Design, synthesis, structure-activity relationships, and docking studies of pyrazole-containing derivatives as a novel series of potent glucagon receptor antagonists

Article abstract of DOI:10.1016/j.bmc.2016.04.053

Glucagon receptor antagonists possess a great potential for treatment of type 2 diabetes mellitus. A series of pyrazole-containing derivatives were designed, synthesized and evaluated by biological assays as glucagon receptor antagonists. Most of the compounds exhibited good in vitro efficacy. Two of them, compounds 17f and 17k, displayed relatively potent antagonist effects on glucagon receptors with IC₅₀ values of 3.9 and 3.6 μM, respectively. The possible binding modes of 17f and 17k with the cognate receptor were explored by molecular docking simulation.

Full text of DOI:10.1016/j.bmc.2016.04.053

Bioorganic & Medicinal Chemistry 24 (2016) 2852–2863
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Design, synthesis, structure–activity relationships, and docking
studies of pyrazole-containing derivatives as a novel series of potent
glucagon receptor antagonists
Shuangjie Shu a,y, Xiaoqing Cai b,y, Jia Li a,c,y, Yang Feng b, Antao Dai b, Jiang Wang a, Dehua Yang a,b
,
Ming-Wei Wang a,b, , Hong Liu a,c,
⇑
⇑
a CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Shanghai 201203, People’s Republic of China
b The National Center for Drug Screening, 189 Guo Shou Jing Road, Shanghai 201203, People’s Republic of China
c School of Pharmacy, China Pharmaceutical University, Jiangsu, Nanjing 210009, People’s Republic of China
a r t i c l e i n f o
a b s t r a c t
Article history:
Glucagon receptor antagonists possess a great potential for treatment of type 2 diabetes mellitus. A series
of pyrazole-containing derivatives were designed, synthesized and evaluated by biological assays as
glucagon receptor antagonists. Most of the compounds exhibited good in vitro efficacy. Two of them,
compounds 17f and 17k, displayed relatively potent antagonist effects on glucagon receptors with IC50
Received 25 March 2016
Revised 23 April 2016
Accepted 25 April 2016
Available online 26 April 2016
values of 3.9 and 3.6
lM, respectively. The possible binding modes of 17f and 17k with the cognate
receptor were explored by molecular docking simulation.
Keywords:
Ó 2016 Published by Elsevier Ltd.
Glucagon receptor
Antagonist
Molecular docking
Pyrazole
Type 2 diabetes mellitus
1. Introduction
Glucagon is a polypeptide hormone consisting of 29 amino
acids that counteracts the action of insulin9–13 and binds to gluca-
gon receptor (GCGR) to trigger two transduction cascade signals
resulting in stimulation of hepatic glucose production.14–18 In
patients with T2DM, inappropriately elevated glucagon levels lead
to excessive hepatic glucose output, which is the main contributing
factor to hyperglycemia. Therefore, blockage of glucagon-induced
hyperglycemia by means of GCGR antagonism is theoretically pru-
dent to treat T2DM.19–25 Previous studies have validated this
approach using glucagon-specific antibodies as well as peptidic
and non-peptidic GCGR antagonists in animal models of dia-
betes.22,25 Recently, a number of publications have claimed several
chemical scaffolds as GCGR antagonists.26–32 The biaryl containing
compound BAY-27-995521 (Fig. 1) was the first active agent shown
to decrease glucose output upon the administration of exogenous
glucagon. Compounds MK-357733 and MK-089334,35 (Fig. 1) with
b-alanine acid motif were discontinued in phase II clinical trials
for treating T2DM.
Type 2 diabetes mellitus (T2DM), a chronic metabolic disorder
characterized by fasting hyperglycemia consequential to dysfunc-
tion of pancreatic b-cells coupled with elevated insulin resistance
and reduced insulin secretion, is affecting approximately 415 mil-
lion people in 2015.1 According to International Diabetes Federa-
tion (IDF), the estimated number of people suffering from
diabetes will rise to 642 million in less than 25 years.2 Despite
many potential drug targets are continuously pursued and multi-
ple therapeutic strategies exist for the treatment of T2DM, there
is still a significant need for additional anti-diabetic agents to
improve safety and efficacy.3–6 In recent years, activation of gluca-
gon-like peptide-1 receptor (GLP-1R) and inhibition of dipeptidyl
peptidase-4 (DPP-4) to stimulate insulin secretion7 and inhibition
of sodium-glucose cotransporter-2 (SGLT-2) to regulate glucose
reabsorption8 have gained prominence in the management of
T2DM, while glucagon receptor as drug target appeared to be less
attractive.
In this paper, we adopted bioisosteric and conformational
restraint strategies to synthesize a series of novel pyrazole-contain-
ing derivatives based on the structure of MK-0893. These com-
pounds exhibited sound GCGR binding affinities and cAMP
responses. Follow-up design, synthesis, structure–activity relation-
ship (SAR) and docking studies report our efforts toward a novel
series of GCGR antagonists.
⇑
Corresponding authors. Tel./fax: +86 21 50807042 (H.L.).
E-mail addresses: mwwang@simm.ac.cn (M.-W. Wang), hliu@simm.ac.cn
(H. Liu).
y
These authors contributed equally to this work.
http://dx.doi.org/10.1016/j.bmc.2016.04.053
0968-0896/Ó 2016 Published by Elsevier Ltd.

S. Shu et al. / Bioorg. Med. Chem. 24 (2016) 2852–2863
2853
Cl
F
O
O
O
O
N
H
OH
HN
N
N
OH
O
HN
Cl
H
O
Cl
F
MK-0893
MK-3577
BAY-27-9955
Figure 1. Structures of BAY-27-9955, MK-3577 and MK-0893.
2.2. Chemical synthesis
2. Materials and methods
The synthetic route of compound 9 was shown in Scheme 1.
Briefly, esterification of commercially available compound 1 easily
afforded 2, which was subsequently transformed to compound 3
by reaction with tert-butyl carbazate and then reduced by sodium
cyanoborohydride. Deprotection of compound 3 provided hydra-
zine 4, which was smoothly reacted with ethyl 3-(3,5-dichlorophe-
nyl)-3-oxopropanoate 5 to afford compound 6. Triflate 7 was
generated using trifluoromethanesulfonic anhydride (Tf2O) and
experienced a classical Suzuki coupling reaction with different
boric acid (Z-B(OH)2) to provide compound 8. Compound 9 was
then generated by hydrolysis of compound 8, condensation with
substituted 3-aminopropanoate and hydrolysis of ester.
The synthesis of compound 17 was shown in Scheme 2. Esteri-
fication of commercially available compound 10 easily afforded
compound 11, which was subsequently transformed to compound
12 by reaction with tert-butyl carbazate and then reduced by
sodium cyanoborohydride. Deprotection of compound 12 provided
hydrazine 13, which was smoothly reacted with ethyl 3-(3,5-
dichlorophenyl)-3-oxopropanoate 5 to afford compound 14. Tri-
flate 15 was generated using trifluoromethanesulfonic anhydride
(Tf2O) and experienced a classical Suzuki coupling reaction with
boric acid (R2-B(OH)2) to provide pyrazoles 16. Compound 17
was then generated by hydrolysis of 16, condensation with
b-alanine tert-butyl ester and removing of tert-butyl ester.
2.1. Molecular design
In 2013, Fai et al. reported the crystal structure of the seven-
transmembrane helical domain of human GCGR.36 Comparison of
the ligand-binding pocket of GCGR with that of class A GPCRs
shows that GCGR has a larger binding cavity, and glucagon is cap-
able of inducing this domain to the active state.37 The crystal struc-
ture of GCGR (PDB ID: 4l6r) and Glide program were used for
docking to study the interactions between GCGR and MK-0893.38
The modeling results indicated that MK-0893 occupied the S1
and S2 pockets and formed two hydrogen bonds in hydrogen bind-
ing districts (HB) with GCGR (Fig. 2A). In the well-defined
hydrophobic S1 pocket, the 6-methoxy-2-naphthalenyl ring of
MK-0893 formed extensive hydrophobic interactions with resi-
dues Tyr149, Asp195, Ile194, Met231 and Gln232. In the S2 pocket,
the 3,4-dichloro-phenyl ring formed hydrophobic interactions
with residues Leu307, Val311, Tyr239, Ile235, Glu362 and
Gln392. The 6-methoxy-2-naphthalenyl ring and two phenyl rings
in the MK-0893 are stacked against the side chain of Trp295 and
Phe365. Moreover, 6-methoxyl and b-alanine carboxylic acid of
MK-0893 appear to form hydrogen bonds with Tyr149 and
Lys381, respectively, which may account for the excellent antago-
nistic activity. However, the cavity formed by residues Gln232,
Leu307 and Arg308 is a large pocket suggesting that introduction
of steric hindrance on MK-0893 may gain better antagonistic activ-
ity (Fig. 2B). To verify this hypothesis, we replaced b-alanine acid
as substituted and cyclic b-alanine acid to obtain compound 9
(Fig. 3). Since conformational restraint is an established strategy
to improve binding potency to GCGR,39 we introduced an indane
motif to obtain compound 17 (Fig. 3). A series of pyrazole-contain-
ing derivatives were subsequently synthesized and evaluated.
3. Results and discussion
All the synthesized compounds were evaluated for antagonism
of human GCGR and the cAMP response in vitro. Antagonistic
potency was measured by a competitive binding assay with rac-
MK-0893 as the positive control, and the results are reported as
Figure 2. Docking study of MK-0893. All figures were prepared using PyMol (http://www.pymol.org).

2854
S. Shu et al. / Bioorg. Med. Chem. 24 (2016) 2852–2863
O
R1
N
N
Z
Cl
e
c
n
a
r
e
d
in
h
g
O
n
i
c
u
d
o
r
t
f
n
i
Cl
9a-9h
O
HN
N
N
OH
O
Cl
O
O
N
H
OH
Cl
N
N
S2 pocket
c
o
n
o
r
m
a
t
io
n
R2
a
l
r
Cl
e
s
t
r
a
in
S1 pocket
t
rac-MK-0893
GCGR binding IC50 = 0.6 0.3 μΜ
cAMP response IC50 = 0.1 0 μΜ
Cl
17a-17l
Figure 3. Design of novel GCGR antagonists 9 and 17.
O
O
O
O
a
d
OH
O
O
b
c
H
N
O
N
H
O
O
1
2
3
O
O
O
O
O
N
N
f
Cl
O
e
O
O
H
N
Cl
+
H2N
Cl
g
Cl
4
5
6
O
O
O
R1
O
O
N
N
N
9
N
N
N
h
Z
Z
Cl
Cl
OTf
Cl
i, j
Cl
Cl
Cl
7
8
Scheme 1. Reagents and conditions: (a) EtOH, H2SO4, reflux, 3 h; (b) cat. HOAc, toluene, 80 °C, overnight; (c) NaBH3CN, p-toluene sulfonic acid, THF, rt, 3 h; (d) TFA, DCM, rt,
1 h; (e) HOAc, reflux, 4 h; (f) Tf2O, TEA, THF, ꢀ78 °C, 3 h; (g) Z-B(OH)2, Pd(PPh3)4, TEA, DME, 85 °C, 2 h; (h) NaOH, MeOH, 1,4-dioxane, 60 °C, 1 h; (i) substituted
3-aminopropanoate, PyBOP, DIEA, DMF, rt, overnight; (j) NaOH, MeOH, 1,4-dioxane, 60 °C, 1 h.
concentration for 50% inhibition (IC50). SAR for these compounds is
summarized in Tables 1 and 2.
Firstly, we focus on the important pharmacophore Z with 3-
aminobutanoic acid (Table 1). Compound 9c containing 6-
amino acids. Compound 9e thus appeared to be a potent GCGR
antagonist. However, the activities of all the compounds in this ser-
ies were poorer than rac-MK-0893, implying that the steric effect
of b-alanine acid might have significantly reduced their antagonis-
tic activity on GCGR.
methoxynaphthalen-2-yl was able to bind GCGR (IC50 = 20.2
lM)
and elicit cAMP responses (IC50 = 1.9 M). Compared with 6-
methoxynaphthalen-2-yl (9c), the 4-tert-butylphenyl (9a) and
2,3-dihydrobenzofuran-5-yl (9b) groups were not preferred for Z
l
Next, multiple substituents were introduced to understand the
effect of conformational restraint (Table 2). By introducing large
alkoxyl groups, compounds 17a and 17b were synthesized first
and showed lower potencies as compared with rac-MK-0893. Com-
in the pyrazole series. Substituent of methyl at the
b-amino acid (9e) exhibited better binding (IC50 = 8.4
cAMP activities (IC50 = 1.6 M) than that of methyl (9c) and phenyl
a-position of
lM) and
pound 17a showed better GCGR binding activity (IC50 = 5.7
lM) and
l
cAMP response (IC50 = 1.2 M) than compound 17b. According to
l
(9d) at the b-position. Cyclic amino acid substitution was also
investigated and the six-membered heterocyclic amino acid substi-
tution (9g and 9h) showed better binding affinity than the five-
membered heterocyclic amino acid (9f). In addition, the cAMP
the results of 17a and 17b, we investigated the substitutions on phe-
nyl ring by non-substitution and introduction of methyl group to
make compounds 17c and 17d but both displayed low potency.
However, 17c exhibited a better cAMP response (IC50 = 3.5
17d (IC50 = 6.9 M). Compound 17f containingchloro-substituted at
the meta-position of the phenyl ring demonstrated good GCGR bind-
ing (IC50 = 3.9 M) and cAMP (IC50 = 1.4 M) activities, while a flu-
oro-substituted (17e) was less active for GCGR binding
lM) than
response of 9h (IC50 = 4.5
lM) was better than that of 9f and 9g
l
(IC50 = 6.9 M and 26.5 M, respectively). The compounds contain-
l
l
ing methyl substitution on b-amino acid induced better cAMP
responses than those substituted by phenyl and heterocyclic
l
l

S. Shu et al. / Bioorg. Med. Chem. 24 (2016) 2852–2863
2855
O
O
O
O
O
a
O
O
O
b
d
OH
O
O
c
NH
N
O
H
O
11
O
12
10
O
N
N
Cl
O
O
O
Cl
f
e
+
NH
13
H2N
Cl
5
14
Cl
O
O
O
O
O
O
N
H
OH
N
h,i
N
g
N
N
N
N
OTf
Cl
R2
Cl
R2
j
Cl
Cl
Cl
15
Cl
16
17
Scheme 2. Reagents and conditions: (a) EtOH, H2SO4, reflux, 4 h; (b) cat. HOAc, toluene, 80 °C, overnight; (c) NaBH3CN, p-toluene sulfonic acid, THF, rt, 3 h; (d) TFA, DCM, rt,
2 h; (e) HOAc, reflux, 4 h; (f) Tf2O, TEA, THF, ꢀ78 °C to rt, 1.5 h; (g) R2-B(OH)2, Pd(PPh3)4, TEA, DME, microwave, 100 °C, 25 min; (h) NaOH, MeOH, 1,4-dioxane, 60 °C, 1 h;
(i) b-alanine tert-butyl ester, PyBOP, DIEA, rt, overnight; (j) TFA, DCM, rt, 1 h.
(IC50 = 8.9
(IC50 = 1.4
l
l
M) and displayed an equivalent cAMP response
M) as 17f. In addition, heterocyclic ring substituted
modulators, the hydrogen binding district is a promising pocket
for in-depth investigation.
compounds17g and 17h were also studied and showed poor binding
activity (IC50 = 71.8 M and 14.5 M, respectively). Meanwhile, it
l
l
4. Conclusions
was observed that (1) thiophene-substitution (17h) was somewhat
more favored than furan-substitution (17g) in the pyrazole scaffold;
(2) naphthyl ring especially naphthyl-2-yl ring (17k) possessed
In conclusion, we designed, synthesized and evaluated a series
of pyrazole derivatives as novel GCGR antagonists. SAR studies
have identified compounds 17f, 17k and 17l as potential candi-
dates for further development. Among these, compound 17k was
found to display comparatively better GCGR antagonistic binding
activity with an IC50 value of 3.6 lM. Molecular modeling demon-
strated that this series of compounds occupied the large binding
cavity of GCGR in three directions.
good binding (IC50 = 3.6 lM) and cAMP (IC50 = 1.2 lM) activities,
while 6-methoxy-substitution (17l) on the naphthyl ring modestly
reduced the activity (Table 2). Like 9a–9h, this series of compounds
did not improve the bioactivity beyond that of rac-MK-0893, sug-
gesting that the conformational restraint had a negative impact on
GCGR antagonism.
Structures of all synthesized pyrazole-derivatives were docked
against the binding domain of human GCGR to investigate the
binding modes and gain insights into the interactions between
these compounds and the receptor.
5. Experimental section
5.1. Biological evaluation
Docking results of compounds 17f and 17k indicated that these
two compounds occupied the S1 and S2 pockets and showed bind-
ing modes similar to MK-0893 (Fig. 4). They also formed extensive
hydrophobic interactions in S1 pocket with residues Tyr145,
Ile194, Met231 and Gln232. In S2 pocket, they both formed
hydrophobic interactions with residues Leu307, Glu362 and
Gln392.
5.1.1. Construction of GCGR vector
The wild-type human GCGR was cloned into the EcoRI and
HindIII sites of the pcDNA3.1/V5-His-TOPO vector (Invitrogen,
Carlsbad, CA, USA). Sequences of receptor clones were confirmed
by sequencing with ABI 3730xl DNA Analyzer (Applied Biosystems,
Foster City, CA, USA).
The observable differences in the binding site originate from the
interactions between different residues and moieties. Compared to
the methyl group in MK-0893, the indane ring of compounds 17f
(Fig. 4A) and 17k (Fig. 4B) is a relatively big group which may
result in change of the direction of b-alanine carboxylic acid group
and finally form a hydrogen bond with residue Arg308 leading to
dramatic decrease of potential hydrophobic interactions. In addi-
tion, the three aryl rings heading for three directions in the MK-
0893 are all stacked against the side chain of Trp295 and
5.1.2. Binding assay
The whole cell binding was conducted according to previously
reported method.36 Briefly, cells were harvested 24 h after trans-
fections with GCGR and incubated with blocking buffer (F12 sup-
plemented with 33 mM HEPES and 0.1% bovine serum albumin
(BSA), pH 7.4) for 2 h. Then the binding assay were performed by
incubating the cells with constant concentration of [125I]-glucagon
(40 pM, PerkinElmer, Boston, MA, USA) and different concentra-
Phe365, however, only one
p–p stacking interaction remained
tions of unlabeled compounds (1.28 nM–100 lM) at room temper-
with residue Trp295 was observed in compounds 17f and 17k. This
may be the main reason why all the compounds of this series were
not as potent as MK-0893. Therefore, we propose that the modifi-
cation of MK-0893 should take flexibility of introducing group at
benzyl position into consideration. Furthermore, for other GCGR
ature for 3 h. After washing cells with ice-cold PBS for three times,
the cells were subsequently lysed and counted for radioactivity
(counts per minute, CPM) in a scintillation counter (MicroBeta2
Plate Counter, PerkinElmer) using
(OptiPhase SuperMix, PerkinElmer).
a scintillation cocktail

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S. Shu et al. / Bioorg. Med. Chem. 24 (2016) 2852–2863
Table 1
In vitro GCGR binding and functional cAMP activities of compounds 9a–9h
O
R1
N
N
Z
Cl
Cl
Compd
R1
Z
GCGR bindinga
IC50 M)
cAMP responsea
IC50 M)
(
l
(l
O
H2N
9a
41.0 6.3
6.3 0.9
OH
O
H2N
H2N
9b
9c
18.1 4.7
20.2 5.1
13.0 1.9
1.9 0.4
OH
O
O
O
OH
O
Ph
O
H2N
H2N
HN
9d
NAc
20.8 3.8
OH
O
O
9e
9f
8.4 1.5
1.6 0.4
6.9 1.2
OH
O
O
131.1 49.9
OH
O
O
9g
112.2 55.1
26.5 1.5
HN
OH
O
O
HN
9h
41.7 2.1
0.6 0.3
4.5 1.1
0.1 0b
OH
O
O
H2N
1
rac-MK-0893
OH
a
b
c
Activities are reported as means SEM (N P 3).
SEM < 0.05.
NA: not active.
5.1.3. cAMP assays
center residue). All compounds were minimized by OPLS2001 force
field to generate low energy 3D structure using LigPrep under its
default parameters. Docking studies were carried out using Glide
in standard precision mode, with up to 40 conformers saved per
molecule. Glide score was obtained to assess the most favorable
ligand and its conformer, which was then exported to a Maestro-
formatted output file.
cAMP accumulation was measured using HTRF-cAMP dynamic
kit (Cisbio International, Gif sur Yvette Cedex, France) according
to manufacturer’s instructions and previous literature.40 Briefly,
HEK293T cells were transfected with GCGR. Cells were then trans-
ferred to 384-well plates at a density of 3000 cells per well and
incubated for a further 24 h at 37 °C. After incubation for 30 min
in assay buffer (DMEM, 1 mM 3-isobutyl-1-methylxanthine) with
1 nM glucagon and different concentration of compounds at
37 °C, the reactions were stopped by addition of lysis buffer con-
taining HTRF reagents. Plates were then incubated for 60 min at
room temperature, and time-resolved FRET signals were measured
after excitation at 620 nm and 650 nm by EnVision (PerkinElmer).
5.3. Synthesis
All commercially available compounds and solvents were used
without further purification. All target products were characterized
by their NMR and MS spectra. 1H spectra were recorded in deute-
rochloroform (CDCl3) and dimethylsulfoxid-d6 (DMSO-d6) on
400 MHz instrument. Chemical shifts were reported in parts per
million (ppm, d) downfield from tetramethylsilane. Proton cou-
pling patterns are described as singlet (s), doublet (d), triplet (t),
quartet (q), multiplet (m), and broad (br). Low- and high-resolution
mass spectra (LRMS and HRMS) were measured on a spectrometer.
All compounds (exclude compound 17f) were confirmed with over
95% purity which were determined by Agilent-1100 HPLC with
binary pump, photodiode array detector (DAD), using Agilent
5.2. Molecular docking
The Glide program from Schrӧdinger Suite41 was employed for
the docking simulation. The receptor protein was selected and
downloaded from worldwide Data Bank (PDB ID: 4l6r), which
was then imported, optimized and minimized to add hydrogen
and to delete unwanted molecules using protein preparation wiz-
ard.42 The receptor grid was sized to 15 Å in each direction and
centered using Centroid of selected residues (Specify Met231 as
Extend-C18 column (150 ꢁ 4.6 mm, 5
lm). Compound 17f was

S. Shu et al. / Bioorg. Med. Chem. 24 (2016) 2852–2863
2857
Table 2
In vitro GCGR binding and functional cAMP activities of compounds 17a–17l
O
O
N
H
OH
N
N
R2
Cl
Cl
Compd
R2
GCGR bindinga
IC50 M)
cAMP responsea
IC50 M)
(
l
(l
O
17a
5.7 1.2
1.2 0.2
O
O
17b
17c
17d
31.9 7.7
11.1 1.3
14.5 2.3
6.3 0.7
3.5 0.5
6.9 1.3
O
F
17e
17f
8.9 1.1
3.9 0.3
1.4 0.3
1.4 0.5
Cl
17g
17h
71.8 16.5
14.5 0.3
10.8 0.9
5.3 1.7
O
S
17i
12.7 3.9
2.6 0.6
17j
5.9 1.4
3.6 0.5
2.1 0.8
1.2 0.3
17k
O
17l
4.7 0.7
0.6 0.3
1.5 0.5
0.1 0b
rac-MK-0893
a
Activities are reported as means SEM (N P 3).
SEM < 0.05.
b
Figure 4. Docking structures of compounds 17f (A) and 17k (B).

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S. Shu et al. / Bioorg. Med. Chem. 24 (2016) 2852–2863
analyzed by Agilent Eclipse XDB-C18 column (150 ꢁ 4.6 mm,
m), using CH3CN/H2O = 70:30 (v/v) (7 min, 1 mL/min), then
HOAc (80 mL) for 4 h. The solvent was removed under reduced
pressure and the residue taken up with ethyl acetate, washed with
saturated NaHCO3 and brine, and dried over Na2SO4. Flash column
chromatography gave 2.6 g (60.7%) of ethyl 4-{1-[3-(3,5-dichloro-
5
l
CH3CN/H2O = 85:15 (v/v) (5 min, 1 mL/min) and calculated the
peak areas at 254 nM. Compounds 9a–9h were analyzed using
CH3CN/H2O = 85:15 (v/v) at 1 mL/min and calculated the peak
areas at 254 nM. Compounds 17a–e, 17g–l were analyzed using
CH3CN/H2O = 65:35 (v/v) at 1 mL/min and calculated the peak
areas at 254 nM.
phenyl)-5-oxo-4,5-dihydro-1H-pyrazol-1-yl]ethyl}benzoate as
a
yellow oil. 1H NMR (400 MHz, CDCl3) d 8.02 (d, J = 8.3 Hz, 2H),
7.51 (dd, J = 10.4, 5.0 Hz, 4H), 7.38 (dd, J = 4.8, 3.0 Hz, 1H),
5.56 (q, J = 7.1 Hz, 1H), 4.40–4.32 (m, 2H), 3.66–3.50 (m, 2H),
1.78 (d, J = 7.1 Hz, 3H), 1.37 (t, J = 7.1 Hz, 3H). MS (ESI, m/z):
404.8 [M+H]+.
5.3.1. General procedure for the preparation of compound 9
5.3.1.1. Ethyl 4-acetylbenzoate (2).
A solution of 4-acetyl-
benzoic acid (10 g, 60.9 mmol) in ethanol (100 mL) was stirred in
ice bath. After H2SO4 was slowly added, the mixture was refluxed
at 80 °C for 3 h. The mixture was extracted with EtOAc, washed
with saturated brine, dried (Na2SO4), and concentrated. The resi-
due was purified by chromatography to afford ethyl 4-acetylben-
zoate as a white solid (10 g, 85.4%). 1H NMR (400 MHz, CDCl3) d
8.13–8.09 (m, 2H), 7.99 (dt, J = 6.7, 1.0 Hz, 2H), 4.39 (q, J = 7.1 Hz,
2H), 2.63 (s, 3H), 1.40 (t, J = 7.1 Hz, 3H). MS (ESI, m/z): 193.1 [M
+H]+.
5.3.1.6. Ethyl 4-{1-[3-(3,5-dichlorophenyl)-5-{[(trifluoromethyl)-
sulfonyl]oxy}-1H-pyrazol-1-yl]ethyl}benzoate
(7).
Ethyl
4-{1-[3-(3,5-dichlorophenyl)-5-oxo-4,5-dihydro-1H-pyrazol-1-yl]
ethyl}benzoate (2.6 g, 6.4 mmol) and TEA (1.9 g, 19.2 mmol) were
dissolved in THF (100 mL) at ꢀ78 °C. Triflic anhydride (2.7 g,
9.6 mmol) was added. The reaction mixture was stirred for 3 h.
The reaction was quenched by adding ethyl acetate and water.
The organic layer was washed with brine, dried over Na2SO4 and
concentrated. The residue was purified by flash column
chromatography to afford ethyl 4-{1-[3-(3,5-dichlorophenyl)-
5-{[(trifluoromethyl)sulfonyl]oxy}-1H-pyrazol-1-yl]ethyl}benzoate
(2.5 g, 72.5%) as a colorless oil. 1H NMR (400 MHz, CDCl3) d 8.02 (d,
J = 8.3 Hz, 2H), 7.70–7.66 (m, 2H), 7.40–7.33 (m, 3H), 6.43 (s, 1H),
5.55 (q, J = 7.1 Hz, 1H), 4.36 (q, J = 7.1 Hz, 2H), 1.98 (d, J = 7.0 Hz,
3H), 1.37 (t, J = 7.1 Hz, 3H). MS (ESI, m/z): 536.3 [MꢀH]ꢀ.
5.3.1.2.
hydrazine carboxylate (3).
tert-Butyl
2-{1-[4-(ethoxycarbonyl)-phenyl]ethyl}
A solution of tert-butyl carbazate
(5.5 g, 41.6 mmol) and ethyl 4-acetylbenzoate (10 g, 52.0 mmol) in
toluene (60 mL) with catalytic HOAc was stirred at 80 °C overnight.
tert-Butyl-2-{1-[4-(ethoxycarbonyl)phenyl]-ethylidene}hydrazine
carboxylate separated as a crystalline solid (10 g, 65.7%) and was
collected by filtration. NaBH3CN (2.4 g, 38.2 mmol) and tert-butyl
2-{1-[4-(ethoxycarbonyl)phenyl]ethylidene}-hydrazinecarboxylate
(10 g, 32.6 mmol) were dissolved in THF (100 mL). A solution
of p-toluene sulfonic acid (4.3 g, 22.6 mmol) in THF (25 mL) was
slowly added. After stirring the reaction for 3 h, the mixture was
extracted with EtOAc and washed with brine, dried (Na2SO4), and
concentrated to give a white solid. The solid was separated and
washed with 1 N HCl twice and brine twice, dried (Na2SO4), and
concentrated. Product precipitated as white solid and was washed
with petroleum ether/ethyl acetate (4:1) to yield 5.6 g (55.6%) of
tert-butyl 2-{1-[4-(ethoxycarbonyl)-phenyl]ethyl}hydrazine car-
boxylate. 1H NMR (400 MHz, CDCl3) d 8.01 (d, J = 8.3 Hz, 2H),
7.41 (d, J = 8.3 Hz, 2H), 4.37 (q, J = 7.1 Hz, 2H), 4.22 (d, J = 6.4 Hz,
1H), 1.41 (d, J = 2.7 Hz, 9H), 1.38 (d, J = 7.1 Hz, 3H), 1.35 (d,
J = 6.6 Hz, 3H). MS (ESI, m/z): 307.1 [MꢀH]ꢀ.
5.3.1.7. 4-{1-[3-(3,5-Dichlorophenyl)-5-[4-(tert-butyl)phenyl]-
1H-pyrazol-1-yl]ethyl}benzoate (8).
Ethyl 4-{1-[3-(3,5-
dichlorophenyl)-5-{[(trifluoromethyl)sulfonyl]oxy}-1H-pyrazol-1-
yl]ethyl}benzoate (1.0 g, 1.9 mmol), 4-(tert-butyl)phenyl boronic
acid (430.9 mg, 2.4 mmol), and TEA (1.0 g, 10.2 mmol) were dis-
solved in dimethoxyethane. The catalyst Pd(PPh3)4 (129.2 mg,
0.1 mmol) was added, and the mixture was deoxygenated before
refluxed in 85 °C for 2 h. The mixture was extracted with EtOAc,
washed with saturated brine, dried (Na2SO4), and concentrated.
The residue was purified by chromatography to afford ethyl 4-{1-
[3-(3,5-dichlorophenyl)-5-[4-(tert-butyl)phenyl]-1H-pyrazol-1-yl]
ethyl}benzoate as a colorless oil (632.8 mg, 65.2%). 1H NMR
(400 MHz, CDCl3) d 8.00 (d, J = 8.5 Hz, 2H), 7.77 (t, J = 3.0 Hz, 2H),
7.48–7.40 (m, 2H), 7.31–7.24 (m, 3H), 7.22–7.16 (m, 2H), 6.56 (d,
J = 4.1 Hz, 1H), 5.58 (q, J = 6.9 Hz, 1H), 4.41–4.32 (m, 2H), 1.95 (d,
J = 7.0 Hz, 3H), 1.41–1.34 (m, 12H). MS (ESI, m/z): 518.8 [MꢀH]ꢀ.
5.3.1.3. {1-[4-(Ethoxycarbonyl)phenyl]ethyl}hydrazinium-tri-
5.3.1.8. 3-[4-{1-[3-(3,5-Dichlorophenyl)-5-[4-(tert-butyl)phenyl]-
fluoroacetate (4).
A solution of tert-butyl 2-{1-[4-(ethoxycar-
1H-pyrazol-1-yl]ethyl}-benzamido]butanoic acid (9a).
Ethyl
bonyl)-phenyl]ethyl}hydrazine carboxylate in DCM (10 mL) was
treated with TFA (10 mL) at room temperature for 1 h. The mixture
was concentrated under reduced pressure without further purifica-
tion. MS (ESI, m/z): 209.0 [M+H]+.
4-{1-[3-(3,5-dichlorophenyl)-5-[4-(tert-butyl)phenyl]-1H-pyrazol-
1-yl]ethyl}benzoate (632.8 mg, 1.2 mmol) was dissolved in MeOH–
dioxane (1:1). A solution of NaOH (0.7 g/15 mL) was added. The
mixture was heated to 60 °C for 1 h. This was acidified with 1 N HCl,
extracted with EtOAc, washed with saturated brine, dried (Na2SO4),
and concentrated to give a yellow solid. This solid was suspended in
DMF, followed with addition of DIEA (2.4 mL), methyl 3-aminobu-
tanoate (569.6 mg, 4.9 mmol). A solution of benzotriazol-1-yl-
oxytripyrrolidino-phosphonium hexafluorophosphate (PyBOP)
(760.0 mg, 1.5 mmol) in DMF was then added. After stirring at room
temperature for 3 h, more PyBOP (380.0 mg, 0.75 mmol) was added,
and the reaction mixture was stirred overnight. After addition of
water (5 mL), the mixture was heated to 60 °C for 30 min. Ethyl acet-
ate was added, and the organic layer was washed with 0.5 N HCl
twice, 5% K2CO3 twice, and brine twice. Evaporation of solvent gave
an oily residue. The oily residue was dissolved in MeOH–dioxane
(1:1). A solution of NaOH (0.7 g/15 mL) was added. The mixture
was heated to 60 °C for 1 h. This was acidified with 1 N HCl, extracted
with EtOAc, washed with saturated brine, dried (Na2SO4), and
concentrated to give a yellow solid, which after flash column
5.3.1.4. 3-(3,5-Dichlorophenyl)-3-oxopropanoate (5).
Ethyl
3,5-dichlorobenzoate (5 g, 22.8 mmol) and sodium hydride
(1.10 g, 27.4 mmol) were dissolved in 35 mL THF under N2. Then
the solution was added 7.8 mL ethyl acetate slowly at room tem-
perature and refluxed overnight. The solvent was removed under
reduced pressure. The residue was extracted with ethyl acetate,
washed with water and brine, and dried over Na2SO4. Flash column
chromatography gave 5.5 g (92%) of 3-(3,5-dichlorophenyl)-3-oxo-
propanoate 5 as orange oil. 1H NMR (400 MHz, DMSO-d6) d 7.98–
7.91 (m, 3H), 4.27 (s, 2H), 4.12 (q, J = 7.1 Hz, 2H), 1.18 (t,
J = 7.1 Hz, 3H). MS (ESI, m/z): 262.0 [M+H]+.
5.3.1.5. Ethyl 4-{1-[3-(3,5-dichlorophenyl)-5-oxo-4,5-dihydro-
1H-pyrazol-1-yl]ethyl}benzoate (6).
(3.0 g, 11.6 mmol) and {1-[4-(ethoxycarbonyl)phenyl]ethyl}
hydrazinium trifluoroacetate (2.2 g, 10.6 mmol) was refluxed in
A solution of compound
5

S. Shu et al. / Bioorg. Med. Chem. 24 (2016) 2852–2863
2859
chromatography afforded 526.5 mg (75.0%) of 3-[4-{1-[3-(3,5-
dichlorophenyl)-5-[4-(tert-butyl)phenyl]-1H-pyrazol-1-yl]ethyl}-
benzamido]butanoic acid as a white solid. Mp 112–115 °C. 1H NMR
(400 MHz, CDCl3) d 7.73 (dd, J = 17.5, 4.9 Hz, 4H), 7.43 (d, J = 8.2 Hz,
2H), 7.28 (d, J = 1.9 Hz, 2H), 7.19 (d, J = 8.3 Hz, 2H), 6.76 (d,
J = 8.4 Hz, 1H), 6.55 (s, 1H), 5.56 (d, J = 7.0 Hz, 1H), 4.55 (s, 1H), 2.68
(s, 2H), 1.92 (d, J = 6.9 Hz, 3H), 1.35 (s, 12H). 13C NMR (125 MHz,
CDCl3) d 166.1, 151.7, 147.7, 145.9, 145.2, 136.2, 134.6, 132.9, 128.3,
126.9, 126.8, 126.6, 126.1, 125.2, 123.5, 103.2, 57.1, 41.9, 39.2, 34.3,
30.8, 21.8, 19.5. LRMS (ESI, m/z): 578.0 [M+H]+. HRMS (ESI, m/z): calcd
for C32H34O3N3Cl2, 578.1972 [M+H]+; found 578.1961, purity: 97.3%.
general procedure for the preparation of compounds 9. Mp
131–133 °C. 1H NMR (400 MHz, DMSO-d6)
8.47 (s, 1H),
d
7.96–7.78 (m, 4H), 7.72 (d, J = 8.4 Hz, 2H), 7.56 (d, J = 1.8 Hz, 1H),
7.46–7.34 (m, 2H), 7.25–7.13 (m, 4H), 5.83–5.67 (m, 1H), 3.88
(s, 3H), 3.43–3.37 (m, 1H), 3.23 (dd, J = 13.7, 6.7 Hz, 1H), 2.56
(d, J = 7.0 Hz, 1H), 1.90 (d, J = 6.9 Hz, 3H), 1.02 (d, J = 7.0 Hz, 3H).
13C NMR (125 MHz, CDCl3) d 167.4, 158.5, 148.2, 146.4, 145.8,
136.7, 135.2, 134.5, 129.7, 128.4, 128.4, 127.4, 127.3, 126.9,
126.6, 125.1, 124.0, 119.9, 105.7, 104.0, 57.8, 55.4,
31.6, 22.4, 14.2. LRMS (ESI, m/z): 602.0 [M+H]+. HRMS (ESI, m/z):
calcd for
purity: 98.2%.
C
33H30O4N3Cl2, 602.1608 [M+H]+; found 602.1599,
5.3.1.9. 3-[4-{1-[3-(3,5-Dichlorophenyl)-5-(2,3-dihydrobenzofu-
ran-5-yl)-1H-pyrazol-1-yl]ethyl}-benzamido]butanoic
acid
5.3.1.13.
1-[4-{1-[3-(3,5-Dichlorophenyl)-5-(6-methoxynaph-
(9b). 3-[4-{1-[3-(3,5-Dichlorophenyl)-5-(2,3-dihydrobenzo-
thalen-2-yl)-1H-pyrazol-1-yl]ethyl}-benzoyl]pyrrolidine-3-car-
furan-5-yl)-1H-pyrazol-1-yl]ethyl}-benzamido]butanoic acid (9b)
was prepared as 9a according to the general procedure for the
preparation of compounds 9. Mp 114–116 °C. 1H NMR (400 MHz,
CDCl3) d 7.72 (dd, J = 21.0, 5.1 Hz, 4H), 7.27 (t, J = 1.9 Hz, 1H),
7.24 (s, 1H), 7.04 (s, 1H), 7.00–6.93 (m, 1H), 6.82–6.74 (m, 2H),
6.51 (s, 1H), 5.51 (q, J = 6.9 Hz, 1H), 4.63 (t, J = 8.7 Hz, 2H), 4.54
(d, J = 6.8 Hz, 1H), 3.20 (dd, J = 20.7, 12.1 Hz, 2H), 2.70–2.64 (m,
2H), 1.91 (d, J = 7.0 Hz, 3H), 1.34 (dd, J = 6.8, 1.8 Hz, 3H). 13C NMR
(125 MHz, CDCl3) d 174.2, 166.0, 160.4, 147.5, 145.8, 145.4,
136.1, 134.6, 132.9, 128.8, 127.3, 126.8, 126.1, 125.4, 123.5,
121.6, 109.0, 103.3, 71.1, 57.1, 41.8, 39.0, 29.0, 21.7, 19.6. LRMS
(ESI, m/z): 564.1 [M+H]+. HRMS (ESI, m/z): calcd for C30H28O4N3Cl2,
564.1451 [M+H]+; found 564.1455, purity: 98.0%.
boxylic
acid
(9f).
1-[4-{1-[3-(3,5-Dichlorophenyl)-5-(6-
methoxynaphthalen-2-yl)-1H-pyrazol-1-yl]ethyl}-benzoyl]pyrro-
lidine-3-carboxylic acid (9f) was prepared as 9a according to the
general procedure for the preparation of compounds 9. Mp 123–
125 °C. 1H NMR (400 MHz, CDCl3) d 7.78 (dd, J = 12.0, 5.1 Hz, 3H),
7.70–7.63 (m, 2H), 7.46 (d, J = 8.0 Hz, 2H), 7.33–7.28 (m, 2H),
7.26–7.15 (m, 4H), 6.65 (s, 1H), 5.58 (q, J = 6.9 Hz, 1H), 3.95 (s,
3H), 3.81–3.43 (m, 3H), 3.14 (d, J = 40.7 Hz, 1H), 2.22 (dd, J = 24.7,
16.8 Hz, 2H), 1.94 (d, J = 7.0 Hz, 3H). 13C NMR (125 MHz, CDCl3) d
169.1, 158.1, 147.5, 145.6, 144.1, 135.7, 134.9, 134.7, 134.1,
129.2, 128.0, 127.9, 127.2, 127.1, 126.9, 126.4, 126.0, 124.4,
123.7, 119.4, 105.2, 103.8, 57.7, 55.0, 29.2, 21.8. LRMS (ESI, m/z):
614.0 [M+H]+. HRMS (ESI, m/z): calcd for C34H29O4N3Cl2Na,
636.1427 [M+Na]+; found 636.1428, purity: 99.7%.
5.3.1.10.
thalen-2-yl)-1H-pyrazol-1-yl]ethyl}-benzamido]butanoic acid
(9c). 3-[4-{1-[3-(3,5-Dichlorophenyl)-5-(6-methoxynaph-
3-[4-{1-[3-(3,5-Dichlorophenyl)-5-(6-methoxynaph-
5.3.1.14.
thalen-2-yl)-1H-pyrazol-1-yl]ethyl}-benzoyl]piperidine-3-car-
boxylic acid (9g). 1-[4-{1-[3-(3,5-Dichlorophenyl)-5-(6-
1-[4-{1-[3-(3,5-Dichlorophenyl)-5-(6-methoxynaph-
thalen-2-yl)-1H-pyrazol-1-yl]ethyl}-benzamido]butanoic acid (9c)
was prepared as 9a according to the general procedure for the
preparation of compounds 9. Mp 134–136 °C. 1H NMR (400 MHz,
CDCl3) d 7.80–7.58 (m, 7H), 7.30 (s, 1H), 7.25–7.12 (m, 4H), 6.84–
6.54 (m, 2H), 5.58 (s, 1H), 4.55 (s, 1H), 3.95 (d, J = 8.9 Hz, 3H),
2.65 (s, 2H), 2.03–1.79 (m, 3H), 1.41–1.28 (m, 3H). 13C NMR
(125 MHz, CDCl3) d 174.6, 166.1, 158.1, 147.7, 145.8, 145.4,
136.1, 134.7, 134.0, 133.0, 129.2, 127.9, 126.9, 126.8, 126.4,
126.1, 124.5, 123.6, 119.4, 105.2, 103.6, 57.5, 54.9, 41.8, 39.0,
21.8, 19.5. LRMS (ESI, m/z): 602.1 [M+H]+. HRMS (ESI, m/z): calcd for
C33H30O4N3Cl2, 602.1608 [M+H]+; found 602.1600, purity: 97.5%.
methoxynaphthalen-2-yl)-1H-pyrazol-1-yl]ethyl}-benzoyl]piperi-
dine-3-carboxylic acid (9g) was prepared as 9a according to
the general procedure for the preparation of compounds 9. Mp
128–131 °C. 1H NMR (400 MHz, CDCl3) d 7.78 (dd, J = 11.9, 5.1 Hz,
3H), 7.73–7.63 (m, 2H), 7.32 (dd, J = 17.1, 7.2 Hz, 4H), 7.25 (d,
J = 3.7 Hz, 2H), 7.22–7.15 (m, 2H), 6.65 (s, 1H), 5.58 (q, J = 6.9 Hz,
1H), 4.77–4.06 (m, 1H), 3.95 (s, 3H), 3.64 (s, 1H), 3.21 (s, 1H),
3.05 (s, 1H), 2.61 (s, 1H), 2.13 (d, J = 9.1 Hz, 1H), 1.94
(d, J = 7.0 Hz, 3H), 1.75 (dd, J = 21.7, 10.6 Hz, 2H), 1.49 (s, 1H).
13C NMR (125 MHz, CDCl3) d 177.9, 169.7, 158.1, 147.6, 145.4,
143.9, 136.2, 134.68, 134.4, 134.0, 129.2, 127.9, 126.8, 126.8,
126.4, 126.1, 124.6, 123.6, 119.3, 105.2, 103.5, 57.5, 54.9, 40.0,
22.0. LRMS (ESI, m/z): 628.1 [M+H]+. HRMS (ESI, m/z): calcd for
5.3.1.11.
thalen-2-yl)-1H-pyrazol-1-yl]ethyl}-benzamido]-3-phenyl-
propanoic acid (9d). 3-[4-{1-[3-(3,5-Dichlorophenyl)-5-(6-
3-[4-{1-[3-(3,5-Dichlorophenyl)-5-(6-methoxynaph-
C
35H31O4N3Cl2Na, 650.1584 [M+Na]+; found 650.1589, purity:
methoxynaphthalen-2-yl)-1H-pyrazol-1-yl]ethyl}benzamido]-3-
phenylpropanoic acid (9d) was prepared as 9a according to the
general procedure for the preparation of compounds 9. Mp 123–
125 °C. 1H NMR (400 MHz, CDCl3) d 7.80–7.60 (m, 8H), 7.30 (d,
J = 15.7 Hz, 7H), 7.21–7.13 (m, 3H), 6.63 (s, 1H), 5.60 (dd, J = 14.3,
7.3 Hz, 2H), 3.94 (s, 3H), 3.13–2.88 (m, 2H), 1.93 (d, J = 6.9 Hz,
3H). 13C NMR (125 MHz, CDCl3) d 174.5, 166.5, 158.6, 148.2,
146.5, 145.9, 140.1, 136.5, 135.2, 134.5, 133.2, 129.7, 128.9,
128.4, 127.9, 127.5, 127.4, 127.3, 126.9, 126.7, 126.3, 125.0,
124.1, 119.9, 105.7, 104.1, 58.0, 55.5, 49.8, 39.3, 22.3. LRMS (ESI,
m/z): 664.0 [M+H]+. HRMS (ESI, m/z): calcd for C38H32O4N3Cl2,
664.1764 [M+H]+; found 664.1758, purity: 98.4%.
97.5%.
5.3.1.15.
1-[4-{1-[3-(3,5-Dichlorophenyl)-5-(6-methoxynaph-
thalen-2-yl)-1H-pyrazol-1-yl]ethyl}-benzoyl]piperidine-4-car-
boxylic acid (9h). 1-[4-{1-[3-(3,5-Dichlorophenyl)-5-(6-
methoxynaphthalen-2-yl)-1H-pyrazol-1-yl]ethyl}-benzoyl]piperi-
dine-4-carboxylic acid (9h) was prepared as 9a according to the
general procedure for the preparation of compounds 9. Mp 129–
130 °C. 1H NMR (400 MHz, CDCl3) d 7.78 (dd, J = 11.5, 5.2 Hz, 3H),
7.72–7.65 (m, 2H), 7.36–7.28 (m, 4H), 7.27 (s, 1H), 7.25 (s, 2H),
7.22–7.16 (m, 2H), 6.65 (s, 1H), 5.59 (q, J = 6.8 Hz, 1H), 3.95 (s,
3H), 3.77 (s, 1H), 3.06 (s, 2H), 2.68–2.56 (m, 1H), 2.04 (s, 1H),
1.94 (d, J = 7.0 Hz, 3H), 1.73 (s, 4H). 13C NMR (125 MHz, CDCl3) d
175.9, 170.0, 158.0, 147.6, 145.4, 144.0, 136.1, 134.7, 134.3,
134.0, 129.2, 127.9, 127.0, 126.9, 126.8, 126.5, 126.0, 124.6,
123.6, 119.3, 105.2, 103.6, 57.5, 54.9, 29.2, 26.9, 22.0. LRMS (ESI,
m/z): 628.0 [M+H]+. HRMS (ESI, m/z): calcd for C35H32O4N3Cl2,
628.1764 [M+H]+; found 628.1760, purity: 97.0%.
5.3.1.12.
thalen-2-yl)-1H-pyrazol-1-yl]ethyl}-benzamido]-2-methyl-
propanoic acid (9e). 3-[4-{1-[3-(3,5-Dichlorophenyl)-5-(6-
3-[4-{1-[3-(3,5-Dichlorophenyl)-5-(6-methoxynaph-
methoxynaphthalen-2-yl)-1H-pyrazol-1-yl]ethyl}-benzamido]-2-
methylpropanoic acid (9e) was prepared as 9a according to the

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S. Shu et al. / Bioorg. Med. Chem. 24 (2016) 2852–2863
5.3.2. General procedure for the preparation of compounds 17
5.3.2.1. Ethyl 1-indanone-5-carboxylate (11). A solution of
5.3.2.5. Ethyl 1-{3-(3,5-dichlorophenyl)-5-{[(trifluoromethyl)-
sulfonyl]oxy}-1H-pyrazol-1-yl}-2,3-dihydro-1H-indene-5-car-
1-indanone-5-carboxylic acid 10 (5.0 g, 28.4 mmol) in 30 mL anhy-
drous ethanol was added 3.00 mL sulfuric acid slowly at 0 °C. After
stirring at 80 °C for 4 h, the mixture was extracted with ethyl acet-
ate and washed sequentially with water and saturated sodium
bicarbonate and brine, dried over sodium sulfate, and concentrated
as white solid (5.3 g, 90%). The crude product ethyl 1-indanone-5-
carboxylate 11 was used without further purification. 1H NMR
(400 MHz, DMSO-d6) d 8.13 (s, 1H), 7.95 (d, J = 7.9 Hz, 1H), 7.73
(dd, J = 7.9, 2.5 Hz, 1H), 4.35 (qd, J = 7.1, 1.4 Hz, 2H), 3.22–3.09
(m, 3H), 2.75–2.63 (m, 2H), 1.34 (t, J = 7.1 Hz, 3H). MS (ESI, m/z)
205.1 [M+H]+.
boxylate (15).
Compound 14 (2.5 g, 6.0 mmol) and TEA
(2.5 mL, 17.9 mmol) were stirred in 50 mL anhydrous THF at
ꢀ78 °C. The trifluoromethanesulfonic anhydride (1.5 mL,
9.1 mmol) was added over half an hour. Then the temperature
was slowly rose to room temperature and the mixture was stirred
for 1 h. The reaction was quenched by adding ethyl acetate and
water. The organic layer was washed by 0.5 N HCl and brine, dried
over sodium sulfate. The flash chromatography (SiO2, 10% ethyl
acetate in petroleum ether) gave 2.3 g (70%) of ethyl 1-{3-(3,5-
dichlorophenyl)-5-{[(trifluoromethyl)sulfonyl]oxy}-1H-pyrazol-1-
yl}-2,3-dihydro-1H-indene-5-carboxylate 15 as pale yellow oil. 1H
NMR (400 MHz, CDCl3) d 8.03 (s, 1H), 7.90 (d, J = 8.0 Hz, 1H), 7.55
(d, J = 1.9 Hz, 2H), 7.28 (t, J = 1.9 Hz, 1H), 7.11 (d, J = 7.9 Hz, 1H),
6.46 (s, 1H), 5.88 (t, J = 7.5 Hz, 1H), 4.38 (q, J = 7.1 Hz, 2H), 3.37
(ddd, J = 16.1, 8.8, 4.4 Hz, 1H), 3.07 (dt, J = 16.0, 7.9 Hz, 1H), 2.85–
2.55 (m, 2H), 1.39 (t, J = 7.1 Hz, 3H). MS (EI, m/z) 572.1 [M+Na]+.
5.3.2.2. tert-Butyl 2-[5-(ethoxycarbonyl)-2,3-dihydro-1H-inden-
1-yl]-hydrazin-1-carboxylate (12).
A solution of compound
11 (5.0 g, 24.5 mmol) and tert-butyl carbazate (4.9 g, 36.7 mmol)
in 40 mL toluene was added catalytic acetic acid and was stirred
at 80 °C overnight. The tert-butyl-2-[5-(ethoxycarbonyl)-2,3-dihy-
dro-1H-inden-1-ylidene]hydrazine-1-carboxylate separated as a
crystalline solid and was collected by filtration (5.5 g, 17.3 mmol,
70%). Then the crude product and sodium cyanoborohydride
(1.2 g, 19.0 mmol) were dissolved in 30 mL THF under N2 atmo-
sphere. A solution of p-toluene sulfonic acid (3.6 g, 20.7 mmol) in
10 mL THF was added dropwise. After stirring at room temperature
for 3 h, the mixture was diluted with ethyl acetate and washed by
saturated sodium bicarbonate, brine and dried over sodium sulfate,
concentrated under reduced pressure to afford tert-butyl 2-[5-
(ethoxycarbonyl)-2,3-dihydro-1H-inden-1-yl]-hydrazine-1-car-
boxylate 12 as white solid (5.1 g, 92%). 1H NMR (400 MHz, CDCl3) d
7.92–7.87 (m, 2H), 7.41 (d, J = 7.7 Hz, 1H), 6.08 (s, 1H), 4.54 (d,
J = 6.0 Hz, 1H), 4.36 (q, J = 7.1 Hz, 2H), 3.13–3.01 (m, 1H), 2.91–
2.80 (m, 1H), 2.29 (dd, J = 14.4, 7.5 Hz, 1H), 2.06–1.95 (m, 1H),
1.54 (s, 1H), 1.46 (s, 9H), 1.38 (t, J = 7.1 Hz, 3H). MS (ESI, m/z)
319.2 [MꢀH]ꢀ.
5.3.2.6. Ethyl 1-[3-(3,5-dichlorophenyl)-5-(6-methoxynaph-
thalen-2-yl)-1H-pyrazol-1-yl]-2,3-dihydro-1H-indene-5-car-
boxylate (16).
Compound 15 (181.3 mg, 0.3 mmol), TEA
(233.3 L, 1.7 mmol), and 6-methoxy-2-naphthylboronic acid
l
(84.7 mg, 0.4 mmol) were dissolved in 6 mL dimethoxyethane
and deoxygenated by vacuum-N2 fill cycles. Tetrakis(triph-
enylphosphine) palladium [Pd(PPh3)4] (38.1 mg, 10% mmol) was
add quickly as catalyst and deoxygenated again before heated in
microwave reactor at 100 °C for 25 min. The mixture was
quenched by saturated ammonium chloride, extracted by ethyl
acetate, and washed by brine, dried over sodium sulfate. The flash
chromatography (SiO2, 10% ethyl acetate in petroleum ether) gave
166 mg (90%) of ethyl 1-[3-(3,5-dichlorophenyl)-5-(6-methoxy-
naphthalen-2-yl)-1H-pyrazol-1-yl]-2,3-dihydro-1H-indene-5-car-
boxylate 16 as colorless oil. 1H NMR (400 MHz, DMSO-d6) d 8.10 (s,
1H), 7.98 (dd, J = 19.0, 8.8 Hz, 2H), 7.89 (s, 1H), 7.79 (d, J = 1.9 Hz,
2H), 7.75 (d, J = 7.9 Hz, 1H), 7.68 (dd, J = 8.5, 1.6 Hz, 1H), 7.52 (t,
J = 1.9 Hz, 1H), 7.43 (d, J = 2.5 Hz, 1H), 7.26 (dd, J = 9.0, 2.5 Hz,
1H), 7.19 (s, 1H), 7.04 (d, J = 8.0 Hz, 1H), 6.06 (t, J = 7.9 Hz, 1H),
4.29 (q, J = 7.1 Hz, 2H), 3.91 (s, 3H), 3.23 (dt, J = 16.1, 6.3 Hz, 1H),
2.98 (dt, J = 16.1, 8.3 Hz, 1H), 2.76–2.64 (m, 2H), 1.30 (t,
J = 7.1 Hz, 3H). MS (ESI, m/z) 558.8 [M+H]+.
5.3.2.3. Ethyl 1-hydrazinyl-2,3-dihydro-1H-indene-5-carboxy
late (13).
Compound 12 (5.1 g, 15.9 mmol) was treated with
40 mL of TFA/DCM (1:1) at room temperature for 2 h. The mixture
was concentrated under reduced pressure to provide crude pro-
duct hydrazine 13 (2.6 g, 75%) as colorless oil. 1H NMR (400 MHz,
CDCl3) d 8.02–7.88 (m, 2H), 7.56 (d, J = 8.1 Hz, 1H), 4.95–4.84 (m,
1H), 4.38 (q, J = 7.1 Hz, 2H), 3.16 (dt, J = 25.6, 8.4 Hz, 1H), 3.09–
2.84 (m, 1H), 2.56 (dd, J = 15.4, 7.8 Hz, 1H), 2.50–2.33 (m, 1H),
1.40 (t, J = 6.7 Hz, 3H). MS (ESI, m/z) 221.2 [M+H]+.
5.3.2.7. 3-{1-[3-(3,5-Dichlorophenyl)-5-(6-methoxynaphthalen-
2-yl)-1H-pyrazol-1-yl]-2,3-dihydro-1H-indene-5-carboxamido}
propanoic acid (17l).
Compound 16 (166 mg, 0.3 mmol) was
dissolved in MeOH–dioxane (1:1, 3 mL). A solution of NaOH
(0.7 g/15 mL) was added at room temperature. After stirring at
60 °C for 1 h, the mixture was acidified by 2 N HCl and extracted
by DCM, dried over sodium sulfate and concentrated under reduced
pressure to give 150 mg (95%) of 1-[3-(3,5-dichlorophenyl)-5-
(6-methoxynaphthalen-2-yl)-1H-pyrazol-1-yl]-2,3-dihydro-1H-
indene-5-carboxylic acid as pale yellow solid. The solid was directly
dissolved in DMF (3 mL), followed by addition of DIEA (0.2 mL,
1.3 mmol), b-alanine tert-butyl ester hydrochloride (154.4 mg,
0.9 mmol) and a solution of PyBOP (165.1 mg, 0.3 mmol) in 2 mL
DMF. After stirring at room temperature for 3 h, 53 mg PyBOP was
added directly. The mixture was stirred at room temperature over-
night and quenched by water before heated to 60 °C for 30 min.
Ethyl acetate was added, the organic layer was washed sequentially
by 0.5 N HCl, 5% potassium carbonate, brine and then dried over
sodium sulfate. The flash chromatography (SiO2, 25% ethyl acetate
in petroleum ether) gave 149 mg (80%) of tert-butyl 3-{1-[3-(3,5-
dichlorophenyl)-5-(6-methoxynaphthalen-2-yl)-1H-pyrazol-1-yl]-
2,3-dihydro-1H-indene-5-carboxamido}propanoate as white solid.
5.3.2.4. Ethyl 1-[3-(3,5-dichlorophenyl)-5-oxo-4,5-dihydro-1H-
pyrazol-1-yl]-2,3-dihydro-1H-indene-5-carboxylate
(14).
Without further purification, hydrazine 13 (2.6 g,
11.9 mmol) and ethyl 3-(3,5-dichlorophenyl)-3-oxopropanoate 5
(3.4 g, 13.1 mmol) were refluxed in 80 mL acetic acid for 4 h. The
solvent was removed under reduced pressure, then redissolved in
ethyl acetate, washed sequentially by saturated sodium carbonate,
brine and dried over sodium sulfate. The flash chromatography
(SiO2, 10% to 20% ethyl acetate in petroleum ether) gave 2.5 g
(50%) of ethyl 1-[3-(3,5-dichlorophenyl)-5-oxo-4,5-dihydro-1H-
pyrazol-1-yl]-2,3-dihydro-1H-indene-5-carboxylate 14 as orange
solid. 1H NMR (400 MHz, CDCl3) d 7.98 (s, 1H), 7.88 (d, J = 7.9 Hz,
1H), 7.42 (d, J = 1.9 Hz, 2H), 7.33 (t, J = 1.9 Hz, 1H), 7.21 (d,
J = 7.9 Hz, 1H), 5.89 (t, J = 7.6 Hz, 1H), 4.36 (q, J = 7.1 Hz, 2H), 3.65
(d, J = 2.2 Hz, 2H), 3.26 (ddd, J = 16.0, 8.9, 4.5 Hz, 1H), 3.00 (dt,
J = 15.9, 7.9 Hz, 1H), 2.66–2.50 (m, 1H), 2.41 (ddt, J = 13.1, 8.9,
7.2 Hz, 1H), 1.38 (t, J = 7.1 Hz, 3H). MS (ESI, m/z) 415.1 [MꢀH]ꢀ.

S. Shu et al. / Bioorg. Med. Chem. 24 (2016) 2852–2863
2861
The solid was then treated with TFA–DCM (1:2, 3 mL) for 1 h. The
solvent was removed under reduced pressure. The flash chromatog-
raphy (SiO2, 5% MeOH in DCM) gave 130 mg (95%) of 3-{1-[3-(3,5-
dichlorophenyl)-5-(6-methoxynaphthalen-2-yl)-1H-pyrazol-1-yl]-
2,3-dihydro-1H-indene-5-carboxamido}propanoic acid 17l as
white solid. Mp 129–130 °C. 1H NMR (400 MHz, DMSO-d6) d 12.22
(s, 1H), 8.48 (t, J = 5.5 Hz, 1H), 8.11 (s, 1H), 7.98 (dd, J = 18.8,
8.8 Hz, 2H), 7.79 (d, J = 1.9 Hz, 2H), 7.76 (s, 1H), 7.68 (dd, J = 8.4,
1.9 Hz, 1H), 7.60 (d, J = 8.1 Hz, 1H), 7.52 (t, J = 1.9 Hz, 1H), 7.43 (d,
J = 2.5 Hz, 1H), 7.26 (dd, J = 9.0, 2.5 Hz, 1H), 7.19 (s, 1H), 6.98 (d,
J = 7.9 Hz, 1H), 6.04 (t, J = 7.7 Hz, 1H), 3.91 (s, 3H), 3.43 (q,
J = 6.7 Hz, 2H), 3.24–3.10 (m, 1H), 2.97 (d, J = 8.0 Hz, 1H), 2.74–
2.60 (m, 2H), 2.47 (t, J = 7.1 Hz, 2H). 13C NMR (125 MHz, DMSO-d6)
d 173.3, 166.7, 158.6, 147.9, 146.6, 145.9, 143.9, 137.1, 135.0,
134.9, 134.7, 130.4, 128.7, 128.6, 128.0, 127.4, 127.3, 126.4, 125.2,
124.1, 123.9, 123.8, 120.0, 106.4, 105.0, 63.3, 56.5, 55.8, 36.0, 34.2, 33.3,
30.4, 19.0. LRMS (ESI, m/z): 602.2 [M+H]+. HRMS (ESI, m/z): calcd
for C33H26N3O4Cl2, 598.1300 [M+H]+; found 598.1312, purity: 95.4%.
1.78–1.68 (m, 1H). 13C NMR (125 MHz, DMSO-d6) d 172.9, 166.2,
147.4, 146.0, 145.3, 143.4, 136.5, 134.5, 130.0, 129.1, 129.0,
126.9, 126.0, 123.7, 123.4, 123.3, 104.4, 62.7, 45.9, 35.6, 33.8,
32.7, 30.0, 25.9. LRMS (ESI, m/z): 519.1 [M+H]+. HRMS (ESI, m/z):
calcd for C28H22N3O3Cl2, 518.1038 [M+H]+; found 518.1047, purity:
98.2%.
5.3.2.11. 3-{1-[3-(3,5-Dichlorophenyl)-5-(o-tolyl)-1H-pyrazol-1-
yl]-2,3-dihydro-1H-indene-5-carboxamido}propanoic
acid
(17d). 3-{1-[3-(3,5-Dichlorophenyl)-5-(o-tolyl)-1H-pyrazol-
1-yl]-2,3-dihydro-1H-indene-5-carboxamido}propanoic acid (17d)
was prepared as 17l according to the general procedure for prepa-
ration of compounds 17. Mp 124–126 °C. 1H NMR (400 MHz,
DMSO-d6) d 12.20 (s, 1H), 8.47 (t, J = 5.5 Hz, 1H), 7.81–7.71 (m,
3H), 7.63–7.55 (m, 1H), 7.50 (t, J = 1.9 Hz, 1H), 7.48–7.41 (m, 3H),
7.40–7.31 (m, 1H), 7.02 (s, 1H), 6.92 (d, J = 7.9 Hz, 1H), 5.51 (t,
J = 7.7 Hz, 1H), 3.43 (q, J = 7.0 Hz, 2H), 3.18 (dt, J = 15.7, 6.6 Hz,
1H), 2.93 (dt, J = 16.0, 8.1 Hz, 1H), 2.65–2.54 (m, 2H), 2.48 (t,
J = 7.2 Hz, 2H), 2.27 (s, 3H). 13C NMR (125 MHz, DMSO-d6) d
172.9, 166.3, 147.5, 145.2, 144.5, 143.5, 137.3, 136.7, 134.6,
130.5, 129.6, 129.5, 126.9, 126.2, 126.0, 123.7, 123.4, 123.4,
104.5, 62.7, 35.6, 33.8, 32.7, 30.0, 19.8. LRMS (ESI, m/z): 535.2 [M
+H]+. HRMS (ESI, m/z): calcd for C29H24N3O3Cl2, 532.1195 [M
+H]+; found 532.1188, purity: 97.6%.
5.3.2.8. 3-{1-[3-(3,5-Dichlorophenyl)-5-(3,5-dimethoxyphenyl)-
1H-pyrazol-1-yl]-2,3-dihydro-1H-indene-5-carboxamido}pro-
panoic
acid
(17a).
3-{1-[3-(3,5-Dichlorophenyl)-5-(3,5-
dimethoxyphenyl)-1H-pyrazol-1-yl]-2,3-dihydro-1H-indene-5-
carboxamido}propanoic acid (17a) was prepared as 17l according
to the general procedure for preparation of compounds 17. Mp
118–120 °C. 1H NMR (400 MHz, DMSO-d6) d 12.23 (s, 1H), 8.48 (t,
J = 5.5 Hz, 1H), 7.77 (s, 1H), 7.75 (d, J = 2.0 Hz, 2H), 7.66–7.54 (m,
1H), 7.50 (t, J = 1.9 Hz, 1H), 7.11 (s, 1H), 6.97 (d, J = 7.9 Hz, 1H),
6.74 (d, J = 2.3 Hz, 2H), 6.65 (t, J = 2.2 Hz, 1H), 6.01 (t, J = 7.7 Hz,
1H), 3.81 (s, 6H), 3.44 (q, J = 6.8 Hz, 2H), 3.19 (m, 1H), 2.98 (m,
1H), 2.65 (m, 2H), 2.48 (t, J = 7.2 Hz, 2H). 13C NMR (125 MHz,
DMSO-d6) d 172.9, 166.2, 160.8, 147.4, 145.8, 145.4, 143.5, 136.5,
134.5, 134.5, 131.5, 126.9, 126.0, 123.7, 123.4, 123.3, 107.1,
104.4, 100.8, 62.9, 55.4, 35.60, 33.8, 32.8, 30.0. LRMS (ESI, m/z):
582.2 [M+H]+. HRMS (ESI, m/z): calcd for C30H26N3O5Cl2,
578.1250 [M+H]+; found 578.1234, purity: 99.3%.
5.3.2.12.
pyrazol-1-yl]-2,3-dihydro-1H-indene-5-carboxamido}propanoic
acid (17e). 3-{1-[3-(3,5-Dichlorophenyl)-5-(4-fluorophenyl)-
3-{1-[3-(3,5-Dichlorophenyl)-5-(4-fluorophenyl)-1H-
1H-pyrazol-1-yl]-2,3-dihydro-1H-indene-5-carboxamido}propanoic
acid (17e) was prepared as 17l according to the general procedure for
preparation of compounds 17. Mp 154–156 °C. 1H NMR (400 MHz,
DMSO-d6) d 12.23 (s, 1H), 8.49 (t, J = 5.5 Hz, 1H), 7.78 (s, 1H), 7.76
(d, J = 1.9 Hz, 2H), 7.73–7.65 (m, 2H), 7.63–7.58 (m, 1H), 7.51 (t,
J = 1.9 Hz, 1H), 7.47–7.36 (m, 2H), 7.12 (s, 1H), 6.95 (d, J = 8.0 Hz,
1H), 5.91 (t, J = 7.7 Hz, 1H), 3.44 (q, J = 7.0 Hz, 2H), 3.20 (m, 1H),
2.97 (dt, J = 16.0, 8.1 Hz, 1H), 2.71–2.59 (m, 2H), 2.48 (t, J = 7.2 Hz,
2H). 13C NMR (125 MHz, DMSO-d6) d 172.9, 166.3, 163.5, 161.6,
147.5, 145.3, 145.0, 143.5, 136.5, 134.6, 134.6, 131.5, 131.4, 127.0,
126.3, 126.2, 126.0, 123.7, 123.5, 123.4, 116.2, 116.1, 104.6, 62.8,
35.6, 33.8, 32.9, 30.0. LRMS (ESI, m/z): 540.0 [M+H]+. HRMS (ESI,
m/z): calcd for C28H21N3O3FCl2, 536.0944 [M+Na]+; found 536.0956,
purity: 97.5%.
5.3.2.9.
[1,4]dioxin-6-yl)-1H-pyrazol-1-yl]-2,3-dihydro-1H-indene-5-
carboxamido}propanoic acid (17b). 3-{1-[3-(3,5-Dichloro-
3-{1-[3-(3,5-Dichlorophenyl)-5-(2,3-dihydrobenzo[b]
phenyl)-5-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1H-pyrazol-1-
yl]-2,3-dihydro-1H-indene-5-carboxamido}propanoic acid (17b)
was prepared as 17l according to the general procedure for prepa-
ration of compounds 17. Mp 118–120 °C. 1H NMR (400 MHz,
DMSO-d6) d 12.23 (s, 1H), 8.48 (t, J = 5.5 Hz, 1H), 7.77 (s, 1H),
7.74 (d, J = 1.9 Hz, 2H), 7.60 (dd, J = 7.8, 1.6 Hz, 1H), 7.49 (t,
J = 1.9 Hz, 1H), 7.12–6.99 (m, 4H), 6.92 (d, J = 7.9 Hz, 1H), 5.93 (t,
J = 7.8 Hz, 1H), 4.30 (s, 4H), 3.44 (q, J = 7.0 Hz, 2H), 3.24–3.13 (m,
1H), 2.97 (dt, J = 16.0, 8.2 Hz, 1H), 2.70–2.58 (m, 2H), 2.48 (t,
J = 7.2 Hz, 2H). 13C NMR (125 MHz, DMSO-d6) d 172.9, 166.2,
147.3, 145.6, 145.4, 144.2, 143.6, 143.4, 136.6, 134.5, 126.8,
126.0, 123.7, 123.4, 123.2, 122.7, 122.0, 117.7, 104.1, 64.2, 64.1,
62.6, 35.6, 33.8, 32.8, 29.9. LRMS (ESI, m/z): 579.0 [M+H]+. HRMS
(ESI, m/z): calcd for C30H24N3O5Cl2, 576.1093 [M+H]+; found
576.1078, purity: 99.4%.
5.3.2.13.
pyrazol-1-yl]-2,3-dihydro-1H-indene-5-carboxamido}propanoic
acid (17f). 3-{1-[5-(4-Chlorophenyl)-3-(3,5-dichlorophenyl)-
3-{1-[5-(4-Chlorophenyl)-3-(3,5-dichlorophenyl)-1H-
1H-pyrazol-1-yl]-2,3-dihydro-1H-indene-5-carboxamido}propanoic
acid (17f) was prepared as 17l according to the general procedure for
preparation of compounds 17. Mp 169–171 °C. 1H NMR (400 MHz,
CDCl3) d 7.68 (s, 1H), 7.60 (d, J = 1.8 Hz, 2H), 7.53 (d, J = 7.9 Hz, 1H),
7.45 (dd, J = 23.3, 8.5 Hz, 4H), 7.22 (t, J = 1.8 Hz, 1H), 6.94 (d,
J = 7.7 Hz, 1H), 6.81 (t, J = 6.1 Hz, 1H), 6.59 (s, 1H), 5.84 (t, J = 7.9 Hz,
1H), 3.74–3.65 (m, 2H), 3.32–3.20 (m, 1H), 3.01–2.88 (m, 1H), 2.83–
2.74 (m, 1H), 2.67 (t, J = 5.7 Hz, 2H), 2.64–2.51 (m, 1H). 13C NMR
(125 MHz, CDCl3) d 175.5, 167.8, 149.0, 145.9, 145.1, 144.1, 136.3,
135.5, 135.2, 134.6, 130.5, 129.5, 128.8, 127.6, 126.0, 124.2, 124.0,
123.9, 104.1, 63.5, 35.4, 33.64, 33.4, 30.6. LRMS (ESI, m/z): 555.8 [M
+H]+. HRMS (ESI, m/z): calcd for C28H21N3O3Cl3, 552.0648 [M+H]+;
found 556.0754, purity: 94.5%.
5.3.2.10. 3-{1-[3-(3,5-Dichlorophenyl)-5-phenyl-1H-pyrazol-1-
yl]-2,3-dihydro-1H-indene-5-carboxamido}propanoic
acid
(17c). 3-{1-[3-(3,5-Dichlorophenyl)-5-phenyl-1H-pyrazol-1-
yl]-2,3-dihydro-1H-indene-5-carboxamido}propanoic acid (17c)
was prepared as 17l according to the general procedure for prepa-
ration of compounds 17. Mp 205–207 °C. 1H NMR (400 MHz,
DMSO-d6) d 12.24 (s, 1H), 8.49 (t, J = 5.3 Hz, 1H), 7.77 (s, 2H),
7.76 (s, 1H), 7.66–7.49 (m, 7H), 7.12 (s, 1H), 6.93 (d, J = 8.0 Hz,
1H), 5.94 (t, J = 7.7 Hz, 1H), 3.43 (q, J = 7.0 Hz, 2H), 3.20 (m, 1H),
3.03–2.97 (m, 1H), 2.73–2.60 (m, 2H), 2.47 (t, J = 7.1 Hz, 2H),
5.3.2.14.
zol-1-yl]-2,3-dihydro-1H-indene-5-carboxamido}propanoic acid
(17g). 3-{1-[3-(3,5-Dichlorophenyl)-5-(furan-2-yl)-1H-pyra-
zol-1-yl]-2,3-dihydro-1H-indene-5-carboxamido}propanoic acid
(17g) was prepared as 17l according to the general procedure
for preparation of compounds 17. Mp 128–130 °C. 1H NMR
3-{1-[3-(3,5-Dichlorophenyl)-5-(furan-2-yl)-1H-pyra-

2862
S. Shu et al. / Bioorg. Med. Chem. 24 (2016) 2852–2863
(400 MHz, DMSO-d6) d 12.25 (s, 1H), 8.49 (t, J = 5.1 Hz, 1H), 7.92 (d,
J = 1.8 Hz, 1H), 7.80 (s, 1H), 7.76 (d, J = 1.9 Hz, 2H), 7.61 (d,
J = 7.9 Hz, 1H), 7.51 (t, J = 1.9 Hz, 1H), 7.29 (s, 1H), 7.06–6.90 (m,
2H), 6.73 (dd, J = 3.4, 1.8 Hz, 1H), 6.30 (t, J = 7.3 Hz, 1H), 3.44 (d,
J = 6.6 Hz, 2H), 3.26 (dt, J = 8.3, 3.7 Hz, 1H), 3.03 (dt, J = 15.6,
7.6 Hz, 1H), 2.75–2.60 (m, 2H), 2.47 (t, J = 7.1 Hz, 2H). 13C NMR
(125 MHz, DMSO-d6) d 166.2, 147.5, 145.0, 144.2, 143.7, 143.2,
136.1, 135.8, 134.6, 134.6, 127.0, 126.0, 123.8, 123.6, 123.5,
112.0, 110.3, 103.5, 63.9, 35.6, 33.8, 32.4, 30.2. LRMS (ESI, m/z):
511.1 [M+H]+. HRMS (ESI, m/z): calcd for C26H20N3O4Cl2,
508.0831 [M+H]+; found 508.0816, purity: 97.4%.
acid (17k) was prepared as 17l according to the general procedure for
preparation of compounds 17. Mp 202–204 °C. 1H NMR (400 MHz,
DMSO-d6) d 12.26 (s, 1H), 8.48 (t, J = 5.4 Hz, 1H), 8.20 (s, 1H), 8.11
(d, J = 8.5 Hz, 1H), 8.09–7.99 (m, 2H), 7.87–7.71 (m, 4H), 7.66–7.58
(m, 3H), 7.51 (t, J = 1.9 Hz, 1H), 7.23 (s, 1H), 6.99 (d, J = 8.0 Hz, 1H),
6.06 (t, J = 7.7 Hz, 1H), 3.44 (q, J = 7.1 Hz, 2H), 3.21 (dt, J = 15.9,
6.3 Hz, 1H), 2.96 (dt, J = 16.0, 8.2 Hz, 1H), 2.78–2.67 (m, 2H), 2.49 (t,
J = 6.4 Hz, 2H). 13C NMR (125 MHz, DMSO-d6) d 172.9, 166.2, 147.5,
146.0, 145.4, 143.4, 136.5, 134.6, 134.5, 132.8, 132.7, 128.7, 128.3,
127.7, 127.2, 127.1, 126.9, 126.5, 126.0, 123.7, 123.5, 123.4, 104.8,
62.9, 62.2, 35.6, 33.8, 32.8, 30.0. LRMS (ESI, m/z): 571.1 [M+H]+. HRMS
(ESI, m/z): calcd for
C
32H24N3O3Cl2, 568.1195 [M+H]+; found
5.3.2.15.
pyrazol-1-yl]-2,3-dihydro-1H-indene-5-carboxamido}propanoic
acid (17h). 3-{1-[3-(3,5-Dichlorophenyl)-5-(thiophen-2-yl)-
3-{1-[3-(3,5-Dichlorophenyl)-5-(thiophen-2-yl)-1H-
568.1201, purity: 96.9%.
Acknowledgments
1H-pyrazol-1-yl]-2,3-dihydro-1H-indene-5-carboxamido}propanoic
acid (17h) was prepared as 17l according to the general procedure for
preparation of compounds 17. Mp 203–205 °C. 1H NMR (400 MHz,
DMSO-d6) d 12.24 (s, 1H), 8.50 (t, J = 5.5 Hz, 1H), 7.84–7.74 (m, 4H),
7.65–7.59 (m, 1H), 7.51 (t, J = 1.9 Hz, 1H), 7.47 (dd, J = 3.6, 1.2 Hz,
1H), 7.27 (dd, J = 5.1, 3.6 Hz, 1H), 7.22 (s, 1H), 6.97 (d, J = 7.9 Hz,
1H), 6.15 (t, J = 7.6 Hz, 1H), 3.44 (q, J = 6.7 Hz, 2H), 3.23 (ddd,
J = 15.9, 8.0, 4.7 Hz, 1H), 3.07–2.92 (m, 2H), 2.66 (m, 2H), 2.48 (t,
J = 7.2 Hz, 2H). 13C NMR (125 MHz, DMSO-d6) d 172.9, 166.3, 147.5,
145.1, 143.6, 138.6, 136.2, 134.6, 134.6, 129.6, 128.5, 128.4, 128.4,
127.1, 126.0, 123.8, 123.5, 123.5, 105.3, 63.0, 45.9, 35.6, 33.8, 32.7,
30.1, 25.9. LRMS (ESI, m/z): 525.0 [M+H]+. HRMS (ESI, m/z): calcd for
We gratefully acknowledge the financial support from the
National Natural Science Foundation of China (81220108025,
81302633, 2147220 and 91229204), SA-SIBS scholarship program,
the National Health and Family Planning Commission
(2012ZX09304-011, 2013ZX09401003-005, 2013ZX09507001,
2013ZX09507-002 and 2014ZX09507002-001), Shanghai Science
and Technology Development Fund (15DZ2291600) and the
Thousand Talents Program in China.
Supplementary data
C
26H20N3O3Cl2S, 524.0602 [M+H]+; found 524.0605, purity: 97.2%.
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.bmc.2016.04.053.
5.3.2.16. 3-{1-[5-([1,10-Biphenyl]-4-yl)-3-(3,5-dichlorophenyl)-
References and notes
1H-pyrazol-1-yl]-2,3-dihydro-1H-indene-5-carboxamido}pro-
panoic
acid
(17i).
3-{1-[5-([1,10-Biphenyl]-4-yl)-3-(3,5-
1. Saltiel, A. R. Cell 2001, 104, 517.
dichlorophenyl)-1H-pyrazol-1-yl]-2,3-dihydro-1H-indene-5-car-
boxamido}propanoic acid (17i) was prepared as 17l according to
the general procedure for preparation of compounds 17. Mp
142–144 °C. 1H NMR (400 MHz, DMSO-d6) d 8.49 (t, J = 5.5 Hz,
1H), 7.87 (d, J = 8.0 Hz, 2H), 7.83–7.68 (m, 7H), 7.62 (d, J = 8.0 Hz,
1H), 7.57–7.46 (m, 3H), 7.42 (d, J = 7.4 Hz, 1H), 7.16 (s, 1H), 6.97
(d, J = 8.0 Hz, 1H), 6.02 (t, J = 7.8 Hz, 1H), 3.44 (q, J = 6.9 Hz, 3H),
3.25–3.16 (m, 1H), 2.98 (dt, J = 15.6, 8.0 Hz, 1H), 2.68 (q,
J = 7.4 Hz, 2H), 2.49 (t, J = 7.2 Hz, 2H). 13C NMR (125 MHz, DMSO-
d6) d 172.9, 166.2, 147.5, 145.7, 145.4, 143.4, 140.7, 139.2, 136.5,
134.5, 129.6, 129.1, 128.7, 127.9, 127.3, 126.9, 126.8, 126.0,
123.7, 123.4, 123.3, 104.4, 62.9, 35.6, 33.8, 32.9, 29.9. LRMS (ESI,
m/z): 597.1 [M+H]+. HRMS (ESI, m/z): calcd for C34H26N3O3Cl2,
594.1351 [M+H]+; found 594.1352, purity: 97.4%.
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3-{1-[3-(3,5-Dichlorophenyl)-5-(naphthalen-1-yl)-
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acid (17j) was prepared as 17l according to the general procedure for
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C
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acid (17k).
3-{1-[3-(3,5-Dichlorophenyl)-5-(naphthalen-2-yl)-
1H-pyrazol-1-yl]-2,3-dihydro-1H-indene-5-carboxamido}propanoic

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