Discovery of a potent and orally bioavailable dual antagonist of CC chemokine receptors 2 and 5

Article abstract of DOI:10.1021/ml500505q

We describe the hybridization of our previously reported acyclic and cyclic CC chemokine receptor 2 (CCR2) antagonists to lead to a new series of dual antagonists of CCR2 and CCR5. Installation of a γ-lactam as the spacer group and a quinazoline as a benz

Full text of DOI:10.1021/ml500505q

Letter
pubs.acs.org/acsmedchemlett
Discovery of a Potent and Orally Bioavailable Dual Antagonist of CC
Chemokine Receptors 2 and 5
Percy H. Carter,*,† Gregory D. Brown,† Robert J. Cherney,† Douglas G. Batt,† Jing Chen,‡
Cheryl M. Clark,† Mary Ellen Cvijic,‡ John V. Duncia,† Soo S. Ko,† Sandhya Mandlekar,§ Ruowei Mo,†
David J. Nelson,† Jian Pang,⊥ Anne V. Rose,§ Joseph B. Santella, III,† Andrew J. Tebben,∥
Sarah C. Traeger,§ Songmei Xu,‡ Qihong Zhao,⊥ and Joel C. Barrish†
†Departments of Discovery Chemistry, ‡Lead Discovery & Optimization, §Preclinical Candidate Optimization, ∥Molecular Discovery
Technologies, and ⊥Disease Sciences & Biology, Research and Development, Bristol-Myers Squibb Company, Princeton, New Jersey
08543, United States
S
* Supporting Information
ABSTRACT: We describe the hybridization of our previously reported acyclic and cyclic CC
chemokine receptor 2 (CCR2) antagonists to lead to a new series of dual antagonists of CCR2 and
CCR5. Installation of a γ-lactam as the spacer group and a quinazoline as a benzamide mimetic
improved oral bioavailability markedly. These efforts led to the identification of 13d, a potent and
orally bioavailable dual antagonist suitable for use in both murine and monkey models of
inflammation.
KEYWORDS: Chemokine, GPCR, dual antagonist, inflammation
ver the past 15 years, both the academic and
Opharmaceutical communities have directed a tremendous
amount of research effort toward CC chemokine receptor 2
(CCR2) and its primary ligand (CCL2, which historically was
referred to as monocyte chemoattractant protein-1, or MCP-1).
This has primarily been motivated by the central role that these
two proteins play in the migration and activation of
inflammatory monocytes,1 which results in their implication
as key players in a number of preclinical disease models.2−4
A
Figure 1. Cyclic and acyclic CCR2 antagonists from our laboratories.
number of companies have advanced CCR2 antagonists from
different chemical series into human clinical studies,5−7 but
none of these compounds appear to have yet advanced to
pivotal Phase 3 trials.
exhibited IC50 values of 3 and 24 nM in binding and chemotaxis
assays, and was modestly orally bioavailable in mouse (F =
29%).13 Despite these similarities, there were key differences in
the structure−activity relationships in the two series, such that
compound 2 exhibited reasonable activity at mouse CCR2
(mCCR2, IC50 = 3 nM), whereas 1 did not (IC50 = 440 nM).
This trend paralleled their activity in human CCR5, where
binding IC50 values of 86 and 1000 nM were observed for 2 and
1, respectively.
We were intrigued by the possibility of hybridizing these two
series, in order to benefit from structure−activity relationship
(SAR) trends evident in each. We thus turned to our earlier
receptor mutagenesis and homology modeling experiments14
for insights into how this might be accomplished. Three key
observations guided our hypothesis (Figure 2):
Much has been written about the unspectacular results
observed to date with chemokine receptor antagonists in
autoimmune and inflammatory diseases,8−10 and we have also
commented on the nuances of the clinical results in the CCR2
field.5 In this context, we have recently become attracted to the
growing body of literature that supports a role for dual
antagonism of CCR2 and the closely related CCR5 for the
treatment of cardiovascular and autoimmune diseases.11 Herein,
we describe an important extension of our earlier work on
CCR2 antagonism, which has led to the discovery of a potent
and orally bioavailable CCR2/5-dual antagonist.
As described in recent publications from our laboratories, we
have pursued both cyclic and acyclic antagonists of CCR2. Our
early efforts led to the discovery of compounds in both series
that exhibited potent activity in binding and functional assays,
but only modest oral bioavailability (Figure 1). For example,
cyclic compound 1 showed IC50 values of 0.3 and 1.9 nM for
blockade of monocyte binding and chemotaxis, and modest oral
bioavailability in mouse (F ≈ 10%).12 Acyclic compound 2
1. Installation of nitrogen into the cyclic chemotype to give
a piperidine subseries (Figure 2, Cyclic-1) yielded an
Received: December 6, 2014
Accepted: March 4, 2015
Published: March 4, 2015
© 2015 American Chemical Society
439
DOI: 10.1021/ml500505q
ACS Med. Chem. Lett. 2015, 6, 439−444

ACS Medicinal Chemistry Letters
Letter
an N-benzyl substituent (see 3h, CCR2 binding IC50 = 1.1 nM;
chemotaxis IC50 < 0.45 nM). The analogous neopentyl and
cyclopropylmethyl groups were also effective N-substituents,
and all of the N-alkyl analogues compared favorably with the
benchmark N-benzyl compound (cf. 3h−3j and 3k, Table 1).
These three N-alkyl compounds also exhibited activity against
CCR5, with binding IC50 values of 137, 321, and 1242 nM for
compounds 3h, 3i, and 3j, respectively. Compound 3i was also
dosed in an abbreviated mouse pharmacokinetic study and
found to be orally bioavailable (4 h AUC ≈ 1500 nM·h at 10
mpk PO).
The data shown in Table 1 gave us confidence that the
alignment shown in Figure 2 was plausible. We next sought to
capitalize on the hybridization model to enhance the properties
of the cyclic series, which already had both greater potency
(e.g., compare chemotaxis activity of 1 vs that of 3a) and
scaffold rigidity. We specifically focused on replacing the
phenylsulfonylmethyl of Cyclic-2 with the propyl group of
Acyclic. From the perspective of physicochemical properties,
this switch should reduce the polar surface area in the cyclic
series and lead to improved bioavailability. From a pharmaco-
logical perspective, we knew that the propyl group was a key
determinant of both mouse CCR2 and human CCR5 binding
in the acyclic series, and hypothesized that incorporating it into
the cyclic series could endow this series with greater potency
for these two receptors.
In order to synthesize the cyclic analogues with nonsulfone
side chains, we capitalized on our recently reported synthesis of
(1R,2S,5R)-tert-butyl 2-(benzyl-oxycarbonylamino)-7-oxo-6-
azabicyclo[3.2.1]octane-6-carboxylate.18 As shown in Scheme
1,19 compound 5 could be partially reduced with DIBAL-H.
The subsequent hemiaminal was not purified, but rather
reacted with an ylide to give the cis-olefin 6. Hydrogenation and
deprotection provided the free amine, which was readily
coupled with a hippuric acid to afford 7. Deprotection of the t-
Bu carbamate with TFA provided the penultimate amine, which
could be subjected to sequential reductive amination with
acetone and formaldehyde to yield the target product 4e. Other
alkyl side chains could be installed in a similar fashion.
Alternatively, ether side chains were accessed from 8, the
Figure 2. Potentially analogous binding interactions exist between
acyclic and cyclic series.
enhancement in CCR2 binding. Such compounds were
shown to interact with Glu291 in transmembrane
domain 7 (TMD7) of CCR2, whereas the parent
cyclohexyl derivatives did not.15
2. Both the amide16 and hydroxyethyl isostere (e.g., 2)13
variants of the acyclic series appeared to strongly engage
this same glutamic acid in TMD7.
3. The stereochemical preferences in the cyclic and acyclic
series were identical. Given that the stereochemistry of
the trisubstituted series (e.g., 1) pointed to a low energy
conformer (Figure 2, Cyclic-2),12 the conformation of the
acyclic series could be inferred (Figure 2, Acyclic).
The proposed model could be tested readily in the acyclic
series: if the alignment was as illustrated in Figure 2, then the
propyl in the acyclic series could be replaced by phenethyl; or
alternatively, the N-benzyl substituent could be replaced by an
alkyl group. We thus synthesized the analogues in Table 1 using
the methods described in our earlier work.13,17 As described
previously,13 benzyl was a poor replacement for n-propyl on the
side chain (cf. 3c and 3a). Remarkably, when we extended the
side chain by one carbon to phenethyl (3d), the potency was
improved ∼100-fold, bringing it within range of the previously
described isobutyl side chain (3b). Given that this was
consistent with the model, we moved to replace the N-benzyl
substituent. Relative to our expectations based on earlier CCR2
SAR (prior to 2003), we were surprised to see that the benzyl
group could be replaced with a larger branched alkyl substituent
(see 3e−3g, Table 1). The isobutyl group was studied further
in the context of the more potent ureidobenzamide group,
which confirmed that high potency could be achieved without
a
Table 1. Binding and Functional Activity of Acyclic Molecules Designed to Test the Hypothesis in Figure 2
compd
X
Y
Z
CCR2 binding IC50 (nM)
monocyte chemotaxis IC50 (nM)
240
3a
3b
3c
3d
3e
3f
2,4-Me2Ph
2,4-Me2Ph
2,4-Me2Ph
2,4-Me2Ph
i-Pr
CH2Et
CH2i-Pr
CH2Ph
CH2CH2Ph
CH2Et
CH2Et
CH2Et
CH2Et
CH2Et
CH2Et
CH2Et
H
H
H
H
H
H
H
5
19
6400
62
15
i-Bu
337
567
1.1
0.8
4.9
4.7
3g
3h
3i
c-Hex
i-Pr
NHC(O)NHi-Pr
NHC(O)NHi-Pr
NHC(O)NHi-Pr
NHC(O)NHi-Pr
<0.45
<0.45
3.3
t-Bu
3j
c-Pr
3k
2,4-Me2Ph
9.9
a
Binding was performed with 0.3 nM [125I]MCP-1 and human peripheral blood mononuclear cells at room temperature (RT) (ref 17). IC50 values
reported as the average of two or more determinations. Antagonism of chemotaxis of human peripheral blood mononuclear cells was induced by 10
nM MCP-1 at 37 °C (ref 17). Compounds 3a, 3b, 3c, and 3k were described in our earlier paper on the acyclic series (ref 13).
440
DOI: 10.1021/ml500505q
ACS Med. Chem. Lett. 2015, 6, 439−444

ACS Medicinal Chemistry Letters
Letter
a
groups (4e, 4f), but in each case, this resulted in a decrease in
both metabolic stability and off-target selectivity (Table 2).
Replacement of one of the side chain methylenes with an
oxygen provided compounds that exhibited lower hERG and
CYP inhibition than their matched analogues but that were also
less potent at CCR2 and sometimes also less metabolically
stable (cf. 4a/4b; 4c/4d; and 4f/4g). Given the overall profile
of the compounds, we elected to focus the bulk of our follow-
up efforts on the n-propyl side chain. Our research on the
optimization of phenethyl 4f is described separately,22 as is our
work on alkyl sulfone analogues.23
Scheme 1. Synthesis of Novel Cyclic Analogues
Our attempts to improve the mouse oral bioavailability and
CCR2 potency of 4a capitalized on our previous structure−
activity relationship studies, which suggested two approaches:
replacement of the glycinamide motif with a γ-lactam20 and
replacement of the benzamide with anthranilic amide
derivatives.13 Chart 1 shows a pair-wise comparison of the
a
Reagents and conditions: (a) i. DIBAL-H, CH2Cl2, −55 → 0 °C; ii.
BrPh3PCH2i-Pr, KHMDS, THF, 0 °C. (b) i. H2, MeOH, 5% Pd/C,
Degussa; ii. HO2CCH2NH(O)C(3-CF3Ph), BOP, i-Pr2NEt, CH2Cl2/
DMF. (c) i. TFA, CH2Cl2; ii. MeOH, Me2CO, NaCNBH3, 4 h, then
aq. CH2O. (d) THF, H2O, NaBH4. (e) MeI, Ag2O, DMF. (f) PhOH,
Ph3P, DEAD, THF.
Chart 1. Pairwise Comparison of in Vitro CCR2 Binding
Activity and Dose-Normalized Mouse AUC (PO) Observed
with Glycinamides and γ-Lactams in Phenylsulfonylmethyl
and n-Propyl Subseries
product of complete reduction from bicycle 5.18 The
hydroxylmethyl group could either be methylated directly or
subjected to Mitsonobu displacement to yield intermediates 9
and 10, respectively. These could be advanced to final products
in a manner similar to the sequence 6 → 7 → 4e.
The structure−activity relationships are shown in Table 2 for
this novel series of compounds lacking either amide or sulfone
functionality in the side chain. The characterization of
phenylsulfone 4h20 in the same assay suite is provided for
comparison. The targeted n-propyl compound 4a was indeed
active at CCR2, albeit with lower potency than sulfone 4h. It
displayed relatively low turnover in rat liver microsomes and
∼1000-fold selectivity for CCR2 antagonism relative to hERG
or CYP2D6 inhibition. Furthermore, compound 4a was orally
bioavailable in the mouse and showed exposures in 4 h PK
studies21 (AUC/D = 2.4 μM·h) that were lower than those
achieved with the acyclic compound 3i (AUC/D = 3.2 μM·h)
but higher than those with the cyclic sulfone 4h (AUC/D = 1.1
μM·h).
effect of γ-lactam substitution of the glycinamide in both
phenylsulfonylmethyl and n-propyl subseries. In both instances,
the γ-lactam had only a modest effect on the CCR2 binding
It proved possible to improve the potency of 4a through
simple extension of the chain (4c) or addition of hydrophobic
a
Table 2. Binding, Functional, and Selectivity Data for Cyclic CCR2 Antagonists with Nonsulfone Side Chains
CCR2 binding IC50
(nM)
monocyte chemotaxis IC50
(nM)
Rat LM (rate,
nmol/min/mg)
hERG flux (IC50,
μM)
CYP 2D6 (IC50,
compd
R
μM)
4a
4b
4c
4d
4e
4f
CH2Et
13
25
60
2.7
23
16
1.8
76
1.3
0.01
0.01
0.17
0.27
0.08
0.26
0.29
0.16
32
20
CH2OMe
33
>80
0.36
>80
2.7
>40
0.32
>40
0.01
0.02
0.73
2.7
CH2CH2Et
CH2CH2OMe
CH2CH2i-Pr
CH2CH2Ph
CH2OPh
2.5
20
4.7
2.5
41
1.6
4g
4h
5.3
CH2SO2Ph
0.8
>80
a
See footnote to Table 1 for description of binding and chemotaxis assays. Also shown are the rate of metabolism in rat liver microsomes, inhibition
of hERG activity in a high throughput ion flux assay, and inhibition of CYP2D6 activity.
441
DOI: 10.1021/ml500505q
ACS Med. Chem. Lett. 2015, 6, 439−444

ACS Medicinal Chemistry Letters
Letter
Table 3. Structure−Activity Relationships of Compounds Bearing Alkyl Side Chains and γ-Lactam Linkers, with Different
a
Anthranilamide Derivatives
monocyte
chemotaxis IC50
(nM)
CCR2 binding
IC50 (nM)
CCR5 binding
IC50 (nM)
mouse PO PK
(AUC/D, μM·h)
hERG patch clamp
(IC50 or %Inh)
compd
R
RHS
13a
13b
13c
13d
13e
13f
CH2CH2CH3
CH2CH2CH3
CH2CH2CH3
CH2CH2CH3
CH2CH3
U (X = Et)
1.0
1.7
2.2
0.7
0.5
2.2
0.4
3.0
0.6
3.3
2.7
0.24
0.5
3.4
6.2
4.8
16
22
13
11
15
0.7
14 μM
U (X = i-Pr)
Q (Y = Cl)
72% @ 10 μM
87% @ 10 μM
1.8 μM
22
Q (Y = CF3)
Q (Y = CF3)
Q (Y = CF3)
Q (Y = CF3)
Q (Y = CF3)
2.4
3.0
CH3
10% @ 1 μM
81% @ 10 μM
13g
13h
CH(CH3)2
CH2OCH3
10
18
a
CCR2 binding and chemotaxis assays were performed as described in Table 1. CCR5 binding was assessed through antagonism of MIP-1β binding
to HT1080 cells stably expressing CCR5. For mouse PK, the dose-adjusted 4 h AUC is listed from experiments in which mice were dosed orally with
the indicated compounds (ref 21). The final column shows activity in the hERG patch clamp assay, expressed either as estimated IC50 or % inhibition
at a single test concentration.
affinity but increased the exposure achieved in mouse after oral
dosing.
The synthesis of compound 13d is provided as representative
of the series (Scheme 2).24 Partial reduction of compound 518
The high level of oral exposure observed with compounds
having the combination of n-propyl side chain and γ-lactam
spacer was observed in multiple instances, as highlighted in
Table 3.24 Urea analogues 13a and 13b were more potent than
benzamide 12 but had lower exposure in 4 h mouse
pharmacokinetic studies. The introduction of the quinazoline
as an alternative anthranilic acid mimetic13,25 afforded
compounds 13c and 13d, both of which exhibited potent
CCR2 activity with improved exposure. In the quinazoline
series, the use of other side chain alkyl groups (methyl, ethyl,
iso-propyl, methoxymethyl) did not offer a clear advantage over
the n-propyl analogue 13d.
a
Scheme 2. Synthesis of Key Compound 13d
Compound 13d was profiled further in vitro. The potent
binding to CCR2 (IC50 = 0.7 0.3 nM) was confirmed in two
functional assays: monocyte chemotaxis (IC50 = 0.24
0.16
nM) and CD11b upregulation (whole blood IC50 = 4.7 0.9
nM). The potent binding of 13d to CCR5 in HT1080 cells
(Table 3) was confirmed in human T-cells (IC50 = 2.3 1.8
nM) and whole blood CCR5-mediated CD11b upregulation
(IC50 = 4.3 4.4 nM). The dual activity at CCR2 and CCR5
was maintained against the mouse receptors, where it likewise
exhibited affinities <2 nM.
Compound 13d was studied further and found to exhibit
high selectivity against other chemokine receptors and GPCRs,
showing >1000-fold separation for everything but members of
the muscarinic family. The Ki values for M1, M2, and M4
receptors were 341, 4010, and 865 nM, respectively. As
indicated in Table 3, compound 13d was ca. 2000-fold selective
for binding CCR2 over hERG. It exhibited a similar level of
selectivity (ca. 5000-fold) relative to the Na+ channel. In
Sprague−Dawley rats, it displayed a clearance of 40 mL/min/
kg (2.5 mg/kg, IV) and an oral bioavailability of 68% (AUC =
9294 nM·h after a 25 mg/kg PO dose). In Cynomolgus
monkey, compound 13d was cleared at 25 mL/min/kg (1.0
mg/kg, IV) and had an oral bioavailability of 46% (AUC = 862
nM·h after a 1.4 mg/kg PO dose).
a
Reagents and conditions: (a) i. DIBAL-H, CH2Cl2, 0 °C; ii.
BrPh3PCH2Et, KHMDS, THF, 0 °C. (b) i. H2, MeOH, 5% Pd/C,
Degussa; ii. (S)-Cbz-methionine, BOP, i-Pr2NEt, CH2Cl2/DMF. (c) i.
Neat MeI; ii. Cs2CO3, DMF. (d) i. TFA, CH2Cl2; ii. MeOH, Me2CO,
NaCNBH3, 4 h, then aq. CH2O; iii. neat 30% HBr/AcOH. (e) 4-Cl-6-
CF3-quinazoline, Et3N, EtOH, 75 °C.
with DIBAL-H produced the hemiaminal, which was reacted
with the propyl ylide to produce olefin 14. Introduction of the
γ-lactam followed a 4-step procedure analogous to that
reported by us previously.20 Conversion of the bis-carbamate
16 to the diamine 17 was accomplished in good yield through
sequential TFA-mediated Boc removal, one-pot double
reductive amination, and HBr-mediated Cbz removal. The
primary amine was reacted with the quinazoline chloride24 in
refluxing ethanol to obtain 13d in 52% yield after
crystallization.
442
DOI: 10.1021/ml500505q
ACS Med. Chem. Lett. 2015, 6, 439−444

ACS Medicinal Chemistry Letters
Letter
recent developments. Expert Opin. Invest. Drugs 2011, 20 (6), 745−
In order to secure the stereochemistry of 13d and explore its
solution conformation, we conducted a detailed NMR study.
We were able to confirm the relative stereochemical relation-
ships using coupling constant analysis and nuclear Overhauser
effect spectroscopy (NOESY) studies (see Supporting In-
formation). Furthermore, these studies showed the axial/
equatorial disposition of cyclohexyl substituents to be
analogous to that previously hypothesized12 (Figure 2, Cyclic-
2): the γ-lacatm was oriented axially, while the propyl and
amine groups were equatorial.
756.
(3) Charo, I. F.; Ransohoff, R. M. The many roles of chemokines and
chemokine receptors in inflammation. N. Engl. J. Med. 2006, 354 (6),
610−621.
́
(4) Feria, M.; Díaz-Gonzalez, F. The CCR2 receptor as a therapeutic
target. Expert Opin. Ther. Pat. 2006, 16 (1), 49−57.
(5) Carter, P. H. Progress in the discovery of CC chemokine receptor
2 antagonists, 2009 - 2012. Expert Opin. Ther. Pat. 2013, 23 (5), 549−
568.
(6) Struthers, M.; Pasternak, A. CCR2 antagonists. Curr. Top. Med.
In summary, we have described the hybridization of our
previously reported acyclic and cyclic CCR2 antagonists. These
studies confirmed our binding model and led to the
introduction of potent CCR5 activity into our CCR2-selective
cyclic series. The oral bioavailability of the new series was
improved through the combined replacement of the glycine
spacer and benzamide capping group with a γ-lactam and
quinazoline, respectively. These studies led to the identification
of 13d, a potent and orally bioavailable dual antagonist of
CCR2 and CCR5. Subsequent manuscripts from our
laboratories will detail the characterization of the activity of
this molecule in multiple animal models of acute and chronic
inflammation.
Chem. 2010, 10 (13), 1278−1298.
(7) Xia, M.; Sui, Z. Recent developments in CCR2 antagonists.
Expert Opin. Ther. Pat. 2009, 19 (3), 295−303.
(8) Schall, T. J.; Proudfoot, A. E. I. Overcoming hurdles in
developing successful drugs targeting chemokine receptors. Nat. Rev.
Immunol. 2011, 11 (5), 355−363.
(9) Gladue, R. P.; Brown, M. F.; Zwillich, S. H. CCR1 antagonists:
what we have learned from clinical trials. Curr. Top. Med. Chem. 2010,
10 (13), 1268−1277.
(10) Horuk, R. Chemokine receptor antagonists: overcoming
developmental hurdles. Nat. Rev. Drug Discovery 2009, 8 (1), 23−33.
(11) Zhao, Q. Dual targeting of CCR2 and CCR5: therapeutic
potential for immunologic and cardiovascular diseases. J. Leukoc. Biol.
2010, 88 (1), 41−55.
(12) Cherney, R. J.; Mo, R.; Meyer, D. T.; Voss, M. E.; Lo, Y. C.;
Yang, G.; Miller, P. B.; Scherle, P. A.; Tebben, A. J.; Carter, P. H.;
Decicco, C. P. Novel sulfone-containing di- and trisubstituted
cyclohexanes as potent CC chemokine receptor 2 (CCR2)
antagonists. Bioorg. Med. Chem. Lett. 2009, 19 (13), 3418−3422.
(13) Carter, P. H.; Brown, G. D.; King, S. R.; Voss, M. E.; Tebben, A.
J.; Cherney, R. J.; Mandlekar, S.; Lo, Y. C.; Yang, G.; Miller, P. B.;
Scherle, P. A.; Zhao, Q.; Decicco, C. P. Discovery of an orally-
bioavailable CC chemokine receptor 2 antagonist derived from an
acyclic diaminoalcohol backbone. Bioorg. Med. Chem. Lett. 2012, 22
(9), 3311−3316.
(14) Carter, P. H.; Tebben, A. J. Chapter 12. The use of receptor
homology modeling to facilitate the design of selective chemokine
receptor antagonists. Methods Enzymol. 2009, 461, 249−279.
(15) Cherney, R. J.; Nelson, D. J.; Lo, Y. C.; Yang, G.; Scherle, P. A.;
Jezak, H.; Solomon, K. A.; Carter, P. H.; Decicco, C. P. Synthesis and
evaluation of cis-3,4-disubstituted piperidines as potent CC chemokine
receptor 2 (CCR2) antagonists. Bioorg. Med. Chem. Lett. 2008, 18
(18), 5063−5065.
(16) Carter, P. H.; Brown, G. D.; Friedrich, S. R.; Cherney, R. J.;
Tebben, A. J.; Lo, Y. C.; Yang, G.; Jezak, H.; Solomon, K. A.; Scherle,
P. A.; Decicco, C. P. Capped diaminopropionamide-glycine dipeptides
are inhibitors of CC chemokine receptor 2 (CCR2). Bioorg. Med.
Chem. Lett. 2007, 17 (19), 5455−5461.
(17) For the details on the synthesis and biological testing of
compounds 3e−3j, see: Carter, P. H. PCT Int. Appl WO 2005021499.
(18) Campbell, C. L.; Hassler, C.; Ko, S. S.; Voss, M. E.; Guaciaro, M.
A.; Carter, P. H.; Cherney, R. J. Enantioselective synthesis of benzyl
(1S,2R,4R)-4-(tert-butoxycarbonylamino)-2-(hydroxymethyl)-
cyclohexylcarbamate using an iodolactam-ization as the key step. J.
Org. Chem. 2009, 74 (16), 6368−6370.
ASSOCIATED CONTENT
* Supporting Information
Synthetic procedures and complete characterization data for
compound 13d; NMR study on 13d; and protocols for assays
listed in Tables 1−3. This material is available free of charge via
the Internet at http://pubs.acs.org.
■
S
AUTHOR INFORMATION
Corresponding Author
*Tel: 609-252-4144. E-mail: percy.carter@bms.com.
Author Contributions
■
The manuscript contains contributions from all authors.
Funding
This research was funded by Bristol-Myers Squibb Company.
Notes
The authors declare no competing financial interest.
ACKNOWLEDGMENTS
■
We gratefully acknowledge other team members who
synthesized analogues in related chemical series, as well as
Dr. Arvind Mathur and the Department of Chemical Synthesis
and the Biocon/BMS Research Center for their synthesis of
13d to support large scale in vivo studies that will be disclosed
separately. The support of our colleagues in Lead Evaluation,
Lead Profiling, and Bioanalytical Sciences is also much
appreciated.
(19) For more synthetic details around the preparation of
compounds 4a−4g, see: Carter, P. H.; Cherney, R. J.; Batt, D. G.;
Brown, G. D.; Duncia, J. V.; Gardner, D. S.; Yang, M. G. PCT Int.
Appl. WO 2005020899.
(20) Cherney, R. J.; Mo, R.; Meyer, D. T.; Voss, M. E.; Yang, M. G.;
Santella, J. B., 3rd; Duncia, J. V.; Lo, Y. C.; Yang, G.; Miller, P. B.;
Scherle, P. A.; Zhao, Q.; Mandlekar, S.; Cvijic, M. E.; Barrish, J. C.;
Decicco, C. P.; Carter, P. H. gamma-Lactams as glycinamide
replacements in cyclohexane-based CC chemokine receptor 2
(CCR2) antagonists. Bioorg. Med. Chem. Lett. 2010, 20 (8), 2425−
2430.
ABBREVIATIONS
■
AUC, area under the curve; CCR2, CC chemokine receptor 2;
CCL2, CC chemokine ligand 2; CCR5, CC chemokine
receptor 5; PK, pharmacokinetics
REFERENCES
■
(1) Gordon, S.; Taylor, P. R. Monocyte and macrophage
heterogeneity. Nat. Rev. Immunol. 2005, 5 (12), 953−964.
(2) Kang, Y. S.; Cha, J. Jo.; Hyun, Y. Y.; Cha, D. R. Novel C-C
chemokine receptor 2 antagonists in metabolic disease: a review of
(21) Compound was administered orally to mice at a nominal
concentration of 10 mg/kg. The area under the compound
443
DOI: 10.1021/ml500505q
ACS Med. Chem. Lett. 2015, 6, 439−444

ACS Medicinal Chemistry Letters
Letter
concentration curve from 0 to 4 h was normalized to the actual dose,
as determined by measurement of the dosing solution. AUC/D =
AUC(10/actual dose), where AUC has units of μM·h and the actual
dose has units of mg/kg.
(22) Batt, D. G.; Clark, C. M.; et al. Carboxylic acid incorporation
improves the selectivity of potent CC Chemokine Receptor 2 (CCR2)
antagonists against inhibition of potassium and sodium ion channels.
Bioorg. Med. Chem. Lett. To be submitted.
(23) Cherney, R. J.; Mo, R.; Yang, M. G.; Xiao, Z.; Zhao, Q.;
Mandlekar, S.; Cvijic, M. E.; Charo, I. F.; Barrish, J. C.; Decicco, C. P.;
Carter, P. H. Alkylsulfone-containing trisubstituted cyclohexanes as
potent and bioavailable chemokine receptor 2 (CCR2) antagonists.
Bioorg. Med. Chem. Lett. 2014, 24 (7), 1843−1845.
(24) For more synthetic details around the preparation of
compounds 12a and 13a−13h, see: Carter, P. H.; Cherney, R. J.;
Batt, D. G.; Duncia, J. V.; Gardner, D. S.; Ko, S. S.; Srivastava, A. S.;
Yang, M. G. PCT Int. Appl. WO 2005021500.
(25) Subasinghe, N. L.; Lanter, J.; Markotan, T.; Opas, E.;
McKenney, S.; Crysler, C.; Hou, C.; O’Neill, J.; Johnson, D.; Sui, Z.
A novel series of N-(azetidin-3-yl)-2-(heteroarylamino)-acetamide
CCR2 antagonists. Bioorg. Med. Chem. Lett. 2013, 23 (4), 1063−1069.
444
DOI: 10.1021/ml500505q
ACS Med. Chem. Lett. 2015, 6, 439−444