BAY-850

Protein target name:

ATAD2

Mechanism of action:

Inhibitor

Recommended Concentration for use in cells:

~~up to 10 uM~~

Reviewer suggested Concentration for use in cells:

up to 1 uM

Probe Availability

MedChemExpress

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

ATPase

HUGO ID:

HGNC:30123

Entrez gene ID:

29028

UNIPROT ID:

Q6PL18

Target alias:

ANCCA; CT137; PRO2000

Probe Details

Chemical Probes Portal ID:

CPP2297

PubChem CID:

129196931

ChEMBL ID:

CHEMBL4536031

SMILES string:

O=C(N[C@H](CC1=CC=C(C#N)C=C1)CN[C@H]2CC[C@@H](N)CC2)C3=CC(C4=CC=C(CN[C@@H](C5=CC=C(C)C=C5)C)O4)=C(OC)C=C3Cl

InChi Key:

BSISGUIVBKDTQO-JLXKDNNHSA-N

Control compound:

BAY-460

Orthogonal probe:

GSK8814

Physico-chemical properties

NMR:

1H-NMR (500 MHz, DMSO-d6) δ [ppm] = 1.25 (d, 3H), 1.38-1.47 (m, 5H), 1.50-1.60 (m, 3H), 2.26 (s, 3H), 2.57 (m, 1H), 2.63-2.66 (m, 2H), 2.71 (m, 1H), 2.78 (dd, 1H), 3.09 (dd, 1H), 3.53 (d, 1H), 3.60 (d, 1H), 3.73 (q, 1H), 3.94 (s, 3H), 4.23 (m, 1H), 6.31 (d, 1H), 6.86 (d, 1H), 7.11 (d, 2H), 7.17 (s, 1H), 7.24 (d, 2H), 7.48 (d, 2H), 7.51 (s, 1H), 7.74 (d, 2H), 8.25 (d, 1H).

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50Kd

22-166 nM (T), 157 nM (A), 115 nM (B), 85 nM (M)

Potency assay:

TR-FRET (T), Alphascreen (A), BROMOscan (B), MST (M)

Off target activity

Probe Selectivity in Vitro:

The selectivity profile of BAY-850 was initially investigated in the BROMOscan panel, where it hits ATAD2 but not the closely related ATAD2B and confirmed with MST @ 10 uM.

Outside Target Family: BAY-850 showed no inhibitory activity in a panel of 354 kinases and had only modest effects at high concentrations on a few GPCRs.

In cell validation

On target activity in cells

Potency in cells:

pGI50

5.8-5.9

Potency assay (cells):

Growth Inhibition Assay

Target engagement assay (cells):

FRAP assay was used to to measure compound-related displacement of labeled full-length ATAD2 from chromatin in MCF7 cells

Primary Reference

Reference (doi):

10.1021/acschembio.7b00708

Reference (PMID):

29043777

Endorsed probe

The Chemical Probes Portal endorses this compound as chemical probe for use in cells.

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

no rating

SAB Members

SAB Comments

The discovery of BAY-850 was facilitated by optimisation of hits from a DNA encoded library screen. It shows remarkable specificity for the ATAD2 BD and does not interact with the closely related ATAD2B BD. This may be explained by a non-canonical binding mode which results in ATAD2 protein dimer induction.

Jul 26 2021 - 11:28am

BAY-850 was identified from a DNA Encoded Library screen and optimised to what appears to be a useful ATAD2 probe with impressive selectivity both within the target class and more broadly. Notably, BAY850 shows selectivity over ATAD2B which differentiates it from GSK8814. Data are presented which show that BAY-850 induces dimerization of ATAD2 and prevents association with chromatin. A Fluorescence Recovery After Photobleaching (FRAP) assay was used to confirm displacement of FL ATAD2 from chromatin in cells. Single-digit uM activity was observed in antiproliferative cellular assays, however, the cytotoxic effects could not be unambiguously linked to ATAD2 BD inhibition.

Aug 26 2021 - 9:48am

It is unclear whether the cytotoxicity observed between 5-10 uM is a result of on-target effects of ATAD2 inhibition. Thus, concentrations, where cell toxicity is observed, should not be the primary determinant for recommended concentrations for use in cells as is listed above. Rather, the 1 uM dose used in the FRAP experiments effectively displaces the ATAD2 BD from chromatin, displays no cytotoxicity in HMEC cells, and maybe a better concentration for cellular studies.
I recommend starting with a 1 uM dose of the compound in cell systems. If increasing dose, take care to assess viability effects. Especially in cancer cell lines, genetic loss of ATAD2 has only modest effects on cell proliferation (see depmap.org), so considerable viability effects in a given cell type at doses higher than 1 uM may be indicative of off-target effects.

Sep 21 2021 - 11:28am

BAY-850

Probe Availability

Probe Information

Target Details

ANCCA; CT137; PRO2000

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

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BAY-850

Probe Availability

Probe Information

Target Details

ANCCA; CT137; PRO2000

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?