Afatinib

Reagent Authentication Certificate:

Chemical Probes Portal Afatinib.pdf

Protein target name:

EGFR, ERBB2

Mechanism of action:

ATP-competitive inhibitorcovalent inhibitor

Recommended Concentration for use in cells:

Up to 100 nM

Probe Availability

MedChem Express (850140-72-6)

Abcam (439081-18-2)

Cayman (439081-18-2)

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

RTK

HUGO ID:

EGFR ERBB2

Entrez gene ID:

1956 2064

UNIPROT ID:

P00533 P04626

Target alias:

ERBB, ERBB1, HER1, NISBD2, PIG61, mENA and CD340, HER-2, HER-2/neu, HER2, MLN 19, NEU, NGL, TKR1

Probe Details

Chemical Probes Portal ID:

CPP135

Probe alias:

439081-18-2, Tovok, BIBW2992, BIBW 2992, Tomtovok

PubChem CID:

10184653

ChEMBL ID:

CHEMBL1173655

SMILES string:

CN(C)C/C=C/C(=O)NC1=C(C=C2C(=C1)C(=NC=N2)NC3=CC(=C(C=C3)F)Cl)O[C@H]4CCOC4

InChi Key:

ULXXDDBFHOBEHA-CWDCEQMOSA-N

Orthogonal probe:

CanertinibSJF1528Lapatinib

PAINs evaluation:

PAINs free

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50 (app)

EGFR 0.5 nM, ERBB2 14 nM

Potency assay:

In vitro kinase assays

Off target activity

Potency (off target):

IC50

Off target protein and potency:

LYN 736 nM

Potency assay (off target):

Afatinib was assessed against 52 off-target kinases, and it was most potent against LYN.

In cell validation

On target activity in cells

Potency in cells:

EC50

13-71 nM

Potency assay (cells):

In cell autophosphorylation detected by ELISA. Cells assayed include: A431, NIH-3T3, BT-474, NCI-N87; IC50 varied by cell type.

Target engagement assay (cells):

Indirect, inhibition of autophosphorylation in cells

In vivo validation

Organism:

nude mice

Dose:

20 mg/kg

Route of delivery:

Oral

Cmax:

83-309 nM

Organ of Interest

Organ (O):

xenograft

Primary Reference

Reference (doi):

10.1038/onc.2008.109

Reference (PMID):

18408761

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

As a probe for ERBB2 this would not be recommended as it exhibits greater potency for EGFR, more suitable selective probes for ERBB2 exist. Treat the quoted biochemical potency with caution as it is an irreversible inhibitor. Hits WT and activating mutant forms of EGFR with similar potency.

Jun 11 2016 - 4:18am

Potent irreversible inhibitor of EGFR and HER2. Some selectivity data are reported - ideally, a full kinase profile would be required to better understand higher dose range limit.

Jun 11 2016 - 4:19am

Treat the quoted biochemical potency with caution as it is an irreversible inhibitor. Afatinib hits wild-type and activating-mutant forms of EGFR with similar potency.

Aug 17 2016 - 6:28pm

Afatinib

Probe Availability

Probe Information

Target Details

ERBB, ERBB1, HER1, NISBD2, PIG61, mENA and CD340, HER-2, HER-2/neu, HER2, MLN 19, NEU, NGL, TKR1

Probe Details

In vitro validation

In cell validation

In vivo validation

Primary Reference

External Resources

Supporting Organizations

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Afatinib

Probe Availability

Probe Information

Target Details

ERBB, ERBB1, HER1, NISBD2, PIG61, mENA and CD340, HER-2, HER-2/neu, HER2, MLN 19, NEU, NGL, TKR1

Probe Details

In vitro validation

In cell validation

In vivo validation

Primary Reference

External Resources

Supporting Organizations

Interested in supporting the portal?