Design, synthesis, and evaluation of N-phenyl-4-(2-phenylsulfonamido)-benzamides as microtubule-targeting agents in drug-resistant cancer cells, displaying HDAC inhibitory response

Article abstract of DOI:10.1016/j.ejmech.2020.112158

Microtubule-targeting agents (MTA) have enjoyed significant clinical success for decades. However, several mechanisms may cause inactivation of such drugs, leading to acquired resistance in patients treated with them. Therefore, drugs containing a stilbene-like skeleton and possessing dual inhibitory activity may provide a new and differentiated treatment for patients to overcome challenging acquired resistance. A new compound (16c) displays promising anticancer activity with GI₅₀ of 22 ± 2 and 12 ± 0.1 nM in vincristine-resistant nasopharyngeal (KB-Vin) cancer cells and etoposide-resistant nasopharyngeal (KB-7D) cancer cells and is better than vincristine, etoposide, ABT-751, and MS-275. A mechanistic study revealed that 16c interferes with the cell cycle distribution and induces cell cycle arrest at the G2/M phase and severe mitotic spindle defects followed by apoptosis. In addition, it produces much more significant cytotoxicity than vincristine and etoposide in the corresponding resistant cells, indicating that it may be a promising candidate to overcome drug resistance in cancer cells. Compound 16c also displays inhibitory activity against HDAC 1 and HDAC 2 with IC₅₀ values of 1.07 μM, and 1.47 μM, respectively. These findings may lead to a new type of structural motif for future development of drugs that could overcome acquired resistance to MTAs.

Full text of DOI:10.1016/j.ejmech.2020.112158

Journal Pre-proof
Design, synthesis, and evaluation of N-phenyl-4-(2-phenylsulfonamido)-benzamides
as microtubule-targeting agents in drug-resistant cancer cells, displaying HDAC
inhibitory response
Wei-Cheng Wu, Yi-Min Liu, Mei-Hsiang Lin, Yu-Hsuan Liao, Mei-Jung Lai, Hsun-Yueh
Chuang, To-Yu Hung, Chun-Han Chen, Jing-Ping Liou
PII:
S0223-5234(20)30125-2
DOI:
https://doi.org/10.1016/j.ejmech.2020.112158
EJMECH 112158
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 28 November 2019
Revised Date: 17 February 2020
Accepted Date: 17 February 2020
Please cite this article as: W.-C. Wu, Y.-M. Liu, M.-H. Lin, Y.-H. Liao, M.-J. Lai, H.-Y. Chuang, T.-Y.
Hung, C.-H. Chen, J.-P. Liou, Design, synthesis, and evaluation of N-phenyl-4-(2-phenylsulfonamido)-
benzamides as microtubule-targeting agents in drug-resistant cancer cells, displaying HDAC
inhibitory response, European Journal of Medicinal Chemistry (2020), doi: https://doi.org/10.1016/
j.ejmech.2020.112158.
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Graphical Abstract

Revised manuscript for Article
Design, Synthesis, and Evaluation of
N-phenyl-4-(2-phenylsulfonamido)-benzamides as Microtubule-targeting agents
in Drug-resistant cancer cells, displaying HDAC inhibitory response
a,1
b,f,1
a
a
b
Wei-Cheng Wu, Yi-Min Liu, Mei-Hsiang Lin, Yu-Hsuan Liao, Mei-Jung Lai, Hsun-Yueh
a
a
*c,e
*,a,b,d
Chuang, To-Yu Hung, Chun-Han Chen, Jing-Ping Liou
1
Contributed equally to this work.
To whom correspondence should be addressed. For J. P. Liou: (Phone) 886-2-2736-1661 ext 6130;
*
(e-mail) jpl@tmu.edu.tw. For C. H. Chen: (Phone) 886-2-27361661 ext 3195, (e-mail)
brianchc@tmu.edu.tw
a
b
c
School of Pharmacy, College of Pharmacy, Taipei Medical University, Taiwan.
TMU Biomedical Commercialization Center, Taipei Medical University, Taiwan.
Department of Pharmacology, School of Medicine, College of Medicine, Taipei Medical University,
Taipei, Taiwan
d
School of Pharmacy, National Defense Medical Center, Taipei, Taiwan.
e
Cell Physiology and Molecular Image Research Center, Wan Fang Hospital, Taipei Medical
University, Taipei, Taiwan.
f
Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
1

Abstract
Microtubule-targeting agents (MTA) have enjoyed significant clinical success for decades. However,
several mechanisms may cause inactivation of such drugs, leading to acquired resistance in patients
treated with them. Therefore, drugs containing a stilbene-like skeleton and possessing dual inhibitory
activity may provide a new and differentiated treatment for patients to overcome challenging
acquired resistance. A new compound (16c) displays promising anticancer activity with GI of 22 ±
5
0
2
and 12 ± 0.1 nM in vincristine-resistant nasopharyngeal (KB-Vin) cancer cells and
etoposide-resistant nasopharyngeal (KB-7D) cancer cells and is better than vincristine, etoposide,
ABT-751, and MS-275. A mechanistic study revealed that 16c interferes with the cell cycle
distribution and induces cell cycle arrest at the G2/M phase and severe mitotic spindle defects
followed by apoptosis. In addition, it produces much more significant cytotoxicity than vincristine
and etoposide in the corresponding resistant cells, indicating that it may be a promising candidate to
overcome drug resistance in cancer cells. Compound 16c also displays inhibitory activity against
HDAC 1 and HDAC 2 with IC values of 1.07 µM, and 1.47 µM, respectively. These findings may
5
0
lead to a new type of structural motif for future development of drugs that could overcome acquired
resistance to MTAs.
2

1. Introduction
Numerous templates and structures have been identified through drug discovery as potent
1
inhibitors of tumor cells. Combination of two different mechanisms by the use of suitable scaffolds or
linkers in multi-target drug designs may be more efficacious than drugs that depend on a single
2
mechanism. Stilbene-based compounds, including (E)-stilbene (1), (Z)-stilbene (2) and
dihydrostilbene (3) which occur widely in nature products, have attracted the interest of biologists and
3
chemists. Resveratrol (4), a natural trihydroxy stilbene which was isolated from Veratrum
grandiflorum, has shown anti-inflammatory and anti-oxidative effects, and other promising
4
pharmacological properties relevant to various diseases. However, the major obstacle of resveratrol
5
-6
in translating its effects in patients was poor bioavailability due to its rapid metabolism.
7
Histone deacetylases (HDAC) are drug targets and have been highly explored for decades for their
significant roles in different human diseases. SAHA (5, vorinostat), PXD101 (6, belinostat) and
LBH-589 (7, panobinostat) all bearing a hydroxamic acid functional group, have been approved for
use in the treatment of cutaneous T cell lymphoma, peripheral T cell lymphoma and multiple
8
myeloma, respectively. In addition, benzamide containing HDAC inhibitors, such as MS-275 (8,
3

Entinostat) and MGCD0103 (9, Mocetinostat), have also been entered into clinical trials for treatment
8
of various cancers (Figure 1).
Microtubules are protein polymers which play important roles in several cellular processes, such as
cell division, cell migration and cell shape maintenance. Anti-mitotic agents, including taxans, vinca
alkaloids, and colchicine (10), achieve potent activity against cancer cells by interrupting cell cycle
9
-10
progression. However, drug resistance caused by different mechanisms has created a great amount
of variation between individual patients. The best-known mechanism is P-glycoprotein
(P-gp)-mediated multidrug resistance which provides an opportunity for tumor cells to escape cell
1
1-12
death induced by chemotherapy.
In an effort to develop more structurally diverse inhibitors
towards microtubules, many effective compounds from natural sources have been examined.
Combretastatin A-4 (11, CA-4), isolated from the South African tree Combretum caffrum, contains a
stilbene skeleton and three methoxy groups, binds to the colchicine-binding site and shows strong
cytotoxicity against a variety of types of cancer (Figure 1). Similar to combretastatin, ABT-751 (12,
E7010), an orally bioavailable sulfonamide, also binds to the same colchicine-binding site and shows a
9
,13
marked anticancer effect.
Notably, neither of these compounds is a transport-related substrate of
membrane-bound P-glycoprotein, and this suggests that stilbene could be used as the linker to combine
a benzamide with a sulfonamide to generate a series of compounds capable of inhibiting HDAC and
microtubules as a therapeutic strategy in drug-resistant nasopharyngeal cancer cells (Figure 2). Our
aim therefore, is to optimize an appropriate stilbene as a linker to combine multiple active binding
4

groups and generate a series of N-phenyl-4-(2-(phenylsulfonamido)phenethyl)benzamides and related
substituted compounds as shown in Figure 3.
2. Results and Discussion
2.1 Chemistry
Scheme 1 shows the synthetic routes to compounds 13a-13d. Commercially available
nitrobenzaldehydes
(19a-19c)
underwent
a
Wittig
reaction
with
(4-(methoxycarbonyl)benzyl)-triphenylphosphonium bromide to produce compounds 20a-20c, which
were then hydrogenated with a catalytic amount of 10% Pd/C to afford compounds 21a-21c. These
compounds were reacted with 4-methoxybenzensulfonyl chloride to produce 22a-22c which were
hydrolyzed by LiOH to afford 18a-18c. The acid products were reacted with o-phenylenediamine to
get target compounds 13a-13c. Compound 23c also reacted with NH OTHP to protect the amide
2
group, and this product was hydrolyzed by 10% TFA, yielding the designed compound 13d. The
synthesis of compounds 14a-14g is shown in Scheme 2. Various substituted benzenesulfonamide
groups were introduced into compound 21c to give compounds 24a-24g. Compounds 24a-24g were
hydrolyzed by LiOH to afford acid intermediates (25a-25g) which were then reacted with
o-phenylenediamine to get the target compounds 14a-14g. In addition, compound 23c was reacted
5

with various substituted anilines to produce the designed compounds 15a-15e, which are shown in
Scheme 3. Scheme 4 shows the synthesis of compound 16a from compound 20c. Compound 20c was
reduced by iron powder to get an amine product (26). Compound 16a was produced from 26 using the
similar reaction in Scheme 1.
Compounds 29a and 29b were synthesized from compounds 30a and 30b respectively, by a SN2
reaction, reduction and imine reduction as shown in Scheme 6. The acid compounds 31a and 31b were
produced by LiOH hydrolysis and then reacted with coupling reagents to give the designed compounds
16b and 16c, respectively. A biphenyl compound 16d was synthesized from compound 32 using
tetrakis(triphenylphosphine)palladium and phenylboronic acid under Suzuki coupling reaction
conditions, as described in Scheme 6. The biphenyl carboxylate was treated using a method similar to
that in Scheme 1 to obtain the desired target compound 16d. The target compound 17 without a
benzamide group was synthesized from compound 19c as shown in Scheme 7. The target compound
18, lacking a sulfonamide group, was produced from compound 39 as displayed in Scheme 8. Both of
these two designed compounds were afforded by synthetic routes similar to those in Scheme 1.
6

2
2
.2 Biological evaluation
.2.1 In vitro cell growth inhibitory activity
The synthetic compounds (13-18) were evaluated for their antiproliferative activity against human
nasopharyngeal cancer KB cells. Compound 17 and 18 were used to examine the importance of the
effect of sulfonamide and benzamide functional groups on the cytotoxicity. We found compound 18
lacking the sulfonamide group had diminished antiproliferative activity in an SRB assay (Table 1),
indicating that the sulfonamide group is required for anticancer activity against KB cells.
In order to investigate which of the HDAC inhibitors with various functional groups tends to
increase the antiproliferative activity toward cancer cells, we introduced a hydroxamic acid and a
benzamide group, giving compounds 13a-13d and examined the anticancer activity in KB cells. As
shown in Table 1, 13d failed to show obvious growth inhibition activity in KB cells, but 13c showed
significant growth inhibitory activity in KB cells, suggesting that the benzamide group may offer an
opportunity to improve the anticancer activity. In addition, the GI values of compounds 13a-13c
5
0
indicate the sulfonamide group at the ortho position of stilbene together with a benzamide may
provide the most potent activity against KB cells. The substituted functional group of the sulfonamide,
as in 13c was also considered. As shown in Table 1, removal or translocation of the methoxy group
7

(14a, 14b) or its replacement with electron-donating groups (14c-14f) resulted in decreased activity.
Further, the substituted groups of the benzamide show that one fluorine atom at the 5’-position has an
antiproliferative activity similar to that of 13c.
Variations in the stilbene linker, including a methylene linkage, direct bonding or substitution (16a
and 16d) have been explored and heteroatoms have also been introduced (16b and 16c). It was found
16a and 16d displayed decreased activity while 16c exhibited the best antiproliferative activity of
these compounds, indicating that nitrogen substitution of the linker has the most potent
antiproliferative activity against the KB cells.
2.2.2 In vitro cell growth inhibitory activity in drug-resistant KB cells
We selected several compounds (13c, 15c, 16b and 16c) with significant antiproliferative activities
against KB cells and tested their growth inhibitory activity in vincristine-resistant (KB-vin) and
etoposide-resistant (KB-7D) KB cancer cell lines. As shown in Table 2, the GI50 values of 12
(
ABT751) dramatically increased in KB-vin cells while 8 shows a similar cell growth inhibitory
activity compared to the parent KB cells shown in Table 1. The selected compounds (13c, 15c, 16b and
6c) also exhibit similar or better potency in inhibition of cell growth in two drug-resistant cancer
1
cells. In a clear result for instance, 15c shows improved anticancer activity in both KB-Vin and KB-7D
cells compared to KB cells. Among the four compounds tested, 16c exhibits the most potent cell
growth inhibitory activity in KB-Vin and KB-7D cells.
8

2.2.3 Inhibition of HDAC isoforms
In an attempt to evaluate the influence of compounds 13c, 15c, 16b and 16c on HDAC isoforms,
these compounds and the reference compound 8 (MS-275) were assayed for inhibitory activity against
HDAC class I isoforms (Table 3). We found all the synthesized compounds showed no inhibitory
activity against HDAC 8 while compounds 13c, 15c, and 16b selectively inhibit the activity of HDAC
3
. Compound 16c displays strong inhibitory activity with IC values of 1.07 µM, and 1.47 µM against
50
HDAC 1 and 2 respectively.
2.2.4 Cell death evaluation in drug-resistant cells
Based on the results in Table 1-3, we selected three compounds (13c, 15c, and 16c) to examine
multiple apoptosis markers in KB-vin and KB-7D cells. As shown in Figure 4, low concentrations of
16c (0.01 µM) induce significant activation of caspase-3, -8, -9, and PARP in KB-vin cells. In
addition, 16c shows better potency than 15c and 13c in activation of apoptotic death markers,
indicating 16c may be a candidate to overcome resistance to vincristine.
2.2.5 Evaluation of cell cycle progression and cell death response in drug-resistant KB cells
We performed flow cytometry and western blot analysis to investigate the effect of 16c on cell cycle
progression and cell death in comparison with 12 (ABT-751), 8 (MS-275), and vincristine or etoposide
in KB-vin and KB-7D cells. Compound 16c failed to generate much more significant cellular subG1
phase accumulation and severe apoptosis than the other drugs in both cell lines after 24 h treatment
9

(Figure 5A-5D). Although treatment with 0.5 µM 16c induces more significant subG1 elevation than
vincristine (Figure 5A), the activated expression levels of caspase-3, -8 and -9, PARP, and γH2AX
were almost identical in vincristine-treated cells (Figure 5C). In addition, the expression of γH2AX
was increased significantly in response to 16c and reference compounds (8, 12, and etoposide) in
KB-7D cells (Figure 5D). These results show that there was not much difference between 16c and
those reference compounds. However, 16c leads to more dramatic apoptotic cell death than the other
compounds after 48 h treatment (Figure 6A-6D). It is notable that treatment with 0.05 µM 16c induces
a much more significant subG1 population of cells (Figure 6A) and cell apoptotic biomarkers in
KB-vin cells (Figure 6C) than 0.1 µM vincristine. A similar effect can also be detected in KB-7D cells
(Figures 6B, 6D). These results confirm that 16c can effectively overcome resistance in both of
vincristine- and etoposide-resistant KB cells.
2.2.6 Compound 16c influences microtubule dynamics in cells
Paclitaxel and vincristine have been recognized as mitotic arrest inducers which induce stabilization or
destabilization of microtubules, respectively. We found that treatment with 16c triggers obvious G2/M
(mitotic) arrest between 18-24 h time points in KB-Vin and KB-7D cells (Figures 7A, 7B). In addition,
we observed using deconvolution fluorescence microscopy that paclitaxel-induced aggregation of
microtubule (round dots) and vincristine-induced abnormal spindles (reticular structure) spread in
cells after 24 h treatment. A similar effect of 16c and vincristine on tubulin polarization change was
detected (Figures 8A and 8B), suggesting 16c may have the same effect as vincristine on disruption of
1
0

microtubule dynamics. Further, we carried out intracellular tubulin polymerization assay to directly
determine the potency of inhibitory activity of three compounds (paclitaxel, vincristine, and 16c) on
blocking dynamics of tubulins in KB-Vin and KB-7D cells. As shown in Figure 9A and 9B,
paclitaxel showed significant induction of polymerized tubulin in cells. Compound 16c shows
significant inhibition of tubulin polymerization in KB-Vin and KB-7D cells. Notably, significant
decrease of the polymerized tubulin (P) was detected in cells treated with 0.1 µM of 16c, indicating
compound 16c is much more potent than vincristine in both cell lines. Taken together, our results
demonstrated a consistent outcome by using two different experiments to confirm 16c shows
promising activity in inhibiting tubulin polymerization in cells.
3. Conclusion
In this
paper
we
have
described
the
synthesis
of
a
series
of
N-phenyl-4-(2-(phenylsulfonamido)phen-ethyl)benzamides (13-18). Of all the synthesized
compounds, compound 16c was identified as a potent tubulin inhibitor demonstrating much better
cellular cytotoxicity than reference compounds 8 or 12 with a mean GI value against the KB cell line
5
0
of 10 nM. In addition, compound 16c also demonstrated over 100- and 40-fold antitumor efficacy
against KB-Vin and KB-7D cancer cells with mean GI values of 22 nM and 12 nM respectively. It
5
0
also displayed selective suppression of the function of class I HDAC isoforms. Although 16c did not
exhibit remarkable inhibitory activity against HDACs, its overcoming drug resistance in KB-Vin and
KB-7D cells may be caused by the presence of the benzamide functional group. This study provides a
1
1

lead compound for development of novel tubulin inhibitors with HDAC inhibition and may provide a
new strategy for the treatment of drug-resistance cancer cells.
1
2

4. Experimental section
4.1 Chemistry
1
13
Nuclear magnetic resonance ( H and C NMR) spectra were obtained with a Bruker DRX-500
spectrometer operating at 500 and 125 MHz and Bruker Fourier 300 and 75MHz. Chemical shifts are
reported in parts per million (ppm, δ) downfield from TMS as an internal standard. High-resolution
mass spectra (HRMS) were measured with an AB SCIE X (QSTAR® XL) High Resolution
Electrospray (ESI) Mass Spectrometry spectrometer. Melting points were measured with Buchi B-545
(Büchi, Switzerland). Purity of the final compounds was achieved with a Hitachi 2000 series HPLC
system using C-18 column (Agilent ZORBAX Eclipse XDB-C18 5 µm. 4.6 mm × 150 mm). Flash
column chromatography used silica gel: SILICYCLE (SilicaFlash Irregular Silica Gel P60, 40 - 63
µm, 60 Å (R12030B)
N-(2-Aminophenyl)-4-(4-((4-methoxyphenyl)sulfonamido)phenethyl)benzamide (13a)
A mixture of 23a (0.97 mmole), N,N,N’,N’-tetramethyl-O-(1H-benzotriazol-1-yl)uronium
hexafluorophosphate (HBTU, 1.26 mmole), N,N-diisopropylethylamine (DIPEA, 1.26 mmole),
o-phenylenediamine (1.16 mmole) and DMF (2 mL) was stirred at rt overnight. The reaction was
quenched with water and extracted with EtOAc. The residue was purified by flash chromatography
1
over silica gel to afford compound 13a in 56% yield. H-NMR (500 MHz, DMSOd ) δ (ppm):
6
2
.78-2.80 (m, 2H), 2.85-2.87 (m, 2H), 3.77 (s, 3H), 4.85 (s, 2H), 6.58 (t, J = 7.5Hz, 1H), 6.76 (d, J =
.5Hz, 1H), 6.93-6.97 (m 3H), 7.02-7.07 (m, 4H), 7.14 (d, J = 7.5Hz, 1H), 7.26 (d, J = 8Hz, 2H), 7.64
7
1
3
(
d, J = 8.5Hz, 2H), 7.85 (d, J = 8Hz, 2H), 9.56 (s, 1H) 9.98 (s, 1H). C-NMR (125 MHz, DMSO-d ) δ
6
1
3

(ppm): 36.43, 37.09, 56.08, 114.75, 116.61, 116.75, 120.80, 123.91, 126.87, 127.11, 128.19, 128.75,
1
1
29.31, 129.49, 131.77, 132.67, 136.25, 137.35, 143.57, 145.57, 162.78, 165.64. mp = 191.7–
+
92.4°C. HRMS (ESI) for C H N O S [M+H] : Calcd., 502.1795; Found, 502.1801.
2
8
28
3
4
N-(2-Aminophenyl)-4-(3-((4-methoxyphenyl)sulfonamido)phenethyl)benzamide (13b)
The title compound was obtained as a white solid in 54% yield from compound 23b in a manner
1
similar to that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.822.88
6
(m, 4H), 3.77 (s, 3H), 4.86 (s, 2H), 6.59 (t, J = 7.5Hz, 1H), 6.78 (d, J = 8Hz, 1H) 6.83-6.88 (m, 2H),
6
2
3
1
.95-6.99 (m, 2H), 7.04-7.10 (m, 3H), 7.16 (d, J = 8Hz, 1H), 7.26 (d, J = 8Hz, 2H), 7.67 (d, J = 8.5Hz,
1
3
H), 7.86 (d, J = 8Hz, 2H), 9.58 (s, 1H), 10.01 (s, 1H). C-NMR (125 MHz, DMSO-d ) δ (ppm):
6
6.85, 56.08, 114.76, 116.63, 116.76, 118.08, 120.38, 123.92, 124.56, 126.88, 127.12, 128.20, 128.73,
29.38, 131.73, 132.68, 138.47, 142.57, 143.58, 145.34, 162.83, 165.61. mp = 175.4–176.1°C. HRMS
+
(
ESI) for C H N O S [M+H] : Calcd., 502.1795; Found, 502.1802.
2
8
28
3
4
N-(2-Aminophenyl)-4-(2-((4-methoxyphenyl)sulfonamido)phenethyl)benzamide (13c)
The title compound was obtained as a white solid in 55% yield from compound 23c in a manner
1
similar to that described for the preparation of 13a H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.72,
6
2.76 m, 2H), 2.79-2.81 (m, 2H), 3.79 (s, 3H), 4.85 (s, 2H), 6.60 (t, J = 7.5Hz, 1H), 6.78 (dd, J = 1, 8Hz,
H), 6.89 (d, J = 7.5Hz, 1H), 6.96 (t, J = 7.5Hz, 1H), 7.05-7.11 (m, 3H), 7.13 (d, J = 7.5Hz, 1H), .22 (d,
J = 6.5Hz, 1H), 7.27 (d, J = 8Hz, 1H), 7.61 (d, J = 9Hz, 2H), 7.89 (d, J = 7.5Hz, 2H), 9.49 s, 1H), 9.59
1
3
(
s, 1H). C-NMR (125 MHz, DMSO-d ) δ (ppm): 32.32, 35.72, 56.12, 114.78, 116.65, 116.82,
6
123.91, 126.92, 126.98, 127.08, 127.13, 128.20, 128.63, 129.26, 130.24, 132.65, 132.83, 135.10,
1
4

+
1
38.36, 134.56, 145.87, 162.77, 165.71. mp = 195.2-196.0°C HRMS (ESI) for C H N O S [M+H] :
28 28 3 4
Calcd., 502.1795; Found, 502.1803.
N-Hydroxy-4-(2-((4-methoxyphenyl)sulfonamido)phenethyl)benzamide (13d)
A mixture of 18c (1.13 mmole), HBTU (1.47 mmole), DIPEA (1.47 mmole) and DMF(3 mL) was
stirred briefly then o-(tetrahydro-2H-pyran-2-yl)hydroxylamine (1.36 mmole) was added at rt and the
mixture was stirred overnight. The residue was purified by flash column over silica gel to afford an
oily product which was dissolved in MeOH (4 mL) and 10% TFA(aq) (4 mL) was added at rt. After
stirring for 4 h, the mixture was purified by flash chromatography over silica gel to afford compound
1
1
3
7
3d in 35% yield. H-NMR (500 MHz, DMSO-d ) δ (ppm): .70-2.73 (m, 2H), 2.76-2.80 (m, 2H),
6
.79 (s, 3H), 6.89 (dd, J = 1, 7.5Hz, 1H), 7.05-7.11 (m, 4H), .20-7.23 (m, 3H), 7.61 (d, J = 9Hz, 2H),
1
3
.67 (d, J = 8Hz, 2H), 8.96 (s, 1H), 9.48 (s, 1H), 11.13 (s, 1H). C-NMR (125MHz, DMSO-d ) δ
6
(ppm): 32.27, 35.70, 56.11, 114.77, 126.95, 127.28, 128.67, 129.24, 130.18, 130.84, 132.90, 135.22,
+
1
4
38.26, 145.54, 162.74. mp = 164.4-164.9°C. HRMS (ESI) for C H N O S [M+H] : Calcd.,
2
2
23
2
5
27.1322; Found, 427.1328.
N-(2-Aminophenyl)-4-(2-(phenylsulfonamido)phenethyl)benzamide (14a)
The title compound was obtained as a solid in 60% yield from compound 25a in a manner similar to
1
that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.72-2.79 (m, 4H),
6
4
7
7
.88 (s, 2H), 6.61 (t, J = 7Hz, 1H), 6.78 (dd, J = 1, 8Hz, 1H), 6.88 (dd, J = 1, 8Hz, 1H), 6.95 (t, J =
Hz, 1H), 7.08-7.10 (m, 1H), 7.12-7.16 (m, 2H), 7.23 (dd, J = 1, 7.5Hz, 1H), 7.27 (d, J = 8.5Hz, 2H),
13
.54-7.70 (m, 5H), 7.89 (d, J = 8Hz, 2H), 9.60 (s, 1H), 9.70 (s, 1H). C-NMR (125MHz, DMSO-d ) δ
6
1
5

(ppm): 32.37, 35.76, 116.60, 116.74, 123.88, 126.90, 127.05, 127.15, 127.17, 128.24, 128.63, 128.96,
129.72, 130.32, 132.68, 133.21, 134.81, 138.48, 141.08, 143.61, 145.80, 165.65. mp = 158.7-159.4°C.
+
HRMS (ESI) for C H N O S [M+H] : Calcd., 472.1689; Found, 472.1696.
2
7
26
3
3
N-(2-Aminophenyl)-4-(2-((3-methoxyphenyl)sulfonamido)phenethyl)benzamide (14b)
The title compound was obtained as solid in 49% yield from compound 25b in a manner similar to
1
that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.72-2.79 (m, 4H),
6
3
6
2
1
1
.73 (s, 3H), 4.87 (s, 2H), 6.61 (t, J = 7Hz, 1H), 6.78 (dd, J = 1, 8Hz, 1H), 6.93 (dd, J = 1.5, 7.5Hz, H),
.95 (t, J = 7Hz, 1H), 7.11-7.19 (m, 5H), 7.21-7.28 (m, 4H), 7.46 (t, J = 8.5Hz, 1H), 7.90 (d, J = Hz,
1
3
H), 9.58 (s, 1H), 9.67 (s, 1H). C-NMR (125MHz, DMSO-d ) δ (ppm): 32.34, 35.77, 55.99, 11.94,
6
16.63, 116.76, 119.09, 119.14, 123.93, 126.89, 127.05, 127.13, 127.18, 128.22, 128.59, 130.27,
30.90, 132.69, 134.85, 138.43, 142.32, 143.59, 145.81, 159.84, 165.64. mp = 169.1169.9°C. HRMS
+
(
ESI) for C H N O S [M+H] : Calcd., 502.1795; Found, 502.1800.
2
8
28
3
4
N-(2-Aminophenyl)-4-(2-((4-fluorophenyl)sulfonamido)phenethyl)benzamide (14c)
The title compound was obtained as solid in 52% yield from compound 25c in a manner similar to
1
that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.77-2.81 (m, 4H),
6
4
.86 (s, 2H), 6.60 (t, J = 7.5Hz, 1H), 6.78 (d, J = 8Hz, 1H), 6.85 (d, J = 7.5Hz, 1H), 6.96 (t, J = 7.5Hz,
H), 7.09 (t, J = 7Hz, 1H), 7.14-7.17 (m, 2H), 7.25 (d, J = 7.5Hz, 1H), 7.28 (d, J = 7.5Hz, 2H), 7.40 (t,
1
1
3
J = 8.5Hz, 2H), 7.73-7.76 (m, 2H), 7.90 (d, J = 8Hz, 2H), 9.59 (s, 1H), 9.72 (s, 1H). C-NMR (125
MHz, DMSO-d ) δ (ppm): 32.39, 35.78, 116.65, 116.78, 116.81, 116.96, 123.91, 126.92, 127.12,
6
127.31, 127.27, 128.24, 128.61, 130.09, 130.17, 130.38, 132.70, 134.66, 137.44, 138.70, 143.57,
1
6

+
1
4
45.79, 163.71, 165.70. mp = 178.9-179.6°C. HRMS (ESI) for C H FN O S [M+H] : Calcd.,
27 25 3 3
90.1595; Found, 490.1603.
N-(2-Aminophenyl)-4-(2-((4-chlorophenyl)sulfonamido)phenethyl)benzamide (14d)
The title compound was obtained as solid in 47% yield from compound 25d in a manner similar to
1
that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.77-2.79 (m, 4H),
6
4.86 (s, 2H), 6.60 (t, J = 7.5Hz, 1H), 6.78 (d, J = 8Hz, 1H), 6.86 (d, J = 8Hz, 1H), 6.96 (t, J = 7.5Hz,
H), 7.10 (t, J = 7.5Hz, 1H), 7.14-7.16 (m, 2H), 7.25-7.28 (m, 3H), 7.64 (d, J = 8.5Hz, 2H), 7.68 (d, J =
1
3
8
3
1
1
.5Hz, 2H), 7.89 (d, J = 8Hz, 2H), 9.59 (s, 1H), 9.79 (s, 1H). C-NMR (125 MHz, DMSO-d ) δ ppm):
6
2.38, 35.74, 116.65, 116.81, 123.91, 126.92, 127.13, 127.15, 127.33, 127.40, 128.25, 128.60, 129.02,
29.88, 130.36, 132.70, 134.59, 137.52, 138.06, 138.69, 139.98, 143.57, 145.77, 165.69. mp =
+
98.3-199.0°C. HRMS (ESI) for C H ClN O S [M+H] : Calcd., 506.1300; Found, 506.1305.
2
7
25
3
3
N-(2-Aminophenyl)-4-(2-((4-bromophenyl)sulfonamido)phenethyl)benzamide (14e)
The title compound was obtained as a solid in 49% yield from compound 25e in a manner similar to
1
that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.69-2.81 (m, 4H),
6
4.87 (s, 2H), 6.59 (td, J = 1, 7.5Hz, 1H), 6.78 (dd, J = 1, 7.5Hz, 1H), 6.87 (dd, J = 1, 8Hz, 1H), 6.96 (td,
J = 1.5, 8Hz, 1H), 7.11 (td, J = 2, 7.5Hz, 1H), 7.16 (t, J = 7.5Hz, 2H), 7.27 (t, J = 8Hz, 3H), 7.63 (dd,
J = 2.5, 9.5Hz, 2H), 7.78 (dd, J = 1.5, 6.5Hz, 2H), 7.90 (d, J = 8Hz, 2H), 9.58 (s, 1H), 9.79 s, H).
1
3
C-NMR (125 MHz, DMSO-d ) δ (ppm): 31.25, 32.36, 36.25, 116.62, 116.76, 123.92, 126.88, 27.02,
6
127.11, 127.15, 127.33, 127.39, 128.26, 128.59, 129.10, 130.35, 132.73, 132.83, 134.58, 138.65,
1
7

+
1
40.42, 143.58, 145.75, 165.64. mp = 189.5-190.2°C. HRMS (ESI) for C H BrN O S [M+H] :
27 25 3 3
Calcd., 550.0795; Found, 550.0801.
N-(2-Aminophenyl)-4-(2-((4-cyanophenyl)sulfonamido)phenethyl)benzamide (14f)
The title compound was obtained as a solid in 57% yield from compound 25f in a manner similar to
1
that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.75-2.82 (m, 4H),
6
4
.86 (s, 2H), 6.60 (t, J = 7.5Hz, 1H), 6.78 (d, J = 8Hz, 1H), 6.81 (d, J = 7.5Hz, 1H), 6.96 (t, J = 7.5Hz,
H), 7.10 (t, J = 7Hz, 1H), 7.15-7.19 (m, 3H), 7.27 (d, J = 8.1Hz, 3H), 7.85 (d, J = 8.5Hz, 2H), 7.90 (d,
1
1
3
J = 7.5Hz, 2H), 8.05 (d, J = 8.5Hz, 2H), 9.59 (s, 1H), 10.00 (s, 1H). C-NMR (125 MHz, DMSO-d )
6
δ (ppm): 32.40, 36.26, 115.59, 116.63, 116.80, 118.12, 123.90, 126.93, 127.14, 127.25, 127.45,
1
1
4
27.67, 127.85, 128.27, 128.58, 130.47, 132.73, 133.95, 134.25, 138.91, 143.58, 145.18, 145.71,
+
65.68. mp = 132.6-133.4°C. HRMS (ESI) for C H N O S [M+H] : Calcd., 497.1642; Found,
2
8
25
4
3
97.1648.
N-(2-Aminophenyl)-4-(2-((3,4-dimethoxyphenyl)sulfonamido)phenethyl)benzamide (14g)
The title compound was obtained as a white solid in 49% yield from compound 25g in a manner
1
similar to that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.722.80
6
(m, 4H), 3.68 (s, 3H), 3.79 (s, 3H), 4.87 (s, 2H), 6.60 (t, J = 7Hz, 1H), 6.78 (d, J = 7Hz, 1H), .92-6.98
(m, 2H), 7.02 (d, J = 8.4Hz, 1H), 7.09-7.17 (m, 4H), 7.24 (dd, J = 2.4, 8.7Hz, 2H), 7.27 (d, J = 8.4Hz,
1
3
2
H), 7.89(d, J = 8.1Hz, 2H), 9.49 (s, 1H), 9.59 (s, 1H). C-NMR (125 MHz, DMSO-d ) δ (ppm):
6
31.15, 32.34, 35.74, 58.07, 56.29, 109.83, 111.52, 116.63, 116.76, 120.80, 123.92, 126.74, 126.88,
1
8

126.97, 127.12, 128.20, 128.57, 130.15, 132.67, 138.31, 143.59, 145.87, 149.02, 152.51, 165.64. mp
+
=
171.9-172.4°C. HRMS (ESI) for C H N O S [M+H] : Calcd., 532.1901; Found, 532.1906.
2
9
30
3
5
4-(2-((4-Methoxyphenyl)sulfonamido)phenethyl)-N-phenylbenzamide (15a)
The title compound was obtained as a white solid in 41% yield from compound 23c in a manner
1
similar to that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.762.78
6
(m, 2H), 2.81-2.83 (m, 2H), 3.79 (s, 3H), 6.89 (dd, J = 1, 8Hz, 1H), 7.05-7.12 (m, 5H), 7.22 (dd, = 1,
7
7
3
1
Hz, 1H), 7.30 (d, J = 8.5Hz, 2H), 7.34 (t, J = 8Hz, 2H), 7.62 (d, J = 9Hz, 2H), 7.77 (d, J = 8Hz, 2H),
1
3
.87 (d, J = 8.5Hz, 2H), 9.49 (s, 1H), 10.14 (s, 1H). C-NMR (125 MHz, DMSO-d ) δ (ppm): 32.29,
6
5.74, 56.12, 114.79, 120.82, 124.03, 126.99, 127.03, 127.09, 128.11, 128.71, 129.05, 129.28, 130.25,
32.80, 133.02, 135.04, 138.35, 139.71, 146.06, 162.79, 165.87. mp = 166.4167.3°C. HRMS (ESI)
+
for C H N O S [M+H] : Calcd., 487.1686 ; Found, 487.1693.
2
8
27
2
4
N-(2-Hydroxyphenyl)-4-(2-((4-methoxyphenyl)sulfonamido)phenethyl)benzamide (15b)
A mixture of 23c (2.21 mmole), HBTU (2.87 mmole), DIPEA (2.87 mmole), 2-benzyloxyaniline
(2.65 mmole) and DMF (4 mL) was stirred at rt overnight. The reaction was quenched with water and
extracted by EtOAc. The residue was purified by flash chromatography over silica gel to afford an
intermediate compound. Then a mixture of the intermediate compound, 10% palladium on carbon (0.4
g) in MeOH (40 mL) was stirred at rt under hydrogen overnight. The organic layer was filtrated and
the residue was purified by flash chromatography over silica gel to afford compound 15b in 32%
1
yield. H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.76-2.81 m, 4H), 3.79 (s, 3H), 6.83 (t, J = 7.5Hz,
6
1H), 6.91 (t, J = 7.5Hz, 2H), 7.01-7.11 (m, 5H), 7.21 (d, J = .5Hz, 1H), 7.29 (d, J = 8Hz, 2H), 7.61 (d,
1
9

1
3
J = 9Hz, 2H), 7.68 (d, J = 8Hz, 1H), 7.88 (d, J = 8Hz, H), 9.45 (s, 1H), 9.74 (s, 1H). C-NMR (125
MHz, DMSO-d ) δ (ppm): 32.27, 35.74, 56.11, 69.62, 79.15, 114.77, 116.51, 119.54, 124.42, 126.05,
6
126.46, 126.97, 127.96, 128.86, 129.24, 130.22, 132.38, 146.25, 149.68, 162.72, 165.63. mp =
+
1
75.2-175.9°C. HRMS (ESI) for C H N O S [M+H] : Calcd., 503.1635; Found, 503.1642.
2
8
27
2
5
N-(2-Amino-5-fluorophenyl)-4-(2-((4-methoxyphenyl)sulfonamido)phenethyl)benzamide (15c)
The title compound was obtained as a white solid in 56% yield from compound 23c in a manner
1
similar to that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.742.77
6
(m, 2H), 2.79-2.81(m, 2H), 3.80 (s, 3H), 5.19 (s, 2H), 6.35 (td, J = 2.5, 8.5Hz, 1H), 6.54 (dd, J 3,
1
1
1Hz, 1H), 6.89 (dd, J = 1, 7.5Hz, 1H), 7.06 (d, J = 9Hz, 2H), 7.05-7.14 (m, 3H), 7.22 (d, J = .5Hz,
H), 7.27 (d, J = 8Hz, 2H), 7.61 (d, J = 9Hz, 2H), 7.89 (d, J = 8Hz, 2H), 9.49 (s, 1H), 9.51 (s, 1H).
1
3
C-NMR (125 MHz, DMSO-d ) δ (ppm): 32.34, 35.73, 56.12, 101.85, 102.05, 102.43, 102.61, 14.79,
6
1
1
19.86, 126.97, 127.08, 128.23, 128.59, 128.93, 129.01, 129.26, 130.24, 132.56, 132.83, 138.36,
45.87, 145.96, 160.51, 162.41, 162.77, 165.93. mp = 216.5-217.3°C. HRMS (ESI) for
+
C H FN O S [M+H] : Calcd., 520.1701; Found, 520.1707.
28
27
3
4
N-(3-Aminophenyl)-4-(2-((4-methoxyphenyl)su lfonamido)phenethyl)benzamide (15d)
A mixture of 23c (1.67 mmole), HBTU (2.17 mmole), DIPEA (2.17 mmole), m-phenylenediamine
(2.00 mmole) and DMF (3 mL) was stirred at rt overnight. The reaction was quenched with water and
extracted with EtOAc. The residue was purified by flash chromatography over silica gel to afford an
intermediate compound. Then a mixture of the intermediate compound, 10% palladium on carbon (0.5
g) in MeOH (50 mL) was stirred at rt under hydrogen overnight. The organic layer was filtrated and
2
0

the residue was purified by flash chromatography over silica gel to afford compound 15d in 45%
1
yield. H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.74-2.76 (m, 2H), 2.79-2.81 (m, 2H), 3.78 (s, 3H),
6
5
8
2
1
1
5
.03 (s, 2H), 6.30 (dt, J = 2, 8Hz, 1H), 6.84, (d, J = 8Hz, 1H), 6.88 (dd, J = 1.5, 8Hz, 1H), 6.94 (t, J =
Hz, 1H), 7.04-7.11 (m, 5H), 7.21 (dd, J = 1.5, 7.5Hz, H), 7.27 (d, J = 8Hz, 2H), 7.61 (dd, J = 2, 7Hz,
13
H), 7.83 (d, J = 8Hz, 2H), 9.83 (s, 1H). C-NMR 125 MHz, DMSO-d ) δ (ppm): 32.32, 35.71, 56.11,
6
06.66, 108.94, 110.21, 114.77, 126.97, 128.04, 128.62, 129.25, 130.21, 133.30, 138.30, 140.22,
+
45.81, 149.33, 162.74, 165.63. mp 210.3-210.9°C. HRMS (ESI) for C H N O S [M+H] : Calcd.,
2
8
28
3
4
02.1795; Found, 502.1801.
N-(4-Aminophenyl)-4-(2-((4-methoxyphenyl)sulfonamido)phenethyl)benzamide (15e)
The title compound was obtained as a white solid in 42% yield from compound 23c in a manner
1
similar to that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.732.75
6
(m, 2H), 2.78-2.81 (m, 2H), 3.79 (s, 3H), 4.88 (s, 2H), 6.53 (d, J = 9Hz, 2H), 6.89 (dd, J = 1, 7.5Hz,
1
H), 7.06 (dd, J = 2, 7Hz, 2H), 7.07-7.11 (m, 2H), 7.21 (dd, J = 1.5, 7Hz, 1H), 7.25 (d, J = 8Hz, 2H),
.35 (d, J = 8.5Hz, 2H), 7.61 (dd, J = 1.5, 7Hz, 2H), 7.83 (d, J = 8Hz, 2H), 9.49 (s, 1H), 9.77 (s, 1H).
7
1
3
C-NMR (125 MHz, DMSO-d ) δ (ppm): 32.30, 35.71, 56.11, 79.17, 79.43, 79.70, 114.15, 114.78,
6
1
1
5
22.70, 126.90, 127.86, 128.59, 128.65, 129.25, 130.23, 132.82, 133.36, 135.07, 138.33, 145.51,
+
45.59, 162.77, 165.00. mp = 184.9-185.5°C. HRMS (ESI) for C H N O S [M+H] : Calcd.,
2
8
28
3
4
02.1795; Found, 502.1805.
2
1

(E)-N-(2-Aminophenyl)-4-(2-((4-methoxyphenyl)sulfonamido)styryl)benzamide (16a)
The title compound was obtained as a white solid in 48% yield from compound 28 in a manner
1
similar to that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 3.63 (s,
6
3
7
4
1
1
1
H), 4.89 (s, 2H), 6.61 (t, J = 7.5Hz, 1H), 6.79 (d, J = 8Hz, 1H), 6.91 (d, J = 8.5Hz, 2H), 6.96 (t, J =
Hz, 1H), 7.00 (d, J = 8.5Hz, 2H), 7.15 (dd, J = 2.5, 6.5Hz, 1H), 7.18 (d, J = 7.5Hz, 1H), 7.24 (t, J =
Hz, 2H), 7.27 (d, J = 16.5Hz, 1H), 7.52 (d, J = 1.5Hz, 2H), 7.54 (d, J = 1.5Hz, 2H), 7.70 (t, J = 9.5Hz,
1
3
H), 7.99 (d, J = 8Hz, 2H), 9.66 (s, 1H), 9.76 (s, 1H). C-NMR (125 MHz, DMSO-d ) δ (ppm): 55.90,
6
14.69, 116.64, 116.78, 123.83, 125.96, 126.07, 126.72, 126.98, 127.19, 128.56, 128.79, 128.88,
29.13, 133.44, 133.75, 140.54, 143.66, 162.70, 165.37. mp = 210.7-211.4°C. HRMS (ESI) for
+
C H N O S [M+H] : Calcd., 500.1639; Found, 500.1645.
28
26
3
4
N-(2-Aminophenyl)-4-((2-((4-methoxyphenyl)sulfonamido)phenoxy)methyl)benzamide (16b)
The title compound was obtained as a white solid in 54% yield from compound 31a in a manner
1
similar to that described for the preparation of 13a. H-NMR (500MHz, DMSO-d ): 3.74 (s, 3H), 4.87
6
s, 2H), 5.00 (s, 2H), 6.60 (t, J = 7.5Hz, 1H), 6.78 (d, J = 8Hz, 1H), 6.86 (t, J = 7.5Hz, 1H), 6.90-6.92 m,
3H), 6.97 (t, J = 9Hz, 1H), 7.05 (t, J = 7.5Hz, 1H), 7.15 (d, J = 7.5Hz, 1H), 7.25 (d, J = 8Hz, H), 7.41
1
3
(
d, J = 8Hz, 2H), 7.59 (d, J = 6Hz, 2H), 7.92 (d, J = 8Hz, 2H), 9.38 (s, 1H), 9.65 (s, 1H). C-NMR
(
125 MHz, DMSO-d ) δ (ppm): 56.00, 69.21, 113.42, 114.42, 116.62, 116.78, 121.15, 123.75, 125.85,
6
1
1
5
26.80, 127.02, 127.23, 128.12, 129.25, 132.76, 134.23, 139.60, 140.79, 143.64, 151.41, 162.64,
+
65.57. mp = 144.6-145.4°C. HRMS (ESI) for C H N O S [M+H] : Calcd., 504.1588; Found,
2
7
26
3
5
04.1593.
2
2

N-(2-Aminophenyl)-4-(((2-((4-methoxyphenyl)sulfonamido)phenyl)amino)methyl)benzamide (16c)
The title compound was obtained as a solid in 57% yield from compound 31b in a manner similar
1
to that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 3.81 (s, 3H),
6
4
1
7
9
1
1
.32 (s, 2H), 4.86 (s, 2H), 5.76 (s, 1H), 6.36-6.41 (m, 2H), 6.59 (t, J = 7.5Hz, 1H), 6.65 (d, J = 7Hz,
H), .77 (d, J = 7Hz, 1H), 6.89 (t, J = 7.5Hz, 1H), 6.96 (td, J = 1, 7.5Hz, 1H), 7.06 (d, J = 9Hz, 2H),
.15 d, J = 7.5Hz, 1H), 7.32 (d, J = 8Hz, 2H), 7.36 (d, J = 9Hz, 2H), 7.88 (d, J = 8Hz, 2H), 9.20 (s, 1H),
1
3
.58 (s, 1H). C-NMR (125 MHz, DMSO-d ) δ (ppm): 46.42, 56.10, 111.57, 114.62, 116.09, 116.59,
6
16.75, 121.76, 123.84, 126.91, 127.10, 127.20, 127.83, 128.20, 128.25, 129.59, 132.27, 133.59,
43.56, 143.88, 144.57, 162.83, 165.67. mp = 206.8-207.4°C. HRMS (ESI) for C H N O S
2
7
27
4
4
+
[
M+H] : Calcd., 503.1748; Found, 503.1752.
N-(2-Aminophenyl)-2'-((4-methoxyphenyl)sulfonamido)-[1,1'-biphenyl]-4-carboxamide (16d)
The title compound was obtained as a solid in 49% yield from compound 36 in a manner similar to
1
that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 3.81 (s, 3H), 4.90
6
(s, 2H), 6.62 (t, J = 7.5Hz, 1H), 6.80 (d, J = 8Hz, 1H), 6.96-7.00 (m, 3H), 7.06 (t, J = 5Hz, 1H), 7.19 (d,
J = 7.5Hz, 1H), 7.29-7.30 (m, 3H), 7.35 (d, J = 8.5Hz, 2H), 7.47 (d, J = 9Hz, 2H), 7.95 (d, J = 8Hz,
1
3
2
H), 9.42 (s, 1H), 9.71 (s, 1H). C-NMR (125 MHz, DMSO-d ) δ (ppm): 56.07, 114.64, 116.67,
6
116.81, 123.85, 127.01, 127.24, 127.53, 127.92, 128.92, 129.09, 129.59, 132.77, 133.45, 133.99,
+
1
38.51, 142.29, 143.67, 162.65, 165.55. mp = 71.9-72.4°C. HRMS (ESI) for C H N O S [M+H] :
26 24 3 4
Calcd., 474.1482; Found, 474.1490.
2
3

4-Methoxy-N-(2-phenethylphenyl)benzenesulfonamide (17)
A mixture of 38 (2.00 mmole), 4-methoxybenzenesulfonyl chloride (2.20 mmole) and pyridine (4
mL) was stirred at rt overnight. The reaction was quenched with water and extracted with CH Cl . The
2
2
residue was purified by flash chromatography over silica gel to afford compound 17 in 61% yield.
1
H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.64-2.68 (m, 2H), 2.76-2.79 (m, 2H), 3.79 (s, 3H), 6.88 (d,
6
J = 1, 8Hz, 1H), 7.04-7.21 (m, 8H), 7.25-7.28 (m, 2H), 7.61 (dd, J = .5, 7Hz, 2H), 9.46 (s, 1H).
1
3
C-NMR (125 MHz, DMSO-d ) δ (ppm): 32.63, 35.95, 56.10, 114.77, 126.25, 126.87, 126.95,
6
127.00, 128.64, 128.71, 129.25, 130.17, 132.88, 135.05, 138.60, 142.15, 162.76. mp = 131.9-132.3°C.
+
HRMS (ESI) for C H NO S [M+H] : Calcd., 368.1315; Found, 368.1321.
2
1
22
3
N-(2-Aminophenyl)-4-phenethylbenzamide (18)
The title compound was obtained as a white solid in 54% yield from compound 39 in a manner
1
similar to that described for the preparation of 13a. H-NMR (500 MHz, DMSO-d ) δ (ppm): 2.912.96
6
(m, 4H), 4.84 (s, 2H), 6.59 (t, J = 7.5Hz, 1H), 6.77 (d, J = 7.5Hz, 1H), 6.95 (td, J = 0.5, 8Hz, H),
7.13-7.18 (m, 2H), 7.22-7.28 (m, 4H), 7.33 (d, J = 8Hz, 2H), 7.87 (d, J = 8Hz, 2H), 9.58 (s, 1H).
1
3
C-NMR (125 MHz, DMSO-d ) δ (ppm): 37.12, 37.20, 116.64, 116.80, 123.91, 126.36, 126.90,
6
127.12, 128.22, 128.72, 128.79, 128.88, 132.65, 141.68, 143.56, 145,68, 165.69. mp = 185.1-185.7°C.
+
HRMS (ESI) for C H N O [M+H] : Calcd., 317.1648; Found, 317.1655.
2
1
21
2
Methyl-4-(4-nitrostyryl)benzoate (20a)
A mixture of 19a (16.5 mmole), (4-(methoxycarbonyl)benzyl)triphenylphosphonium bromide
(16.5 mmole), NaOH (21.5 mmole) and THF (40 mL) was stirred at rt overnight. The reaction was
2
4

quenched with water and extracted with CH Cl . The residue was purified by flash chromatography
2
2
1
over silica gel to afford compound 20a in 67% yield. H-NMR (300 MHz, DMSOd ) δ (ppm): 3.86 (s,
6
3
H), 7.52 (s, 1H), 7.60 (s, 1H), 7.81 (d, J = 8.4Hz, 2H), 7.91 (d, J = 9Hz, 2H), 7.99 (d, J = 8.4Hz, 2H),
.25 (d, J = 9Hz, 2H).
8
Methyl-4-(3-nitrostyryl)benzoate (20b)
The title compound was obtained as a white solid in 54% yield from compound 19b in a manner
1
similar to that described for the preparation of 20a. H-NMR (300 MHz, CD OD) δ (ppm): 3.89 (s,
3
3
H), 6.82 (d, J = 12.3Hz, 1H), 6.88 (d, J = 12.3Hz, 1H), 7.31 (d, J = 8.4Hz, 2H), 7.48 (d, J = 7.8Hz,
H), 7.57 (d, J = 7.8Hz, 1H), 7.90 (d, J = 4.8Hz, 2H), 8.04-8.06 (m, 2H).
1
Methyl-4-(2-nitrostyryl)benzoate (20c)
The title compound was obtained as a white solid in 56% yield from compound 19c in a manner
1
similar to that described for the preparation of 20a. H-NMR (300 MHz, CDCl ) δ (ppm): 3.86 (s, 3H),
3
6.78 (d, J = 12Hz, 1H), 7.00 (d, J = 12Hz, 2H), 7.10 (d, J = 8.1Hz, 2H), 7.17-7.20 (m, 1H), 7.35-7.44
(m, 2H), 7.82 (dd, J = 1.5, 6.6Hz, 2H), 8.09-8.12 (m, 1H).
Methyl 4-(4-aminophenethyl)benzoate (21a)
A mixture of methyl-4-(4-nitrostyryl)benzoate (12.4 mmole), 10% palladium on carbon (0.5 g) in
MeOH (50 mL) was stirred at rt under hydrogen overnight. The organic layer was filtered and the
residue was purified by flash chromatography over silica gel to afford compound 0a in 88% yield.
1
H-NMR (300 MHz, DMSO-d )) δ (ppm): 2.68-2.71 (m, 2H), 2.81-2.84 (m, 2H), 3.81 (s, 3H), 4.79 (s,
6
2H), 6.43 (d, J = 8.4Hz, 2H), 6.82 (d, J = 8.4Hz, 2H), 7.31 (d, J = 8.4Hz, 2H), 7.83 (d, J = 8.4Hz, 2H).
2
5

Methyl 4-(3-aminophenethyl)benzoate (21b)
The title compound was obtained as solid in 81% yield from compound 20b in a manner similar to
1
that described for the preparation of 21a. H-NMR (300 MHz, DMSO-d ) δ (ppm): 2.69-2.74 (m, 2H),
6
.
86-2.92 (m, 2H), 3.82 (s, 3H), 4.90 (s, 2H), 6.34-6.41 (m, 3H), 6.88 (t, J = 7.5Hz, 1H), 7.35 (d, J =
.4Hz, 2H), 7.85 (dd, J = 1.8, 6.6Hz, 2H).
8
Methyl 4-(2-aminophenethyl)benzoate (21c)
The title compound was obtained as a white solid in 77% yield from compound 20c in a manner
1
similar to that described for the preparation of 21a. H-NMR (300 MHz, DMSO-d ) δ (ppm): 2.682.74
6
(m, 2H), 2.84-2.89 (m, 2H), 3.82 (s, 3H), 4.87 (s, 2H), 6.44 (td, J = 1.2, 7.2Hz, 1H), 6.60 (dd, J = 0.9,
8.1Hz, 1H), 6.84-6.89 (m, 2H), 7.41 (d, J = 8.1Hz, 2H), 7.85 (d, J = 8.1Hz, 2H).
Methyl 4-(4-((4-methoxyphenyl)sulfonamido)phenethyl)benzoate (22a)
The title compound was obtained as a solid in 82% yield from compound 21a in a manner similar to
1
that described for the preparation of 17. H-NMR (300 MHz, DMSO-d ) δ (ppm): 2.75-2.78 (m, 2H),
6
2.83-2.85 (m, 2H), 3.77 (s, 3H), 3.81 (s, 3H), 6.92 (d, J = 8.4Hz, 2H), 7.01 (d, J = 9.3Hz, 4H), 7.25 (d,
J = 8.4Hz, 2H), 7.62 (d, J = 9Hz, 2H), 7.80 (d, J = 8.4Hz, 2H).
Methyl 4-(3-((4-methoxyphenyl)sulfonamido)phenethyl)benzoate (22b)
The title compound was obtained as a solid in 88% yield from compound 21b in a manner similar
1
to that described for the preparation of 17. H-NMR (300 MHz, DMSO-d ) δ (ppm): 2.79-2.84 (m,
6
4H), .77 (s, 3H), 3.82 (s, 3H), 6.80 (d, J = 7.5Hz, 1H), 6.85 (dd, J = 2.1, 7.2Hz, 1H), 6.94 (t, J = 1.8Hz,
2
6

1H), 7.01-7.09 (m, 3H), 7.24 (d, J = 8.4Hz, 2H), 7.64 (d, J = 9Hz, 2H), 7.81 (d, J = 9.3Hz, 2H), 10.04
(s, 1H).
Methyl 4-(2-((4-methoxyphenyl)sulfonamido)phenethyl)benzoate (22c)
The title compound was obtained as a solid in 91% yield from compound 21c in a manner similar to
1
that described for the preparation of 17. H-NMR (300 MHz, CDCl ) δ (ppm): 2.74-2.80(m, 4H),
3
3
7
4
.83(s, 3H), 3.91(s, 3H), 5.99(s, 1H), 6.88(d, J = 8.7Hz, 2H), 7.08-7.13(m, 6H), 7.63(d, J = 9Hz, 2H),
.93(d, J = 8.4Hz, 2H).
-(4-((4-Methoxyphenyl)sulfonamido)phenethyl)benzoic acid (23a)
The mixture of 22a (1.88mmole), 1N lithium hydroxide solution (4ml) and 1,4-dioxane (8 ml) was
stirred at 40°C overnight. The reaction was concentrated and quenched with 3N hydrogen chloride
solution and water. The water layer was filtered to obtain compound 23a in 84% yield without further
1
purification. H-NMR (300 MHz, DMSO-d ) δ (ppm): 2.75-2.77 (m, 2H), 2.81-2.84 (m, 2H), 3.77 (s,
6
3H), 6.93 (d, J = 8.4Hz, 2H), 7.00-7.04 (m, 4H), 7.21 (d, J = 8.4Hz, 2H), 7.62 (d, J = 9.0Hz, 2H), 7.77
(d, J = 8.4Hz, 2H).
4-(3-((4-Methoxyphenyl)sulfonamido)phenethyl)benzoic acid (23b)
The title compound was obtained as a solid in 76% yield from compound 22b in a manner similar
1
to that described for the preparation of 23a. H-NMR (300 MHz, DMSO-d ) δ (ppm): 2.80 (s, 4H),
6
3
.77 (s, 3H), 6.79-6.87 (m, 2H), 6.95 (s, 1H), 7.01-7.09 (m, 3H), 7.17 (d, J = 8.1Hz, 2H), 7.64 (d, J =
.7Hz, 2H), 7.78 (d, J = 8.1Hz, 2H).
8
2
7

4-(2-((4-Methoxyphenyl)sulfonamido)phenethyl)benzoic acid (23c)
The title compound was obtained as a solid in 89% yield from compound 23c in a manner similar to
1
that described for the preparation of 23a. H-NMR (300 MHz, CD OD) δ (ppm): 2.81-2.85 (m, 4H),
3
.
83 (s, 3H), 6.94 (d, J = 7.5Hz, 1H), 6.99 (dd, J = 2.1, 7.2Hz, 2H), 7.01-7.15 (m, 3H), 7.21 (d, J =
.1Hz, 2H), 7.63 (d, J = 9Hz, 2H), 7.90 (d, J = 8.1Hz, 2H).
8
Methyl 4-(2-(phenylsulfonamido)phenethyl)benzoate (24a)
The title compound was obtained as a solid in 82% yield from compound 21c in a manner similar to
1
that described for the preparation of 17. H-NMR (300 MHz, DMSO-d ) δ (ppm): 2.73-2.74 (m, 4H),
6
3
4
.82 (s, 3H), 6.89 (dd, J = 1.5, 7.2Hz, 1H), 7.05-7.12 (m, 5H), 7.22 (d, J = 6.9Hz, 1H), 7.29-7.37 (m,
H), 7.61 (dd, J = 1.8, 6.6Hz, 2H), 7.77 (dd, J = 1.2, 8.7Hz, 2H), 7.87 (d, J = 8.4Hz, 2H), 9.50 (s, 1H).
Methyl 4-(2-((3-methoxyphenyl)sulfonamido)phenethyl)benzoate (24b)
The title compound was obtained as a solid in 86% yield from compound 21c in a manner similar to
1
that described for the preparation of 17. H-NMR (300 MHz, CDCl3) δ (ppm): 2.70-2.77 (m, 4H),
3.71 (s, 3H), 3.91 (s, 3H), 6.02 (s, 1H), 7.07-7.16 (m, 8H), 7.17-7.34 (m, 2H), 7.93 (d, J = 8.4Hz, 2H).
Methyl 4-(2-((4-fluorophenyl)sulfonamido)phenethyl)benzoate (24c)
The title compound was obtained as solid in 85% yield from compound 21c in a manner similar to
1
that described for the preparation of 17. H-NMR (300 MHz, DMSO-d ) δ (ppm): 2.77 (s, 4H), 3.83 (s,
6
3
H), 6.84-6.87 (m, 1H), 7.07-7.10 (m, 2H), 7.18-7.22 (m, 1H), 7.29 (d, J = 8.1 Hz, 2H), 7.37 (t, J =
.7Hz, 2H), 7.70-7.75 (m, 2H), 7.87 (d, J = 8.4Hz, 2H).
8
2
8