The discovery of novel cyclohexylamide CCR2 antagonists

Article abstract of DOI:10.1016/j.bmcl.2011.09.113

As a result of further SAR studies on a piperidinyl piperidine scaffold, we report the discovery of compound 44, a potent, orally bioavailable CCR2 antagonist. While having some in vitro hERG activity, this molecule was clean in an in vivo model of QT pro

Full text of DOI:10.1016/j.bmcl.2011.09.113

Bioorganic & Medicinal Chemistry Letters 21 (2011) 7496–7501
Contents lists available at SciVerse ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
The discovery of novel cyclohexylamide CCR2 antagonists
⇑
James C. Lanter , Thomas P. Markotan, Xuqing Zhang, Nalin Subasinghe, Fu-An Kang, Cuifen Hou,
Monica Singer, Evan Opas, Sandra McKenney, Carl Crysler, Dana Johnson, Christopher J. Molloy,
Zhihua Sui
Johnson & Johnson Pharmaceutical Research and Development, Welsh & McKean Roads, Spring House, PA 19002, USA
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 15 June 2011
Revised 23 September 2011
Accepted 27 September 2011
Available online 8 October 2011
As a result of further SAR studies on a piperidinyl piperidine scaffold, we report the discovery of com-
pound 44, a potent, orally bioavailable CCR2 antagonist. While having some in vitro hERG activity, this
molecule was clean in an in vivo model of QT prolongation. In addition, it showed excellent efficacy when
dosed orally in a transgenic murine model of acute inflammation.
Ó 2011 Published by Elsevier Ltd.
Keywords:
CCR2 Antagonist
MCP-1
hERG
Chemokine CC Motif Receptor 2 (CCR2) belongs to the G
protein-coupled seven-transmembrane receptor super family and
is primarily expressed on the surface of monocytes in the circula-
tory system. Interaction of the CC chemokine monocyte
chemoattractant protein-1 (MCP-1) with CCR2 leads to migration
of monocytes to sites of inflammation as part of the immune re-
sponse.1 Interruption of this protein–protein interaction has been
pursued as a means of addressing the inflammation component
of diseases such as rheumatoid arthritis,2 multiple sclerosis3 and
atherosclerosis.4 In addition to these obvious uses, literature
reports, corroborated by internal research efforts, have indicated
the potential utility of CCR2 antagonists for the treatment of obes-
ity5 and diabetes.6 Finally, there is evidence that CCR2 antagonists
could be effective in pain management.7
lations available, possibly due to its zwitterionic nature. The ter-
tiary nature of the basic amine rendered it difficult to moderate
its basicity by simple substitutions.
One frequently encountered difficulty in pursuing basic CCR2
pharmacophores is the potential for off target cardiovascular ef-
fects mediated by the human Ether-à-go-go Related Gene (hERG)
channel.11 Our previous experience with compounds in the same
series as 1 indicated that it was an issue for this scaffold as well;
indeed a key feature of the carboxylic acid moiety of this com-
pound was its mitigation of hERG activity. As a result, we typically
counter-screened all potent CCR2 binders for affinity to this chan-
nel. Further SAR studies on this scaffold determined that the basic
moiety could be moved out of the piperidine ring, yielding a
cyclohexyl amino acid (3, Table 1). In principle, this would allow
easy modification of the basicity of the nitrogen through substitu-
tion. Unfortunately, simple methyl substitution of the secondary
amine (4) caused a dramatic drop in potency, invalidating this
strategy. Esterification (8) also led to a much less potent compound
as did replacement of the carboxylic acid with either hydroxy-
methyl (7) or methyl (6). Interestingly, not only did these latter
two modifications dramatically reduce activity in contrast to the
original series, they also increased hERG binding by an order of
magnitude. As the SAR of this new lead was further examined, a
critical divergence in the SAR of the two series emerged: in the
cyclohexylamine series, the acid could be replaced by a primary
amide (9) without loss of activity or increased hERG binding. The
primary amide could not be substituted (10–12) so attention was
turned to other parts of the molecule.
With such a wide range of potential utility, it is unsurprising
that a great deal of interest for the target has been displayed. Both
large and small molecule efforts have been undertaken to attack
this target. In the large molecule arena, both anti-MCP-1 and
anti-CCR2 approaches have been pursued, with the latter demon-
strating target engagement in some clinical studies.8 Small mole-
cule research has yielded a wide range of chemotypes with
potent CCR2 antagonist activity.9 Several of these molecules have
progressed into clinical evaluation for indications including meta-
bolic disease, neuropathic pain, allergic rhinitis and multiple
sclerosis.
Among this sampling, internal compound 110 represented our
entry into the arena (Table 1). During further studies, it was
discovered that this molecule had a narrow range of soluble formu-
Investigation of the left side SAR was initiated by reducing the
indole (9) to the corresponding indoline (13), resulting in a drop
in potency (Table 2). Indole aza substitution (14–17) showed some
⇑
Corresponding author.
E-mail address: jlanter@its.jnj.com (J.C. Lanter).
0960-894X/$ - see front matter Ó 2011 Published by Elsevier Ltd.
doi:10.1016/j.bmcl.2011.09.113

J. C. Lanter et al. / Bioorg. Med. Chem. Lett. 21 (2011) 7496–7501
7497
Table 1
Divergent SAR
R2
R2
F
R1
F
F
N
N
F
N
N
HN
F
O
O
F
A
B
N
H
Compd
Struc
R1
—
R2
hCCR2 (nM)
hERG (nM)a
1
2
3
4
5
6
7
8
A
A
B
B
B
B
B
B
B
B
B
B
CO2H
CONH2
CO2H
CO2H
H
5
1500
nt
31,000
nt
nt
290
1800
50
H
Me
H
H
H
H
H
H
H
H
8100
ꢀ25,000
8600
9200
900
25
980
980
670
CH3
CH2OH
CO2Me
CONH2
CONHMe
CONMe2
CONHOH
520
2200
3300
5400
4400
41,000
9
10
11
12
a
nt = not tested.
interesting trends. For CCR2 activity, the order from most to least
potent was 4->5->7->6-azaindole while for hERG activity the order
was 6->5->7->4-; ultimately the 4-azaindole substitution (17) was
the best in terms of balancing the two factors. Replacement of the
indole with phenyl (18) caused a large drop in potency which could
partially be recovered by appropriate substitution (represented by
19). Finally removal of the aryl ring lead to a nearly inactive com-
pound (20).
Using 9 as a starting point, a number of modifications were also
made to the cyclohexyl-piperidine core (Fig. 1). Cyclization of the
primary amide onto the cyclohexane ring (21) reduced potency
about two orders of magnitude while insertion of a phosphine
oxide (22) into the cyclohexane ring abolished activity as did con-
traction of the piperidine to an azetidine (23). Ring contractions of
the cyclohexane moiety (24–26) failed to increase potency much
relative to the reference compound (18) and increased stereo-
chemical complexity (25 and 26). With the sensitivity of the core
established, attention was turned to the right chain.
Removal of the fluorines of 9 to establish a baseline for activity
produced a compound with ꢀ200 nM activity (27, Table 3). Satura-
tion of the cinnamide (28) abolished activity and shortening of the
chain (30 and 31) failed to return it. Replacement of the alkene
with cyclopropane (29, a mixture of isomers) dropped activity
10-fold and a urea replacement (32) was barely tolerated. Though
the basic compound (33) was inactive in CCR2 binding, it was
fairly active in hERG binding, a source of constant concern for
this program. In general cinnamide aryl fluorine substitution
was favored over other substituents (34 vs 37–41) and for
O
HN
NH
HN
N
R
O
H
H
O
H
H
21
5200 nM
O
P
N
H
N
N
N
H
HN
N
R
N
O
R
H
22
23
N
H
N
>25,000 nM
>25,000 nM
H
HN
O
O
H
H
N
R
9
N
N
H
N
H
N
16 nM
H
H
N
R
N
R
24
25
2300 nM
O
H
1400 nM
N
H
H
N
O
R
=
F
F
N
26
R
F
1300 nM
Figure 1. Core SAR.

7498
J. C. Lanter et al. / Bioorg. Med. Chem. Lett. 21 (2011) 7496–7501
Table 2
Left side SAR
monosubstituted fluorobenzenes, the order of activity was para > -
meta > ortho relative to the alkene. Following this trend, the disub-
stituted systems favored meta and para placement (44 and 43 vs
42). Regarding cyclohexane stereochemistry, trans substituted sys-
tems were more active than cis (44 vs 45), possibly due to its better
approximation of the lowest energy piperidine conformation. After
chiral chromatography, it was determined that one enantiomer
(absolute stereochemistry not determined) held most of the
CCR2 activity (46 vs 47). Due to its slightly lower molecular weight,
relative to 9, racemate 44 was selected for further characterization.
In both calcium mobilization and chemotaxis functional assays,
44 was a potent CCR2 antagonist with IC50 values of 6 and 4 nM,
respectively; these values are in a similar range to those of many re-
CONH2
H
N
R
F
F
N
O
F
Compd
R
hCCR2 (nM)
hERG (nM)
4200
13
950
25
N
H
ported CCR2 antagonists. Both oral (dose: 10 mpk in HPMC; Cmax
:
531 ng/mL; AUC 2419 ng⁄h/mL; T1/2: 2.4 h; Bioavailability = 50%)
and intravenous (dose: 2 mpk in HPbCD, C0: 1016 ng/mL; Vss:
3.71 L/kg; CL: 31 mL/min/kg; T1/2: 1.9 h) pharmacokinetic parame-
ters in the mouse were reasonable. Rat PK parameters were similar
to the murine data. Although there was some observed hERG activ-
ity in both binding (8000 nM) and functional (600 nM, Patch Clamp)
assays, evaluation in a standard guinea pig hemodynamic model did
not show any effect on relevant parameters (Fig. 2). While we did
not obtain blood levels for this study, the good IV PK characteristics
across two other rodent species gave us confidence that sufficient
exposure of 44 was achieved in the study.
9
3300
N
H
14
130
8300
N
H
N
15
16
2200
83
<200
970
N
N
N
H
Another challenge associated with working on small molecule
CCR2 antagonists is the lack of homology between the human and
rodent receptors;12 as a result, many compounds, including 44
showed poor potency as antagonists of murine CCR2. For this rea-
son, efficacy studies were carried out using GM animals expressing
human CCR2 in C57Bl6 mice.13 When 44 was dosed orally to
animals in an acute inflammation model (thioglycolate-induced
periotinitis), excellent efficacy was observed at an oral dose of
30 mpk (Fig. 3).
N
H
N
17
18
35
6300
3400
N
H
6000
The synthesis of 44 and related molecules was fairly convergent
shown in Scheme 1. Intermediate ketone C was prepared in three
steps from cyclohexadionemonoethylene ketal by literature
methods.14 Key aminoamide D was prepared in several steps in
analogy to previously reported chemistry.15 Generally, when done,
separation of the cis and trans isomers was carried out late in the
synthesis as was chromatographic resolution of the enantiomers.
19
20
640
11,700
6200
N
O
15,800
Table 3
Right side SAR
CONH2
HN
N
R5
Y
X
R4
R3
R1
N
H
R2
Compd
X
Y
R1
R2
R3
R4
R5
hCCR2 (nM)
hERG (nM)
27
28
29
30
31
32
33
34
CO
CO
CO
CO
CO
CO
—
CH@CH
CH2CH2
C3H4
CH2
—
NH
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
F
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
190
>25,000
1300
>25,000
>25,000
9572
>25,000
36
7350
nt
nt
17,636
5471
14,784
1769
2020
CH2
CH@CH
CO

J. C. Lanter et al. / Bioorg. Med. Chem. Lett. 21 (2011) 7496–7501
7499
Table 3 (continued)
Compd
X
Y
R1
R2
R3
R4
R5
hCCR2 (nM)
hERG (nM)
35
36
37
38
39
40
41
42
CO
CO
CO
CO
CO
CO
CO
CO
CO
CO
CO
CO
CO
CO
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
CH@CH
H
F
F
H
H
H
H
H
H
H
H
H
H
H
F
H
H
H
H
H
H
H
F
H
H
H
H
H
H
64
2076
4128
870
H
H
H
H
H
H
H
F
F
F
F
F
270
130
11,600
140
490
1890
>25,000
38
12
740
7
187
25
H
H
H
H
H
F
H
H
H
H
H
H
Cl
SO2Me
CF3
OCF3
NMe2
H
F
H
H
H
>50,000
1080
1390
>50,000
3734
1714
4906
2200
6600
20,375
3300
43
44 (trans)
45 (cis)
46 (+)-trans
47 (À)-trans
9
F
F
F
F
H
F
F
80
70
40
30
mean arterial pressure
heart rate
60
50
20
40
30
10
20
10
0
0
-10
-20
-30
-40
-50
-60
-10
-20
-30
-40
-50
-60
0.1
0.3
1
3
10 mg/kg i.v.
0.1
0.3
1
3
10 mg/kg i.v.
-70
-80
0
10
10
10
20
30
30
30
40
50
60
70
80
90
100
100
100
0
10
20
30
40
50
60
70
80
90
100
minutes
minutes
60
50
40
30
20
10
0
-10
-20
-30
-40
60
50
40
30
20
QT interval
QTc
10
0
-10
-20
-30
-40
-50
-60
-50
-60
0
0.1
0.3
1
3
10 mg/kg i.v.
80 90 100
0.1
0.3
1
3
10 mg/kg i.v.
80 90
0
10
20
30
40
50
60
70
20
40
50
60
70
minutes
minutes
60
50
40
30
20
10
0
-10
-20
-30
-40
80
70
QRS duration
PR interval
60
50
40
30
20
10
0
-10
-20
-30
-40
-50
-60
-70
-80
-50
-60
0
0.1
0.3
1
3
10 mg/kg i.v.
80 90
0.1
0.3
1
3
10 mg/kg i.v.
80 90
20
40
50
60
70
0
10
20
30
40
50
60
70
100
minutes
minutes
Figure 2. Guinea pig CV study results for 44.
It is also possible to separate the enantiomers earlier by resolution
of intermediate D.16
Structural investigation of an acyl piperidinylpiperidine CCR2
antagonist scaffold lead to the discovery that the basic portion
could be replaced with a cyclohexylamine. A key discovery in this
process was the divergence of SAR between the two series result-
ing in non-zwitterionic lead structures. Systematic modification
of this novel lead culminated in the discovery of a compound with

7500
J. C. Lanter et al. / Bioorg. Med. Chem. Lett. 21 (2011) 7496–7501
Effect of JNJ-40856894 on thioglycollate induced
peritonitis in hCCR2KI mice
3
2
**
1
0
Lymphocytes
Monocytes/macrophages
50
40
30
20
10
0
140
120
100
80
60
40
20
0
Basal
Vehicle
JNJ-40856894 po
Basal
Vehicle
JNJ-40856894 po
Figure 3. Efficacy model results for 44.
(N)
HN
O
a-c
d-h
(N)
+
O
O
HN
CONH2
O
H
N
i-k
HN
C
N
O
CO2Me
N
CONH2
H2N
Ar
N
Boc
Boc
D
Scheme 1. General synthesis of cyclohexylamide CCR2 antagonists. Reagents and conditions: aKOH/MeOH heat; bH2, 10% Pd/C; cHCl/ACN-H2O; dLHMDS; TMSCl; eBr2/THF;
i
j
fNaN3/DMF; gH2, 5% Pd/C; hNH3/MeOH; NaBH(OAc)3/dcm; HCl/MeOH; kRCO2H, PyBroP, TEA.
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in an acute CCR2-mediated inflammation model. Furthermore, de-
spite in vitro activity in hERG binding and functional assays, the
molecule showed no effects on hemodynamic parameters in a
guinea pig CV model.
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Acknowledgments
The authors thank the High Output Synthesis, ADME/PK,
Secondary Pharmacology and Lead Generation Biology Teams and
the Cardiovascular Center of Excellence for their contributions to
this work.
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