Portal ID:

CPP44

Probe alias:

GTPL8231, OICR 9429, AKOS025147341

PubChem CID:

91623360

SMILES string:

CN1CCN(CC1)C2=C(C=C(C=C2)C3=CC(=CC=C3)CN4CCOCC4)NC(=O)C5=CNC(=O)C=C5C(F)(F)F

InChi Key:

DJOVLOYCGXNVPI-UHFFFAOYSA-N

Control compound:

OICR-0547

PAINs evaluation:

PAINs free

Physico-chemical properties

Solubility - preferred solvent:

DMSO

Solubility - concentration:

50 mM

NMR:

1H NMR (500 MHz, DMSO-d6) δ 8.25 (s, 1H), 7.99 (s, 1H), 7.61 (s, 1H), 7.54 (d, J = 7.7 Hz, 1H), 7.47 (dd, J = 1.6, 8.3 Hz, 1H), 7.40 (t, J = 7.6 Hz, 1H), 7.27-7.37 (m, 2H), 6.92 (s, 1H), 3.70 (t, J = 4.5 Hz, 4H), 3.54-3.65 (m, 2H), 3.02 (t, J = 4.6 Hz, 4H), 2.74 (br s, 4H), 2.51 (br. s., 4H), 2.42 (s, 3H)

13C NMR (125 MHz, DMSO-d6) δ 163.6, 162.9, 143.2, 140.4, 140.0 (d, 2JC-F = 33.6 Hz, 1C), 138.4, 137.5, 137.4, 132.4, 128.5, 128.3, 127.7, 125.6, 124.1, 121.2, 120.7, 119.0 (q, 3JC-F = 5.5 Hz, 1C), 122.0 (q, 1JC-F = 274.6 Hz,1C), 113.3, 66.3, 62.9, 54.8 (2), 53.2, 51.0 (2), 44.4.

Storage Recommendations

Storage (dry powder):

2 years -20 oC

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

93 nM

Potency assay:

ITC OICR-9429 disrupts WDR5 interaction with a peptide from WDR5-interacting protein Kdisp = 64 nM.

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

4E5G

Off target activity

Potency assay (off target):

No activity detected against 9 WDR40 or epigenetic reader proteins.

Probe Selectivity in Vitro:

No binding or inhibition was detected against 22 protein methyltransferases. Negligible activity was detected against >250 human kinases, GPCRs, ion channels (Cerep) at 1 uM.

In cell validation

On target activity in cells

Potency in cells:

IC50

223 nM

Potency assay (cells):

OICR-9429 disrupted WDR5-MLL interaction as well as C/EBPalpha p30-WDR5 interaction in cells.

Target engagement assay (cells):

A biotinylated version of OICR-9429 selectively pulled down WDR5 in cells.

Primary Reference

Reference (doi):

10.1038/nchembio.1859

Reference (PMID):

26167872

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

Recommended concentration is based on the finding that the IC50s for the disruption of WDR5-MLL and WDR5-RbBP5 complexes in co-IPs from HEK293 expressing Flag-WDR5 are 223 nM and 458 nM, respectively.

Jun 12 2016 - 2:45pm

OICR-9429 is a small-molecule antagonist of the WDR5-MLL protein-protein interaction, a notoriously difficult class of molecular targets. There is solid evidence showing a direct in vitro interaction between OICR-9429 and its intended target, WDR5. This includes ITC and X-ray crystallography. This compound displays high affinity (Kd <100 nM) for WDR5. It has well-characterized, reversible binding kinetics. The authors also show (at least in vitro) that it does not significantly inhibit other many other HMTs, kinases or other pharmacologically relevant targets. There are convincing target-engagement data in cells via biotinylated pull-down proteomics and gene-expression profiling consistent with such engagement. It is effective in vitro versus p30-expressing cells. Note, there is <1-log difference is cytotoxicity of OICR-9429 to p30-expressing cells versus other cells, as reported in the original publication. Therefore, dosage of this probe should be carefully considered, perhaps even taking care to perform dose-response controls to rule out non-specific effects that may occur at higher compound concentrations. Users should take advantage of a close structural analog, OICR-0547, as a negative control compound. The authors present evidence showing the negative control compound does not disrupt the WDR5-MLL interaction in vitro. It has similar cytotoxoicity as OICR-9429 versus K562 cells. Users should also note that there is only ~1-log difference in cytotoxicity between the active compound and inactive analog.

Jun 30 2016 - 4:17am

OICR-9429

Probe Availability

Probe Information

Target Details

BIG-3, CFAP89, SWD3

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

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OICR-9429

Probe Availability

Probe Information

Target Details

BIG-3, CFAP89, SWD3

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?