BAY-1816032

Protein target name:

BUB1

Mechanism of action:

Inhibitor

Recommended Concentration for use in cells:

~~100 nM~~

Reviewer suggested Concentration for use in cells:

300 nM

Probe Availability

MedChemExpress

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

Ser/Thr Protein Kinase

HUGO ID:

HGNC:1148

Entrez gene ID:

699

UNIPROT ID:

O43683

Target alias:

BUB1A, BUB1L, hBUB1

Probe Details

Chemical Probes Portal ID:

CPP1969

Probe alias:

BAY 1816032

PubChem CID:

118958833

SMILES string:

COC1=CN=C(N=C1NC2=C(C=NC=C2)OC)C3=NN(C4=CC=CC=C43)CC5=C(C=C(C=C5F)OCCO)F

InChi Key:

QVOGVAVHOLLLAZ-UHFFFAOYSA-N

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50, KD

6.1 nM @ 10 uM ATP (F), 2.1 nM (S), 1.0 nM (e)

Potency assay:

Biochemical TR- FRET assay (F), SPR (S), ePCA binding competition (e)

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

6F7B

Structure-activity relationship:

yes

Off target activity

Probe Selectivity in Vitro:

Kinase selectivity was determined at 0.1 and 1 uM, in an active site-directed competition-binding assay measuring 403 human kinases (Lead Hunter, DiscoverX Kinome Scan) followed by Kd determination for those kinases inhibited by >60% at 100 nmol/L. LOK/STK10, DMPK2, and DDR1 were hit at a compound concentration of 100 nmol/L with respective KDs of 57, 850, and 2,300 nM

In cell validation

On target activity in cells

Potency in cells:

IC50

20 nM

Potency assay (cells):

Inhibition of histone H2A-Thr120 phosphorylation and of histone H3-Ser10 phosphorylation assay

Off target activity in cells

Probe Selectivity in Cell:

BAY 1816032 did not inhibit ezrin/radixin/moesin phosphorylation in Jeg-3 and HCT116 cells, did not block TGFβ-stimulated SMAD2/3 phosphorylation in A549 cells even at concentrations up to 30 μmol/L

In vivo validation

Organism:

male CD1 Mouse

Dose:

0.5 mg/Kg (d)

Route of delivery:

OralIntravenous

Plasma half life:

1.8 (m), 3.9 (r), 4.2 (d) h

Systemic clearance:

3.7 (m), 1.7 (r) 0.56 (d) L/h/Kg

Fraction of probe bound to plasma protein:

22 (m), 60 (r), 59 (d) %

Cmax:

0.024 (m), 0.054 (r), 0.17 (d) ug/L

Tmax:

0.25 (m), 7.0 (r), 1.0 (d) h

Primary Reference

Reference (doi):

10.1158/1078-0432.CCR-18-0628

Reference (PMID):

30429199

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

2 review(s)

SAB Members

SAB Comments

BAY 1816032 is an exquisitely selective ATP-competitive inhibitor of BUB1 kinase with long residence time (t1/2 of 87 or 203 minutes, measured by kPCA or SPR, respectively). In the DiscoveRx panel of 403 kinases, BAY 1816032 gives a Kd of 3.3 nM, whereas the next inhibited kinases are LOK (Kd =57 nM) and DMPK2 (Kd = 850 nM). The authors went further to demonstrate that BAY 1816032 at 10 uM does not inhibit pERM, the physiological substrate of LOK, whereas erlotinib (Kd of 19 nM for LOK) does at compound concentration of 10 uM. The cellular potency of BAY 1816032 has been demonstrated by inhibition of histone H2A-Thr120 phosphorylation with IC50 of 29 +/- 23 nM. In nude mice, BAY 1816032 dosed up to 150 mg/kg orally BID has demonstrated dose-dependent inhibition of phospho-Thr120 histone H2A in skin tissues. This is also the dose where combination efficacy has been demonstrated with BAY 1816032 added to paclitaxel in SUM-149 TNBC model, or BAY 1816032 added to olaparib in BRCA-1 mutated MDA-MB-436 TNBC model. Please note that this compound has moderate half-life (1.8 h) in mice, thus BID (twice daily) dosing is likely required. This probe is useful to dissect kinase catalytic and scaffolding function of BUB1.

Sep 15 2020 - 8:39am

BAY-1816032 is a useful chemical tool to investigate the effects of inhibition of human BUB1 in human cancer cells lines, in vitro and in murine xenograft studies. Suitable in vivo doses in murine models are 25 or 50 mg/kg p.o., given twice daily. The compound appears generally well tolerated in toxicity studies, so it is suitable for investigating combinations with other oncology therapeutics.

Oct 12 2020 - 10:32am

BAY-1816032

Probe Availability

Probe Information

Target Details

BUB1A, BUB1L, hBUB1

Probe Details

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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BAY-1816032

Probe Availability

Probe Information

Target Details

BUB1A, BUB1L, hBUB1

Probe Details

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?