PF-477736

Reagent Authentication Certificate:

Chemical Probes Portal PF-477736.pdf

Protein target name:

CHEK1

Mechanism of action:

ATP-competitive inhbitor

Recommended Concentration for use in cells:

50 nM - 1 uM

Probe Availability

Tocris (1188296-52-7)

MedChem Express (952021-60-2)

MilliporeSigma (952021-60-2)

Cayman (952021-60-2)

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

CAMK

HUGO ID:

HGNC:1925

Entrez gene ID:

1111

UNIPROT ID:

O14757

Target alias:

CHK1

Probe Details

Chemical Probes Portal ID:

CPP138

Probe alias:

952021-60-2, PF-00477736, 9DB

PubChem CID:

16750408

ChEMBL ID:

CHEMBL3545137

SMILES string:

CN1C=C(C=N1)C2=NC3=C4C2=CNNC(=O)C4=CC(=C3)NC(=O)[C@@H](C5CCCCC5)N

InChi Key:

YFNWWNRZJGMDBR-LJQANCHMSA-N

Orthogonal probe:

CCT244747

PAINs evaluation:

PAINs free

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

0.49 nM

Potency assay:

In vitro kinase assay with the catalytic domain of CHEK1

Off target activity

Potency (off target):

IC50

Off target protein and potency:

AURA 23 nMFGFR3 23 nMFLT3 25 nMCSF1R 10 nMRET 39 nMYES 14 nMVEGFR2 8 nM (Ki)CHEK2 47 nM (Ki)

Potency assay (off target):

>100 kinases were screened, and PF-477136 was >100-fold selective for all other than those indicated.

In cell validation

On target activity in cells

Potency in cells:

EC50

45 nM

Potency assay (cells):

Abrogation of G2 checkpoint induced by DNA damage (detected by pSer10 H3 in a dot blot assay) in CA46 cells. In HeLa cells, EC50 = 38 nM; in HT29 cells, 42 nM.

Toxicity

Cytoxicity assay:

Yes

Notes on cytotoxicity:

In HT29 cells, PF-477136 was not cytotoxic up to 360 nM.

In vivo validation

Organism:

nu/nu mouse (Sprague-Dawley rat)

Dose:

4, 20, and 40 mg/kg (4 mg/kg)

Route of delivery:

Intraperitoneal

Other route of delivery:

IV in rats

Plasma half life:

2.9 h (rats)

Systemic clearance:

11.8 mL/min/kg (rats)

Cmax:

0.27-6.78 ug/mL (mice)

Primary Reference

Reference (doi):

10.1158/1535-7163.MCT-07-2391

Reference (PMID):

18723486

Endorsed probe

The Chemical Probes Portal endorses this compound as chemical probe for use in cells and in model organisms.

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

This probe is a potent inhibitor of CHK1 in biochemical and cellular assays.

Jun 16 2016 - 4:00pm

The compound abrogates cell cycle arrest induced by DNA damage and enhances cytotoxicity of clinically important chemotherapeutic agents, including gemcitabine and carboplatin. It also inhibits VEGFR2, Aurora-A, FGFR3, FLT3, FMS (CSF1R), RET and YES. It shows ~100-fold selectivity for CHK1 over CHK2. PF-477736 (128 nM) abrogates the camptothecin-induced DNA damage checkpoint in CA46 and HeLa cells. PF-477736 (540 nM) enhances gemcitabine-induced cytotoxicity in HT29 cells. PF-477736 (360 nM) suppresses docetaxel-induced phosphorylation of histone H3 (Ser10) and Cdc25C (Ser216) and potentiates apoptosis in COLO205 cells. PF-477736 (250 nM) combined with MK-1775 have synergistic cytotoxic activity in OVCAR-5 cells.

Jun 29 2016 - 7:47pm

PF-477736

Probe Availability

Probe Information

Target Details

CHK1

Probe Details

In vitro validation

In cell validation

Toxicity

In vivo validation

Primary Reference

Supporting Organizations

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PF-477736

Probe Availability

Probe Information

Target Details

CHK1

Probe Details

In vitro validation

In cell validation

Toxicity

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?