Discovery of BMS-753426: A Potent Orally Bioavailable Antagonist of CC Chemokine Receptor 2

Article abstract of DOI:10.1021/acsmedchemlett.1c00082

To improve the metabolic stability profile of BMS-741672 (1a), we undertook a structure-activity relationship study in our trisubstituted cyclohexylamine series. This ultimately led to the identification of 2d (BMS-753426) as a potent and orally bioavaila

Full text of DOI:10.1021/acsmedchemlett.1c00082

pubs.acs.org/acsmedchemlett
Letter
Discovery of BMS-753426: A Potent Orally Bioavailable Antagonist
of CC Chemokine Receptor 2
Michael G. Yang,* Zili Xiao, Rulin Zhao, Andrew J. Tebben, Bei Wang, Robert J. Cherney,
Douglas G. Batt, Gregory D. Brown, Mary Ellen Cvijic, John V. Duncia, Michael A. Gallela,
Daniel S. Gardner, Purnima Khandelwal, Mary F. Malley, Jian Pang, Anne V. Rose, Joseph B. Santella, III,
Amy A. Sarjeant, Songmei Xu, Arvind Mathur, Sandhya Mandlekar, Ragini Vuppugalla, Qihong Zhao,
and Percy H. Carter*
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ABSTRACT: To improve the metabolic stability profile of BMS-741672 (1a), we undertook a structure−activity relationship study
in our trisubstituted cyclohexylamine series. This ultimately led to the identification of 2d (BMS-753426) as a potent and orally
bioavailable antagonist of CCR2. Compared to previous clinical candidate 1a, the tert-butyl amine 2d showed significant
improvements in pharmacokinetic properties, with lower clearance and higher oral bioavailability. Furthermore, compound 2d
exhibited improved affinity for CCR5 and good activity in models of both monocyte migration and multiple sclerosis in the hCCR2
knock-in mouse. The synthesis of 2d was facilitated by the development of a simplified approach to key intermediate (4R)-9b that
deployed a stereoselective reductive amination which may prove to be of general interest.
KEYWORDS: CC chemokine receptor 2 (CCR2), CCR2 antagonist, Monocyte chemoattractant protein-1, Multiple sclerosis
he CC chemokine receptor 2 (CCR2) is a G-protein
Tcoupled receptor that is mainly expressed on monocytes,
macrophages, and a subset of T cells.1,2 A number of ligands
interact with CCR2, including monocyte chemoattractant
protein-1 (MCP-1 or CCL2), MCP-2 (CCL8), MCP-3
(CCL7), and MCP-4 (CCL13). Of these, MCP-1 is the
primary ligand for CCR2, and CCR2 is considered to be the
exclusive receptor for MCP-1.3 CCR2 plays a nonredundant
role in directing the emigration of monocytes from the bone
marrow into circulation and a context-dependent role in their
extravasation into resting and inflamed tissue.4−6 Given its
fundamental importance in monocyte biology,7 numerous
preclinical studies have been conducted in models of
cardiovascular disease,8 cancer,9 fibrotic conditions,10 and a
wide range of other diseases.11−13
In light of these data, it is not surprising that many different
institutions have pursued the discovery of CCR2 antago-
nists.14,15 Recently, we reported our discovery of BMS-741672
(1a) as a potent, selective, and orally bioavailable antagonist of
CCR2.16 In a pharmacokinetic study of 1a in cynomolgus
monkeys, we found high levels of the N-demethylated
metabolite 2a in the blood circulation. Indeed, the total
AUC0−24h exposure of 2a was 1.4-fold higher than that of the
parent 1a (Figure 1, equiv 1).17 Unfortunately, the functional
inhibition activity of 2a was ∼20× weaker than its parent 1a
(vide infra). The propensity for this demethylation was
confirmed in a similar study with N,N-dimethyl 1b (Figure
1, equiv 2), and the percent of metabolite formation (2b
formation) was even more substantial. In this publication, we
describe our efforts to address the issue of N-demethylation,
which led to the discovery of N-tert-butyl amine as an effective
replacement for the previous dialkylamines.
With the goal of finding a potent CCR2 antagonist with
improved in vivo metabolic stability, we began by varying the
N-alkylamine group (Table 1, compounds 1a−2d). Note that
the assays used to evaluate the different potencies of these
molecules, including CCR2 binding, CCR5 binding, and
monocyte chemotaxis, have all been reported by us
Received: February 5, 2021
Accepted: May 20, 2021
Published: May 25, 2021
https://doi.org/10.1021/acsmedchemlett.1c00082
© 2021 American Chemical Society
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demonstrated good CCR2 potency in the binding and
chemotaxis assays, and the tert-butyl amine 5b showed ca.
26-fold improved affinity for CCR5. The bicyclic analogue of
5b, that is, 6-tert-butyl-pyrido[3,2-d]pyrimidine (6), retained
similar binding and functional antagonism at CCR2, with still
further improved CCR5 affinity.
We explored the basis for the increase in CCR5 potency
with the tert-butyl amine via structural modeling21,22 and
solvent mapping23 of compounds 2c and 2d (Table 1),
capitalizing on the availability of the previously reported crystal
structures of CCR2 with BMS-687681 (RCSB 5T1A)24 and
CCR5 with maraviroc (RCSB 4MBS).25 The predicted
binding mode of 2c and 2d in CCR5 orients the ligand as
previously observed in the CCR2/BMS-687681 crystal
structure, with the quinazoline moiety sandwiched between
TM1 and TM7 and the trisubstituted cyclohexyl projecting
toward TM2 and TM3. The cyclohexyl 2-amino substituents
are predicted to occupy a hydrophobic pocket formed by the
top of TM3 and bottom of EC2 (Figure 2). WaterMap analysis
of this pocket in the absence of ligand predicted the presence
of entropically unfavorable waters whose displacement into
bulk solvent would be required to achieve high potency. Two
of these waters became trapped in the presence of the amino
(Wat1 and Wat2 in Figure 2A) and aminomethyl substituents
(data not shown), increasing their entropic unfavorability. The
tert-butyl amine (Figure 2B) is predicted to fill this pocket,
thereby displacing the unfavorable waters with the resulting
free energy gain leading to increased binding affinity.
WaterMap analysis of the equivalent pocket CCR2 crystal
structure suggested the unfavorable waters would be displaced
by the amino moiety or accessible to bulk solvent (Figure 2C),
consistent with the relative insensitivity of CCR2 to amino
substitution (Table 1).
Figure 1. In vivo cyno PK studies of 1a and 1b.
previously.16,18 Compared to 1a, the demethylated 2a was 9- to
25-fold less active in inhibition of CCR2, as shown in the
binding and chemotaxis assays, respectively. We found that a
smaller R group had little effect on CCR2 potency, as the
binding IC50 values of 2b (R = Me) and 2c (R = H) were 4.6
and 2.9 nM, respectively; however, the primary amine 2c had
markedly reduced affinity for CCR5 (Table 1). In previous
studies, we had explored a range of exocyclic amines but had
not found any that were superior to N-isopropyl, N-methyl.19
It was therefore of interest when we determined that tert-butyl
amine 2d was 20 times more potent than 2a in the monocyte
chemotaxis assay and also exhibited 60-fold higher affinity for
CCR5 (CCR5 binding IC50: 2a = 4959 vs 2d = 83 nM). The
CCR5 binding for compound 2d was also assessed in primary
T-cells, wherein it exhibited a binding value of 6.3 1.5 nM. A
brief survey of replacements of the 6-trifluoromethyl-
quinazolin-4-yl moiety confirmed the SAR finding with the
tert-butyl amine 2d (Table 1, compounds 3−6). The 6-
trifluoromethoxyquinazoline analogue 3 retained potent bind-
ing to both CCR2 and CCR5. The simple 6-trifluoromethyl-
benzamide 4 exhibited weaker affinity for both CCR2 and
CCR5 in the binding assays (Table 1), consistent with
previous SAR studies.20 Notably, the picolinamides 5a and 5b
All of the compounds shown in Table 1 were also profiled
for their binding to ion channels (a well-described issue for
chemokine receptor antagonists), passive permeability, and
metabolic stability. As shown in the Supporting Information,
all of the tested compounds exhibited low Na+ channel
binding and excellent liver microsome metabolic stability. As a
a
Table 1. In Vitro Characterization of CCR2 Antagonists 1−6
b
compd
CCR2 binding IC50 (nM)
CCR5 binding IC50 (nM)
CCR5/2 ratio
chemotaxis IC50 (nM)
Pampa PC (nm/s)
1a
1b
2a
2b
2c
2d
3
1.1 0.7 (n = 18)
3.9 1.5 (n = 3)
15 7.2 (n = 6)
4.6 1.4 (n = 31)
2.9 0.8 (n = 4)
2.7 1.3 (n = 15)
7.6 3.6 (n = 9)
15
780 92 (n = 4)
8056 6212 (n = 3)
4959 2258 (n = 3)
4631 1758 (n = 9)
18745 7497 (n = 3)
83 43 (n = 9)
∼ 710
∼ 2100
∼ 330
∼ 1000
∼ 6500
∼ 31
0.7
4.5
20
443
557
208
235
30
560
507
ND
30
1.7
3.0
0.8
c
34 19 (n = 6)
∼ 4.5
ND
4
2930
∼ 190
∼ 1300
∼ 110
∼ 2.7
ND
1.6
0.3
0.6
5a
5b
6
3.0 0.3 (n = 4)
1.3 0.5 (n = 5)
6.6 1.0 (n = 6)
3867 1406 (n = 2)
147 52 (n = 3)
18 18 (n = 3)
332
251
a
b
IC50 values reported as the average of two or more determinations, with the exception of compound 4. Antagonism of chemotaxis of human
c
peripheral blood mononuclear cells was induced by 10 nM MCP-1 at 37 °C (see ref 18). Not determined.
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Figure 2. Structural models of compounds 2c (A) and 2d (B) in CCR5. Unstable waters occupying the hydrophobic pocket between TM3 and
EC2 are shown as spheres in (A). (C) WaterMap predicted waters in the hydrophobic pocket between EC2 and TM3 from the CCR2/BMS-
687681 crystal structure (RCSB 5T1A).
Table 2. Rat Pharmacokinetic Properties of 1a, 2b, 2d, 3, 5b, and 6
compd/route/dose
Cmax (nM)
AUC0−24 (μM·h)
T1/2 (h)
Cl (mL/min/kg)
43
Vss (L/kg)
11.3
F (%)
1a/iv/2 mg/kg
1a/po/10 mg/kg
2b/iv/2 mg/kg
2b/po/10 mg/kg
2d/iv/2 mg/kg
2d/po/10 mg/kg
3/iv/2 mg/kg
3/po/11.5 mg/kg
5b/iv/2 mg/kg
5b/po/10 mg/kg
6/iv/2 mg/kg
5.1
500
1632
1765
1868
720
3.79
10.1
10.2
11.9
3.10
2.56
51
88
3.6
4.5
5.8
2.7
3.7
31
25
5.4
7.1
79
36
13.5
10
100
66
75
157
20.7
6/po/13 mg/kg
478
100
result, it was difficult to triage compounds based on those data.
We were able to disqualify two of the compounds based on
their low passive permeability (2c, 5a; see Table 1), and
elected to advance compounds 2b, 2d, 3, 5b, and 6 were
further for in vivo PK profiling, as summarized in Table 2.
Compounds 2b, 2d, and 3 showed low plasma clearances (25−
36 mL/min/kg) and good oral bioavailability (79−100 F%) in
their respective PK studies. The oral exposures of 2b, 2d, and 3
(both Cmax and AUC0−24) were consistent with high intestinal
absorption and agreed well with the PAMPA permeability
(Table 1). Compared to 2b, 2d, and 3, the clearance of 5b was
high (75 mL/min/kg) after a single 2 mg/kg intravenous dose,
and the oral bioavailability was moderate (F% = 66) after a
single 10 mg/kg oral dose. Compound 6 exhibited a still higher
clearance (157 mL/min/kg) and volume of distribution (20.7
L/kg) after a single 2 mg/kg intravenous dose; although its
oral bioavailability was 100%, the oral exposure (both Cmax and
AUC0−24) was the lowest of the compounds tested (Table 2).
Although all five compounds exhibited excellent in vitro liver
microsome stability profiles, the in vivo clearance data of 5b
and 6 were high and not consistent with the in vitro
observation. On the basis of the profiles of high exposures
(both Cmax and AUC0−24) and low clearance shown in Table 2,
2b, 2d, and 3 became compounds of interest and were chosen
for further advancement.
Table 3. In Vitro and In Vivo Characterization of 2b, 2d,
and 3
2b
2d
3
a
CYP IC50 (μM)
3A4/2D6
2C9/2C19
> 40 > 40
> 40 > 40
24
> 40 > 40
> 40 > 40
19
> 40 > 40
> 40 > 40
36
hERG patch clamp: % Inh @
30 μM
Na+ patch clamp: % Inh @ 30
2.5
13
65
μM, 4 Hz
protein binding %free:
human/cyno/dog
rat/mouse
50/65/78
53/51
59/49/51
53/47
mouse PK data (IV: 2 mg/kg; P
O: 10 mg/kg)
AUC0−24h (μM·h)/F (%)
Cl (mL/min/kg)/T1/2 (h)
2.0/38
70/2.6
5.4/93
63/4.1
a
CYP = cytochrome P450.
Compounds 2b, 2d, and 3 were profiled further (Table 3).
None of the compounds exhibited meaningful inhibition of
CYP450 isoforms 2D6, 3A4, 2C9, or 2C19 at concentrations
up to 40 μM. Accordingly, drug interactions involving the
inhibition of the major drug-metabolizing CYPs are not
anticipated with these compounds at clinically relevant
concentrations. Likewise, none of the three compounds
showed meaningful inhibition in a hERG patch clamp assay
at 30 μM (2d = 19% < 2b = 24% < 3 = 36%). However,
compound 3 showed significantly higher % inhibition in the
Na+ channel patch clamp assay at 30 μM compared to
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compounds 2b and 2d (2b = 2.5% < 2d = 13% < 3 = 65%).
Relative to its comparators, the t-Bu amine 2d also showed a
favorable PK profile in mouse and monkey. After a single 10
mg/kg oral dose to mice, the oral bioavailability of 2d was
superior to that of 2b (F% = 93 vs 38%, respectively), and the
oral exposure of 2d was almost three times higher than that of
2b (AUC0−24: 5.4 vs 2.0 μM·h). In addition, examination of
results from a cyno PK study of 2d, orally dosed at 1 mg/kg,
clearly indicated that 2d had an excellent oral exposure in
terms of AUC0−24h and oral bioavailability (AUC0−24h = 2352
nM·h and F% = 95); these values are substantially better than
our previous lead compound 1a (AUC0−24h = 836 nM·h and F
% = 47, Figure 1). Importantly, the level of undesired primary
N-dealkylated metabolite (M1) was significantly diminished in
the case of 2d: the M1 (2c) to parent ratio was <0.01 after a
single 1 mg/kg oral dose of 2d in cyno monkeys (vs 1a:M1
(2a)/P ratio = 1.4, Figure 1).
Figure 4. Study the effect of 1a and 2d in EAE model of multiple
sclerosis.
demonstrate a significant difference in total inflammatory
cellular infiltrate relative to vehicle.
On the basis of its overall profile, compound 2d was
evaluated further in pharmacodynamic and efficacy models. On
account of its weaker affinity for the mouse homologue of
CCR2, we conducted these studies in hCCR2 knock-in (KI)
mice.26 The thioglycolate (TG) peritonitis model was used to
determine the activity of 2d in inhibiting monocyte/macro-
phage infiltration. Mice were administered TG and dosed
orally with 2d at 1, 25, or 100 mg/kg, twice daily. Then 48 h
post TG treatment, the peritoneal lavage was obtained for
cellular infiltrate analysis. When administered at doses of 1, 25,
and 100 mg/kg, compound 2d inhibited 28%, 74%, and 78% of
monocyte/macrophage influx, respectively (Figure 3), at
The key intermediate 7, which we have previously
described,16 was used for the preparation of all the final
compounds studied in this study. We initially investigated a
direct approach to synthesizing t-butyl amines from 7, even
though the available literature on the subject of converting a
primary aliphatic amine (H2N-R) to a tert-butylamine (t-
BuHN-R) at the time was limited. Our two initial attempts at
making the tert-butylamine adduct from 7 met with only partial
success (Scheme 1): the route through aziridine 8 proceeded
in <20% yield and required five transformations, and the path
through N-alkylated oxime 10 took seven steps and provided
(4R)-9b in <10% yield.
In light of the poor results with direct routes, we pivoted to a
counterintuitive approach in which we first destroyed the C4-
stereocenter only to reintroduce the stereochemistry via a
reductive amination step with tert-butylamine (Table 4).
Conversion of carbamate 7 to ketone 11 proceeded through
simple Boc deprotection, followed by treatment of the
resulting primary amine with 3,5-di-tert-butylcyclohexa-3,5-
diene-1,2-dione in MeOH and Dowex-50WX8-200 resin.27
The crude solid isolated from the reaction was triturated with a
mixture of hexane and ethyl acetate to provide the pure ketone
11 in 90% yield.
With the ketone 11 in hand, we were in a position to explore
the Titanium catalyst-based asymmetric reductive amination
reactions with tert-butylamine. Representative reactions with
selected reagents are summarized in Table 4. Neither reaction
of 11 with Ti(OPri)4 followed by NaBH3CN nor with TiCl4
followed by Pd/C/H2 gave any appreciable level of
diastereoselectivity (entries 1 and 2, R:S = 1:1). Using the
combination of PtO2 and H2 as the reducing reagents, the
TiCl4-catalyzed reactions provided moderate diastereoselectiv-
ity, especially when CH2Cl2 was used as the solvent (entries 3
and 4, Table 4). The diastereoselectivity was much better for
the reaction using the combination of TiCl4 and BH3·SMe2
compared to the combination of TiCl4 and BH3·THF (entry 5
vs 6). After significant efforts to optimize the reaction
conditions, we found that the combination of 0.6 equiv
TiCl4 and 0.6 equiv Ti(OPri)4 in CH2Cl2 followed by BH3·
SMe2 as the reducing reagent gave (4R)-9b in 78% yield with
8:1 (entry 8). After aqueous workup, the crude product was
treated with an equal amount of CH2Cl2 and 2% aqueous
NH4Cl. The isomer (4R)-9b was found exclusively in the
organic fraction, with the diastereomer (4S)-9b in the aqueous
fraction, presumably because of the modulation of exposed
polarity via conformational switching.16 Evaporation of the
Figure 3. Compound 2d in the hCCR2 KI mouse TG peritonitis
model.
plasma concentrations corresponding to 51%, 91%, and 98%
estimated receptor occupancy.18 On the basis of three separate
studies with multiple doses, the average EC50 for inhibition of
monocyte/macrophage infiltration was estimated as 3.9 nM.
This can be compared with the activity of compound 1a in the
same model, which exhibited an EC50 of 2.2 nM.
To assess the effect of 2d on chronic models of disease, we
used the experimental autoimmune encephalomyelitis (EAE)
model of multiple sclerosis in hCCR2 KI mice and included
the earlier clinical compound 1a as a comparator. Oral dosing
of 2d at 25 mg/kg (BID) and 1a at 55 mg/kg was initiated on
day 1. Compound 2d reduced the area under curve (AUC) of
the clinical score by 49% (p < 0.05) (Figure 4) and provided a
comparable efficacy profile to 1a at the doses studied.
Although the antagonist 2d did reduce clinical score,
histological evaluation of the spinal cord on day 22 did not
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Scheme 1a
a
Reagents and conditions. (a) TFA, CH2Cl2, rt, 1 h, 93%. (b) Step 1: Me2CHCHO, MgSO4, CH2Cl2, rt, 1 h; step 2: NCS, MeCN, rt; step 3:
NaBH4, EtOH, rt, 20% for three steps. (c) H2, Pd(OH)2, EtOAc, 15 psi, 95%. (d) Step 1: TFA, CH2Cl2, rt, 1 h, 93%; step 2: BrCH2CN, DIEA,
MeCN, 40 °C, 16 h; step 3: m-CPBA, CH2Cl2, rt, 0.5 h, 55% for 3 three steps. (e) Step 1: NH2OH·HCl, MeOH, 60 °C, 2 h; step 2: acetone,
CH2Cl2, rt, 8 h, 58% for two steps. (f) Step 1: MeMgBr, PhH, rt; step 2: CS2C, MeCN, rt, 30% for two steps.
a
Table 4. Reductive Amination Approach to (4R)-9b
entry
(b) step 1
(b) step 2
(b) solvent
(4R)-9b:(4S)-9b
yield of (4R)-9b (%)
1
2
3
4
5
6
7
8
2.5 equiv Ti(OPri)4
0.6 equiv TiCl4
0.6 equiv TiCl4
0.6 equiv TiCl4
0.7 equiv TiCl4
1.3 equiv NaBH3CN
0.2 equiv Pd/C, H2
0.2 equiv PtO2, H2
MeOH
MeOH
MeOH
CH2Cl2
CH2Cl2
CH2Cl2
CH2Cl2
CH2Cl2
1:1
1:1
2:1
4:1
1:1
6:1
1:1
8:1
30
50
0.2 equiv PtO2, H2
1.1 equiv BH3 (1 M THF)
1.1 equiv BH3·SMe2
1.1 equiv BH3·SMe2
1.1 equiv BH3·SMe2
0.7 equiv TiCl4
1.1 equiv Ti(OEt)4
0.6 equiv TiCl4 and 0.6 equiv Ti(OPri)4
70
78
a
Reagents and conditions. (a) step 1: TFA, CH2Cl2, rt, 1 h, 93% yield; step 2: 3,5-di-tert-butylcyclohexa-3,5-diene-1,2-dione, Dowex-50WX8−200
resin, MeOH, 90% yield; (b) step 1 :2 equiv t-BuNH2; step-2: rt 16 h.
organic fraction provided (4R)-9b in >98% diastereomeric
excess without any column chromatography purification
(Supporting Information). Triethylamine-promoted coupling
reaction of (4R)-9b and 4-chloro-6-(trifluoromethyl)-
quinazoline gave 2d in 88% yield with 99.7% purity after a
simple recrystallization protocol. The absolute and relative
stereochemistry of 2d was confirmed by single X-ray crystal
structure analysis (see Supporting Information).
Our efforts to improve the in vivo metabolic stability of
clinical candidate BMS-741672 (1a), a tertiary amine subject
to N-demethylation, led to the SAR study of 2−6 described
herein. The use of tert-butylamine as a hydrophobic secondary
amine facilitated the identification of potent, permeable, and
metabolically stable compounds. Pharmacokinetic studies in
monkeys confirmed that key molecule 2d (C4 substituent =
N(H)t-Bu) was metabolically stable in vivo, unlike 1a (C4
substituent = N(Me)i-Pr). To prepare some of the key
compounds tested in this study, we developed a diaster-
eoselective reductive amination protocol for the preparation of
key intermediate (4R)-9; the diastereomeric purity was
enriched further by capitalizing on the modulation of exposed
polarity via conformational switching.16 As a result, we were
able to prepare more than 100 g of (4R)-9d (de > 98%) from
common intermediate 7 without a single column chromatog-
raphy purification.
Unfortunately, as for so many of the chemokine
receptors,28−30 the pursuit of CCR2 antagonists has not led
to the registration of a novel drug, yet. Several authors,
including us,31 have proposed that dual inhibition of both
CCR2 and CCR5 could provide superior results, and studies in
both fibrosis and cancer continue toward this end.32−34 As
such, we note that an additional aspect associated with the
discovery of clinical candidate BMS-753426 (2d) was the
identification of a novel vector for enhancing CCR5 affinity in
our series of trisubstituted cyclohexane derivatives. Indeed, the
ability to improve CCR5 affinity while retaining the conforma-
tional plasticity engendered by the C2-acetamide16 and
avoiding C2-hydrophobes20 represented an important advance
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for our overall efforts. These studies will be reported in due
course.
Purnima Khandelwal − Research and Development, Bristol
Myers Squibb Company, Princeton, New Jersey 08543-4000,
United States
Mary F. Malley − Research and Development, Bristol Myers
Squibb Company, Princeton, New Jersey 08543-4000,
United States
Jian Pang − Research and Development, Bristol Myers Squibb
Company, Princeton, New Jersey 08543-4000, United States
Anne V. Rose − Research and Development, Bristol Myers
Squibb Company, Princeton, New Jersey 08543-4000,
United States
Joseph B. Santella, III − Research and Development, Bristol
Myers Squibb Company, Princeton, New Jersey 08543-4000,
United States
Amy A. Sarjeant − Research and Development, Bristol Myers
Squibb Company, Princeton, New Jersey 08543-4000,
United States
ASSOCIATED CONTENT
■
sı
* Supporting Information
The Supporting Information is available free of charge at
https://pubs.acs.org/doi/10.1021/acsmedchemlett.1c00082.
Analytic experimental HPLC methods; synthetic proce-
dures and analytical data of 1−6; synthetic references for
intermediates 7, 9, and compounds 11−16; reference for
the biological methods; SMILES representation of
compounds with key data (PDF)
Accession Codes
Accession code CCDC 2055911 for compound 2d contains
the supplementary crystallographic data for this paper. These
data can be obtained free of charge via www.ccdc.cam.ac.uk/
data_request/cif.
Songmei Xu − Research and Development, Bristol Myers
Squibb Company, Princeton, New Jersey 08543-4000,
United States
AUTHOR INFORMATION
Arvind Mathur − Research and Development, Bristol Myers
Squibb Company, Princeton, New Jersey 08543-4000,
United States
Sandhya Mandlekar − Research and Development, Bristol
Myers Squibb Company, Princeton, New Jersey 08543-4000,
United States
Ragini Vuppugalla − Research and Development, Bristol
Myers Squibb Company, Princeton, New Jersey 08543-4000,
United States
Qihong Zhao − Research and Development, Bristol Myers
Squibb Company, Princeton, New Jersey 08543-4000,
United States
■
Corresponding Authors
Michael G. Yang − Research and Development, Bristol Myers
Squibb Company, Princeton, New Jersey 08543-4000, United
States; orcid.org/0000-0003-2908-4060; Phone: 609-
252-3234; Email: michael.yang@bms.com
Percy H. Carter − Research and Development, Bristol Myers
Squibb Company, Princeton, New Jersey 08543-4000, United
States; Phone: 617-844-8249; Email: pcarter@
blueprintmedicines.com
Authors
Zili Xiao − Research and Development, Bristol Myers Squibb
Company, Princeton, New Jersey 08543-4000, United States
Rulin Zhao − Research and Development, Bristol Myers
Squibb Company, Princeton, New Jersey 08543-4000,
United States
Complete contact information is available at:
https://pubs.acs.org/10.1021/acsmedchemlett.1c00082
Notes
The authors declare no competing financial interest.
Andrew J. Tebben − Research and Development, Bristol
Myers Squibb Company, Princeton, New Jersey 08543-4000,
United States
ABBREVIATIONS
■
AUC, area under curve; CCR2, chemokine receptor 2; CYPs,
cytochromes P450; EAE, experimental autoimmune encepha-
lomyelositis; PAMPA, parallel artificial membrane permeability
assay; PK, pharmacokinetics; TNFα, tumor necrosis factor α;
MS, multiple sclerosis; MCP-1, monocyte chemoattractant
protein-1; RA, rheumatoid arthritis; SAR, structure−activity
relationships; TG, thioglycollate.
Bei Wang − Research and Development, Bristol Myers Squibb
Company, Princeton, New Jersey 08543-4000, United States
Robert J. Cherney − Research and Development, Bristol Myers
Squibb Company, Princeton, New Jersey 08543-4000,
United States; orcid.org/0000-0002-6642-2834
Douglas G. Batt − Research and Development, Bristol Myers
Squibb Company, Princeton, New Jersey 08543-4000,
United States
Gregory D. Brown − Research and Development, Bristol
Myers Squibb Company, Princeton, New Jersey 08543-4000,
United States
Mary Ellen Cvijic − Research and Development, Bristol Myers
Squibb Company, Princeton, New Jersey 08543-4000,
United States
John V. Duncia − Research and Development, Bristol Myers
Squibb Company, Princeton, New Jersey 08543-4000,
United States
Michael A. Gallela − Research and Development, Bristol
Myers Squibb Company, Princeton, New Jersey 08543-4000,
United States
Daniel S. Gardner − Research and Development, Bristol Myers
Squibb Company, Princeton, New Jersey 08543-4000,
United States
REFERENCES
■
(1) Griffith, J. W.; Sokol, C. L.; Luster, A. D. Chemokines and
Chemokine Receptors: Positioning Cells for Host Defense and
Immunity. Annu. Rev. Immunol. 2014, 32, 659−702.
(2) Luther, S. A.; Cyster, J. G. Chemokines as regulators of T cell
differentiation. Nat. Immunol. 2001, 2, 102−107.
(3) Deshmane, S. L.; Kremlev, S.; Amini, S.; Sawaya, B. E. Monocyte
Chemoattractant Protein-1 (MCP-1): An Overview. J. Interferon
Cytokine Res. 2009, 29, 313−326.
(4) Ingersoll, M. A.; Platt, A. M.; Potteaux, S.; Randolph, G. J.
Monocyte trafficking in acute and chronic inflammation. Trends
Immunol. 2011, 32 (10), 470−477.
(5) Dyer, D. P.; Medina-Ruiz, L.; Bartolini, R.; Schuette, F.; Hughes,
C. E.; Pallas, K.; Vidler, F.; Macleod, M. K. L.; Kelly, C. J.; Lee, K. M.;
Hansell, C. A. H.; Graham, G. J. Chemokine Receptor Redundancy
and Specificity Are Context Dependent. Immunity 2019, 50 (2), 378−
389.
974
https://doi.org/10.1021/acsmedchemlett.1c00082
ACS Med. Chem. Lett. 2021, 12, 969−975

ACS Medicinal Chemistry Letters
pubs.acs.org/acsmedchemlett
Letter
(6) Ginhoux, F.; Jung, S. Monocytes and macrophages: devel-
opmental pathways and tissue homeostasis. Nat. Rev. Immunol. 2014,
14 (6), 392−404.
(22) Mason, J. S.; Bortolato, A.; Congreve, M.; Marshall, F. H. New
insights from structural biology into the druggability of G protein-
coupled receptors. Trends Pharmacol. Sci. 2012, 33 (5), 249−260.
(23) Yang, Y.; Lightstone, F. C.; Wong, S. E. Approaches to
efficiently estimate solvation and explicit water energetics in ligand
binding: the use of WaterMap. Expert Opin. Drug Discovery 2013, 8
(3), 277−287.
(24) Zheng, Y.; Qin, L.; Zacarías, N. V.; de Vries, H.; Han, G. W.;
Gustavsson, M.; Dabros, M.; Zhao, C.; Cherney, R. J.; Carter, P.;
Stamos, D.; Abagyan, R.; Cherezov, V.; Stevens, R. C.; IJzerman, A.
P.; Heitman, L. H.; Tebben, A.; Kufareva, I.; Handel, T. M. Structure
of CC chemokine receptor 2 with orthosteric and allosteric
antagonists. Nature 2016, 540 (7633), 458−461.
(7) Wynn, T. A.; Chawla, A.; Pollard, J. W. Macrophage biology in
development, homeostasis, and disease. Nature 2013, 496, 445−455.
̂
(8) Franca̧ , C. N.; Izar, M. C. O.; Hortencio, M. N. S.; do Amaral, J.
B.; Ferreira, C. E. S.; Tuleta, I. D.; Fonseca, F. A. H. Monocyte
subtypes and the CCR2 chemokine receptor in cardiovascular disease.
Clin. Sci. 2017, 131 (12), 1215−1224.
(9) Mollica Poeta, V.; Massara, M.; Capucetti, A.; Bonecchi, R.
Chemokines and Chemokine Receptors: New Targets for Cancer
Immunotherapy. Front. Immunol. 2019, 10, 379.
(10) Braga, T. T.; Correa-Costa, M.; Silva, R. C.; Cruz, M. C.;
Hiyane, M. I.; da Silva, J. S.; Perez, K. R.; Cuccovia, I. M.; Camara, N.
O. S. CCR2 contributes to the recruitment of monocytes and leads to
kidney inflammation and fibrosis development. Inflammopharmacology
2018, 26 (2), 403−411.
(11) Raghu, H.; Lepus, C. M.; Wang, Q.; Wong, H. H.; Lingampalli,
N.; Oliviero, F.; Punzi, L.; Giori, N. J.; Goodman, S. B.; Chu, C. R.;
Sokolove, J. B.; Robinson, W. H. CCL2/CCR2, but not CCL5/
CCR5, mediates monocyte recruitment, inflammation and cartilage
destruction in osteoarthritis. Ann. Rheum. Dis. 2017, 76 (5), 914−922.
(12) Feria, M.; Diaz-Gonzalez, F. The CCR2 receptor as a
therapeutic target. Expert Opin. Ther. Pat. 2006, 16, 49−57.
(13) O’Connor, T.; Borsig, L.; Heikenwalder, M. CCL2-CCR2
signaling in disease pathogenesis. Endocr., Metab. Immune Disord.:
Drug Targets 2015, 15 (2), 105−18.
(14) Carter, P. H. Progress in the discovery of CC chemokine
receptor 2 antagonists, 2009−2012. Expert Opin. Ther. Pat. 2013, 23
(5), 549−568.
(15) Qin, L. H.; Wang, Z. L.; Xie, X.; Long, Y. Q. Discovery and
synthesis of 6,7,8,9-tetrahydro-5H-pyrido[4,3-c]azepin-5-one-based
novel chemotype CCR2 antagonists via scaffold hopping strategy.
Bioorg. Med. Chem. 2018, 26 (12), 3559−3572.
(16) Yang, M. G.; Xiao, Z.; Cherney, R. J.; Tebben, A. J.; Batt, D. G.;
Brown, G. D.; Chen, J.; Cvijic, M. E.; Dabros, M.; Duncia, J. V.;
Galella, M. A.; Gardner, D. S.; Khandelwal, P.; Ko, S. S.; Malley, M.
F.; Mo, R.; Pang, J.; Rose, A. V.; Santella, J. B.; Shi, H.; Srivastava, A.;
Traeger, S. C.; Wang, B.; Xu, S.; Zhao, R.; Barrish, J. C.; Mandlekar,
S.; Zhao, Q.; Carter, P. H. Use of a conformational-switching
mechanism to modulate exposed polarity: discovery of CCR2
antagonist BMS-741672. ACS Med. Chem. Lett. 2019, 10 (3), 300−
305.
(25) Tan, Q.; Zhu, Y.; Li, J.; Chen, Z.; Han, G. W.; Kufareva, I.; Li,
T.; Ma, L.; Fenalti, G.; Li, J.; Zhang, W.; Xie, X.; Yang, H.; Jiang, H.;
Cherezov, V.; Liu, H.; Stevens, R. C.; Zhao, Q.; Wu, B. Structure of
the CCR5 chemokine receptor-HIV entry inhibitor maraviroc
complex. Science 2013, 341 (6152), 1387−1390.
(26) 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.
(27) Corey, E. J.; Achiwa, K. Oxidation of primary amines to
ketones. J. Am. Chem. Soc. 1969, 91 (6), 1429−1432.
(28) Schall, T. J.; Proudfoot, A. E. I. Overcoming hurdles in
developing successful drugs targeting chemokine receptors. Nat. Rev.
Immunol. 2011, 11 (5), 355−363.
(29) Solari, R.; Pease, J. E.; Begg, M. Chemokine receptors as
therapeutic targets: why aren’t there more drugs? Eur. J. Pharmacol.
2015, 746, 363−367.
(30) Pease, J.; Horuk, R. Chemokine receptor antagonists. J. Med.
Chem. 2012, 55 (22), 9363−9392.
(31) Zhao, Q. Dual targeting of CCR2 and CCR5: therapeutic
potential for immunologic and cardiovascular diseases. J. Leukocyte
Biol. 2010, 88, 41−55.
(32) Tacke, F. Cenicriviroc for the treatment of non-alcoholic
steatohepatitis and liver fibrosis. Expert Opin. Invest. Drugs 2018, 27
(3), 301−311.
(33) Gale, J. D.; Gilbert, S.; Blumenthal, S.; Elliott, T.; Pergola, P. E.;
Goteti, K.; Scheele, W.; Perros-Huguet, C. Effect of PF-04634817, an
oral CCR2/5 chemokine receptor antagonist, on albuminuria in
adults with overt diabetic nephropathy. Kidney Int. Rep. 2018, 3 (6),
1316−1327.
(34) Le, D.; Gutierrez, M. E.; Saleh, M.; Chen, E.; Mallick, A. B.;
Pishvaian, M. J.; Krauss, J.; O’Dwyer, P.; Garrido-Laguna, I.; Zhao,
Q.; Kippel, A.; Wang, X.; Liu, Y.; Pollack, A.; Sama, A.; Lenz, H.-J. A
phase Ib/II study of BMS-813160, a CC chemokine receptor (CCR)
2/5 dual antagonist, in combination with chemotherapy or nivolumab
in patients (pts) with advanced pancreatic or colorectal cancer. In:
Proceedings of the American Association for Cancer Research Annual
Meeting 2018; 2018 Apr 14−18, Chicago, IL, Philadelphia, PA; Cancer
Research; AACR, 2018; Vol. 78 (13 Suppl), Abstract CT124.
(17) Yang, M. G.; Cherney, R. J. Modulators of chemokine receptor
activity, crystalline forms and process. U.S. patent US7,671,062B2,
2010.
(18) Carter, P. H.; Duncia, J. V.; Mudryk, B. M.; Randazzo, M. E.;
Xiao, Z.; Yang, M. G.; Zhao, R. N((1R,2S,5R)-5-(tert-butylamino)-2-
((S)-2-oxo-3-(6-(trifluoromethyl)quinazolin-4-ylamino)pyrrolidin-1-
yl)cyclohexyl)acetamide and other modulators of chemokine receptor
activity, crystalline forms, and process. U.S. patent , US7,629,351B2,
2009.
(19) Cherney, R. J.; Mo, R.; Meyer, D. T.; Voss, M. E.; Yang, M. G.;
Santella, J. B.; 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. γ-Lactams as glycinamide replacements in
cyclohexane-based CC chemokine receptor 2 (CCR2) antagonists.
Bioorg. Med. Chem. Lett. 2010, 20 (8), 2425−2430.
(20) Carter, P. H.; Brown, G. D.; Cherney, R. J.; Batt, D. G.; Chen,
J.; Clark, C. M.; Cvijic, M. E.; Duncia, J. V.; Ko, S. S.; Mandlekar, S.;
Mo, R.; Nelson, D. J.; Pang, J.; Rose, A. V.; Santella, J. B., III; Tebben,
A. J.; Traeger, S. C.; Xu, S.; Zhao, Q.; Barrish, J. C. Discovery of a
potent and orally bioavailable dual antagonist of CC chemokine
receptors 2 and 5. ACS Med. Chem. Lett. 2015, 6 (4), 439−444.
(21) Carter, P. H.; Tebben, A. J. The use of receptor homology
modeling to facilitate the design of selective chemokine receptor
antagonists. Methods Enzymol. 2009, 461, 249−279.
975
https://doi.org/10.1021/acsmedchemlett.1c00082
ACS Med. Chem. Lett. 2021, 12, 969−975