A Chemical Probe for the ATAD2 Bromodomain

Article abstract of DOI:10.1002/anie.201603928

ATAD2 is a cancer-associated protein whose bromodomain has been described as among the least druggable of that target class. Starting from a potent lead, permeability and selectivity were improved through a dual approach: 1) using CF₂as a sulfone bio-isostere to exploit the unique properties of fluorine, and 2) using 1,3-interactions to control the conformation of a piperidine ring. This resulted in the first reported low-nanomolar, selective and cell permeable chemical probe for ATAD2.

Full text of DOI:10.1002/anie.201603928

Angewandte
Communications
Chemie
International Edition: DOI: 10.1002/anie.201603928
German Edition: DOI: 10.1002/ange.201603928
Epigenetics
A Chemical Probe for the ATAD2 Bromodomain
Paul Bamborough,* Chun-wa Chung, Emmanuel H. Demont,* Rebecca C. Furze,
Andrew J. Bannister, Ka Hing Che, Hawa Diallo, Clement Douault, Paola Grandi,
Tony Kouzarides, Anne-Marie Michon, Darren J. Mitchell, Rab K. Prinjha, Christina Rau,
Samuel Robson, Robert J. Sheppard, Richard Upton, and Robert J. Watson
Abstract: ATAD2 is a cancer-associated protein whose
Table 1: Micromolar lead to nanomolar ATAD2 inhibitors. For statistics
bromodomain has been described as among the least drug-
gable of that target class. Starting from a potent lead,
permeability and selectivity were improved through a dual
see Table S1a, Supporting Information.
approach: 1) using CF as a sulfone bio-isostere to exploit the
2
unique properties of fluorine, and 2) using 1,3-interactions to
control the conformation of a piperidine ring. This resulted in
the first reported low-nanomolar, selective and cell permeable
chemical probe for ATAD2.
H
igh expression levels of ATAD2 (ATPase family, AAA
domain containing 2), also called ANCCA (AAA + nuclear
coregulator cancer-associated), correlate with poor outcomes
in several cancers, and its knockdown modulates multiple
[
1–3]
2
3
4
5
6
7
8
9
tumor cell growth factors.
have focused on competitive binding to the acetyl-lysine
KAc) site of its bromodomain, but the role of the bromo-
Efforts to target this protein
ATAD2 TR-FRET pIC50
5.6 7.2 6.9 6.7 6.9 6.5 7.4 7.1
(
ATAD2 Bromosphere pIC50 5.4 7.4 7.3 6.5 7.5 7.0 7.2 7.1
domain in the biology of ATAD2 is unclear. We have
developed chemical tools to try to understand this, and
recently reported the discovery of quinolinones and naph-
thyridones such as 1–3 and 5–7 that bind to the KAc site of
BRD4 BD1 TR-FRET pIC50 5.4 5.4 5.0 5.8 4.8 4.1 5.2 5.1
TR-FRET selectivity (logs)
Chrom logD (pH 7.4)
Polar surface area [ꢀ ]
0.2 1.8 1.9 0.9 2.1 2.4 2.2 2.0
3.3 2.1 0.4 4.0 1.6 2.3 3.0 4.1
79 113 152 92 126 117 92 83
130 <3 <3 138 <3 <10 86 395
2
Artificial membrane per-
[
4,5]
ATAD2.
À1
meability (nms , pH 7.4)
The profound biological effects associated with BET
inhibition complicate interpretation of phenotypes observed
[6]
with unselective inhibitors of other bromodomains. While
potent against ATAD2, naphthyridone 5 had only modest
selectivity over the BETs, represented in Table 1 by the first
bromodomain of BRD4 (BRD4 BD1). We recently reported
selectivity improvements from occupying an electrostatically
positive site near the KAc pocket, the RVF shelf, with
negative polarity (compare 5 to 6 to see the effect of
introducing cyclic sulfones at the C3’ position of the
[
5]
piperidine ring with (R,R) stereochemistry). An X-ray
structure of ATAD2 bound to 3 showed that its sulfone
oxygen atoms displace a weakly-bound water molecule and
accept two hydrogen bonds from the guanidinium group of
Arg1077 (Figure 1a). The WPF shelf of the BET bromodo-
mains (the analogous subsite to the ATAD2 RVF shelf)
contains lipophilic amino acids Trp81 and Met149 in place of
ATAD2 residues Arg1007 and Arg1077, so is less tolerant of
the sulfone groupꢀs polarity. Compounds with hydrophobic
C3’-substituents such as the cyclohexylmethylenes 2 and 5 are
more potent against the BETs, which is consistent with the
crystal structure of 2 bound to BRD4 BD1, where the C3’-
cyclohexyl ring rests on the WPF shelf (Figure 1b). Unfortu-
nately, while the sulfone C3’ group improved selectivity it also
impacted passive permeability, raising doubts over the suit-
ability of 6 and 7 as tool compounds for chromatin-bound
ATAD2. Here, we describe the conclusion of our optimiza-
tion of the series to overcome this.
[
*] Dr. P. Bamborough, Dr. C. Chung, Dr. E. H. Demont, Dr. R. C. Furze,
Dr. H. Diallo, C. Douault, D. J. Mitchell, Dr. R. K. Prinjha,
R. J. Sheppard, Dr. R. Upton, Dr. R. J. Watson
GlaxoSmithKline
Gunnels Wood Road, Stevenage, SG1 2NY (UK)
E-mail: Paul.A.Bamborough@gsk.com
Emmanuel.H.Demont@gsk.com
Dr. P. Grandi, Dr. A.-M. Michon, Dr. C. Rau
Cellzome GmbH, GlaxoSmithKline
Meyerhofstrasse 1, 69117 Heidelberg (Germany)
Dr. A. J. Bannister, Dr. K. H. Che, Prof. T. Kouzarides, Dr. S. Robson
Gurdon Institute
Tennis Court Road, Cambridge CB2 1QN (UK)
R. J. Sheppard
Current address: AstraZeneca
Milton Road, Cambridge CB4 OWG (UK)
Supporting information (X-ray, ITC, assay, molecular modelling,
LNCaP biology, NMR and chemistry supplementary methods) and
the ORCID identification numbers for the authors of this article can
be found under http://dx.doi.org/10.1002/anie.201603928.
Alternative C3’ functional groups of intermediate polarity
had been found inferior to the sulfone in potency or
Angew. Chem. Int. Ed. 2016, 55, 1 – 6
ꢀ 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
1
These are not the final page numbers!

Angewandte
Communications
Chemie
[
5]
selectivity. We therefore considered the possibilities offered
by fluorine substitution. The effect of fluorine on features of
organic molecules such as conformation, pK , permeability
a
[
7]
and metabolism has recently been reviewed. Fluorine
combines unique properties of polarity, lipophilicity and low
[
8–10]
polarisability.
Regarding noncovalent interactions, the
ability of organofluorine to participate in hydrogen bonds has
[10–15]
been debated.
While the lone pairs of organic fluorine
seem too tightly held to hydrogen-bond, strong charge–dipole
[15]
interactions can be formed with cations.
The bi-lobal negative charge patches characteristic of the
molecular surface of the SO group of 3 are well oriented to
2
interact with the ATAD2 RVF shelf residue Arg1077 (Fig-
ure 1a). The CF2 group possesses geometrically similar
electronegative patches (Figure S1, Supporting Information)
and should be compatible with polar interactions with
Arg1077. Indeed, the guanidinium group of arginine has
[
14]
been described as highly fluorophilic. We reasoned that CF2
may provide an isosteric replacement for SO , that its large
2
dipole moment relative to CH (2.57 D in 1,1-difluorocyclo-
2
[
16]
hexane ) might disfavour interaction with the hydrophobic
BRD4 BD1 WPF shelf, and that the expected increase in
lipophilicity should have a positive effect on permeability.
Fluorinated derivatives 8 and 9 were made according to
[5]
previously reported procedures and their profile compared
to the direct sulfone analogues 6 and 7 (Table 1). The higher
logD and lower polar surface area of the difluoromethylene
compounds relative to the sulfones resulted in dramatically
improved artificial membrane permeability, comparable to
the cyclohexyl analogues (compare 5, 6 and 8). The difluoro-
methylenes 8 and 9 had greater TR-FRET ATAD2 potency
than the sulfones (6 and 7), which we rationalize by
a decreased desolvation penalty. While the selectivity of 8
and 9 for ATAD2 over BRD4 BD1 was slightly lower than
the analogous sulfones 6 and 7, it was significantly better than
the cyclohexyl 5 and matched our probe critera of > 2 logs.
Overall, in this series the CF group proved to be an excellent
2
SO isostere.
2
In parallel, we sought ways to increase the selectivity
window by better understanding the binding mode of
analogues of 6 to BRD4 BD1. We assumed that this would
be similar to previous analogues such as 1 and 2 (Figure 1b).
The naphthyridone binds in the KAc-pocket, with the 3’
substituent of 2 located on the WPF shelf as outlined above.
We obtained a crystal structure of BRD4 BD1 bound to 4, the
amino-pyridine analogue of 6. The bound position of the
naphthyridone of 4 is very like that of 1. Unexpectedly, rather
than the 3’ substituent lying on the WPF shelf as expected by
analogy with 2, the piperidine ring of 4 adopts a trans di-axial
conformation, positioning the SO -containing 3’-substituent
2
Figure 1. A) 2 views of the X-ray structure of 3 bound to ATAD2 (PDB
in a solvent-exposed position far from the WPF shelf (Fig-
ure 1c). This is in sharp contrast to the trans di-equatorial
conformation seen in ATAD2 for sulfone-containing 3’-
substituted compounds like 3 (Figure 1a). In this axial
conformation, the piperidine nitrogen of 4 makes a salt-
bridge interaction with the sidechain of BRD4 Asp144.
However, as this was not seen with 1 or 2 this cannot be the
main driver for the adoption of the di-axial conformation in
BRD4 BD1. Rather, this conformation allows the polar
5
1
a83). B) Superimposed X-ray structures of BRD4 BD1 bound to
(orange, PDB 5a5s) and 2 (cyan, PDB 5a85). C) Superimposed X-ray
structures of BRD4 BD1 bound to 1 (orange, PDB 5a5s) and 4 (green,
PDB 5lj2) showing the di-axial conformation of the piperidine ring.
D) ATAD2 crystallographic binding modes of 16 (cyan, PDB 5lj0) and 3
(
5
green, PDB 5a83). E) Binding modes in BRD4 BD1 of 16 (cyan, PDB
lj1) and 4 (green, PDB 5lj2). For refinement statistics see Table S3;
for density maps Figure S6, Supporting Information.
2
www.angewandte.org
ꢀ 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Angew. Chem. Int. Ed. 2016, 55, 1 – 6
These are not the final page numbers!

Angewandte
Communications
Chemie
sulfone group to evade the hydrophobic WPF shelf environ-
ment.
Table 2: Comparison of di- and tri-substituted inhibitors. For statistics
see Table S1a, Supporting Information.
Interestingly, gas-phase ab initio calculations on model
systems favor the di-axial conformation, which is stabilised by
a 1,3 interaction between the alkoxy oxygen and the
protonated piperidine nitrogen (Figure S1c, Supporting Infor-
mation). In a continuum water model the strong electrostatic
contribution was diminished and di-equatorial was slightly
preferred. NMR experiments on 4 in water indicated an
equilibrium, which was rapid on the NMR timescale between
these two piperidine chair conformers, in an approximately
6
15
8
16
17
ATAD2 TR-FRET pIC50
6.9
7.5
4.8
2.1
1.6
126
<3
6.9
8.4
3.8
3.1
1.7
135
<3
7.4
7.2
5.2
2.2
3.0
92
7.3
8.0
5.5
ATAD2 Bromosphere pIC50
BRD4 BD1 TR-FRET pIC50
TR-FRET selectivity (logs)
Chrom logD (pH 7.4)
ꢀ4.5
ꢁ2.8
3.2
101
190
ꢀ4.4
ꢁ1.1
3.1
101
156
2
Polar surface area (ꢀ )
Artificial membrane permeability
86
À1
(
nms , pH 7.4)
5
:2 ratio in favour of the trans di-equatorial (see Supporting
Information). Hence, both molecular modelling and exper-
imental data support the trans di-axial conformer as a low
energy form for this structure.
retained the ATAD2 activity of des-methoxy 6, but its BRD4
BD1 activity was significantly weaker, resulting in an
increased selectivity window of > 3 logs. We believe that
this is due to the destabilization of the axial conformation
favored for BRD4 BD1 binding relative to the equatorial
favored by ATAD2. This conclusion was supported by NMR
data for 15 in water which indicated a strong preference for
the tri-equatorial conformation, with no evidence for signifi-
cant presence of tri-axial (see Supporting Information).
Encouraged by this, we also made 16, the equatorially-
biased C5’-OMe analogue of the difluorocyclohexyl 8. By TR-
FRET, this retained the enhanced selectivity of 15, being
equipotent to 8 against ATAD2 with reduced BRD4 BD1
activity (Table 2). As before, replacing the sulfone by
difluoromethyl resulted in improved permeability. Com-
pound 17 (the opposite enantiomer of 16) is significantly
weaker against ATAD2, so represents a useful negative
control for cellular assays.
These observations suggested that selectivity for ATAD2
over BRD4 BD1 might be increased by stabilization of the
equatorial piperidine conformation over axial. Introduction
of a substituent cis to the ether at the C5’ position would
create a 1,3 steric repulsion in the tri-axial conformation,
favouring the tri-equatorial. We tested this idea by introduc-
ing a methoxy substituent in the C5’ position via the route
shown in Scheme 1.
Crystal structures were obtained of 16 bound to both
ATAD2 and to BRD4 BD1. As expected, in ATAD2 the
binding mode of 16 is very similar to that of 3, with the entire
molecules closely superimposable (Figure 1d). The piperidine
binds in a tri-equatorial conformation with the C5’ methoxy
group making no apparent interaction with the protein. The
two fluorine atoms are close to the guanidinium terminal
nitrogen of Arg1077 (3.1 ꢁ, 3.3 ꢁ) suggesting that while
probably not hydrogen-bonding the group shows good
electrostatic complementarity to the ATAD2 RVF shelf. In
BRD4 BD1, 16 showed a very similar binding mode to 4
(Figure 1e). Surprisingly, even in the presence of the C5’-
OMe group, the piperidine ring of 16 adopts a tri-axial
conformation. 15 binds to BRD4 BD1 in a similar tri-axial
conformation (data not shown). These results illustrate that
ligands may adopt higher-energy conformers when their
binding site requires.
Scheme 1. Conditions: a) m-CPBA, CH Cl , 08C; b) NaH, MeI, DMF,
2
2
0
8C, 83% (2 steps); c) LiClO , NaN , CH CN, 808C, 35%; d) tBuOK,
4 3 3
THF, 08C then RCH OTf, 70–72%; e) H , Pd/C, MeOH, room temper-
2
2
ature, 90–91%; f) 14, tBuONa, BrettPhos, Pd(OAc) or Pd(dba) , THF,
2
2
6
08C, 37–43%; g) NBS, CH Cl , À108C, 94–97%; h) ArB(OH) , K CO ,
2
2
2
2
3
Pd(OAc) , cataCXium A, dioxane/H O, microwave, 1008C, 40–49%; i)
2
2
TFA, reflux, >93%.
Epoxide 11 was easily obtained as a single diastereoiso-
mer from alcohol 10 and could be opened with sodium azide
Table 3 shows further characterization of the potency and
selectivity of 16. By isothermal titration calorimetry a pK of
d
in the presence of LiClO to give a 1:1 mixture of regioisom-
ers, which were separable. Following alkylation and reduc-
8.1 (K 8 nm) was estimated for ATAD2, with no interaction
4
d
[17]
detected with the tandem BRD4 bromodomains (Figure S2,
Supporting Information). Whether measured by ITC or TR-
FRET, the potency against the BET bromodomains was close
to the lower limit, giving a selectivity window of over 2.8 logs.
16 has similar potency in the ATAD2 Bromosphere assay,
confirming its activity against endogenous full-length
ATAD2. The potency against ATAD2 and the closely related
ATAD2B bromodomain were comparable by TR-FRET. The
tion of 12, amines 13a,b could be coupled with naphthyridone
[5]
1
4 in moderate yields. Regioselective bromination, Suzuki
coupling and deprotection in acidic conditions provided the
inhibitors 15–17. Chiral purification could be performed at
the final stage or following the Buchwald coupling.
Data for conformationally biased analogues are shown in
Table 2. The C5’-OMe C3’-sulfone-containing piperidine 15
Angew. Chem. Int. Ed. 2016, 55, 1 – 6
ꢀ 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.angewandte.org
3
These are not the final page numbers!

Angewandte
Communications
Chemie
Table 3: Profile of 16 (GSK8814) and its enantiomer 17 (GSK8815). For
statistics see Table S1b, Supporting Information.
compound concentrations (Figure S5, Supporting Informa-
tion). Therefore, we cannot be sure that the effects were
caused by ATAD2 inhibition, although they were not seen to
such an extent with the negative control 17. Considering the
in vitro potency and permeability of 16, these results suggest
that the opportunity to regulate proliferation in a therapeutic
context via the ATAD2 bromodomain may be limited. There
is presently little data associating the anti-proliferative effects
seen upon ATAD2 knockdown with its bromodomain,
1
6
17
ATAD2 pKd ITC
8.1
8.9
7.3
7.7
5.5
n.d.
5.5
5.5
[
a]
ATAD2 BROMOScan pKi
ATAD2 TR-FRET pIC50
ATAD2B TR-FRET pIC50
BRD4 tandem pKd ITC
<5.3
<5.3
BRD2 BD1/ BD2 TR-FRET pIC50
ꢀ4.4/<4.3
<4.3/<4.3
ꢀ4.5/<3.3
ꢀ4.3/<4.3
>439
ꢀ4.9/ꢀ4.6
ꢀ4.6/ꢀ4.7
ꢀ4.4/ꢀ4.7
ꢀ4.5/ꢀ4.4
326
BRD3 BD1/ BD2 TR-FRET pIC50
BRD4 BD1/ BD2 TR-FRET pIC50
although an intact bromodomain is required for binding
[18,19]
acetylated histones.
Another group recently concluded
BRDT BD1/ BD2 TR-FRET pIC50
CLND solubility [mm]
that the ATPases of the SMARCA2/4 proteins provide better
[
20]
oncology opportunities than their bromodomains.
The
[a] n.d.=not determined.
RNA knockdown data may be consistent with reports that
other ATAD2 domains are necessary for its function: the N-
terminal region is required for oligomerization and binding of
[
1,21]
enantiomeric compound 17 has a similar profile, but is
consistently around 2 logs weaker against ATAD2.
acetylated histone H4 and chromatin,
E2-stimulated gene expression.
and the ATPase for
[18]
Perhaps the oligomeric
The selectivity of 7 and 16 against the wider bromodomain
family was assessed in the BROMOscan panel (Table S2,
Figure S3, Supporting Information). 16 is one of the most
selective bromodomain inhibitors so far reported in this
panel, with ATAD2 pKd of 8.9, displaying ꢁ 3.0 logs
selectivity over 30/33 bromodomains tested, the exceptions
being ATAD2B, TAF1 BD2 (2.2 logs) and TAF1L BD2 (2.5
logs). As a further assessment of possible off-target activity,
state of ATAD2 results in chromatin binding with such avidity
that far more potent bromodomain inhibitors would be
needed. However, it is possible that the bromodomain of
ATAD2 exerts a more subtle influence and that systems other
than those reported here may be more sensitive to inhibitors.
For instance, it has recently been proposed that ATAD2 has
a more critical function in differentiating embryonic stem
cells. Hence, we hope that reporting this probe, GSK8814
(16) and its less active control GSK8815 (17), to the scientific
community will enable others to investigate the biology of
ATAD2 further. We recommend not exceeding low-micro-
molar compound concentrations in cells to avoid any possible
unknown off-target effects.
[
22]
1
6 was inactive against an internal GSK panel of 40 targets
considered as potential liabilities (data not shown).
With compound 16 showing improved passive permeabil-
ity over the sulfone analogue 7, we sought additional evidence
for increased cellular permeability. For this, we used a Nano-
BRET assay measuring displacement of labelled ATAD2
bromodomain from histone H3.3 (Promega). As expected,
treatment with 16 but not 7 or the negative control
enantiomer 17 resulted in dose-dependent displacement
To conclude, we report the optimization of ATAD2
inhibitors for improved cell permeability and selectivity
over the BET bromodomains. This was accomplished using
what we believe to be a novel use of CF2 as a polar
(
Figure 2; Figure S4, Supporting Information). Interestingly,
hydrophobic isostere of SO . This modification retained the
2
1
6 was ineffective at displacing full-length ATAD2 from H3.3.
We attempted to reproduce some of the reported
favourable ATAD2 interactions of the sulfone and selectivity
over the BETs, yet gave a dramatic improvement in logD and
passive permeability. In parallel, an unexpected difference in
the bound piperidine conformation in ATAD2 and BRD4
BD1 was exploited to further improve selectivity. The highly
unusual tri-axial conformation adopted by 16 in BRD4 BD1
highlights the difference between the conformational prefer-
ences of substituted piperidines and cyclohexanes. This effort
culminated in the identification of the first reported nano-
molar, selective and cell permeable ATAD2 bromodomain
inhibitor.
antiproliferative effects of ATAD2 knockdown using 16.
This is the first time that this has been evaluated using a small
molecule inhibitor targeting the bromodomain. Disappoint-
ingly, effects on colony formation and inhibition of genes
involved in cell cycle and division were only seen at high
Acknowledgements
We thank Bhumika Karamshi and Laurie Gordon for TR-
FRET data, Emma Jones for protein production, Abigail
Lucas and Fiona Shilliday for crystallization support, and
Jacqui Mꢂndez and Danette Daniels of Promega Corporation
for facilitating NanoBRET assays. A.J.B., K.H.C., S.R. and
T.K. acknowledge grants from Wellcome Trust (092096),
Cancer Research UK (C6946/A14492, RG17001), and
BBSRC (RG69031).
Figure 2. Unlike the impermeable sulfone analog 7 (grey circles), 16
(
black diamonds) displaces the ATAD2 bromodomain construct in
a cellular nanoBRET displacement assay. 16 does not displace full-
length ATAD2 (crosses). (Error bars=SD).
4
www.angewandte.org
ꢀ 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Angew. Chem. Int. Ed. 2016, 55, 1 – 6
These are not the final page numbers!

Angewandte
Communications
Chemie
Keywords: bioisosteres · conformation analysis · epigenetics ·
fluorine · medicinal chemistry
[12] L. Brammer, E. A. Bruton, P. Sherwood, Cryst. Growth Des.
001, 1, 277.
13] J. D. Dunitz, ChemBioChem 2004, 5, 614.
14] P. Zhou, J. Zou, F. Tian, Z. Shang, J. Chem. Inf. Model. 2009, 49,
344.
2
[
[
2
[
[
15] D. OꢀHagan, Chem. Soc. Rev. 2008, 37, 308.
16] M. Loudet, F. Metras, J. Petrissans, J. Deschamps, G. Pfister, J.
Mol. Struct. 1974, 20, 357.
[
1] C. Caron, et al., Oncogene 2010, 29, 5171.
[
2] F. Boussouar, M. Jamshidikia, Y. Morozumi, S. Rousseaux, S.
Khochbin, Biochim. Biophys. Acta Gene Regul. Mech. 2013,
[
[
17] O. Tokuda, T. Aikawa, T. Ikemoto, I. Kurinoto, Tetrahedron Lett.
2010, 51, 2832.
1829, 1010.
18] J. X. Zou, A. S. Revenko, L. B. Li, A. T. Gemo, H. W. Chen,
Proc. Natl. Acad. Sci. USA 2007, 104, 18067.
19] M. Ciro, et al., Cancer Res. 2009, 69, 8491.
20] B. Vangamudi, et al., Cancer Res. 2015, 75, 3865.
21] Y. Zhan, et al., Epigenetics. Chromatin. 2015, 8, 37.
22] Y. Morozumi, et al., J. Mol. Cell Biol. 2016, DOI: 10.1093/jmcb/
mjv060.
[
3] J. X. Zou, L. Guo, A. S. Revenko, C. G. Tepper, A. T. Gemo,
H. J. Kung, H. W. Chen, Cancer Res. 2009, 69, 3339.
[
[
[
[
[
[
[
[
4] E. H. Demont, et al., J. Med. Chem. 2015, 58, 5649.
5] P. Bamborough, et al., J. Med. Chem. 2015, 58, 6151.
6] S. Mꢃller, S. Knapp, MedChemComm 2014, 5, 288.
7] E. P. Gillis, K. J. Eastman, M. D. Hill, D. J. Donnelly, N. A.
Meanwell, J. Med. Chem. 2015, 58, 8315.
[
8] J. C. Biffinger, H. W. Kim, S. G. Dimagno, ChemBioChem 2004,
5, 622.
[
9] K. Muller, C. Faeh, F. Diederich, Science 2007, 317, 1881.
[
[
10] B. E. Smart, J. Fluorine Chem. 2001, 109, 3.
11] J. A. Howard, V. J. Hoy, D. OꢀHagan, G. T. Smith, Tetrahedron
Received: April 22, 2016
Revised: July 8, 2016
1996, 52, 12613.
Published online: && &&, &&&&
Angew. Chem. Int. Ed. 2016, 55, 1 – 6
ꢀ 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.angewandte.org
5
These are not the final page numbers!

Angewandte
Communications
Chemie
Communications
Epigenetics
ANCCA away: ATAD2 or ANCCA is
a cancer-associated bromodomain-con-
taining protein. Starting from a potent
lead, permeability and selectivity over BET
bromodomains were improved by 1)
P. Bamborough,* C. Chung,
E. H. Demont,* R. C. Furze,
A. J. Bannister, K. H. Che, H. Diallo,
C. Douault, P. Grandi, T. Kouzarides,
A.-M. Michon, D. J. Mitchell,
R. K. Prinjha, C. Rau, S. Robson,
R. J. Sheppard, R. Upton,
using CF as a sulfone bio-isostere to
2
exploit the unique properties of fluorine,
and 2) using 1,3-interactions to control
the conformation of a piperidine ring.
GSK8814 is the first low-nanomolar,
selective and cell permeable chemical
probe for ATAD2.
R. J. Watson
&&&& — &&&&
A Chemical Probe for the ATAD2
Bromodomain
6
www.angewandte.org
ꢀ 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Angew. Chem. Int. Ed. 2016, 55, 1 – 6
These are not the final page numbers!