WZ4002

Protein target name:

EGFR

Mechanism of action:

Covalent InhibitorMutant Selective

Recommended Concentration for use in cells:

~~100-1000 nM~~

Reviewer suggested Concentration for use in cells:

up to 100 nM

Probe Availability

MedChemExpress

Selleck Chemicals

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

RTK

HUGO ID:

HGNC:3236

Entrez gene ID:

1956

UNIPROT ID:

P00533

Target alias:

ERBB; HER1; mENA; ERBB1; PIG61; NISBD2

Probe Details

Chemical Probes Portal ID:

CPP1229

Probe alias:

WZ4002, WZ-4002, 0UN

PubChem CID:

44607530

ChEMBL ID:

CHEMBL1229592

SMILES string:

COC1=C(C=CC(N2CCN(CC2)C)=C1)NC3=NC=C(C(OC4=CC(NC(C=C)=O)=CC=C4)=N3)Cl

InChi Key:

ITTRLTNMFYIYPA-UHFFFAOYSA-N

Orthogonal probe:

Rociletinib

Physico-chemical properties

NMR:

1H NMR 600 MHz (DMSO-d6) δ 8.32 (s, 1H), 7.38 (m, 2H), 7.26 (m, 2H), 6.96 (m, 1H), 6.48 (m, 2H), 6.35 (dd, J=10.2 Hz, 17.4 Hz, 1H), 6.21 (m, 1H), 5.75 (d, J=9.6 Hz, 1H), 3.80 (s, 3H), 3.61 (m, 4H), 3.11 (m, 4H), 2.38 (s, 3H)

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

14 WT, 2.2 T790M, 1.5 L858R, 0.13 L858R/T790M nM

Potency assay:

EGFR kinase assays

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

3IKA

Structure-activity relationship:

yes

Off target activity

Probe Selectivity in Vitro:

Profiled against a panel of 400 kinases using the Ambit kinome screening platform. wildetype ERBB2 with an IC50 of 32 nM.BTK (Kd 100 nM in Ambit binding assay) FAK (Kd 100 nM in Ambit binding assay)

In cell validation

On target activity in cells

Potency in cells:

IC50

2 nM EGFR-L858R, 8 nM EGFR-L858R/T790M, 3 nM EGFR (del), 2 nM EGFR (del/T790M)

Potency assay (cells):

pERK1/2, pAKT Western blot in PC9 GR lung cancer cells

In vivo validation

Organism:

Mouse

Dose:

1 mg/Kg IV10-25 mg/Kg PO

Route of delivery:

OralIntravenous

Plasma half life:

2.46 h PO1.76 h IV

Systemic clearance:

17.1 mL/min/Kg

Fraction of probe bound to plasma protein:

24.5 %

Cmax:

1485 ng/mL IV429 ng/mL PO

Tmax:

0.25 h

Primary Reference

Reference (doi):

10.1038/nature08622

Reference (PMID):

20033049

External Resources

Available data:

Nathanael Gray Lab - Probe Collection

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

WZ4002 is a covalent inhibitor of EGFR variants carrying the T790M mutant. The cmpd shows roughly a 10fold selectivity window vs. EGFR wt in cellular studies which also translates into in vivo studies. The data set is limited and does not allow conclusions about selectivity against other kinases. Biochemical selectivity data show off-target activity against certain kinases with a cysteine in a position analogous to C797, e.g. BMX, SNARK, BTK. However, for covalent inhibitors stationary IC50 values do not allow to accurately assess selectivity. For BMX it is shown that the cellular IC50 is within a 10fold range compared to cellular EGFR T790M potency. No data are provided about potential targets beyond kinases. The provided data set does not fulfil quality criteria as would be expected in 2021 for a covalent inhibitor, e.g. a broader assessment of the cellular interactome by a MS proteomics study. Therefore, WZ4002 may only be used for specific cellular studies on EGFR T790M but findings should be confirmed with a second chemically differentiated inhibitor.

Mar 2 2021 - 1:50pm

This is a covalent inhibitor of various EGFR mutants and is potent in biochemical and cellular assays. Depending on the cell line used, the selectivity for EGFR over other cysteine containing kinases, such as ERBB2 and BTK, could start to overlap such that interpretation of the data would benefit from additional inhibitors with different profiles.

Mar 2 2021 - 1:55pm

WZ4002 is identified as a covalent inhibitor potent against EGFR mutants including both primary activating mutations (e.g., L858R, ΔE746-A750) and resistant mutation T790M (e.g., L858R/T790M, ΔE746-A750/T790M double mutants). By design, it is rather weak against EGFR WT. If the intended research is on EGFR wild type, pan-EGFR inhibitor such as afatinib would be more suitable. If the intended research is on EGFR mutants other than those discussed in the primary paper, inhibitor potency needs to be determined. Given its potency requires covalent modification of Cys797, any EGFR kinase construct or cell line bearing C797X mutation would be insensitive. Furthermore, its kinome selectivity is good but not perfect, therefore use of multiple probes against the same target, such as Osimertinib/AZD9291, or counter-screen with selective inhibitors against BTK, ITK, or ERBB4 would be prudent.

Mar 12 2021 - 10:57am

WZ4002

Probe Availability

Probe Information

Target Details

ERBB; HER1; mENA; ERBB1; PIG61; NISBD2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

External Resources

Supporting Organizations

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WZ4002

Probe Availability

Probe Information

Target Details

ERBB; HER1; mENA; ERBB1; PIG61; NISBD2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

External Resources

Supporting Organizations

Interested in supporting the portal?