TBK1 PROTAC 3i

Protein target name:

TBK1

Mechanism of action:

DegraderPROTAC

Recommended Concentration for use in cells:

~~up to 1 uM~~

Reviewer suggested Concentration for use in cells:

up to 100 nM

Probe Availability

TOCRIS

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

Ser/Thr Protein Kinase

HUGO ID:

HGNC:11584

Entrez gene ID:

29110

UNIPROT ID:

Q9UHD2

Target alias:

NAK; T2K; IIAE8; FTDALS4

Probe Details

Chemical Probes Portal ID:

CPP2088

Probe alias:

cpd 3i, PROTAC

PubChem CID:

124108661

ChEMBL ID:

CHEMBL4227907

SMILES string:

CN(C(C1CCC1)=O)CCCNC2=NC(NC3=CC=C(C=C3)OCCCOCCCCOCCCOCC(N[C@@H](C(C)(C)C)C(N4C[C@@H](C[C@H]4C(NCC5=CC=C(C6=C(N=CS6)C)C=C5)=O)O)=O)=O)=NC=C2Br

InChi Key:

QMGHHBHPDDAGGO-IIWOMYBWSA-N

Control compound:

TBK1 control PROTAC 4

Orthogonal probe:

BAY-985

Physico-chemical properties

NMR:

1H NMR (400 MHz, CD3OD): δ 1.05 (s, 9H), 1.60–1.64 (m, 4H), 1.80–2.26 (m, 15H), 2.48 (s, 3H), 2.86–2.91 (2s, 3H), 3.26–3.28 (m, 1H), 3.40–3.54 (m, 9H), 3.59–3.64 (m, 4H), 3.83–3.88 (m, 2H), 3.97–4.06 (m, 4H), 4.34–4.38 (m, 1H), 4.52–4.61 (m, 3H), 4.71–4.72 (m, 1H), 6.87–6.90 (m, 2H), 7.41–7.49 (m, 6H), 7.87–7.90 (2s, 1H), 8.88 (s, 1H).

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

4.6 nM

Potency assay:

Kd determined at DiscoverX, VHL Binding Assay Based on Fluorescence Polarization

In cell validation

On target activity in cells

Potency in cells:

DC50Dmax

12 nM96 %

Off target activity in cells

Potency assay, off target (cells):

Western Blot Assay to Determine TBK1/IKKε Degradation and IRF3 Phosphorylation

Probe Selectivity in Cell:

PROTAC 3i had no effect on the levels of IKKε, at concentrations of more than 50-fold above its TBK1 DC50

Primary Reference

Reference (doi):

10.1021/acs.jmedchem.7b00635

Reference (PMID):

28692295

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

no rating

SAB Members

SAB Comments

A potent VHL-based PROTAC molecule for TBK1 (DC50 = 12 nM & Dmax = 96%). SAR with the TBK1 binding moiety, linker and VHL binding moiety were demonstrated in the paper. Most TBK1 and VHL target engagement data are based on biophysical binding assays. Interrogation of downstream signalling (pIRF3) was also proven. Selectivity was only determined against ikkε. No broader kinase or proteome wide degradation selectivity available in the paper.

Dec 3 2020 - 10:16am

TBK1 PROTAC 3i meets most of the chemical probe criteria for a degrader. The researchers have shown evidence of binding to VHL using an FP assay and that an epimeric negative control compound is unable to degrade TBK1. Proteasomal dependence, quantification of target engagement, and degradation by western blot were also demonstrated. In terms of selectivity, PROTAC 3i had no effect on the degradation of IKKe (65% sequence homology to TBK1) despite poor selectivity of the TBK1 ligand. One criteria for degrader probes is a defined time course for degradation. The researchers in this study only looked at 16 hours. It would be beneficial to see a degradation profile across multiple time points. Additionally, the degradation profile of this PROTAC was not assessed proteome-wide and therefore we can not be fully confident of the selectivity of this PROTAC and that PROTAC 3i has no off-target degradation.

Dec 8 2020 - 10:06am

TBK1 PROTAC 3i

Probe Availability

Probe Information

Target Details

NAK; T2K; IIAE8; FTDALS4

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

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TBK1 PROTAC 3i

Probe Availability

Probe Information

Target Details

NAK; T2K; IIAE8; FTDALS4

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?