Biphenyl Acid Derivatives as APJ Receptor Agonists

Article abstract of DOI:10.1021/acs.jmedchem.9b01513

The APJ receptor and its endogenous peptidic ligand apelin have been implicated as important modulators of cardiovascular function, and APJ receptor agonists may be beneficial in the treatment of heart failure. In this article, we describe the discovery of a series of biphenyl acid derivatives as potent APJ receptor agonists. Following the identification of initial high-throughput screen lead 2, successive optimization led to the discovery of lead compound 15a. Compound 15a demonstrated comparable in vitro potency to apelin-13, the endogenous peptidic ligand for the APJ receptor. In vivo, compound 15a demonstrated a dose-dependent improvement in the cardiac output in male Sprague Dawley rats with no significant changes in either mean arterial blood pressure or heart rate, consistent with the hemodynamic profile of apelin-13 in an acute pressure volume loop model.

Full text of DOI:10.1021/acs.jmedchem.9b01513

Article
pubs.acs.org/jmc
Cite This: J. Med. Chem. XXXX, XXX, XXX−XXX
Biphenyl Acid Derivatives as APJ Receptor Agonists
Shun Su,* Adam Clarke, Ying Han,† Hannguang J. Chao, Jeffrey Bostwick, William Schumacher,
Tao Wang, Mujing Yan, Mei-Yin Hsu, Eric Simmons, Chiuwa Luk, Carrie Xu, Marta Dabros,
Michael Galella, Joelle Onorato, David Gordon, Ruth Wexler, Peter S. Gargalovic,
and R. Michael Lawrence
Research and Development, Bristol-Myers Squibb, Co., P.O. Box 5400, Princeton, New Jersey 08543-5400, United States
S
* Supporting Information
ABSTRACT: The APJ receptor and its endogenous peptidic
ligand apelin have been implicated as important modulators of
cardiovascular function, and APJ receptor agonists may be
beneficial in the treatment of heart failure. In this article, we
describe the discovery of a series of biphenyl acid derivatives as
potent APJ receptor agonists. Following the identification of
initial high-throughput screen lead 2, successive optimization
led to the discovery of lead compound 15a. Compound 15a
demonstrated comparable in vitro potency to apelin-13, the
endogenous peptidic ligand for the APJ receptor. In vivo, compound 15a demonstrated a dose-dependent improvement in the
cardiac output in male Sprague Dawley rats with no significant changes in either mean arterial blood pressure or heart rate,
consistent with the hemodynamic profile of apelin-13 in an acute pressure volume loop model.
INTRODUCTION
■
Heart failure (HF) and related complications have become an
increasing concern to the world’s population. In the US alone,
HF affects approximately six million people and leads to
increased morbidity, premature death, poor quality of life, and
significant health-care expenditure.1 HF is a clinical syndrome
characterized by inadequate performance of the heart and
inability to deliver sufficient blood and oxygen supply to meet
the body’s demands. Despite encouraging advances in HF
management,2 which focus primarily on inhibition and
Figure 1. (Pyr1)apelin-13 and HTS hit.
downregulation of the renin−angiotensin aldosterone system
and co-medications including β-blockers, vasodilators, and
diuretics, the prognosis remains poor. Patients display 10%
mortality at 28 days posthospitalization for HF, and survival
rates are approximately 50% within 5 years of diagnosis.3 As a
been reported as the most stable and most abundant form
present in human plasma and cardiac tissue.6b,c Preclinical
result, the development of novel therapeutic agents is necessary
animal models indicate that APJ receptor agonists can improve
cardiac function7 and this notion is also supported by apelin
studies in humans. Acute infusion of (Pyr1)apelin-13 into
healthy volunteers and into patients with HF resulted in
improved cardiac output (CO) and increased ejection fraction
without significant effects on blood pressure or heart rate.8
Therefore, interest in APJ-R agonism as a potential therapeutic
target for HF management has been widespread and is
evidenced by the development of both peptido-mimetic9 and
small-molecule10 APJ-R agonists. In this paper, the identi-
fication and optimization of a series of biphenyl acid (BPA)-
based APJ agonists are described.
to further improve HF therapy.
The APJ receptor (APJ-R) was identified in 1993 as a
member of the rhodopsin-like G-protein-coupled receptor
family.4 APJ-R is most closely related to the angiotensin AT1
receptor, with 54% homology in the transmembrane domain,
but lacks binding affinity for angiotensin II.5a APJ-R is widely
expressed in various tissues, including the heart, kidney, and
blood vessels. APJ-R was deorphanized following the discovery
of its endogenous peptidic ligand, apelin.5a Recently, the
Elabela peptide was also identified as an endogenous APJ-R
ligand.5b Apelin is produced as a 77 amino acid pre-pro form
and is progressively cleaved to smaller peptidic fragments.6a
Among the biologically active apelin peptides, which include
apelin-13, -17, and -36, as well as the pyroglutamate form of
apelin-13 ((Pyr1)apelin-13, Figure 1), (Pyr1)apelin-13 has
Received: October 8, 2019
© XXXX American Chemical Society
A
DOI: 10.1021/acs.jmedchem.9b01513
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a
Scheme 1. Synthetic Route to Bis-Amides
a
Reagents and conditions: (a) 2-(trimethylsilyl)ethanol, NaH, toluene, 0−50 °C, 62%; (b) Tf2O, pyridine, DCM, 0 °C, 99%; (c)
bis(pinacolato)diborane, PdCl2(dppf)-CH2Cl2, KOAc, 1,4-dioxane, 65 °C, 96%; (d) PdCl2(dppf)−CH2Cl2, Na2CO3, toluene, H2O, 65 °C, 57%;
(e) Zn, sat. NH4Cl solution, THF, rt, 97%; (f) 3-fluoro-4-methoxyaniline, HATU, DIEA, MeCN, rt, 81%; (g) TBAF, THF, rt, quant.; (h) RNH2,
HATU, DIEA, DCM, rt; (i) TFA, DCM, rt, 68−95% for two steps.
a
Table 1. SAR of Bis-Amide Examples
RESULTS AND DISCUSSION
■
In an effort to identify small-molecule APJ-R agonists, a high-
throughput screen (HTS) of 1.2 million compounds was
undertaken using a homogeneous time-resolved fluorescence
assay11 in HEK293 cells stably expressing human APJ-R.
Compounds that demonstrated ≥20% inhibition of forskolin-
stimulated cAMP (Ymax) and ≤20% efficacy against a parental
HEK293 cell line were further evaluated using confirmatory
concentration response analysis. Confirmed hits were then
screened against the AT1 receptor to rule out any compounds
with AT1 activity. This led to the identification of a number of
small-molecule agonists including bis-amide 2 (Figure 1),
which contained a biphenyl core, a privileged structure for
GPCRs,12 and exhibited full agonism13 of APJ-R with an EC50
of 29 nM.
Following the identification of compound 2, an efficient
synthetic sequence was developed and used to generate diverse
BPA bis-amides through a key orthogonally protected biphenyl
tricarboxylic ester intermediate 7 (Scheme 1). Starting with the
known bis-ester 3,14 ester exchange followed by triflate
formation produced intermediate 4. Subsequent palladium-
mediated borylation followed by Suzuki coupling generated tri-
ester 7. Each carboxylic ester on 7 could be cleaved selectively,
providing a versatile starting point for array synthesis. For the
re-synthesis of 2, zinc-mediated reductive trichloroethyl ester
cleavage of 7 and subsequent amide bond formation afforded
bis-ester 8. tetra-n-Butylammonium fluoride (TBAF)-mediated
(trimethylsilyl)ethyl ester deprotection, amide coupling, and
removal of the t-butyl group under acidic conditions generated
bis-amides 9 in excellent yields.
One notable potential liability presented by 2 was the
possible release of aniline as a result of amide hydrolysis under
metabolic activation. Efforts were focused on replacement of
the aniline and identification of analogues with improved
potency. Following the synthetic route depicted in Scheme 1, a
series of structurally diverse aliphatic amines were surveyed on
amide A of the BPA core (Table 1). From these efforts, we
found that the dihydroindene amine could be replaced with 2-
phenylethanolamine to generate 9a with comparable potency
to compound 2. Meanwhile, it was also noted that amides with
lipophilic substituents afforded better potency compared to
polar amides. Thus, the methylated 2-phenylethanolamine was
incorporated resulting in 9b with an EC50 of 0.19 nM, a 310-
fold potency improvement compared to 9a. It was determined
that APJ activity resided primarily in the R enantiomer,
a
Data represent the mean SEM, n = 2 or more determinations.
whereas the S enantiomer 9c showed reduced potency. An in
vitro biotransformation study of an early analogue related to
9b identified the methyl ether as the major site of
metabolism.15 The methoxy phenyl ethylene amine was
replaced with isosteric n-butyl phenyl ethylene amine in an
effort to address this liability, providing 9d with comparable
potency to 9b.
Following the identification of 9d, amide B of the BPA core
was extensively surveyed. Array synthesis was executed by
alternating the order of ester cleavage. However, a narrow
structure−activity relationship (SAR) was observed at this
position and all attempts to replace anilines with aliphatic
amines were unproductive.16 Subsequent efforts to replace the
amide led to the identification of benzimidazole analogue 11a
(Table 2), the synthesis of which is depicted in Scheme 2.
(Trimethylsilyl)ethyl ester deprotection of 7 followed by
amide coupling provided intermediate 10. Reductive cleavage
of the trichloroethyl ester generated the corresponding
B
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a
and incorporation of N at the 6 position of the phenyl ring led
to a significant loss in potency. In addition to benzimidazoles,
heterocycles such as phenylimidazole and benzoxazole were
also investigated. Whereas phenyl imidazole 11g retained nM
level potency, the benzoxazole analogue 11h resulted in micro-
molar potency, indicating that the benzimidazole NH may play
a pivotal role in the interaction with APJ-R.
Table 2. Benzimidazole SAR
Parallel to the exploration of the benzimidazole SAR, the
substitution pattern of the D-ring was investigated (Table 3).
a
Table 3. D-Ring SAR
compd
R
hAPJEC50 (nM)
11a
12a
12b
12c
12d
12e
12f
3-Cl
3-H
1.4 0.47
5.7 2.8
1.5 0.64
5.7 2.2
2100 970
9.0 2.6
0.20 0.15
0.51 0.32
0.18 0.013
3-Me
3-OMe
1-Cl
2-Cl
4-Cl
4-F
12g
12h
4-Me
a
Data represent the mean SEM, n = 2 or more determinations.
Replacing the 3-Cl with a proton (12a) or methoxy group
(12c) led to less potent compounds, whereas the 3-Me (12b)
analogue retained potency. A chlorine walk on the D-ring was
subsequently executed. Whereas 1-Cl (12d)- and 2-Cl (11e)-
substituted analogues showed reduced potency, the 4-Cl
analogue (12f) demonstrated improved potency by sevenfold
compared to 11a. Compound 12f was determined to exist as a
1:1 mixture of atropisomers because of the steric encumbrance
surrounding the biaryl bond. The corresponding 4-F (12g) and
4-Me analogues (12h) showed comparable potency to 12f.
The chlorine substitution was selected to be used for
subsequent studies as it was considered to offer higher
atropisomeric thermal stability compared to the 4-F ana-
logue,17 and afforded superior yield for the biaryl formation
step compared to the 4-Me analogue. It is worth noting that
the Suzuki coupling between boronic ester 5 and bromide 13
was challenging and only moderate yields were initially
obtained. Extensive screening of reaction conditions afforded
14 in satisfactory yield (Scheme 3).15
a
Data represent the mean SEM, n = 2 or more determinations.
carboxylic acid, which was subjected to amide coupling with o-
phenylenediamine. The resulting regiosomeric mixture of
amides was treated with acetic acid under elevated temperature
to effect benzimidazole formation and t-butyl ester cleavage in
one pot to generate 11. Exploration of the benzimidazole
substitution pattern indicated that various substituents at the 5
position were tolerated for potency. Both 5-Cl (11b) and 5-Me
(11c) benzimidazole analogues exhibited slightly improved
potency and the 5-OMe compound (11d) offered an
additional potency boost. In contrast, as demonstrated by
11e and 11f, substitution with a methyl group at the 7 position
The atropisomeric mixture 12f was subjected to chiral
supercritical fluid chromatography (SFC) separation and stable
a
Scheme 2. Syntheses of Benzimidazole-Substituted BPAs
a
Reagents and conditions: (a) TBAF, THF, rt, quant.; (b) (R)-1-phenylbutan-1-amine, HATU, DIEA, DCM, rt, 75%; (c) Zn, sat. NH4Cl solution,
THF, rt, 90%; (d) HATU, DIEA, DCM, rt, then AcOH, 85 °C, 68%−92%.
C
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a
the corresponding R-atropisomer. The dihedral angle of the
biphenyl system was nearly perpendicular in the crystalline
form. The 15-fold potency boost compared to 11a indicated
that conformational restriction of the biphenyl ring system
imposed a favorable bioactive conformation.18
Scheme 3. Synthesis of Tri-Ester 14
The optimized BPA 15a demonstrated agonist potency
comparable to (Pyr1)apelin-13 versus both human and rat APJ-
R (0.093 vs 0.047 nM and 0.12 vs 0.062 nM, respectively). It is
worth noting that, in contrast to the 3-Cl series (Table 2,
compounds 11a vs 11d), installation of a methoxy group on
the benzimidazole (Table 4, 15c) failed to provide further
potency improvement. It is hypothesized that the reinforced
orthogonal orientation of the biphenyl core projected the
methoxy group in a less favorable orientation. To understand
the binding site of 15a, competition studies between 15a and
a
Reagents and conditions: (a) Pd(OAc)2, xantphos, 1 M K3PO4,
toluene, 70 °C, 65−74%.
atropisomers 15a and 15b were obtained (Table 4). The
absolute configurations of 15a and 15b were confirmed by X-
a
Table 4
[
125I]-apelin-13, as well as between [3H]-15a and apelin-13,
were performed (Figure 2). The nearly quantitative displace-
ment of [125I]-apelin-13 by 15a and the displacement of [3H]-
15a by apelin-13 support the assumption that 15a occupied
the orthosteric site of APJ-R. Compared to 15c, 15a appeared
to exhibit improved solubility at pH 7.4 (0.66 vs 0.18 mg/mL,
amorphous) and was advanced to pressure−volume loop
analysis (PV-loop)19 in isoflurane-anesthetized male Sprague
Dawley (SD) rats to evaluate its acute hemodynamic effect.
When dosed via intravenous infusion at 100 μg/min/kg, 15a
induced a 10% increase in CO with minimal impact on blood
pressure (<5% transient blood pressure drop)15 and no effect
on heart rate,15 which was consistent with the hemodynamic
effect of (Pyr1)apelin-13 (1) in this model (Figure 3).
Subsequent dose response studies revealed that 15a exhibited
a dose-dependent increase in CO without a significant effect
on HR, consistent with a (Pyr1)apelin-13 response in this
model.
CONCLUSIONS
■
BPA bis-amide 2 was identified from HTS screening as an APJ-
R agonist. Initial SAR exploration of the bis-amide series led to
optimization of the upper amide, which resulted in an 80-fold
increase in in vitro potency (9d vs 2). The bis-amide series was
further advanced by replacing the lower amide with a
benzimidazole. Optimization of the benzimidazole C5 and
C7 position substitution and subsequent biphenyl core C4
substitution afforded non-interconverting atropisomer 12f.
Separation of the respective atropisomers led to the
a
Data represent the mean SEM, n = 2 or more determinations.
ray crystallography of the individual atropisomers. The S-
atropisomer 15a was found to be 200-fold more potent than
Figure 2. Radioligand binding experiments; 1 μg of the human APJ-R-expressing HEK293 cell membrane was incubated with increasing
concentration of 1 or 15a and 0.1 nM [125I]-(Pyr1)apelin-13 in a final volume of 200 μL assay buffer containing 25 mM 4-(2-hydroxyethyl)-1-
piperazineethanesulfonic acid, 10 mM MgCl2, 2 mM ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, and 0.1% bovine serum
albumin, pH = 7.2. The membrane was separated from free compounds with a 0.3% poly ethyleneimine-soaked UniFilter-96 GF/B plate after
incubation at room temperature for 1 h; inhibition binding reaction was carried out at room temperature for 3 h when 1 nM of [3H]-15a was used.
D
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Figure 3. Rat pressure-volume loop analysis of 1 and 15a. Isoflurane-anesthetized male SD rats with a Millar PV-loop catheter in LV and a Millar
pressure catheter in the abdominal aorta. After establishing a stable baseline, 1 [6 μg/kg/min (3.92 nmol/kg/min), n = 7] or 15a (10, 100, 1000
μg/kg/min (0.0157, 0.157, 1.57 μmol/kg/min), n = 8/dose) or N,N-Dimethylacetamide vehicle (100 μL/kg/min, n = 10) was infused into the
jugular vein for 15 min with continuous monitoring of hemodynamics. Hemodynamics were monitored for an additional 15 min after terminating
infusion. Separate rats were prepared to measure exposures in response to the same 15 min infusions of 15a (n = 3/dose); washout samples were
obtained after stopping the 10 and 1000 μg/kg/min infusions; plasma concentration of 15a (μM) after 15 min of infusion: 0.44 0.18 (10 μg/kg/
min); 4.4 0.28 (100 μg/kg/min); 82 4.2 (1000 μg/kg/min).
5:95 AcCN/H2O (10 mM ammonium acetate) over 3 min and then
100% of 95:5 AcCN/H2O (10 mM ammonium acetate) over 0.75
identification of S antipode, 15a, with in vitro potency
consistent with that observed with the endogenous ligand,
(Pyr1)apelin-13 (1). Compound 15a demonstrated significant
improvement in CO in male SD rats with no significant
changes in either mean arterial blood pressure or heart rate,
which was consistent with the effect of 1 in this acute rat PV-
loop model. The studies described herein demonstrate that the
BPA chemotype shows promise as a novel scaffold for the
development of potent APJ-R agonists for treatment of HF.
1
min wavelength = 220 nm (method B). H NMR and 13C NMR
spectra were recorded on either a Bruker or a JEOL Fourier transform
spectrometer operating at frequencies as follows: 1H NMR, 400 MHz
(Bruker or JEOL) or 500 MHz (JEOL); 13C NMR, 101 MHz (Bruker
or JEOL) or 125 MHz (JEOL). Spectra data are reported in the
format chemical shift (multiplicity, coupling constants, and number of
hydrogens). Chemical shifts are specified in ppm referenced to
deuterated solvent peaks. All 13C NMR spectra were proton
decoupled. Most of the final compounds were confirmed for
molecular weight using accurate mass LCMS (HRMS). A Thermo
Fisher LTQ Orbitrap mass spectrometer in line with a Waters Acquity
UPLC allowed collection of molecular ion data with accuracy of <5
ppm. Optical rotations were obtained on a JASCO P1010
Polarimeter.
EXPERIMENTAL SECTION
■
General Information. All reagents and solvents used, including
anhydrous solvents, were of commercial quality. All reactions were
carried out under a static atmosphere of nitrogen and stirred
magnetically unless otherwise stated. All flash chromatographic
separations were performed using an ISCO RediSep on disposable
silica gel columns, eluting with hexanes/ethyl acetate or methylene
chloride/methanol. For diastereomeric mixtures, the desired com-
pound was obtained via SFC on a PIC solution 200 SFC system using
the following method: Chiralpak AD-H, 21 × 250 mm, 5 μm; 22%
EtOH/78% CO2; 45 mL/min, 150 bar, 40 °C. Final compounds were
purified by reverse phase HPLC on a SunFire C18 preparative column
using appropriate gradients of acetonitrile/water/0.1% trifluoroacetic
acid (TFA) or methanol/water/0.1% TFA as the eluent. Reactions
were monitored either by analytical Liquid chromatography−mass
spectrometry (LCMS) or by thin-layer chromatography using 0.25
mm E. Merck silica gel plates (60 F254) and were visualized with UV
light or by staining with various staining agents. LCMS data were
recorded on a Shimadzu LC-10AT equipped with an SIL-10A
injector, an SPD-10AV detector, running DISCOVERY VP software,
and coupled with a Waters ZQ mass spectrometer running MassLynx,
version 3.5, software using the following method: Phenomenex Luna
C-18 2.0 mm × 30 mm column eluted with a 2 min linear gradient
from 0 to 100% of B and then 1 min at 100% of B wherein A = 10%
methanol/90% water/0.1% TFA and B = 90% methanol/10% water/
0.1% TFA. The purity of all final compounds tested was determined
to be ≥95% using the following orthogonal HPLC conditions:
column-1: SunFire C18 3.5 μm, 3.0 × 150 mm; column-2: Xbridge
Phenyl 3.5 μm, 3.0 × 150 mm; flow rate 1.0 mL/min, gradient 10−
100% 95:5 AcCN/H2O (0.05% TFA) in 5:95 AcCN/H2O (0.05%
TFA) over 12 min and then 100% of 95:5 AcCN/H2O (0.05% TFA)
over 3 min; wavelength 1 = 220 nm, wavelength 2 = 254 nm (method
A); Column: Waters Acquity BEH C18 1.7 μm 2.1 × 50 mm; flow
rate 1.1 mL/min, gradient 0−100% 95:5 AcCN/H2O (0.1% TFA) in
5:95 AcCN/H2O (0.1% TFA) over 3 min and then 100% of 95:5
AcCN/H2O (0.1% TFA) over 0.75 min; and flow rate 1.1 mL/min,
gradient 0−100% 95:5 AcCN/H2O (10 mM ammonium acetate) in
5′-Chloro-4-((2,3-dihydro-1H-inden-5-yl)carbamoyl)-2′-((3-fluo-
ro-4-methoxyphenyl)-carbamoyl)-[1,1′-biphenyl]-2-carboxylic Acid
(2). Into the reaction vessel was added 8 (178 mg, 0.297 mmol),
tetrahydrofuran (THF; 5 mL), and TBAF (0.89 mL, 1 M/L in THF,
0.89 mmol). The reaction was stirred at rt for 12 h, diluted with
EtOAc (20 mL), and washed with 1 N HCl. The aqueous phase was
extracted with additional EtOAc (10 mL × 3). The combined organic
layer was dried over Na2SO4 and concentrated to provide the
corresponding carboxylic acid. m/z obs 499.9 [M + H]+. A portion of
this crude carboxylic acid (20 mg, 0.04 mmol) was dissolved in
dichloromethane (DCM; 2 mL). 2,3-Dihydro-1H-inden-5-amine (16
mg, 0.12 mmol), N,N-diisopropylethylamine (DIEA; 0.021 mL, 0.12
mmol), and HATU (16.7 mg, 0.044 mmol) were added subsequently.
The reaction was stirred at rt for 12 h, concentrated, and treated with
DCM (1 mL) and TFA (0.10 mL). The reaction was stirred at rt for
12 h, concentrated, and subjected to prep-HPLC purification to
1
produce 2 (17.8 mg, 0.032 mmol, 80% yield). H NMR (500 MHz,
methanol-d4): δ 8.47 (d, J = 1.8 Hz, 1H), 8.06 (dd, J = 8.0, 1.9 Hz,
1H), 7.66 (d, J = 8.2 Hz, 1H), 7.57−7.51 (m, 2H), 7.41 (d, J = 7.9
Hz, 1H), 7.39−7.34 (m, 1H), 7.33−7.26 (m, 2H), 7.18 (d, J = 8.1 Hz,
1H), 7.01−6.91 (m, 2H), 3.80 (s, 3H), 2.90 (dt, J = 14.8, 7.5 Hz,
4H), 2.09 (quin, J = 7.4 Hz, 2H). Purity 99% (method A). HRMS
(ESI): calcd for C31H25ClFN2O5 [M + H]+, 559.1431; found,
559.1411.
3-tert-Butyl 1-(2-(Trimethylsilyl)ethyl) 4-(((Trifluoromethyl)-
sulfonyl) oxy)isophthalate (4). Into the reaction vessel was added
2-(trimethylsilyl)ethanol (4.30 g, 36.4 mmol) and toluene (50 mL).
The reaction was cooled to 0 °C and NaH (1.248 g, 31.2 mmol) was
added. After 5 min, the mixture was warmed to rt and stirred for an
additional 50 min. Compound 3 (2.623 g, 10.40 mmol) was then
added and the reaction was heated at 50 °C for 30 min. The reaction
was cooled to rt, washed with sat NH4Cl (containing 31.2 mmol
HCl), and extracted with EtOAc. The organic layer was dried over
E
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Na2SO4 and concentrated. Silica gel chromatography (0−20% EtOAc
in hexanes) afforded the corresponding (trimethylsilyl)ethyl ester
(2.18 g, 6.44 mmol, 62% yield) as a colorless oil. 1H NMR (500 MHz,
chloroform-d): δ 11.52 (s, 1H), 8.47 (d, J = 2.2 Hz, 1H), 8.08 (dd, J =
8.8, 2.2 Hz, 1H), 6.98 (d, J = 8.8 Hz, 1H), 4.44−4.38 (m, 2H), 1.63
(s, 9H), 1.15−1.09 (m, 2H), 0.09 (s, 9H). This intermediate (2.18 g,
6.44 mmol) was mixed with DCM (20 mL) and pyridine (2.60 mL,
32.2 mmol). After cooling to 0 °C, Tf2O (1.63 mL, 9.66 mmol) was
added. The reaction was warmed to rt, stirred at rt for 30 min, cooled
to 0 °C, and 30 mL of DCM and 50 mL of water were added. The
organic phase was collected, dried over Na2SO4, concentrated, and
subjected to silica gel chromatography purification (0−10% EtOAc in
hexanes) to produce 4 (3.00 g, 6.38 mmol, 99% yield) as a colorless
7.6 Hz, 2H), 7.24−7.19 (m, 1H), 7.18−7.04 (m, 4H), 7.02−6.97 (m,
1H), 5.10−5.02 (m, 1H), 4.94 (t, J = 6.0 Hz, 1H), 3.74 (s, 3H),
3.72−3.67 (m, 1H), 3.66−3.59 (m, 1H). Purity >99% (method A).
HRMS (ESI): calcd for C30H25ClFN2O6 [M + H]+, 563.1380; found,
563.1360.
(S)-5′-Chloro-2′-((3-fluoro-4-methoxyphenyl)carbamoyl)-4-((2-
methoxy-1-phenylethyl)-carbamoyl)-[1,1′-biphenyl]-2-carboxylic
Acid (9b). 1H NMR (500 MHz, chloroform-d): δ 8.51−8.40 (m, 2H),
7.96−7.91 (m, 1H), 7.63 (dd, J = 8.3, 1.7 Hz, 1H), 7.44 (dd, J = 8.3,
1.1 Hz, 1H), 7.40−7.28 (m, 7H), 7.17−7.08 (m, 2H), 6.89−6.84 (m,
1H), 6.77−6.70 (m, 1H), 5.42−5.36 (m, 1H), 3.84−3.75 (m, 5H),
3.38 (s, 3H). Purity >99% (method A). HRMS (ESI): calcd for
C31H27ClFN2O6 [M + H]+, 577.1536; found, 577.1526.
1
(R)-5′-Chloro-2′-((3-fluoro-4-methoxyphenyl)carbamoyl)-4-((2-
oil. H NMR (500 MHz, chloroform-d): δ 8.60 (d, J = 2.2 Hz, 1H),
8.22 (dd, J = 8.7, 2.3 Hz, 1H), 7.35 (d, J = 8.5 Hz, 1H), 4.49−4.45
(m, 2H), 1.64 (s, 9H), 1.18−1.13 (m, 2H), 0.11 (s, 9H).
methoxy-1-phenylethyl)carbamoyl)-[1,1′-biphenyl]-2-carboxylic
1
Acid (9c). H NMR (500 MHz, methanol-d4): δ 8.41 (d, J = 1.7 Hz,
1H), 8.00 (dd, J = 8.0, 1.8 Hz, 1H), 7.65 (d, J = 8.2 Hz, 1H), 7.52
(dd, J = 8.3, 2.1 Hz, 1H), 7.44−7.31 (m, 5H), 7.31−7.23 (m, 3H),
6.99−6.91 (m, 2H), 5.34 (dd, J = 8.6, 5.0 Hz, 1H), 3.80 (s, 3H), 3.76
(dd, J = 10.1, 8.6 Hz, 1H), 3.67 (dd, J = 10.2, 5.0 Hz, 1H), 3.39 (s,
3H). Purity >99% (method B). HRMS (ESI): calcd for
C31H27ClFN2O6 [M + H]+, 577.1536; found, 577.1523.
3-tert-Butyl 1-(2-(Trimethylsilyl)ethyl) 4-(4,4,5,5-Tetramethyl-
1,3,2-dioxaborolan-2-yl)isophthalate (5). Into the reaction vessel
was added 4 (2.30 g, 4.89 mmol), 4,4,4′,4′,5,5, 5′,5′-octamethyl-2,2′-
bi(1,3,2-dioxaborolane) (1.34 g, 5.28 mmol), and 1,4-dioxane (30
mL). PdCl2(dppf)-CH2Cl2 (0.20 g, 0.244 mmol) and KOAc (1.20 g,
12.2 mmol) were subsequently added and the reaction mixture was
degassed by bubbling N2 for 10 min. The reaction was stirred at 65 °C
for 3 h, cooled to rt, diluted with 1:1:0.01 EtOAc/hexane/Et3N (50
mL), filtered through SiO2 (15 g, 200 mL 1:1:0.01 EtOAc/hexane/
Et3N as eluent), and concentrated to produce 5 (2.10 g, 4.69 mmol,
96%) as a brown solid. Compound 5 was used in the next step
without further purification. m/z obs 449.4 [M + H]+. 1H NMR (500
MHz, chloroform-d): δ 8.44 (d, J = 0.8 Hz, 1H), 8.12 (dd, J = 7.7, 1.7
Hz, 1H), 7.55 (d, J = 8.0 Hz, 1H), 4.47−4.44 (m, 2H), 1.61 (s, 9H),
1.43 (s, 12H), 1.17−1.14 (m, 2H).
(R)-5′-Chloro-2′-((3-fluoro-4-methoxyphenyl)carbamoyl)-4-((1-
1
phenylbutyl)carbamoyl)-[1,1′-biphenyl]-2-carboxylic Acid (9d). H
NMR (500 MHz, DMSO-d6): δ 8.95 (d, J = 8.3 Hz, 1H), 8.32 (d, J =
1.1 Hz, 1H), 7.99−7.95 (m, 1H), 7.66 (d, J = 8.3 Hz, 1H), 7.57 (dd, J
= 8.3, 2.2 Hz, 1H), 7.45−7.37 (m, 3H), 7.34−7.25 (m, 4H), 7.24−
7.19 (m, 1H), 7.16 (br d, J = 8.5 Hz, 1H), 7.03 (t, J = 9.4 Hz, 1H),
5.03 (td, J = 8.7, 6.2 Hz, 1H), 3.76 (s, 3H), 1.92−1.83 (m, 1H),
1.76−1.67 (m, 1H), 1.44−1.35 (m, 1H), 1.33−1.24 (m, 1H), 0.90 (t,
J = 7.3 Hz, 3H). Purity 99% (method B). m/z obs 575.0 [M + H]+.
(R)-2-tert-Butyl 2′-(2,2,2-Trichloroethyl) 5′-Chloro-4-((1-phenyl-
butyl) carbamoyl)-[1,1′-biphenyl]-2,2′-dicarboxylate (10). Into the
reaction vessel was added 7 (600 mg, 0.986 mmol), THF (9.86 mL),
and TBAF (4.93 mL, 4.93 mmol). The reaction was stirred at rt for 30
min, quenched with sat NH4Cl (20 mL), and extracted with EtOAc
(25 mL × 3). The combined organic phase was dried over sodium
sulfate, filtered, and concentrated to yield the corresponding acid. Into
the reaction vessel containing the crude acid was added (R)-1-
phenylbutan-1-amine (147 mg, 0.986 mmol), DIEA (0.517 mL, 2.96
mmol), DCM (10 mL), and HATU (487 mg, 1.28 mmol). The
reaction was stirred at rt for 12 h and concentrated. Silica gel
chromatography (0−20% EtOAc in hexanes) produced 10 (475 mg,
0.743 mmol, 75% yield) as a white solid. m/z obs 637.8 [M + H]+. 1H
NMR (500 MHz, chloroform-d): δ 8.31 (d, J = 9.4 Hz, 1H), 8.12 (d, J
= 8.5 Hz, 1H), 8.05−7.97 (m, 1H), 7.52−7.48 (m, 1H), 7.38 (d, J =
5.0 Hz, 4H), 7.26−7.22 (m, 3H), 6.45 (br d, J = 6.1 Hz, 1H), 5.31 (s,
2H), 5.20 (q, J = 7.4 Hz, 1H), 4.74−4.69 (m, 2H), 1.99−1.87 (m,
2H), 1.46−1.41 (m, 2H), 1.21 (s, 9H), 0.98 (t, J = 7.3 Hz, 3H).
2′-(1H-1,3-Benzodiazol-2-yl)-5′-chloro-4-{[(1R)-1-phenylbutyl]-
carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (11a). Into the
reaction vessel was added 10 (475 mg, 0.743 mmol), THF (7.43
mL), sat NH4Cl solution (3 mL), and Zn (243 mg, 3.71 mmol). The
reaction was stirred vigorously at rt for 1 h and extracted with EtOAc
(20 mL × 3). The combined organic layers were dried over Na2SO4
and concentrated to produce crude carboxylic acid as a white solid
(339 mg, 0.667 mmol, 90% yield), which was used in the subsequent
step without further purification. m/z obs 508.1 [M + H]+. Into the
reaction vessel was added crude carboxylic acid (18 mg, 0.035 mmol),
4-methoxy-benzene-1,2-diamine (9.79 mg, 0.071 mmol), DIEA
(0.031 mL, 0.177 mmol), DCM (1 mL), and HATU (20.2 mg,
0.053 mmol). The reaction was stirred at rt for 12 h, concentrated,
and dissolved in AcOH (1 mL)/water (0.1 mL). After stirring at 85
°C for 12 h, the reaction was cooled to rt, concentrated, and subjected
to preparative HPLC purification to produce 11a (16.3 mg, 0.024
mmol, 69% yield). 1H NMR (500 MHz, methanol-d4): δ 8.30 (s, 1H),
7.93 (br d, J = 7.5 Hz, 1H), 7.85 (d, J = 8.4 Hz, 1H), 7.70 (dd, J = 8.2,
2.1 Hz, 1H), 7.58 (dd, J = 6.1, 3.2 Hz, 2H), 7.53 (d, J = 2.0 Hz, 1H),
7.46−7.40 (m, 2H), 7.39−7.35 (m, 3H), 7.34−7.29 (m, 2H), 7.26−
7.21 (m, 1H), 5.07 (dd, J = 8.8, 6.5 Hz, 1H), 1.93−1.86 (m, 1H),
2-tert-Butyl 2′-(2,2,2-Trichloroethyl) 4-(2-(Trimethylsilyl)ethyl)
5′-Chloro-[1,1′-biphenyl]-2,2′,4-tricarboxylate (7). To a reaction
vessel containing 5 (1.55 g, 3.46 mmol) was added 6 (1.59 g, 4.33
mmol), toluene (42 mL), PdCl2(dppf)-CH2Cl2 (0.127 g, 0.173
mmol), and Na2CO3 (2 M, 6.61 mL, 13.2 mmol). The reaction
mixture was degassed by bubbling with N2 for 10 min, stirred at 65 °C
for 12 h, cooled to rt, concentrated, and subjected to silica gel
chromatography purification (0−10% EtOAc in hexanes) to produce
7 (1.20 g, 1.97 mmol, 57% yield) as a white solid. m/z obs 606.7 [M +
H]+. 1H NMR (500 MHz, chloroform-d): δ 8.63 (d, J = 1.7 Hz, 1H),
8.16 (dd, J = 8.0, 1.7 Hz, 1H), 8.13 (d, J = 8.3 Hz, 1H), 7.49 (dd, J =
8.5, 2.2 Hz, 1H), 7.25 (d, J = 2.5 Hz, 2H), 4.72 (s, 2H), 4.49−4.45
(m, 2H), 1.26 (s, 9H), 1.19−1.15 (m, 2H), 0.11 (s, 9H).
2-(tert-Butyl) 4-(2-(Trimethylsilyl)ethyl) 5′-chloro-2′-((3-fluoro-4-
methoxyphenyl)-carbamoyl)-[1,1′-biphenyl]-2,4-dicarboxylate (8).
Into the reaction vessel was added 7 (250 mg, 0.411 mmol), THF (5
mL), sat NH4Cl solution (2.5 mL), and Zn (134 mg, 2.06 mmol).
The reaction was stirred vigorously at rt for 1 h and extracted with
EtOAc (10 mL × 3). The combined organic layers were dried over
Na2SO4 and concentrated. This crude product was dissolved in DCM
(10 mL) and filtered to remove insoluble impurities. After
concentration, the crude acid was used in the subsequent step
without further purification. m/z obs 477.1 [M + H]+. The crude acid
was dissolved in DCM (6 mL). 3-Fluoro-4-methoxyaniline (116 mg,
0.822 mmol), DIEA (0.215 mL, 1.23 mmol), and HATU (172 mg,
0.452 mmol) were added subsequently. After stirring at rt for 12 h,
the mixture was concentrated and subjected to silica gel
chromatography purification (0−90% EtOAc in hexanes) to produce
1
8 (178 mg, 0.297 mmol, 72% yield). m/z obs 600.1 [M + H]+. H
NMR (500 MHz, chloroform-d): δ 8.44 (s, 1H), 8.40 (d, J = 1.7 Hz,
1H), 8.08 (dd, J = 8.0, 1.9 Hz, 1H), 7.75 (d, J = 8.3 Hz, 1H), 7.49
(dd, J = 8.3, 1.9 Hz, 1H), 7.32 (d, J = 8.0 Hz, 1H), 7.17−7.12 (m,
2H), 6.79−6.75 (m, 2H), 4.48−4.43 (m, 2H), 3.81 (s, 3H), 1.46 (s,
9H), 1.18−1.13 (m, 2H), 0.09 (s, 9H).
(S)-5′-Chloro-2′-((3-fluoro-4-methoxyphenyl)carbamoyl)-4-((2-
hydroxy-1-phenylethyl)carbamoyl)-[1,1′-biphenyl]-2-carboxylic
Acid (9a). 1H NMR (500 MHz, DMSO-d6): δ 8.87 (br d, J = 8.2 Hz,
1H), 8.23 (br s, 1H), 7.85 (br d, J = 7.3 Hz, 1H), 7.63 (d, J = 8.2 Hz,
1H), 7.54 (dd, J = 8.2, 2.1 Hz, 1H), 7.44−7.36 (m, 3H), 7.31 (t, J =
F
DOI: 10.1021/acs.jmedchem.9b01513
J. Med. Chem. XXXX, XXX, XXX−XXX

Journal of Medicinal Chemistry
Article
1.85−1.79 (m, 1H), 1.48−1.39 (m, 1H), 1.38−1.31 (m, 1H), 0.96 (t,
J = 7.4 Hz, 3H). Purity 95% (method B). HRMS (ESI): calcd for
C31H27ClN3O3 [M + H]+, 524.1735; found, 524.1722.
5′-Chloro-2′-(5-chloro-1H-1,3-benzodiazol-2-yl)-4-{[(1R)-1-
phenylbutyl]carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (11b).
1H NMR (500 MHz, methanol-d4): δ 8.33 (d, J = 1.7 Hz, 1H),
7.98 (br d, J = 7.9 Hz, 1H), 7.83 (d, J = 8.4 Hz, 1H), 7.69 (dd, J = 8.3,
2.1 Hz, 1H), 7.58 (d, J = 1.5 Hz, 1H), 7.55−7.51 (m, 2H), 7.43−7.36
(m, 4H), 7.34−7.29 (m, 2H), 7.26−7.21 (m, 1H), 5.08 (dd, J = 8.9,
6.6 Hz, 1H), 1.96−1.88 (m, 1H), 1.86−1.78 (m, 1H), 1.49−1.40 (m,
1H), 1.39−1.30 (m, 1H), 0.97 (t, J = 7.4 Hz, 3H). Purity 96%
(method B). HRMS (ESI): calcd for C31H26Cl2N3O3 [M + H]+,
558.1346; found, 558.1333.
5′-Chloro-2′-(5-methyl-1H-1,3-benzodiazol-2-yl)-4-{[(1R)-1-
phenylbutyl]carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (11c).
1H NMR (500 MHz, methanol-d4): δ 8.32 (br d, J = 2.4 Hz, 1H),
7.99 (br t, J = 6.0 Hz, 1H), 7.85 (d, J = 8.4 Hz, 1H), 7.73 (dd, J = 8.3,
2.1 Hz, 1H), 7.57 (d, J = 2.1 Hz, 1H), 7.50 (d, J = 8.5 Hz, 1H), 7.46−
7.39 (m, 2H), 7.39−7.30 (m, 5H), 7.26−7.20 (m, 1H), 5.08 (dd, J =
8.9, 6.6 Hz, 1H), 2.48 (s, 3H), 1.94−1.88 (m, 1H), 1.85−1.78 (m,
1H), 1.48−1.40 (m, 1H), 1.39−1.32 (m, 1H), 0.97 (t, J = 7.4 Hz,
3H). Purity 96% (method B). HRMS (ESI): calcd for C32H29ClN3O3
[M + H]+, 538.1892; found, 538.1879.
5′-Chloro-2′-(5-methoxy-1H-1,3-benzodiazol-2-yl)-4-{[(1R)-1-
phenylbutyl]carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (11d).
1H NMR (500 MHz, methanol-d4): δ 8.97 (br d, J = 8.3 Hz, 1H),
8.37 (dd, J = 4.8, 1.5 Hz, 1H), 8.08−8.01 (m, 1H), 7.85 (d, J = 8.3
Hz, 1H), 7.74 (dd, J = 8.4, 2.1 Hz, 1H), 7.59 (s, 1H), 7.53 (d, J = 9.1
Hz, 1H), 7.49 (t, J = 7.6 Hz, 1H), 7.40−7.36 (m, 2H), 7.32 (t, J = 7.6
Hz, 2H), 7.27−7.20 (m, 1H), 7.16 (dd, J = 9.1, 2.2 Hz, 1H), 7.08 (d, J
= 2.2 Hz, 1H), 5.12−5.05 (m, 1H), 3.85 (s, 3H), 1.95−1.88 (m, 1H),
1.86−1.79 (m, 1H), 1.49−1.40 (m, 1H), 1.39−1.31 (m, 1H), 0.97 (t,
J = 7.4 Hz, 3H). Purity 100% (method A). HRMS (ESI): calcd for
C32H29ClN3O4 [M + H]+, 554.1841; found, 554.1827.
5′-Chloro-2′-(4-methyl-1H-1,3-benzodiazol-2-yl)-4-{[(1R)-1-
phenylbutyl]carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (11e).
1H NMR (500 MHz, methanol-d4): δ 8.27 (s, 1H), 7.98 (br d, J =
7.2 Hz, 1H), 7.87 (d, J = 8.4 Hz, 1H), 7.74 (dd, J = 8.3, 2.1 Hz, 1H),
7.60 (d, J = 2.0 Hz, 1H), 7.44 (d, J = 8.1 Hz, 1H), 7.40−7.34 (m,
4H), 7.31 (t, J = 7.6 Hz, 3H), 7.27−7.21 (m, 1H), 5.07 (dd, J = 8.8,
6.5 Hz, 1H), 2.53 (s, 3H), 1.93−1.87 (m, 1H), 1.85−1.78 (m, 1H),
1.48−1.40 (m, 1H), 1.37−1.30 (m, 1H), 0.96 (t, J = 7.4 Hz, 3H).
Purity > 95% (method B). HRMS (ESI): calcd for C32H29ClN3O3 [M
+ H]+, 538.1892; found, 538.1877.
5′-Chloro-2′-{1H-imidazo[4,5-c]pyridin-2-yl}-4-{[(1R)-1-
phenylbutyl]carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (11f). 1H
NMR (500 MHz, methanol-d4): δ 9.02 (s, 1H), 8.43 (d, J = 6.6 Hz,
1H), 8.34 (dd, J = 11.6, 1.7 Hz, 1H), 8.02−7.95 (m, 2H), 7.92 (d, J =
8.2 Hz, 1H), 7.67 (dd, J = 8.4, 2.1 Hz, 1H), 7.48 (d, J = 2.1 Hz, 1H),
7.43−7.37 (m, 3H), 7.33 (br t, J = 6.9 Hz, 2H), 7.27−7.20 (m, 1H),
5.13−5.08 (m, 1H), 1.98−1.90 (m, 1H), 1.88−1.79 (m, 1H), 1.53−
1.43 (m, 1H), 1.42−1.32 (m, 1H), 0.99 (t, J = 7.2 Hz, 3H). Purity >
99% (method B). HRMS (ESI): calcd for C30H26ClN4O3 [M + H]+,
525.1688; found, 525.1679.
J = 7.4 Hz, 3H). Purity 100% (method A). HRMS (ESI): calcd for
C31H26ClN2O4 [M + H]+, 525.1576; found, 525.1562.
2′-(1H-1,3-Benzodiazol-2-yl)-4-{[(1R)-1-phenylbutyl]carbamoyl}-
[1,1′-biphenyl]-2-carboxylic Acid (12a). 1H NMR (500 MHz,
methanol-d4): δ 8.29 (s, 1H), 7.96 (br d, J = 7.8 Hz, 1H), 7.88
(dd, J = 7.6, 1.1 Hz, 1H), 7.82−7.77 (m, 1H), 7.74−7.69 (m, 1H),
7.64 (dd, J = 6.3, 3.2 Hz, 2H), 7.56−7.49 (m, 3H), 7.41 (d, J = 7.9
Hz, 1H), 7.39−7.35 (m, 2H), 7.34−7.29 (m, 2H), 7.26−7.20 (m,
1H), 5.08 (dd, J = 8.8, 6.5 Hz, 1H), 1.96−1.87 (m, 1H), 1.86−1.78
(m, 1H), 1.49−1.40 (m, 1H), 1.39−1.31 (m, 1H), 0.97 (t, J = 7.3 Hz,
3H); purity 97% (method B). HRMS (ESI): calcd for C31H28N3O3
[M + H]+, 490.2125; found, 490.2113.
2′-(1H-1,3-Benzodiazol-2-yl)-5′-methyl-4-{[(1R)-1-phenylbutyl]-
1
carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (12b). H NMR (500
MHz, DMSO-d6): δ 8.96 (d, J = 8.3 Hz, 1H), 8.27 (d, J = 1.9 Hz,
1H), 8.00 (dd, J = 8.0, 1.4 Hz, 1H), 7.79 (d, J = 7.7 Hz, 1H), 7.55
(dd, J = 6.1, 3.0 Hz, 2H), 7.48 (d, J = 7.7 Hz, 1H), 7.41−7.37 (m,
2H), 7.35−7.29 (m, 5H), 7.25−7.19 (m, 2H), 5.07−4.98 (m, 1H),
2.45 (s, 3H), 1.92−1.82 (m, 1H), 1.76−1.67 (m, 1H), 1.43−1.33 (m,
1H), 1.31−1.23 (m, 1H), 0.90 (t, J = 7.3 Hz, 3H). Purity 96%
(method B). HRMS (ESI): calcd for C32H30N3O3 [M + H]+,
504.2282; found, 504.2271.
2′-(1H-1,3-Benzodiazol-2-yl)-5′-methoxy-4-{[(1R)-1-
phenylbutyl]carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (12c).
1H NMR (500 MHz, methanol-d4): δ 8.26 (s, 1H), 7.92 (br d, J =
7.8 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.67−7.59 (m, 2H), 7.53−7.48
(m, 2H), 7.40−7.32 (m, 5H), 7.28−7.23 (m, 2H), 7.06 (d, J = 2.4 Hz,
1H), 5.10 (dd, J = 8.9, 6.4 Hz, 1H), 3.95 (s, 3H), 1.98−1.89 (m, 1H),
1.87−1.78 (m, 1H), 1.50−1.42 (m, 1H), 1.40−1.33 (m, 1H), 0.99 (t,
J = 7.4 Hz, 3H). Purity 96% (method B). HRMS (ESI): calcd for
C32H30N3O4 [M + H]+, 520.2231; found, 520.2224.
(R)-2′-(1H-Benzo[d]imidazole-2-yl)-3′-chloro-4-((1-phenylbutyl)-
1
carbamoyl)-[1,1′-biphenyl]-2-carboxylic Acid (12d). H NMR (500
MHz, DMSO-d6): δ 8.85 (br d, J = 8.3 Hz, 1H), 8.17 (d, J = 1.4 Hz,
1H), 7.83−7.77 (m, 1H), 7.68 (d, J = 8.0 Hz, 1H), 7.63−7.54 (m,
1H), 7.48−7.42 (m, 2H), 7.35−7.26 (m, 5H), 7.24−7.19 (m, 2H),
7.15−7.11 (m, 2H), 5.00−4.88 (m, 1H), 1.85−1.75 (m, 1H), 1.71−
1.62 (m, 1H), 1.39−1.26 (m, 1H), 1.24−1.18 (m, 1H), 0.86 (t, J =
7.4 Hz, 3H). Purity 100% (method B). m/z obs 524.0 [M + H]+.
2′-(1H-1,3-Benzodiazol-2-yl)-4′-chloro-4-{[(1R)-1-phenylbutyl]-
1
carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (12e). H NMR (500
MHz, methanol-d4): δ 8.32 (s, 1H), 7.99 (br d, J = 7.2 Hz, 1H), 7.95
(d, J = 2.1 Hz, 1H), 7.79 (dd, J = 8.4, 2.3 Hz, 1H), 7.65−7.60 (m,
2H), 7.53−7.48 (m, 3H), 7.44 (d, J = 7.9 Hz, 1H), 7.38−7.34 (m,
2H), 7.34−7.28 (m, 2H), 7.25−7.21 (m, 1H), 5.07 (dd, J = 8.9, 6.6
Hz, 1H), 1.95−1.87 (m, 1H), 1.85−1.78 (m, 1H), 1.48−1.39 (m,
1H), 1.38−1.29 (m, 1H), 0.96 (t, J = 7.4 Hz, 3H). Purity 96%
(method B). HRMS (ESI): calcd for C31H27ClN3O3 [M + H]+,
524.1735; found, 524.1724.
2′-(1H-1,3-Benzodiazol-2-yl)-6′-chloro-4-{[(1R)-1-phenylbutyl]-
carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (12f). Into the reac-
tion vessel was added SI14 (301 mg, 0.577 mmol), THF (10 mL),
water (5 mL), and LiOH·H2O (121 mg, 2.88 mmol). The reaction
was stirred at rt for 1 day, diluted with EtOAc (30 mL), and washed
with 20 mL of sat NH4Cl containing 2.88 mmol HCl. The organic
phase was dried over Na2SO4 and concentrated to produce crude 2′-
(tert-butoxycarbonyl)-6-chloro-4′-(((R)-1-phenylbutyl)carbamoyl)-
[1,1′-biphenyl]-2-carboxylic acid (293 mg, 0.577 mmol, 100% yield),
which was used for the next step without further purification. Into the
reaction vessel was added the crude carboxylic acid (293 mg, 0.577
mmol), benzene-1,2-diamine (125 mg, 1.154 mmol), DIEA (0.302
mL, 1.730 mmol), DCM (5 mL), and HATU (285 mg, 0.750 mmol).
The reaction was stirred at rt for 12h, concentrated, and dissolved in
AcOH (4.5 mL)/water (0.5 mL). After stirring at 85 °C for 12 h, the
reaction was cooled to rt and concentrated. Preparative HPLC
purification of the resulting residue produced 12f (300 mg, 0.471
mmol, 82% yield). 1H NMR (500 MHz, methanol-d4): δ 8.34 (dd, J =
3.5, 1.8 Hz, 1H), 7.84−7.80 (m, 1H), 7.77 (d, J = 7.6 Hz, 1H), 7.74
(dd, J = 8.1, 1.1 Hz, 1H), 7.63−7.58 (m, 1H), 7.53−7.48 (m, 2H),
7.37−7.33 (m, 2H), 7.33−7.27 (m, 4H), 7.25−7.20 (m, 1H), 7.17 (d,
5′-Chloro-2′-(5-phenyl-1H-imidazol-2-yl)-4-{[(1R)-1-
phenylbutyl]carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (11g).
1H NMR (500 MHz, methanol-d4): δ 8.96 (br d, J = 8.3 Hz, 1H),
8.39 (br d, J = 1.4 Hz, 1H), 8.09 (br t, J = 7.3 Hz, 1H), 7.79 (d, J =
8.3 Hz, 1H), 7.73−7.69 (m, 2H), 7.62−7.55 (m, 3H), 7.50 (br d, J =
8.3 Hz, 1H), 7.45 (br s, 3H), 7.41−7.36 (m, 2H), 7.32 (t, J = 7.6 Hz,
2H), 7.26−7.21 (m, 1H), 5.15−5.07 (m, 1H), 1.97−1.89 (m, 1H),
1.87−1.80 (m, 1H), 1.50−1.41 (m, 1H), 1.41−1.33 (m, 1H), 0.97 (t,
J = 7.3 Hz, 3H). Purity 98% (method B). m/z obs 550.0 [M + H]+.
(R)-2′-(Benzo[d]oxazol-2-yl)-5′-chloro-4-((1-phenylbutyl)-
1
carbamoyl)-[1,1′-biphenyl]-2-carboxylic Acid (11h). H NMR (500
MHz, methanol-d4): δ 8.45 (d, J = 1.7 Hz, 1H), 8.23 (d, J = 8.4 Hz,
1H), 7.98 (dd, J = 7.9, 1.8 Hz, 1H), 7.60 (dd, J = 8.4, 2.1 Hz, 1H),
7.59−7.56 (m, 1H), 7.47−7.40 (m, 3H), 7.39−7.35 (m, 3H), 7.33−
7.25 (m, 4H), 5.15 (dd, J = 8.9, 6.4 Hz, 1H), 2.03−1.95 (m, 1H),
1.92−1.84 (m, 1H), 1.57−1.47 (m, 1H), 1.47−1.39 (m, 1H), 1.02 (t,
G
DOI: 10.1021/acs.jmedchem.9b01513
J. Med. Chem. XXXX, XXX, XXX−XXX

Journal of Medicinal Chemistry
Article
J = 7.9 Hz, 1H), 5.08−5.02 (m, 1H), 1.93−1.84 (m, 1H), 1.83−1.75
(m, 1H), 1.48−1.39 (m, 1H), 1.36−1.32 (m, 1H), 0.98−0.93 (m,
3H). Purity >95% (method B). m/z obs 524.2 [M + H]+.
(S)-2′-Chloro-6′-(5-methoxy-1H-1,3-benzodiazol-2-yl)-4-{[(1R)-1-
phenylbutyl]carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (15c).
1H NMR (500 MHz, methanol-d4): δ 8.97 (br d, J = 8.0 Hz, 1H),
8.47 (d, J = 1.9 Hz, 1H), 7.95 (dd, J = 8.0, 1.9 Hz, 1H), 7.88 (dd, J =
8.3, 1.1 Hz, 1H), 7.83 (dd, J = 7.7, 1.1 Hz, 1H), 7.74−7.66 (m, 1H),
7.52 (d, J = 9.4 Hz, 1H), 7.37−7.34 (m, 2H), 7.33−7.29 (m, 3H),
7.25−7.20 (m, 1H), 7.15 (dd, J = 9.1, 2.5 Hz, 1H), 7.07 (d, J = 2.2
Hz, 1H), 5.10−5.03 (m, 1H), 3.84 (s, 3H), 1.95−1.86 (m, 1H),
1.85−1.77 (m, 1H), 1.49−1.39 (m, 1H), 1.39−1.30 (m, 1H), 0.96 (t,
J = 7.3 Hz, 3H). Purity 95% (method A). HRMS (ESI): calcd for
C32H29ClN3O4 [M + H]+, 554.1841; found, 554.1828.
2′-(1H-1,3-Benzodiazol-2-yl)-6′-fluoro-4-{[(1R)-1-phenylbutyl]-
1
carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (12g). H NMR (500
MHz, methanol-d4): δ 8.47 (d, J = 1.8 Hz, 0.5H), 8.45 (d, J = 1.8 Hz,
0.5H), 7.93−7.88 (m, 1H), 7.76−7.71 (m, 2H), 7.64−7.60 (m, 2H),
7.57−7.52 (m, 1H), 7.51−7.46 (m, 2H), 7.38−7.34 (m, 2H), 7.33−
7.29 (m, 3H), 7.26−7.20 (m, 1H), 5.10−5.04 (m, 1H), 1.93−1.86
(m, 1H), 1.84−1.77 (m, 1H), 1.48−1.40 (m, 1H), 1.38−1.31 (m,
1H), 0.99−0.93 (m, 3H). Purity 97% (method B). HRMS (ESI):
calcd for C31H27FN3O3 [M + H]+, 508.2031; found, 508.2026.
2′-(1H-1,3-Benzodiazol-2-yl)-6′-methyl-4-{[(1R)-1-phenylbutyl]-
ASSOCIATED CONTENT
* Supporting Information
■
1
carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (12h). H NMR (500
S
MHz, methanol-d4): δ 8.34 (dd, J = 6.0, 1.8 Hz, 1H), 7.93 (ddd, J =
7.9, 5.9, 1.9 Hz, 1H), 7.68 (dd, J = 10.5, 7.5 Hz, 2H), 7.64−7.57 (m,
3H), 7.50 (ddd, J = 6.2, 3.2, 1.1 Hz, 2H), 7.37−7.29 (m, 5H), 7.25−
7.20 (m, 1H), 5.10−5.04 (m, 1H), 2.10 (s, 3H), 1.93−1.85 (m, 1H),
1.84−1.77 (m, 1H), 1.46−1.38 (m, 1H), 1.36−1.29 (m, 1H), 0.96 (t,
J = 7.4 Hz, 3H). Purity >95% (method B). HRMS (ESI): calcd for
C32H30N3O3 [M + H]+, 504.2282; found, 504.2272.
The Supporting Information is available free of charge on the
ACS Publications website at DOI: 10.1021/acs.jmed-
chem.9b01513.
Supplementary synthetic schemes, experimental proto-
cols, synthesis of 12a−h, synthesis of 15a via
intermediate 14, assay condition, summary of PV-loop
study, biotransformation study on the analogue of 9b,
details on HTP screening, and NMR spectra and HPLC
traces (PDF).
Preparation of 15a and 15b via chiral SFC purification: 12f was
subjected to the chiral SFC purification condition below to afford
optically pure compounds 15a (second eluting peak) and 15b (first
eluting peak)instrument: PIC solution 200 SFC; column:
Chiralpak AD-H, 21 × 250 mm, 5 μm; mobile phase: 22% EtOH/
78% CO2; flow conditions: 45 mL/min, 150 bar, 40 °C; detector
wavelength: 220 nm; injection details: 0.5 mL of ∼27 mg/mL in
EtOH/MeCN. 15a, RT = 8.37 min; 15b, RT = 6.28 min.
Molecular formula strings for the final compounds
(CSV)
AUTHOR INFORMATION
Corresponding Author
*E-mail: Shun.Su@bms.com. Phone: (+1) 609-466-5079.
■
(S)-2′-(1H-1,3-Benzodiazol-2-yl)-6′-chloro-4-{[(1R)-1-
phenylbutyl]carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (15a).
1H NMR (500 MHz, methanol-d4): δ 8.43 (d, J = 1.8 Hz, 1H),
7.92 (dd, J = 8.0, 1.9 Hz, 1H), 7.84 (td, J = 8.5, 1.1 Hz, 2H), 7.69−
7.65 (m, 1H), 7.63−7.58 (m, 2H), 7.49−7.46 (m, 2H), 7.36−7.28
(m, 5H), 7.24−7.20 (m, 1H), 5.05 (dd, J = 8.9, 6.6 Hz, 1H), 1.92−
1.85 (m, 1H), 1.84−1.75 (m, 1H), 1.47−1.38 (m, 1H), 1.37−1.30
(m, 1H), 0.96 (t, J = 7.4 Hz, 3H). 13C NMR (126 MHz, methanol-
d4): δ 167.6, 167.0, 148.7, 143.0, 140.3, 139.2, 135.5, 134.2, 132.9,
132.4, 132.3, 131.1, 130.3, 129.4, 129.3, 128.6, 128.1, 126.8, 126.3,
126.0, 125.9, 113.9, 54.1, 37.8, 19.5, 12.6. Purity: >99% (chiral
analytical chromatographic condition: Berger analytical SFC;
Chiralpak AD-H, 4.6 × 250 mm, 5 μm; 20% EtOH/80% CO2; 2
mL/min, 150 bar, 40 °C; injection details: 5 μL of 1 mg/mL in
ACN); HRMS (ESI): calcd for C31H27ClN3O3 [M + H]+, 524.1735;
found, 524.1727. [α]D20 = −38.4° (C = 0.5, MeOH). Full
crystallographic data for compound 15a have been deposited to the
Cambridge Crystallographic Data Center (CCDC reference no.
CCDC 1918519) and can be obtained free of charge via the internet
at http://www.ccdc.cam.ac.uk.
ORCID
Shun Su: 0000-0001-8100-9645
Eric Simmons: 0000-0002-3854-1561
Present Address
†GreenBioChem LLC, 410 Sackett Point Road, Bldg 20 North
Haven, CT 06473.
Notes
The authors declare no competing financial interest.
ACKNOWLEDGMENTS
■
We thank Dr. Atsu Apedo for support on chiral separation,
Gerry G. Everlof for support on solubility determination,
Robert Langish and Linping Wang for support with HRMS,
and Dr. Yi Hsiao for support on high-throughput reaction
screening. We thank Dr. Heather Finlay, Dr. Scott Shaw, and
Vladimir Ladziata for their assistance in manuscript prepara-
tion.
(R)-2′-(1H-1,3-Benzodiazol-2-yl)-6′-chloro-4-{[(1R)-1-
phenylbutyl]carbamoyl}-[1,1′-biphenyl]-2-carboxylic Acid (15b).
1H NMR (500 MHz, methanol-d4): δ 8.30 (d, J = 1.7 Hz, 1H),
7.78 (dd, J = 8.0, 1.9 Hz, 1H), 7.75 (dd, J = 7.6, 1.1 Hz, 1H), 7.69
(dd, J = 8.2, 1.1 Hz, 1H), 7.59−7.54 (m, 1H), 7.49−7.44 (m, 2H),
7.37−7.33 (m, 2H), 7.32−7.28 (m, 2H), 7.26−7.19 (m, 3H), 7.11 (d,
J = 7.9 Hz, 1H), 5.05 (dd, J = 8.9, 6.5 Hz, 1H), 1.92−1.83 (m, 1H),
1.82−1.74 (m, 1H), 1.48−1.38 (m, 1H), 1.36−1.28 (m, 1H), 0.95 (t,
J = 7.4 Hz, 3H). 13C NMR (126 MHz, methanol-d4): δ 169.9, 167.4,
150.6, 143.1, 139.8, 139.7, 136.5, 134.9, 134.5, 134.0, 131.1, 130.6,
130.2, 129.3, 129.0, 128.3, 128.1, 128.1, 126.7, 126.3, 123.3, 114.4,
53.9, 37.9, 19.5, 12.7. Purity: >99% (chiral analytical chromatographic
condition: Berger analytical SFC; Chiralpak AD-H, 4.6 × 250 mm, 5
μm; 20% EtOH/80% CO2; 2 mL/min, 150 bar, 40 °C; injection
details5 μL of 1 mg/mL in ACN); HRMS (ESI): calcd for
C31H27ClN3O3 [M + H]+, 524.1735; found, 524.1725. [α]2D0 = +43.3°
(C = 0.43, MeOH). Full crystallographic data for compound 15b have
been deposited to the Cambridge Crystallographic Data Center
(CCDC reference no. CCDC 1918520) and can be obtained free of
charge via the internet at http://www.ccdc.cam.ac.uk.
DEDICATION
■
In memory of Atsu Apedo (1968−2017) and William
Schumacher (1952−2018).
ABBREVIATIONS
■
HF, heart failure; CO, cardiac output; PV-loop, pressure−
volume loop; SAR, structure−activity relationship; SFC,
supercritical fluid chromatography; BPA, biphenyl acid; APJ-
R, APJ receptor
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DOI: 10.1021/acs.jmedchem.9b01513
J. Med. Chem. XXXX, XXX, XXX−XXX

Journal of Medicinal Chemistry
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