SJF1528

Protein target name:

EGFR, ERBB2

Mechanism of action:

DegraderPROTAC

Recommended Concentration for use in cells:

250 nM

Probe Availability

TOCRIS

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

RTK

HUGO ID:

HGNC:3236 HGNC:3430

Entrez gene ID:

1956 2064

UNIPROT ID:

P00533 P04626

Target alias:

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

Probe Details

Chemical Probes Portal ID:

CPP2106

Probe alias:

PROTAC

PubChem CID:

135157151

SMILES string:

CC1=C(C2=CC=C(C=C2)CNC([C@@H]3C[C@H](CN3C([C@H](C(C)(C)C)NC(COCCOCCOC4=CC=C(C5=CC6=C(N=CN=C6NC7=CC(Cl)=C(C=C7)OCC8=CC(F)=CC=C8)C=C5)C=C4)=O)=O)O)=O)SC=N1

InChi Key:

ZSCOIFSUFMYZEZ-YSWDPXALSA-N

Orthogonal probe:

CanertinibAfatinibLapatinib

Validation in Cells and Model Organisms

In cell validation

On target activity in cells

Potency in cells:

DC50, Dmax

39.2 nM97.6 %

Potency assay (cells):

Immunoblots, Cell proliferation assay, suppression of downstream signalling.

Phenotypic Control Experiment:

Co-treatment with either proteasome inhibitor (epoxomicin) or NEDD8-activating enzyme E1 inhibitor (MLN-4924) was able to restore protein levels to untreated levels.

Off target activity in cells

Probe Selectivity in Cell:

Potent degrader of both EGFR and HER2

Primary Reference

Reference (doi):

10.1016/j.chembiol.2017.09.009

Reference (PMID):

29129716

Under Review

This probe is in the process of SAB review. Please continue to check back for new reviews and commentary.

SAB Rating

Rating for use in Cells

2 review(s)

Rating for use in Model Organisms

no rating

SAB Members

SAB Comments

This probe is highly recommended for the purposes of studying the phenotypic outcomes of degradation of HER-2 (ERBB2), EGFR, L858R or Exon 19 del EGFR in cells. Other than these RTK targets, as a result of the highly selective kinase profile of lapatinib from which this probe is based, this is anticipated that the degradation probe is highly selective against other kinases or off-targets. In terms of its use in cells, the only disadvantage of this probe is that it does not discriminate between EGFR and HER-2 in its degradation characteristics. It should be noted that the pharmacokinetic properties of this probe have not been described and therefore additional experiments would need to be undertaken in order to conduct any in vivo studies.

Feb 9 2021 - 8:47pm

Reviewer notes:

The biochemical potencies listed above are approximately that of lapatinib (Chembl lists the IC50 of lapatinib on EGFR and HER2 to be between 2 < IC50 < 10 nM depending on the assay format). No in vitro biochemical potency was provided for the actual PROTAC.
No ‘hook effect’ was observed with this PROTAC in OVCAR8 even at a top dose of 10 micromolar.
Compound 5 in the article (the same warheads but with a linker extended by 1 extra ethylene glycol unit appears to be a more selective reagent, showing no degradation of HER2, unlike the compound shown here (Compound 1 in article). While an EGFR DC50 for Compound 5 was not quantified, the potency by gel looks similar to Compound 1 (Figure 2A).
Compound 2 is a diastereomer of the hydroxyproline of the VHL-binding motif and ablates VHL binding, making a quality negative control.
Despite evidence of broad RNA expression of VHL, VHL has not been observed in all tissues at the protein-level (https://www.proteinatlas.org/ENSG00000134086-VHL/tissue), so a negative result may require quantification of VHL abundance for interpretation.
See comment above about sensitivity of DC50 to EGFR mutational status.

Feb 10 2021 - 4:09pm

SJF1528

Probe Availability

Probe Information

Target Details

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

Probe Details

In cell validation

Primary Reference

Supporting Organizations

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SJF1528

Probe Availability

Probe Information

Target Details

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

Probe Details

In cell validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?