Structure-activity relationship of novel (benzoylaminophenoxy)phenol derivatives as anti-prostate cancer agents

Article abstract of DOI:10.1016/j.bmc.2018.09.008

The androgen receptor (AR) is a ligand-inducible transcription factor belonging to the nuclear receptor superfamily, and is a target molecule for development of drugs to treat prostate cancer. However, AR antagonists in clinical use, such as flutamide (3a) and bicalutamide (4), encounter resistance after several years of hormone therapy, predominantly due to mutations of AR. Thus, although some new-generation AR antagonists have been developed, novel types of AR antagonists are still required to treat drug-resistant prostate cancer. We previously reported a novel (benzoylaminophenoxy)phenol derivative 10a, which is structurally distinct from conventional AR antagonists. Here, we systematically examined the structure–activity relationship of (benzoylaminophenoxy)phenol derivatives on the inhibitory activity on the prostate cancer cell proliferations. We found that the 4-[4-(benzoylamino)phenoxy]phenol backbone is important for anti-prostate cancer activity. Introduction of a small substituent at the 2 position of the central benzene ring (B ring) increases the activity. Among the synthesized compounds, 19a and 19b exhibited the most potent inhibitory activity toward dihydrotestosterone-induced proliferation of several androgen-dependent cell lines, SC-3 (wild-type AR), LNCaP (T877A AR), and 22Rv1 (H874Y AR), but interestingly also inhibited proliferation of AR-independent PC-3 cells. These compounds, which have a different pharmacophore from conventional AR antagonists, are promising drug candidates for the treatment of prostate cancer.

Full text of DOI:10.1016/j.bmc.2018.09.008

Accepted Manuscript
Structure-Activity Relationship of Novel (Benzoylaminophenoxy)phenol De-
rivatives as Anti-prostate Cancer Agents
Yuko Kazui, Shinya Fujii, Ayumi Yamada, Mari Ishigami-Yuasa, Hiroyuki
Kagechika, Aya Tanatani
PII:
S0968-0896(18)31396-8
https://doi.org/10.1016/j.bmc.2018.09.008
BMC 14533
DOI:
Reference:
Bioorganic & Medicinal Chemistry
To appear in:
Received Date:
Revised Date:
Accepted Date:
4 August 2018
4 September 2018
5 September 2018
Please cite this article as: Kazui, Y., Fujii, S., Yamada, A., Ishigami-Yuasa, M., Kagechika, H., Tanatani, A.,
Structure-Activity Relationship of Novel (Benzoylaminophenoxy)phenol Derivatives as Anti-prostate Cancer
Agents, Bioorganic & Medicinal Chemistry (2018), doi: https://doi.org/10.1016/j.bmc.2018.09.008
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Structure-Activity Relationship of Novel (Benzoylaminophenoxy)phenol Derivatives as
Anti-prostate Cancer Agents
Yuko Kazui,a Shinya Fujii,b,c,* Ayumi Yamada,b Mari Ishigami-Yuasa,b Hiroyuki Kagechika,b
and Aya Tanatania, *
aDepartment of Chemistry, Faculty of Science, Ochanomizu University, 2-1-1 Otsuka,
Bunkyo-ku, Tokyo 112-8610, Japan
bInstitute of Biomaterials and Bioengineering, Tokyo Medical and Dental University (TMDU),
2-3-10 Kanda-Surugadai, Chiyoda-ku, Tokyo 101-0062, Japan
cInstitute for Quantitative Biosciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku,
Tokyo 113-0032, Japan
To whom correspondence should be addressed:
Aya Tanatani, E-mail: tanatani.aya@ocha.ac.jp; Tel/Fax: +81-3- 5978-2716
Abstract
The androgen receptor (AR) is a ligand-inducible transcription factor belonging to the nuclear
receptor superfamily, and is a target molecule for development of drugs to treat prostate
cancer. However, AR antagonists in clinical use, such as flutamide (3a) and bicalutamide (4),
encounter resistance after several years of hormone therapy, predominantly due to mutations
of AR. Thus, although some new-generation AR antagonists have been developed, novel
types of AR antagonists are still required to treat drug-resistant prostate cancer. We previously
reported a novel (benzoylaminophenoxy)phenol derivative 10a, which is structurally distinct
from conventional AR antagonists. Here, we systematically examined the structure-activity
relationship of (benzoylaminophenoxy)phenol derivatives on the inhibitory activity on the
prostate cancer cell proliferations. We found that the 4-[4-(benzoylamino)phenoxy]phenol
backbone is important for anti-prostate cancer activity. Introduction of a small substituent at
the 2 position of the central benzene ring (B ring) increases the activity. Among the
synthesized compounds, 19a and 19b exhibited the most potent inhibitory activity toward
dihydrotestosterone-induced proliferation of several androgen-dependent cell lines, SC-3
(wild-type AR), LNCaP (T877A AR), and 22Rv1 (H874Y AR), but interestingly also
inhibited proliferation of AR-independent PC-3 cells. These compounds, which have a
different pharmacophore from conventional AR antagonists, are promising drug candidates
for the treatment of prostate cancer.
Keywords
prostate cancer, cell proliferation, phenoxyphenol, pharmacophore

1. Introduction
Androgen receptor (AR) is a ligand-inducible transcription factor belonging to the
nuclear receptor superfamily.1,2 The endogenous ligands (androgens) of AR, testosterone (1)
and its active metabolite dihydrotestosterone (DHT, 2) (Figure 1), regulate various
physiological phenomena, including differentiation, growth, and maintenance of male
reproductive organs.3,4 Since androgen plays an important role in progression of prostate
cancer,5 various AR antagonists have been developed for clinical use. In general,
non-steroidal AR antagonists are preferred, because steroidal compounds are considered to
have greater potential for off-target activities. Flutamide (3a)6 and bicalutamide (4, Figure 1)7
are typical nonsteroidal AR antagonists in clinical use to treat prostate cancer, but the
development of resistance after several years of hormone therapy, so-called
castration-resistant prostate cancer (CRPC), is significant problem.8,9 Mutations of AR, such
as T877A, are the major cause of CRPC.10-12 Hydroxyflutamide (3b), the active metabolite of
3a, can activate T877AAR, and exacerbate the prostate cancer.13
Figure 1. Structures of testosterones and typical nonsteroidal AR antagonists.
Some second-generation AR antagonists that are effective against CRPC, such as
enzalutamide (5a) and apalutamide (5b), have recently been developed.14 However, most of
the nonsteroidal AR antagonists so far known have been developed from flutamide (3a) and
bicalutamide (4) as lead compounds, and have the common pharmacophore of an anilide
structure with electron-withdrawing functional groups, such as nitro, cyano, or
trifluoromethyl groups, on the phenyl ring, as shown in blue in Figure 1. Compounds 5a and
5b contain essentially the same pharmacophore, although in these compounds, the bulky aryl
ring on the heterocyclic linker is important for the antagonism. On the other hand, some AR
antagonists bearing a different skeleton or pharmacophore have also been reported, such as

DIMN (6), isoxazolone derivative 7 and (2H)-phthalazinone derivative 8 (Figure 2).15-17
Figure 2. Structures of AR antagonists with a different pharmacophore from flutamide (3a) or
bicalutamide (4)
We have developed and synthesized novel AR antagonists based on curcumin (9) as a
lead compound.18 Curcumin (9) is major pigment in turmeric, and exhibits various biological
activities, including anticancer, antioxidant and anti-inflammatory activities. Further, some
curcumin derivatives show antiandrogenic activities in AR-positive cell lines, as well as
cytotoxic activity toward AR-negative PC-3 cells.19 Curcumin (9) consists of two phenolic
groups linked by an conjugated 1,3-diketone (or its enol form). To reduce the chemical and
metabolic reactivity of curcumin (9), we designed AR antagonist candidates containing
4‑(4-benzoylaminophenoxy)phenol as stable aromatic core structure. Among the resulting
compounds, 10a showed AR antagonistic activities in SC-3 cells (wild-type AR), LNCaP cells
(T877A-mutated AR), and 22Rv1 cells (H874Y-mutated AR).19 Thus, it was considered a
promising lead compound for antiandrogen therapy of prostate cancer. Therefore, in the
present study, we synthesized various derivatives of 10a, and examined the structure-activity
relationship in detail.
2. Results and discussion
2.1. Chemistry
Compound 10a contains three benzene rings named A, B, and C from the left, as shown
in Figure 2, linked by an amide bond (A–B rings) and an oxygen atom (B–C rings). Our
previous work showed that loss of the phenolic hydroxyl group on the C ring (Figure 2)
caused a significant decrease of AR antagonistic activity in SC-3 cell assay.18 The substituent
on the A ring also affected the potency, and a 3-methoxy or 3-dimethylamino group was

effective, while compounds with a nitro group were inactive. Based on these results, we
synthesized compounds 11 – 19, and examined their AR antagonistic activities. The
substituents on the terminal benzene rings are limited to methoxy or dimethylamino for the A
ring and hydroxyl for the C ring, and the positions of the terminal hydroxyl group
(compounds 11 and 14), ether linkage (compounds 12 and 13), amide moiety (compounds 15
and 16), and substituents on the central B ring (compounds 17 – 19) were modified.
Figure 3. Structures of 4-(4-benzoylaminophenoxy)phenol derivatives
Compounds 11 – 19 were synthesized according to Schemes 1 – 4. Reaction of
p-fluoronitrobenzene with resorcinol under basic conditions afforded diphenylether 20, which
was hydrogenated to afford amine 21 (Scheme 1). Compounds 11 were obtained by the
coupling reaction of 21 with various benzoyl chloride derivatives. In the synthesis of
compounds 12 – 14 (Scheme 2), m-fluoronitrobenzenes were reacted with monoprotected
hydroquinone or resorcinol to improve the yields of the 1:1 adducts. The obtained
diphenylethers 22 and 24 were hydrogenated, followed by condensation with various benzoyl
chlorides to afford compounds 12 – 14. Compound 22 was reduced with sodium borohydride
in the presence of copper (II) sulfate to afford amine 26 with a benzyloxy group on the C ring;
this was reacted with benzoyl chloride, followed by debenzylation to afford 12b or 12e.
Scheme 1. Synthesis of compounds 11: (a) resorcinol, NaOH, DMSO; (b) H2, Pd-C, MeOH;

(c) R1-PhCOCl, THF
Scheme 2. Synthesis of compounds 12 - 14: (a) O-Monobenzylhydroquinone, K2CO3,
DMF; (b) H2, Pd(OH)2, MeOH/THF; (c) R-PhCOCl, THF; (d) O-monobenzylresorcinol,
K2CO3, DMF; (e) CuSO4, NaBH4
We have shown that aromatic secondary amides such as benzanilide generally exist in
trans form, while the N-methylated amides exist in folded cis form both in the crystal and in
various solvents.20 In order to clarify the relationship between the amide conformation and the
activity, N-methylated amide derivatives 15 and 16 were synthesized by N-methylation of
O-benzyl-protected (benzoylaminophenoxy)phenols 31 and 27, respectively, followed by
1
debenzylation (Scheme 3). In the H NMR spectra of compounds 15 and 16, the chemical
shifts of aromatic protons were shifted to higher field, compared to those of the corresponding
secondary amides 10a, 12b, and 12c, as observed in the cases of benzanilide and
N-methylbenzanilide, indicating that these compounds existed mainly in cis form in solution.
Finally, we synthesized compounds 17 – 19 bearing various substituents on the central B ring
from 29 (Scheme 3).
Scheme 3. Synthesis of compounds 15 - 19: (a) NaH, DMF; CH3I; (b) H2, Pd(OH)2, THF ; (c)
O-Monobenzylhydroquinone, NaOH, DMSO; (d) CuSO4, NaBH4; (e) m-MeOPhCOCl, THF.

2.2. Inhibitory activity on prostate cancer cell proliferations
The biological activities of the synthesized compounds were first evaluated in terms of
growth-inhibitory activity toward androgen-dependent SC-3 cells bearing wt AR.21
Compounds 11 – 19 alone did not affect the proliferation of SC-3, i.e., they did not act as AR
agonists. The inhibitory activity of the test compounds was examined in terms of their effect
on 1 nM DHT-dependent proliferation of SC-3 cells, and the IC50 values are shown in Table 1.
Table 1. AR antagonistic activity of compounds 11 – 19 in SC-3 cell assaya
Compoundb IC50 (M)
Compoundb IC50 (M)
Compoundb IC50 (M)
10a
11a
11b
11c
11d
11e
11f
0.75
13a
13b
13c
13d
14a
14b
14c
14d
14e
15
>10 d
17a
17b
17c
17d
17e
18a
18b
18c
18d
18e
19a
19b
19c
0.40
0.48
5.7
inactivec
>10 d
>10 d
>10 d
inactivec
inactivec
inactivec
3.2
5.3
4.3
inactivec
>10 d
inactivec
3.0
>10 d
0.65
1.7
12a
12b
12c
12d
12e
12f
>10 d
inactivec
>10 c
inactivec
>10 d
3.4
7.6
inactivec
0.51
0.53
0.48
6.3
16a
16b
inactivec
inactivec
inactivec
7.40
a Inhibitory activity of test compounds toward DHT-induced proliferation of SC-3 cells was examined.
b Structures: see Figure 3.
c‘Inactive’ means that test compound did not inhibit the proliferation of SC-3 cells at concentrations up to 10
M.
d‘>10’ means that test compound at 10 M inhibited the proliferation of SC-3 cells by less than 50% versus
control cells.
Compound 10a, the lead compound in this study, inhibited the DHT-induced
proliferation of SC-3 cells with the IC50 value of 0.75 M. Here, we found that compounds 11
bearing a m-hydroxyl group on the C ring were inactive or showed very weak activity,
compared to 10a. Only compound 11f bearing a p-dimethylamino group on the A ring
exhibited significant potency (IC50: 3.2 M). Since compound 10b without the 2-methyl
group on the B ring also exhibited significant potency (IC50: 2.0 M), the decrease of the
activity in compounds 11 should be due to the inappropriate position of the hydroxyl group on
the C ring. Similarly, a change in the position of the ether linkage, that is, meta to the amide
nitrogen atom in compounds 12 - 14, decreased the activity, independently of the substituents
on the A and B rings. Compounds 15 and 16 bearing an N-methyl amide moiety lacked

activity. These compounds have cis conformation of the amide structure, indicating that the
conformation of the amide bond is critical for the activity. Thus, inhibitory activity towards
SC-3 cell proliferation was observed only with compounds having the
4-[4-(benzoylamino)phenoxy]phenol backbone among the synthesized compounds. Finally,
we examined the effect of the substituent on the B ring of compound 10. The 2-methyl group
on the B ring of 10a is important, since its removal resulted in lower activity. Among
compounds bearing a 2-substituent, 17a (IC50: 0.40 M) with a fluorine atom and 17b (IC50:
0.48 M) with a chlorine atom showed potent activity. Introduction of a larger substituent
such as a bromine atom or trifluoromethyl group was not effective. Among compounds with a
3-substituent on the B ring, only 18b (IC50: 0.65 M) with a 3-fluorine atom was as active as
10a. In this case, the introduction of a chlorine atom decreased the activity. Compounds
bearing multiple substituents on the B ring also exhibited potent activity, and the activities of
19a - c bearing two halogen atoms on the B ring were comparable to those of 17a and 17b
with one halogen atom. Thus, a small 2-substituent, such as methyl, fluorine, or chloride,
increased the activity, and 17a was the most potent of the synthesized compounds.
Next, we examined the activity of selected compounds towards other cell lines.
LNCaP22,23 and 22Rv124 cells are androgen-dependent, having mutated T877A and H874Y
ARs, respectively, while PC-3 shows androgen-independent cell growth. Bicalutamide (4)
inhibited the proliferation of LNCaP cells, but not 22Rv1 or PC-3 cells, in the concentration
range examined. Selected compounds inhibited all three cell lines dose-dependently (Figure
4), in accordance with our previous finding that 10a inhibits the proliferation of these three
cell lines. Here, we found that 19a and 19b showed more potent inhibitory activity than the
lead compound 10a, especially in 22RV1 and PC-3 cell assays. From these results, our
developed compounds would act as AR antagonists toward both wild-type and mutated ARs,
and also that some mechanism of cell growth inhibition other than AR antagonism is involved.
Further structural optimization and the detailed mechanistic studies including AR specificity
are on going.

Figure 4. Inhibitory activities of selected derivatives on the proliferation of three human
prostate cancer cells. Bicalutamide (×), 10a (○), 17a (◆), 17b (●), 18b (▲), 19a (■), 19b
(△), and 19c (□). Cell viability was determined by the WST method and normalized to
vehicle-treated cells as 1.
3. Conclusion
We synthesized a series of (benzoylaminophenoxy)phenol derivatives based on the lead
compound 10a and examined their structure-activity relationship as anti-prostate cancer
agents. The 4-[4-(benzoylamino)phenoxy]phenol backbone is important for anti-prostate
cancer activity in this series. Introduction of a small substituent at the 2-position of the B-ring
increased the activity. Among the synthesized compounds, 19a and 19b exhibited the most
potent growth-inhibitory activity toward androgen-dependent carcinoma cell lines SC-3
(wild-type AR), LNCaP (T877A AR), 22Rv1 (H874Y AR), and also inhibited the growth of
AR-independent PC-3 cells. Since these compounds have a different pharmacophore from
conventional AR antagonists based on flutamide (3a) and bicalutamide (4), they may be
promising candidates for treatment of prostate cancer.
4. Experimental
4.1. Chemistry
All reagents were purchased from Sigma-Aldrich Chemical Co., Tokyo Kasei Kogyo Co.,
Wako Pure Chemical Industries, or Kanto Kagaku Co., Inc. Silica gel for column
1
13
chromatography was purchased from Kanto Kagaku Co., Inc. H and C NMR spectra were
recorded on a JEOL ECA 600, or Bruker 600 spectrometer. Mass spectral data was obtained
on a Bruker Daltonics microTOF-2focus, Thermo Scientific Q-Exactive or Waters Q-TOF
Premier in the positive ion detection mode. Melting points were determined on a RFS-30
melting point apparatus (Round Science).
4.2. Synthesis of 11a (General procedure for compounds 11, Scheme 1)
A 5 M aqueous solution of sodium hydroxide (12.5 ml, 62.5 mmol) was added to DMSO (35
ml) at room temperature under an Ar atmosphere. After 30 min, resorcinol (4.13 g, 37.5
mmol) was added, and the mixture was stirred at 50°C for 30 min. Then, a solution of
4-fluoronitrobenzene (3.53 g, 25.0 mmol) in DMSO (5 ml) was added slowly to the reaction
mixture, and stirring was continued at 50°C for 2.5 h. The reaction mixture was poured into
ice water and filtered. The aqueous layer was acidified with 2 M HCl, and extracted with
ethyl acetate. The organic layer was dried with magnesium sulfate, and evaporated to give 20
(3.82 g, 66%).
20: 1H NMR (600 MHz, CDCl3)  8.21 (d, J = 9.3 Hz, 2 H), 7.28 (t, J = 8.2 Hz, 1 H), 7.04 (d,
J = 7.1 Hz, 2 H), 6.71 (ddd, J = 8.0, 2.3, 0.7 Hz, 1 H), 6.66 (ddd, J = 8.2, 2.2, 0.8 Hz, 1 H),
6.59 (t, J = 2.3 Hz, 1 H), 4.96 (br, 1 H), 13C NMR (150 MHz, CDCl3)  163.01, 157.17,

155.99, 142.82, 130.98, 125.93, 117.40, 112.63, 112.41, 107.87.
A mixture of 20 (2.31 g, 10 mmol) and 10% palladium on carbon (234 mg) in methanol (100
ml) was stirred under a hydrogen atmosphere for 2 h. The reaction mixture was filtered over
Celite, and the filtrate was evaporated. The residue was purified by silica-gel column
chromatography (hexane/ethyl acetate 1:1) to afford 21 (1.93 g, 96%).
21: 1H NMR (600 MHz, CD3OD)  7.09 (t, J = 8.2 Hz, 1 H), 6.84 (d, J = 8.6 Hz, 2 H), 6.76 (d,
J = 9.8 Hz, 2 H), 6.46 (dt, J = 8.0, 1.3 Hz, 1 H), 6.38 (dt, J = 8.7, 1.0 Hz, 1 H), 6.34 (td, J =
13
0.5, 2.2 Hz, 1 H), C NMR (150 MHz, CD3OD) 162.04, 160.09, 150.02, 145.31, 131.24,
122.32, 118.02, 110.42, 109.52, 105.58.
21 (101 mg, 0.50 mmol) was added to a solution of o-anisoyl chloride (271 mg, 1.59 mmol)
in tetrahydrofuran (15 ml). The mixture was stirred at room temperature for 30 min, then
poured into water, and extracted with ethyl acetate. The organic layer was washed
successively with saturated sodium hydrogen carbonate, 2 M HCl, water, and brine, dried
over magnesium sulfate, and evaporated. The residue was purified by silica gel column
chromatography (hexane/ethyl acetate 3:1 to 2:1) to afford 11a (130 mg, 77%).
11a: Colorless prisms (methanol); mp 176.2°C; 1H NMR (600 MHz, CD3OD) 7.95 (dd, J =
7.8, 1.8 Hz, 1 H), 7.70 (dd, J = 6.6, 1.8 Hz, 2 H), 7.57 (td, J = 8.9, 1.7 Hz, 1 H), 7.23 (d, J =
8.4 Hz, 1 H), 7.17 (t, J = 8.4 Hz, 1 H), 7.14 (t, J = 7.2 Hz, 1 H), 7.05 (dd, J = 6.6, 1.8 Hz, 2 H),
6.45 (dd, J = 7.8, 1.8 Hz, 1 H), 6.57 (dd, J = 8.2, 2.2 Hz, 1 H), 6.45 (t, J =2.1 Hz, 1 H), 4.07 (s,
3 H); 13C NMR (150 MHz, CD3OD) 166.87, 160.54, 160.34, 159.15, 155.41, 135.47,
134.52, 132.24, 131.52, 124.43, 123.87, 122.41, 120.81, 113.38, 111.64, 110.76, 107.00,
57.03; HRMS Calcd. for C20H18NO4 [M+H]+ 336.1230; Found 336.1231.
Compound 11b – 11f were synthesized similarly.
11b: Colorless prisms (chloroform); mp 112.4°C; 1H NMR (600 MHz, CD3OD) 7.69 (d, J =
8.9 Hz, 2 H), 7.53 (m, 2 H), 7.44 (t, J = 8.0 Hz, 1 H), 7.17 (m, 2 H), 7.04 (dt, J = 8.9, 2.0 Hz,
2 H), 6.57 (dd, J = 8.2, 1.7 Hz, 1 H), 6.48 (dd, J = 8.2, 1.9 Hz, 1 H), 6.46 (t, J = 7.3 Hz, 1 H),
3.89 (s, 3 H) ; 13C NMR (150 MHz, CD3OD) 168.90, 161.64, 160.48, 160.30, 155.52,
137.87, 135.56, 131.52, 130.99, 124.43, 120.98, 120.67, 119.00, 114.25, 111.65, 110.80,
107.03, 56.24; HRMS Calcd. for C20H18NO4 [M+H]+ 336.1230; Found 336.1234.
1
11c: Colorless prisms (chloroform/ethyl acetate); mp 183.3°C; H NMR (600 MHz, CD3OD)
7.96 (d, J = 8.4 Hz, 2 H), 7.68 (d, J = 9.0 Hz, 2 H), 7.17 (t, J = 7.8 Hz, 1 H), 7.07 (d, J = 8.4
Hz, 2 H), 7.04 (d, J = 8.4 Hz, 2 H), 6.56 (dd, J = 8.1, 2.2 Hz, 1 H), 6.48 (dd, J = 8.0, 2.2 Hz, 1
13
H), 6.45 (t, J = 1.6, Hz 1 H), 3.91 (s, 3 H); C NMR (150 MHz, CD3OD) 168.69, 164.48,
160.56, 160.31, 115.36, 135.79, 131.51, 130.81, 128.49, 124.42, 120.71, 115.16, 111.62,

110.76, 106.99, 56.30; HRMS Calcd. for C20H18NO4 [M+H]+ 336.1230; found 336.1236.
1
11d: Pale brown prisms (methylene chloride/hexane); mp 103.7°C; H NMR (600 MHz,
CD3OD) 8.01 (d, J = 7.8 Hz, 1 H), 7.71 (d, J = 8.4 Hz, 2 H), 7.55 (t, J = 8.4 Hz, 1 H),
7.41(d, J = 8.4 Hz, 1 H), 7.25 (t, J = 7.2 Hz, 1 H), 7.17 (t, J = 8.4 Hz, 1 H), 7.06 (d, J = 8.4 Hz,
2 H), 6.57 (dd, J = 8.4, 1.2 Hz, 1 H), 6.46 (dd, J = 8.4, 1.2 Hz, 1 H), 6.45 (t, J = 2.1 Hz, 1 H),
2.88 (s, 6 H); 13C NMR (150 MHz, CD3OD) 167.66, 160.57, 160.32, 155.16, 154.02,
135.74, 133.90, 131.97, 131.54, 128.85, 125.14, 123.33, 121.52, 121.01, 111.55, 110.64,
106.87, 45.52; HRMS Calcd. for C21H21N2O3 [M+H]+ 349.1547; found 349.1536.
11e: Pale yellow prisms (chloroform); mp 171.0°C; 1H NMR (600 MHz, CD3OD) 7.69 (d, J
= 8.6 Hz, 2 H), 7.37, (t, J = 9.0 Hz, 1 H), 7.34 (s, 1 H), 7.28 (d, J = 6.9 Hz, 1 H), 7.17 (t, J =
8.2 Hz, 1 H), 7.04 (d, J = 8.4 Hz, 2 H), 7.02 (d, J = 9.7 Hz, 1 H), 6.57 (dd, J = 8.2, 1.7 Hz, 1
13
H), 6.48 (dd, J = 8.2, 1.7 Hz, 1 H), 6.45 (t, J = 2.2 Hz, 1 H), 3.05 (s, 6 H); C NMR (150
MHz, CD3OD) 170.02, 160.52, 160.32, 155.38, 152.46, 137.16, 135.72, 131.54, 130.54,
124.42, 120.70, 117.43, 116.90, 113.05, 111.58, 110.71, 106.91, 41.18; HRMS Calcd. for
C21H21N2O3 [M+H]+ 349.1547; found 349.1543.
11f: Pale yellow prisms (chloroform); mp 163.9°C; 1H NMR (600 MHz, CD3OD) 7.88 (dt, J
= 7.0, 2.0 Hz, 2 H), 7.66 (dt, J = 6.8, 2.2 Hz, 2 H), 7.16 (t, J = 8.4 Hz, 1 H), 7.03 (dt, J = 7.0,
2.0 Hz, 2 H), 6.82 (dt, J = 7.0, 2.0 Hz, 2 H), 6.56 (ddd, J= 8.2, 2.3, 0.7 Hz, 1 H), 6.48 (ddd, J=
8.0, 2.3, 0.7 Hz, 1 H), 6.45 (t, J = 2.3 Hz, 1 H), 3.10 (s, 6 H) ; 13CNMR (150 MHz, CD3OD)
169.26, 160.65, 160.27, 155.09, 154.84, 136.09, 131.50, 130.45, 124.39, 122.55, 120.73,
112.52, 111.54, 110.71, 106.91, 40.53; HRMS Calcd. for C21H21N2O3 [M+H]+ 349.1547;
found 349.1547.
4.3. Synthesis of 12a (General procedure for compounds 12 and 13, Scheme 2)
A mixture of p-(benzyloxy)phenol (220 mg, 1.10 mmol), m-fluoronitrobenzene (149 mg, 1.05
mmol), and potassium carbonate (173 mg, 1.25 mmol) in DMF (1.5 ml) was stirred at 150°C
for 3 h, and at 80°C for 24 h. It was allowed to cool to room temperature, then 1 M HCl was
added, and the mixture was extracted with ethyl acetate. The organic layer was dried over
sodium sulfate, and evaporated. The residue was purified by silica gel column
chromatography (hexane/ethyl acetate 19:1) to afford 22a (237 mg, 70%)
22a: 1H NMR (600 MHz, CDCl3)  7.89 (ddd, J = 7.8, 2.4, 0.6 Hz, 1 H), 7.72 (t, J = 2.4 Hz, 1
H), 7.43 (m, 5 H), 7.35 (t, J = 5.4 Hz, 1 H), 7.28 (ddd, J = 8.4, 2.4, 0.6 Hz, 1 H), 7.01 (s, 4 H),
5.08 (s, 2 H).
A mixture of 22a (160 mg, 0.5 mmol) and palladium hydroxide on carbon (17 mg) in

tetrahydrofuran (5 ml) and methanol (5 ml) was stirred under a hydrogen atmosphere for 18 h.
The reaction mixture was filtered over Celite, and the filtrate was evaporated to afford 23a
(106 mg, quant) as a pale brown oil.
23a: 1H NMR (600 MHz, CD3OD)  7.05 (t, J = 8.4 Hz, 1 H), 6.88 (dt, J = 9.0, 2.4 Hz, 2 H),
6.80 (dt, J = 9.0, 2.4 Hz, 2 H), 6.46 (ddd, J = 7.8, 1.8, 0.6 Hz, 1 H), 6.53 (t, J = 2.4 Hz, 1 H),
6.29 (ddd, J = 8.4, 2.4, 0.6 Hz, 1 H).
23a (89 mg, 0.44 mmol) was added to a solution of o-anisoyl chloride (228 mg, 1.34 mmol)
in tetrahydrofuran (15 ml). The mixture was stirred at room temperature for 30 min, then
poured into water, and extracted with ethyl acetate. The organic layer was washed
successively with saturated sodium hydrogen carbonate, 2 M HCl, water, and brine, dried
over magnesium sulfate, and evaporated. The residue was purified by silica gel column
chromatography (chloroform/ethyl acetate 4:1) to afford 12a (63 mg, 43%).
12a: Colorless prisms (chloroform/ethyl acetate); mp 189.4°C; 1H NMR (600 MHz, CD3OD)
7.89 (dd, J= 7.7, 1.6 Hz, 1 H), 7.56 (td, J = 7.8, 1.6 Hz, 1 H), 7.48 (t, J = 1.0 Hz, 1 H), 7.29
(m, 2 H), 7.21 (d, J = 8.4 Hz, 1 H), 7.12 (t, J = 7.4 Hz, 1 H), 6.94 (dt, J = 8.9, 2.2 Hz, 2 H),
13
6.84 (dt, J = 8.8, 2.1 Hz, 2 H), 6.70 (m, 1 H), 4.04 (s, 3 H); C NMR (150 MHz, CD3OD)
167.01, 161.06, 159.09, 155.34, 150.72, 141.26, 134.52, 132.17, 131.04, 124.54, 122.40,
122.34, 117.55, 115.85, 114.51, 113.38, 111.33, 57.02; HRMS Calcd. for C20H18NO4 [M+H]+
336.1230; found 336.1226.
Compound 12c, d, f and 13 were synthesized similarly.
12c: Colorless prisms (ethyl acetate); mp 163.6°C; 1H NMR (600 MHz, CD3OD) 7.92 (d, J
= 8.9 Hz, 2 H), 7.40 (t, J = 2.2 Hz, 1 H), 7.37 (ddd, J = 7.9, 1.7, 0.7 Hz, 1 H), 7.29 (t, J = 8.1
Hz, 1 H), 7.05 (d, J = 8.9 Hz, 2 H), 6.94 (d, J = 8.9 Hz, 2 H), 6.83 (d, J = 8.9 Hz, 2 H), 6.71
(ddd, J = 8.1, 2.3, 0.8 Hz, 1 H), 3.90 (s, 3 H); 13C NMR (150 MHz, CD3OD) 168.75, 164.50,
160.99, 155.33, 150.70, 141.68, 130.90, 130.85, 128.50, 122.39, 117.54, 116.26, 115.12,
114.46, 111.58, 56.27; HRMS Calcd. for C20H18NO4 [M+H]+ 336.1230; found 336.1224.
12d: Pale brown prisms (methanol/ ethyl acetate); mp 196.4°C; 1H NMR (600 MHz, CD3OD)
7.96 (dd, J = 7.9, 1.5 Hz, 1 H), 7.54 (td, J = 8.6, 1.6 Hz, 1 H), 7.48 (t, J = 2.0 Hz, 1 H), 7.40
(d, J = 8.1 Hz, 1 H), 7.31 (m, 1 H), 7.24 (m, 2 H), 6.95 (d, J = 8.9 Hz, 2 H), 6.84 (d, J = 8.8
Hz, 2 H), 6.69 (dd, J = 8.2, 2.2 Hz, 1 H), 2.85 (s, 6H); 13C NMR (150 MHz, DMSO-d6)

; HRMS Calcd. for C21H21N2O3 [M+H]+
349.1547; found 349.1544.
12f: Pale yellow prisms (ethyl acetate); mp 205.8°C; 1H NMR (600 MHz, CD3OD) 7.87 (d,

J = 8.9 Hz, 2 H), 7.43 (t, J = 2.2 Hz, 1 H), 7.39 (d, J = 8.0 Hz, 1 H), 7.31 (t, J = 8.2 Hz, 1 H),
6.97 (d, J = 8.9 Hz, 2 H), 6.86 (d, J = 8.9 Hz, 2 H), 6.83 (d, J = 9.0 Hz, 2 H), 6.71 (dd, J = 8.1,
13
2.0 Hz, 1 H), 3.10 (s, 6 H); C NMR (150 MHz, CD3OD) 169.29, 160.93, 155.27, 154.86,
150.78, 141.97, 130.83, 130.50, 122.55, 122.35, 117.53, 116.26, 114.17, 112.47, 111.60,
40.50; HRMS Calcd. for C21H21N2O3 [M+H]+ 349.1547; found 349.1544.
1
13a: Pale orange prisms (methanol); mp 189.5-190°C; H NMR (600 MHz, acetone-d6)
8.83 (s, 1 H), 8.34 (s, 1 H), 7.95 (d, J = 8.9 Hz, 1 H), 7.34 (dd, J = 11.0, 2.1 Hz, 2 H), 7.17
(d, J = 8.2 Hz, 1 H), 7.01 (dt, J = 8.9, 2.0 Hz, 2 H), 6.89 (dt, J = 9.0, 2.1 Hz, 2 H), 6.83 (dt, J
= 8.9, 2.7 Hz, 2 H), 6.67 (dd, J = 8.3, 2.8 Hz, 1 H), 3.86 (s, 3 H), 3.28 (s, 3 H); 13C NMR (150
MHz, acetone-d6) 165.42, 163.33, 157.84, 154.55, 150.21, 138.73, 131.80, 131.69, 130.18,
128.04, 126.32, 121.54, 116.99, 116.90, 114.90, 114.63, 114.52, 114.44, 55.80, 17.440;
HRMS Calcd. for C21H20NO4 [M+H]+ 350.1387; found 350.1384.
1
13b: Pale brown prisms (methanol); mp 203-204°C; H NMR (600 MHz, acetone-d6) 9.03
(s,1 H), 8.01 (dt, J = 9.0, 2.1 Hz, 2 H), 7.33 (d, J = 8.2 Hz, 1 H), 7.13 (t, J = 8.3 Hz, 1 H),
7.04 (dt, J = 9.0, 2.1 Hz, 2 H), 6.83 (s, 1 H), 6.64 (d, J = 7.5 Hz, 1 H), 3.87 (s, 3 H), 2.21 (s, 3
H); 13C NMR (150 MHz, acetone-d6) 165.61, 163.32, 157.29, 154.16, 150.90, 139.27,
130.23, 126.87, 121.12, 120.37, 116.97, 116.88, 115.57, 114.45, 55.81, 11.23; HRMS Calcd.
for C21H20NO4 [M+H]+ 350.1387; found 350.1383.
1
13c: Colorless prisms (chloroform/ethyl acetate); mp 179.0-179.5°C; H NMR (600 MHz,
acetone-d6) 9.35 (br, 1 H), 8.21 (br, 1 H), 7.91 (d, J = 8.3 Hz, 2 H), 7.54 (dt, J = 8.2, 2.8 Hz,
1 H), 7.29 (t, J = 2.1 Hz, 1 H), 7.19 (d, 8.3 Hz, 1 H), 6.99 (dt, J = 9.0, 2.7 Hz, 2 H), 6.85 (m, 4
13
H), 3.85 (s, 3 H), 2.22 (s, 3 H); C NMR (150 MHz, acetone-d6) 165.47, 163.21, 157.09,
154.19, 150.64, 139.59, 131.72, 130.09, 128.26, 123.93, 120.66, 116.99, 116.90, 115.26,
114.33, 110.14, 55.76, 15.81; HRMS Calcd. for C21H20NO4 [M+H]+ 350.1387; found
350.1383.
1
13d: Colorless needles (methanol); mp 190.0-191.5°C; H NMR (600 MHz, acetone-d6)
9.81 (br, 1 H), 8.19 (s, 1 H), 7.98 (dd, J = 8.3, 2.1 Hz, 1 H), 7.51 (ddd, J = 8.2, 6.9, 1.4 Hz,
1 H), 7.48 (d, J = 2.7 Hz, 1 H), 7.36 (dd, J = 8.3, 2.1 Hz, 1 H), 7.21 (d, J = 8.2 Hz, 1 H), 7.18
(d, J = 8.2 Hz, 1 H), 7.08 (td, J = 7.6, 1.3 Hz, 1 H), 6.84 (s, 4 H), 4.03 (s, 3 H), 2.21 (s, 3H);
13C NMR (150 MHz, CD3OD) 166.65, 158.68, 157.52, 154.37, 151.23, 138.59, 134.12,
132.17, 131.74, 125.87, 124.25, 122.01, 120.60, 117.16, 116.22, 112.92, 111.51, 56.60, 15.93;
HRMS Calcd. for C21H20NO4 [M+H]+ 350.1387; found 350.1385.
4.4. Synthesis of 12b (Scheme 2)

A solution of copper (II) sulfate·5H2O (267 mg, 1.07 mmol) in methanol (4 ml) was added to
a solution of sodium borohydride (111 mg, 2.94 mmol) and 22 (165 mg, 0.5 mmol) in
methylene chloride (5 ml), and the mixture was stirred at 0°C for 3 h, and at room
temperature for 2 h. The mixture was poured into water, and extracted with methylene
chloride. The organic layer was washed with water and brine, dried over sodium sulfate, and
evaporated. The residue was dissolved in methylene chloride, and 1 M hydrochloric acid in
ether was added to it. The resulting precipitate was collected to afford 26 (108 mg, 63%).
26: Pale yellow powder; 1H NMR (600 MHz, CD3OD) 7.43 (d, J = 7.6 Hz, 2 H), 7.36 (t, J =
6.9 Hz, 2 H), 7.29 (t, J = 6.9 Hz, 1 H), 7.26 (d, J = 2.0 Hz, 1 H), 7.12 (dd, J = 8.9, 2.8 Hz, 1
H), 7.01 (dt, J = 9.6, 2.8 Hz), 6.90 (dt, J = 8.9, 2.0 Hz, 2 H), 6.80 (d, J = 8.9 Hz, 1 H), 5.06 (s,
2 H), 2.32 (s, 3 H); 13C NMR (150 MHz, CD3OD)  158.08, 156.78, 151.44, 138.65, 131.96,
129.52, 128.92, 128.58, 126.62, 126.08, 122.65, 121.20, 119.00, 117.32, 71.50, 16.31.
m-Anisoyl chloride (161 mg, 0.18 mmol) was added to a solution of 26 (103 mg, 0.49 mmol)
in pyridine (5 ml). The mixture was stirred at room temperature for 45 min, then poured into
water, and extracted with ethyl acetate. The organic layer was successively washed with
saturated sodium hydrogen carbonate, 2 M hydrochloric acid, water and brine, dried over
magnesium sulfate, and evaporated. The residue was purified by silica gel column
chromatography (hexane/ethyl acetate 2:1) to give 27a (R1 = OMe, 122 mg, 90%).
27a: 1H NMR (600 MHz, CDCl3) 7.74 (br, 1 H), 7.44 (d, J = 7.6 Hz, 2 H), 7.33-7.41 (m, 7
H), 7.29 (t, J = 8.3 Hz, 1 H), 7.01 (ddd, J = 7.6, 2.8, 1.4 Hz, 1 H), 7.01 (dt, J = 8.9, 2.7 Hz, 2
H), 6.97 (dt, J = 9.6, 2.7 Hz, 2 H), 6.75 (ddd, J = 8.3, 2.0, 1.4 Hz, 1 H), 5.05 (s, 2 H), 3.86 (s,
3 H); 13C NMR (150 MHz, CDCl3)165.44, 160.01, 159.15, 155.29, 149.99, 139.13, 136.97,
136.33, 130.06, 129.80, 128.60, 128.00, 127.53, 121.01, 118.58, 118.12, 115.9597, 114.08,
113.73, 112.46, 109.35, 70.52, 55.50.
A mixture of 27a (74 mg, 0.17 mmol) and palladium hydroxide on carbon (5 mg) in
tetrahydrofuran (5 ml) and methanol (2 ml) was stirred at 45°C under a hydrogen atmosphere
for 41 h. The reaction mixture was filtered over Celite, and the filtrate was evaporated. The
residue was purified by silica gel column chromatography (hexane /ethyl acetate 3:2),
followed by recrystallization from chloroform/ethyl acetate to give 12b (33 mg, 57 %).
12b: Colorless prisms (chloroform/ethyl acetate); mp 144.6°C; 1H NMR (600 MHz, CD3OD)
7.50 (t, J = 7.7 Hz, 1 H), 7.48 (d, J = 2.1 Hz, 1 H), 7.43 (t, J = 8.0 Hz, 1 H), 7.42 (d, J = 2.5
Hz, 1 H), 7.39 (d, J = 8.1 Hz, 1 H), 7.30 (t, J = 8.2 Hz, 1 H), 7.16 (ddd, J = 8.0, 2.2, 0.5 Hz, 1
H), 6.94 (d, J = 8.9 Hz, 2 H), 6.84 (d, J = 8.9 Hz, 2 H), 6.72 (dd, J = 8.1, 1.7 Hz, 1 H), 3.89 (s,
3 H); 13C NMR(150 MHz, CD3OD) 169.01, 161.64, 161.00, 155.35, 150.67, 141.49, 137.90,
130.97, 130.94, 122.40, 121.03, 119.12, 117.55, 116.30, 114.66, 114.21, 111.63, 56.25;
HRMS Calcd. for C20H18NO4 [M+H]+ 336.1230; found 336.1226.

Compound 12e were synthesized similarly.
12e: Colorless prisms; mp 167.2°C; 1H NMR (600 MHz, CD3OD) .43 (t, J = 1.8 Hz, 1 H),
7.38 (d, J = 7.6 Hz, 1 H), 7.34 (t, J = 7.9 Hz, 1 H), 7.30 (t, J = 8.2 Hz, 1 H), 7.28 (s, 1 H), 7.22
(d, J = 7.6 Hz, 1 H), 6.98 (ddd, J = 8.2, 2.6, 0.5 Hz, 1 H), 6.94 (d, J = 8.9 Hz, 2 H), 6.83 (d, J
= 8.9 Hz, 2 H), 6.72 (ddd, J = 7.4, 2.6, 1.6 Hz, 1 H), 3.03 (s, 6 H); 13C NMR (150 MHz,
CD3OD) 170.17, 161.00, 155.37, 152.55, 150.56, 141.62, 137.14, 130.92, 130.48, 122.46,
117.51, 117.32, 116.70, 116.21, 114.46, 112.90, 111.49, 41.06; HRMS Calcd. for C21H21N2O3
[M+H]+ 349.1547; found 349.1544.
4.5. Synthesis of 14a (General procedure for compounds 14, Scheme 2)
O-Monobenzylresorcinol (220 mg, 1.10 mmol) was added to
a
solution of
m-fluoronitrobenzene (149 mg, 1.05 mmol) and potassium carbonate (173 mg, 1.25 mmol) in
DMF (1.5 ml) under an argon atmosphere, and the mixture was stirred at 150°C for 3 h and at
80°C for 24 h. It was allowed to cool to room temperature, then poured into 1 M HCl, and
extracted with ethyl acetate. The organic layer was dried over sodium sulfate, and evaporated.
The residue was purified by silica gel column chromatography (hexane/ethyl acetate 19:1) to
afford 24a (273 mg, 80%).
24a: 1H NMR (600 MHz, CDCl3) 7.94 (ddd, J = 8.4, 2.4, 0/6 Hz, 1 H), 7.81 (t, J = 2.4 Hz, 1
H), 7.48 (t, J = 7.8 Hz, 1 H), 7.36 (m, 7 H), 6.84 (dd, J = 8.4, 2.4 Hz, 1 H), 6.67 (m, 2 H),
5.06 (s, 2 H).
A mixture of 24a (166 mg, 0.5 mmol) and palladium hydroxide on carbon (20 mg) in THF (5
ml) and methanol (5 ml) was stirred under a hydrogen atmosphere for 14 h. The reaction
mixture was filtered over Celite, and evaporated to give 25a (107 mg, quant), which was used
for the next reaction without further purification. 25a (115 mg, 0.57 mmol) was added to a
solution of o-anisoyl chloride (295 mg, 1.73 mmol) in tetrahydrofuran (15 ml), and the
mixture was stirred at room temperature for 30 min, then poured into water, and extracted
with ethyl acetate. The organic layer was washed successively with saturated sodium
hydrogen carbonate, 2 M hydrochloric acid, water and brine, dried over magnesium sulfate,
and evaporated. The residue was washed with dichloromethane to give 14a (122 mg, 71%),
which was purified by recrystallization from chloroform.
14a: pale brown prisms (chloroform); mp 166.0°C; 1H NMR (600 MHz, CD3OD) 7.90 (dd,
J = 7.7, 1.7 Hz, 1 H), 7.56 (m, 2 H), 7.37 (m, 2 H), 7.21 (d, J = 8.3 Hz, 1 H), 7.18 (t, J = 8.1
Hz, 1 H), 6.80 (ddd, J = 7.9, 3.3, 1.0 Hz, 1 H), 6.59 (ddd, J = 8.3, 2.2, 0.7 Hz, 1 H), 6.52 (dd,
J = 8.2, 1.7 Hz, 1 H), 6.48 (t, J = 2.3 Hz, 1 H), 4.04 (s, 3 H) ; 13C NMR (150 MHz, CD3OD) 
167.10, 160.36, 160.01, 159.35, 159.04, 141.42, 134.56, 132.12, 131.57, 131.19, 124.50,
122.34, 116.72, 116.04, 113.27, 112.76, 111.85, 111.14, 107.35, 56.95; HRMS; Calcd. for

C20H18NO4 [M+H]+ 336.1230; found 336.1228.
Compound 14b – 14e were synthesized similarly.
1
14b: Pale brown prisms (chloroform); mp 129.5-130.5°C; H NMR (600 MHz, CD3OD)
7.40 (m, 5 H), 7.33 (m, 1 H), 7.13 (m, 2 H), 6.77 (dd, J = 8.3, 2.1 Hz, 1 H), 6.54 (dd, J = 8.3,
2.7 Hz, 1 H), 6.48 (dd, J = 8.2, 2.0 Hz, 1 H), 6.44 (s, 1 H), 3.85 (s, 3 H); 13C NMR (150 MHz,
CD3OD) 168.72, 161.30, 160.07, 159.65, 159.03, 141.36, 137.52, 131.29, 130.81, 130.71,
120.73, 118.84, 116.83, 115.83, 113.82, 112.73, 111.58, 110.93, 107.14, 55.90; HRMS Calcd.
for C20H18NO4 [M+H]+ 336.1230; found 336.1228.
1
14c: Pale brown prisms; mp 132-132.5°C; H NMR (600 MHz, CD3OD) 7.89 (dd, J = 6.8,
2.0 Hz, 2 H), 7.45 (t, J = 2.0 Hz, 1 H), 7.42 (d, J= 9.0 Hz, 1 H), 7.31 (t, J = 8.2 Hz, 1 H), 7.13
(t, J = 8.2 Hz, 1 H), 7.01 (d, J = 8.9 Hz, 2 H), 6.75 (dd, J = 8.6, 2.7 Hz, 1 H), 6.54 (dd, J = 8.6,
2.7 Hz, 1 H), 6.48 (dd, J = 8.3, 2.1 Hz, 1 H), 6.44 (t, J = 2.4 Hz, 1 H), 3.86 (s, 3 H); 13C NMR
(150 MHz, CD3OD) 168.76, 164.51, 160.34, 159.97, 159.30, 141.84, 131.59, 131.08,
130.91, 128.36, 117.11, 115.94, 115.11, 113.01, 111.85, 111.22, 107.43, 56.27; HRMS
Calcd. for C20H18NO4 [M+H]+ 336.1230; found 336.1228.
14d: pale yellow prisms (chloroform); mp 164.8°C; 1H NMR (600 MHz, CD3OD) 7.97 (d, J
= 7.2 Hz, 1 H), 7.58 (s, 1 H), 7.55 (t, J = 8.1 Hz, 1 H), 7.38 (m, 3 H), 7.24 (t, J =7.6 Hz, 1 H),
7.19 (t, J = 8.1 Hz, 1 H), 6.79 (dt, J = 6. 8, 2.3 Hz, 1 H), 6.59 (dd, J = 8.2, 2.3 Hz, 1 H), 6.53
(dd, J = 8.2, 2.2 Hz, 1 H), 6.49 (t, J = 2.2 Hz, 1 H), 2.86 (s, 6 H); 13C NMR (150 MHz,
CDCl3) 167.83, 160.38, 159.95, 159.50, 154.02, 141.64, 134.00, 131.99, 131.58, 131.32,
128.78, 125.15, 121.54, 116.33, 115.81, 112.35, 111.90, 111.20, 107.42, 45.50; HRMS Calcd.
for C20H18NO4 [M+H]+ 349.1547; found 349.1544.
1
14e: Pale brown prisms; mp 179.3°C; H NMR (600 MHz, CD3OD) 7.51 (t, J = 2.3 Hz, 1
H), 7.47 (ddd, J = 8.2, 1.7, 0.8 Hz, 1 H), 7.36 (m. 2 H), 7.30 (t, J = 1.9 Hz, 1 H), 7.23 (dd, J =
6.9, 1.3 Hz, 1 H), 7.18 (t, J = 8.1 Hz, 1 H), 6.99 (dd, J = 6.9, 1.3 Hz, 1 H), 7.18 (t, J = 8.1 Hz,
1 H), 6.99 (dd, J = 8.0, 2.6 Hz, 1 H), 6.81 (ddd, J = 8.2, 2.3, 0.7 Hz, 1 H), 6.58 (ddd, J = 8.2,
2.2, 0.7 Hz, 1 H), 6.53 (ddd, J = 8.3, 2.3, 0.7 Hz, 1 H), 6.49 (t, J= 2.3 Hz, 1 H), 3.04 (s, 6 H);
13C NMR (150 MHz, CDCl3) 170.11, 160.27, 159.98, 159.27, 152.56, 141.71, 137.09,
131.54, 131.03, 130.46, 117.36, 117.27, 116.68, 116.07, 113.20, 112.93, 111.89, 111.26,
107.48, 41.04. HRMS Calcd. for C21H21N2O3 [M+H]+ 349.1547; found 349.1542.
4.6. Synthesis of 15 (Scheme 3)
An aqueous solution of 5 M sodium hydroxide (2.0 ml, 10 mmol) was added to DMSO (8 ml)
at room temperature under an Ar atmosphere. After 10 min, O-monobenzylhydroquinone (961

mg, 4.8 mmol) was added to it. The mixture was stirred at 50°C for 15 min, then
2-fluoro-5-nitrotoluene (620 mg, 4.0 mmol) was added, and stirring was continued at 50°C
for 5.5 h, and at room temperature for 15 h. The reaction mixture was then poured into ice
water and the precipitate was collected by filtration to give 29a (R2 = 2-Me, 1.04 g, 65%).
1
29a (R2 = 2-Me): Yellow powder; H NMR (600 MHz, CDCl3)  8.13 (d, J = 2.1 Hz, 1 H),
7.96 (dd, J = 8.9, 2.7 Hz, 1 H), 7.36 (m, 5 H), 7.02 (dt, J = 9.6, 2.8 Hz, 2 H), 6.99 (dt, J = 8.9,
13
2.7 Hz, 2 H), 6.69 (d, J = 8.9 Hz, 1 H), 5.08 (s, 2 H), 2.42 (s, 3 H); C NMR (150 MHz,
CDCl3)  162.21, 156.04, 148.51, 142.06, 136.67, 128.65, 128.12, 127.48, 126.64, 123.14,
121.50, 116.24, 114.47.
A solution of copper (II) sulfate·5H2O (502 mg, 2.0 mmol) in methanol (4 ml) was added to a
solution of sodium borohydride (189 mg, 5.0 mmol) and 29a (R2 = 2-Me, 234 mg, 0.70
mmol) in methylene chloride (5 ml), and the mixture was stirred at 0°C for 2.5 h. Sodium
borohydride (76 mg, 2.0 mmol) was added to it, and stirring was continued at 0°C for 2 h.
Sodium borohydride (76 mg, 2.0 mmol) was added, and the reaction mixture was stirred at
0°C for 1 h, then poured into water, and extracted with methylene chloride. The organic layer
was washed with water and brine, dried over sodium sulfate, and evaporated. The residue was
dissolved in ether, and 1 M hydrochloric acid in ether was added to the solution. The resulting
precipitate was collected to afford 30 (205 mg, 86%).
30: Pale brown powder; 1H NMR (600 MHz, CD3OD)  7.43 (d, J = 7.6 Hz, 2 H), 7.33 (m, 3
H), 7.26 (d, J = 2.0 Hz, 1 H), 7.12 (dd, J = 8.9, 2.8 Hz, 1 H), 7.01 (dt, J = 9.6, 2.8 Hz, 2 H),
13
6.90 (dt, J = 8.9, 2.0 Hz, 2 H), 6.80 (d, J = 8.9 Hz, 1 H), 5.06 (s, 2 H), 2.33 (s, 3 H) ; C
NMR (150 MHz, CD3OD) 158.08, 156.78, 151.44, 138.65, 131.96, 129.52, 128.92, 128.58,
126.62, 126.08, 122.65, 121.20, 119.00, 117.32, 71.50, 16.31.
m-Anisoyl chloride (61 mg, 0.36 mmol) was added to a solution of 30 (102 mg, 0.30 mmol)
in pyridine (5 ml). The mixture was stirred at room temperature for 30 min, then poured into
water, and extracted with ethyl acetate. The organic layer was washed successively with
saturated sodium hydrogen carbonate, 1 M hydrochloric acid, water and brine, dried over
sodium sulfate, and evaporated to give 31 (113 mg, 86%).
31: 1H NMR (600 MHz, CDCl3)  7.74 (s, 1 H), 7.54 (d, J = 2.7 Hz, 1 H), 7.39 (m, 8 H), 7.08
(dt, J = 6.9, 2.7 Hz, 1 H), 6.93 (dt, J = 9.6, 2.7 Hz, 2 H), 6.75 (dt, J = 9.0, 2.4 Hz, 2 H), 6.56
13
(d, J = 9.0 Hz, 1 H), 5.04 (s, 2 H), 3.87 (s, 3 H), 2.28 (s, 3 H); C NMR (150 MHz, CDCl3)
 165.41, 159.97, 154.37, 152.25, 151.58, 137.03, 136.43, 133.14, 130.19, 129.77, 128.57,
127.96, 127.49,123.37, 119.10, 118.77, 118.55, 117.94, 115.85, 112.45, 70.56, 55.48, 16.38.
Sodium hydride (8.0 mg, 0.30 mmol) was washed with n-hexane twice. A solution of 31 (44
mg, 0.10 mmol) in dry DMF (0.5 ml) was added to a suspension of sodium hydride in dry

DMF (0.3 ml) at 0°C, and the mixture was stirred at room temperature for 30 min.
Iodomethane (43 mg, 0.30 mmol) in dry DMF (0.3 ml) was added at 0°C, and stirring was
continued at room temperature for 30 min. Remaining iodomethane was removed in vacuo.
The residue was poured into water, and extracted with ethyl acetate. The organic layer was
washed with water and brine, dried over sodium sulfate, and evaporated. The residue was
purified by silica gel column chromatography (hexane/ethyl acetate 2:1 to 3:2) to afford 32
(38 mg, 86%).
32: 1H NMR (600 MHz, CDCl3)  7.37 (m, 5 H), 7.08 (t, J = 7.9 Hz, 1 H), 6.95 (br, 1 H), 6.90
(m, 3 H), 6.86 (br d, J = 7.5 Hz, 1 H), 6.80 (m, 3 H), 6.75 (br d, J = 7.5 Hz, 1 H), 6.61 (d, J =
13
9.0 Hz, 1 H), 5.02 (s, 2 H), 3.69 (s, 3 H), 3.46 (s, 3 H), 2.17 (s, 3 H); C NMR (150 MHz,
CDCl3)  170.33, 158.85, 154.63, 154.07, 150.82, 139.72, 137.20, 136.90, 129.39, 128.68,
128.55, 127.96, 127.41, 119.21, 118.25, 115.85, 115.81, 70.50, 55.14, 38.51, 16.15.
A mixture of 32 (38 mg, 0.086 mmol) and palladium hydroxide on carbon (5 mg) in THF (2
ml) was stirred under a hydrogen atmosphere for 21 h. The reaction mixture was filtered over
Celite, and the filtrate was evaporated. The crude product was recrystallized from methanol to
give 15 (9 mg, 30%).
15: Colorless prisms (methanol); mp 172-173°C; 1H NMR (600 MHz, acetone-d6)  7.12 (t, J
= 7.5 Hz, 2 H), 6.90 (br dd, J = 9.0, 2.8 Hz, 1 H), 6.87 (d, J = 7.6 Hz, 2 H), 6.85 (m, 1 H),
6.80 (m, 3 H), 6.74 (dt, J = 9.0, 2.1 Hz, 2 H), 6.60 (d, J = 8.9 Hz, 1 H), 3.66 (s, 3 H) , 3.37 (s,
3 H) , 2.13 (s, 3 H); 13C NMR (150 MHz, acetone- d6)  170.16, 159.80, 155.29, 154.27,
154.17, 150.54, 140.88, 138.99, 130.67, 130.11, 129.47, 126.74, 121.65, 120.35, 118.53,
118.53, 117.00, 115.95, 114.62, 55.45, 38.37, 16.18; HRMS Calcd. for C21H20NO4 [M+H]+
350.1387; found 350.1384.
4.7. Synthesis of 16a (Scheme 3)
Sodium hydride (8.0 mg, 0.30 mmol) was washed with n-hexane twice. A solution of 27a (R1
= m-OMe, 44 mg, 0.10 mmol) in dry DMF (0.7 ml) was added to a suspension of sodium
hydride in dry DMF (0.3 ml) at 0°C, and the mixture was stirred at room temperature for 30
min. Iodomethane (47 mg, 0.33 mmol) was added at 0°C, and stirring was continued at room
temperature for 40 min. Remaining iodomethane was removed in vacuo, then the residue was
poured into water, and extracted with ethyl acetate. The organic layer was washed with water
and brine, dried over sodium sulfate, and evaporated. The residue was purified by silica gel
column chromatography (hexane/ethyl 2:1) to give 28a (37 mg, 82%).
28a: 1H NMR (600 MHz, CDCl3)  7.44 (d, J = 6.9 Hz, 2 H), 7.4 (t, J = 7.6 Hz, 2 H), 7.34 (t,
J = 7.6 Hz, 1 H), 7.20 (t, J = 8.3 Hz, 1 H), 7.06 (t, J = 8.2 Hz, 1 H), 6.84 (m, 5 H), 6.78 (m, 2
H), 6.48 (dt, J = 8.9, 2.1 Hz, 2 H), 6.45 (t, J = 2.1 Hz, 1 H), 5.04 (s, 2 H), 3.67 (s, 3 H), 3.45 (s,
3 H); 13C NMR (150 MHz, CDCl3)  170.31, 159.02, 158.62, 155.16, 149.40, 145.99, 137.02,

136.85, 130.09, 128.77, 128.59, 128.01, 127.41, 121.03, 120.47, 120.11, 116.32, 116.22,
115.98, 115.86, 113.36, 70.41, 55.14, 38.09.
A mixture of 28a (37 mg, 0.085 mmol) and palladium hydroxide on carbon (5 mg) in
tetrahydrofuran (2.5 ml) was stirred under a hydrogen atmosphere for 18.5 h. The reaction
mixture was filtered over Celite, and evaporated. The crude product was recrystallized from
chloroform to give 16a (19 mg, 64%).
16a: Colorless prisms (chloroform); mp 160-160.5°C; 1H NMR (600 MHz, CDCl3)  7.21 (t,
J = 8.3 Hz, 1H), 7.09 (dt, J = 7.5, 1.4 Hz, 1 H), 6.84 (m, 3 H), 6.78 (m, 2 H), 6.72 (dt, J = 9.0,
2.1 Hz, 2 H), 6.59 (dt, J = 9.0, 2.1 Hz, 2 H), 6.42 (br, 1 H), 5.16 (s 1 H), 3.69 (s, 3 H), 3.46 (s,
3 H); 13C NMR (150 MHz, CDCl3)  170.55, 159.07, 158.84, 152.24, 149.01, 145.89, 136.94,
130.14, 128.82, 121.03, 120.80, 119.97, 116.38, 116.33, 116.15, 115.99, 113.36, 55.19, 38.19
HRMS Calcd. for C21H20NO4 [M+H]+ 350.1387; found 350.1385.
Compound 16b was synthesized similarly.
1
16b: Colorless needles; mp 107.5-108.5 °C; H NMR (600 MHz, CDCl3)  7.24 (dt, J = 8.9,
2.1 Hz, 2 H), 7.21 (t, J = 8.2 Hz, 1 H), 6.81 (dd, J = 6.9, 2.1 Hz, 1 H), 6.78(ddd, J = 8.3, 2.7,
1.4 Hz, 1 H), 6.72 (m, 4 H), 6.63 (dt, J = 8.9, 2.1 Hz, 2 H), 6.47 (t, J = 2.1 Hz, 1 H), 5.34 (br d,
J = 10.3 Hz, 1 H), 3.79 (s, 3 H) , 3.46 (s, 3 H); 13C NMR (150 MHz, CDCl3)  170.45, 160.70,
158.80, 152.30, 149.11, 146.39, 130.79, 130.14, 127.74, 120.71, 120.15, 116.37, 116.29,
115.78, 113.08, 55.27, 38.42; HRMS Calcd. for C21H20NO4 [M+H]+ 350.1387; found
350.1383.
4.8. Synthesis of 17a (General procedure for compounds 17 - 19, Scheme 3)
A mixture of 29b (R2 = 2-F, 153 mg, 0.80 mmol) and palladium hydroxide on carbon (36 mg)
in THF (4 ml) and methanol (4 ml) was stirred under a hydrogen atmosphere for 20 h, then
filtered over Celite. The filtrate evaporated to give 33a (R2 = 2-F, 115 mg, 67%).
33a (R2 = 2-F): red brown solid; 1H NMR (600 MHz, CD3OD)  6.78 (t, J = 8.9 Hz, 2 H),
6.70 (m, 4 H), 6.53 (dd, J = 12.4, 2.8 Hz, 1 H), 6.44 (ddd, J = 8.9, 2.8, 1.4 Hz, 1 H); 13C NMR
(150 MHz, CD3OD)  156.37 (d, J = 242.73 Hz), 153.59, 153.14, 146.86 (d, J = 8.63 Hz),
136.46 (d, J = 12.92 Hz), 123.95, 118.27, 116.80, 112.04, 104.48 (d, J = 21.54 Hz).
m-Anisoyl chloride (97 mg, 0.18 mmol) was added to a solution of 33a (R2 = 2-F, 104 mg,
0.47 mmol) in THF (10 ml). The mixture was stirred at room temperature for 50 min, then
poured into water, and extracted with ethyl acetate. The organic layer was washed
successively with saturated sodium hydrogen carbonate, 1 M hydrochloric acid, water and
brine, dried over sodium sulfate, and evaporated. The residue was purified by silica gel
column chromatography (hexane/ethyl acetate 2:1 to 1:1) to give 17a (134 mg, 80%).

17a: Colorless prisms (chloroform); mp 160-161°C; 1H NMR (600 MHz, CD3OD) 7.73 (dd,
J = 13.0, 2.8 Hz, 1 H), 7.49 (dd, J = 7.6, 1.4 Hz, 1 H), 7.47 (t, J = 2.1 Hz, 1 H), 7.41 (t, J = 7.9
Hz, 1 H), 7.36 (dt, J = 8.9, 1.7 Hz, 1 H), 7.14 (ddd, J = 8.3, 2.7, 1.3 Hz, 1 H), 6.95 (t, J = 9.0
13
Hz, 1 H), 6.68 (dt, J = 8.9, 2.1 Hz, 2 H), 6.76 (dt, J = 8.9, 2.4 Hz, 2 H), 3.86 (s, 3 H); C
NMR (150 MHz, CD3OD) 168.55, 161.33, 154.71 (d, J = 244.16 Hz), 154.67, 151.33,
142.94 (d, J = 11.49 Hz), 137.34, 136.04 (d, J = 10.065 Hz), 130.75, 121.52, 120.70, 120.01,
118.81, 118.12, 117.10, 113.89, 111.09 (d, J = 22.98 Hz), 55.91; HRMS Calcd for
C20H17NO4F [M+H]+: 354.1142. Found 354.1142.
Compounds 17b – e, 18, and 19 were synthesized similarly.
17b: Colorless needles (hexane/ethyl acetate); mp: 187-188 °C; 1H NMR (600 MHz, CD3OD)
 7.92 (d, J = 2.8 Hz, 1 H), 7.48 (m, 3 H), 7.41 (t, J = 7.5 Hz, 1 H), 7.13 (dd, J = 8.2, 2.7 Hz,
1 H), 6.87 (d, J = 9.0 Hz, 1 H), 6.82 (dt, J = 9.0, 2.1 Hz, 2 H), 6.77 (dt, J = 8.9, 2.7 Hz, 2 H),
3.86 (s, 3 H); 13C NMR (150 MHz, CD3OD)  167.22, 160.00, 153.58, 150.41, 149.43,
135.97, 134.39, 129.41, 124.24, 122.93, 120.56, 119.37, 118.88, 117.49, 115.87, 112.54,
54.58; HRMS Calcd for C20H17NO4Cl [M+H]+: 370.0846. Found 370.0844.
1
17c: Colorless prisms (hexane/ethyl acetate); mp 192-193°C; H NMR (600 MHz, CD3OD)
8.07 (d, J = 2.2 Hz, 1 H), 7.55 (dd, J = 8.9, 2.8 Hz, 1 H), 7.48 (d, J = 7.5 Hz, 1 H), 7.46 (t, J
= 2.1 Hz, 1 H), 7.41 (t, J = 7.6 Hz, 1 H), 7.13 (dq, J = 6.9, 1.4 Hz, 1 H), 6.83 (m, 3 H), 6.77
(dt, J = 9.0, 2.8 Hz, 2 H), 3.86 (s, 3 H); 13C NMR (150 MHz, CD3OD) 168.56, 161.34,
154.96, 152.99, 150.75, 137.31, 135.91, 130.76, 127.29, 122.64, 120.86, 120.72, 119.91,
118.84, 117.22, 114.35, 113.88, 55.92; HRMS Calcd for C20H17NO4Br [M+H]+: 414.0341.
Found 414.0342.
17d: Colorless prisms (hexane/ethyl acetate); mp 189°C; 1H NMR (600 MHz, CD3OD) 8.09
(d, J = 2.8 Hz, 2 H), 7.78 (dd, J = 8.9, 2.8 Hz, 1 H), 7.50 (dt, J = 8.9, 1.4 Hz, 1 H), 7.47 (dd, J
= 2.8, 1.2 Hz, 1 H), 7.41 (t, J = 8.2 Hz, 1 H), 7.13 (dd, J = 8.3, 2.8 Hz, 1 H), 6.89 (dt, J = 8.9,
2.1 Hz, 2 H), 6.86 (d, J = 8.9 Hz, 1 H), 6.50 (dt, J = 8.9, 2.0 Hz, 2 H), 3.86 (s, 3 H); 13C NMR
(150 MHz, CD3OD) 168.61, 161.35, 155.57, 154.43, 149.98, 137.19, 134.37, 130.77,
127.24, 124.82 (q, J = 270.00 Hz), 121.98, 121.24 (d, J = 31.59 Hz), 120.87 (d, J = 5.75 Hz),
119.25, 118.91, 117.37, 113.87, 55.92; HRMS Calcd for C21H17NO4F3 [M+H]+: 404.1110.
Found 404.1108.
1
17e: Colorless prisms (acetonitrile); mp 229-230°C; H NMR (600 MHz, acetone-d6) 9.74
(br, 1 H), 8.29 (t, J = 3.1 Hz, 1 H), 7.94 (dt, J = 8.9, 2.8 Hz, 1 H), 7.55(dd, J = 6.9, 2.1 Hz, 1
H), 7.41 (t, J = 8.2 Hz, 1 H), 7.14 (ddd, J = 8.2, 2.8, 1.4 Hz, 1 H), 7.01 (dt, J = 9.0, 2.1 Hz, 1
13
H), 6.91 (dt, J = 9.0, 2.8 Hz, 2 H), 6.87 (d, J = 9.0 Hz, 1 H), 3.85 (s, 3 H); C NMR (150

MHz, DMSO-d6) 165.29, 159.21, 156.02, 154.67, 146.87, 135.66, 133.99, 129.67, 127.34,
124.67, 121.07, 119.84, 117.55, 116.02, 112.93, 101.50, 55.37; HRMS: Calcd for C21H17N2O4
[M+H]+: 361.1188. Found 361.1187.
18a: Colorless prisms (chloroform); mp 78.5-79.5°C; 1H NMR (600 MHz, CD3OD) 7.52 (d,
J = 7.5 Hz, 1 H), 7.50 (d, J = 2.6 Hz, 1 H), 7.41 (t, J = 7.9 Hz, 1 H), 7.21 (d, J = 8.2 Hz, 1 H),
7.14 (dd, J = 8.3, 2.1 Hz, 1 H), 6.87 (dt, J = 9.2, 2.7 Hz, 2 H), 6.83 (d, J = 2.8 Hz, 1 H), 6.78
(dt, J = 8.9, 2.1 Hz, 2 H), 6.75 (dd, J = 8.3, 2.8 Hz, 1 H), 3.86 (s, 3 H); 13C NMR (150 MHz,
CD3OD) 169.24, 161.34, 158.98, 155.01, 150.45, 137.69, 136.96, 131.29, 130.75, 129.33,
122.01, 120.72, 120.04, 117.18, 116.06, 113.89, 55.89, 18.35; HRMS Calcd for C21H20NO4
[M+H]+: 350.1387. Found 350.1813.
1
18b: Pale brown prisms (hexane/ethyl acetate); mp: 156-157°C; H NMR (600 MHz,
CD3OD) 7.51 (d, J = 2.7 Hz, 1 H), 7.50 (s, 1 H), 7.48 (d, J = 2.8 Hz, 1 H), 7.41 (t, J = 7.9
Hz, 1 H), 6.91 (dt, J = 8.9, 2.1 Hz, 2 H), 6.81 (dt, J = 8.9, 2.4 Hz, 2 H), 6.73 (m, 2 H) 3.86 (s,
3 H); 13C NMR (150 MHz, CD3OD) 169.06, 161.29, 159.59 (d, J = 10.05 Hz), 159.06,
157.42, 155.50, 149.61, 136.57, 130.83, 129.03, 122.42, 120.83, 120.52 (d, J = 12.93 Hz),
119.07, 117.40, 113.90, 113.51, 105.84 (d, J = 22.98 Hz), 55.95; HRMS Calcd for C20 H17
NO4F [M+H]+: 354.1142. Found 354.1141.
18c: Colorless prisms (chloroform); mp 176-176.5°C; 1H NMR (600 MHz, CD3OD) 7.54 (d,
J = 9.2 Hz, 1 H), 7.53 (d, J = 7.6 Hz, 1 H), 7.50(br, 1 H), 7.42(t, J = 7.7 Hz, 1 H), 7.15 (dd, J
= 8.2, 2.7 Hz, 1 H), 7.00 (d, J = 2.8 Hz, 1 H), 6.92 (dt, J = 9.0, 2.8 Hz, 2 H), 6.90 (dd, J = 8.9,
13
2.8 Hz, 1 H), 6.82 (dt, J = 8.9, 2.8 Hz, 2 H), 3.87 (s, 3 H); C NMR (150 MHz, CD3OD) 
167.65, 160.04, 158.04, 154.34, 148.22, 135.36, 130.68, 129.48, 128.69, 121.059, 119.40,
117.70, 117.47, 116.06, 115.63, 112.57, 54.56; HRMS Calcd for C20H17NO4Cl [M+H]+:
370.0846. Found 370.0844.
1
18d: Colorless prisms (hexane/ethyl acetate); mp 161-161.5°C; H NMR (600 MHz,
DMSO-d6)  9.41 (s, 1 H), 9.34 (s, 1 H), 7.49 (m, 3 H), 7.40 (t, J = 7.5 Hz, 1 H), 7.12 (dd, J =
8.2, 2.1 Hz, 1 H), 6.90 (dt, J = 9.0, 2.1Hz, 2 H), 6.73 (dt, J = 9.0, 2.1 Hz, 2 H), 6.72 (d, J = 2.1
13
Hz, 1 H), 6.38 (dd, J = 8.9, 2.7 Hz, 1 H), 3.80 (s, 3 H), 3.75 (s, 3 H); C NMR (150 MHz,
DMSO-d6)  164.82, 159.22, 156.65, 153.80, 153.54, 148.02, 135.92, 129.57, 126.43, 121.19,
120.75, 119.65, 117.37, 116.22, 112.64, 107.72, 101.70, 55.79, 55.30; HRMS Calcd for
C21H20NO5 [M+H]+: 366.1341. Found 366.1332.
18e: Pale pink prisms (hexane/ethyl acetate); mp 147-148°C; 1H NMR (600 MHz, acetone-d6)
 9.45 (br, 1 H), 8.40 (br, 1 H), 7.57 (m, 3 H), 7.43 (t, J = 8.2 Hz, 1 H), 7.15(dd, J = 7.6, 2.7

Hz, 2 H), 6.91 (dt, J = 9.0, 2.8 Hz, 2 H), 6.85 (dt, J = 8.9, 2.8 Hz, 2 H), 6.50 (d, J = 2.7 Hz, 1
13
H), 6.45 (dd, J = 9.0, 2.8 Hz, 1 H), 3.86 (s, 3 H); C NMR (150 MHz, CD3OD)  168.41,
161.40, 158.76, 155.02, 151.85, 150.38, 137.11, 130.83, 125.73, 122.09, 121.47, 120.47,
118.83, 117.13, 113.81, 109.25, 106.15, 55.90; HRMS Calcd for C20H18NO5 [M+H]+:
352.1185. Found 352.1182.
19a: Colorless prisms (chloroform); mp 169 °C; 1H NMR (600 MHz, acetone-d6)  7.71 (d, J
= 10.3 Hz, 1 H), 7.54 (dd, J = 7.6, 2.7 Hz, 1 H), 7.51(t, J = 2.1 Hz, 1 H), 7.43 (t, J = 7.9 Hz, 1
H), 6.95 (m, 4 H), 3.00 (s, 3 H); 13C NMR (150 MHz, CD3OD)  168.61, 161.36, 158.19 (d, J
= 7.19 Hz), 156.53 (d, J = 5.75 Hz), 154.08, 152.75, 137.48 (t, J = 12.93 Hz), 137.09, 130.81,
129.44 (t, J = 14.37 Hz), 120.74, 119.00, 117.00 (d, J = 37.35 Hz), 113.96, 105.64 (d, J =
27.29 Hz), 55.94; HRMS Calcd for C20H16NO4F2 [M+H]+: 372.1047. Found 372.1046.
1
19b: Pale brown prisms (hexane/ethyl acetate); mp 141-141.5°C; H NMR (600 MHz,
CD3OD)  7.52 (dd, J = 8.2, 1.2 Hz, 1 H), 7.49 (t, J = 2.1 Hz, 1 H), 7.42 (t, J = 8.2 Hz, 1 H),
7.28 (td, J = 8.9, 2.1 Hz, 1 H), 7.15 (dd, J = 8.2, 2.7 Hz, 1 H), 6.91 (dt, J = 8.9, 2.1 Hz, 2 H),
13
6.79 (dt, J = 8.9, 3.1 Hz, 2 H), 6.73 (td, J = 8.9, 2.8 Hz, 1 H), 3.86 (s, 3 H); C NMR (150
MHz, acetone-d6)  166.25 (d, J = 10.05 Hz), 160.70, 155.11, 146.00 (dt, J = 247.04, 10.82
Hz), 144.99, 143.29 (dd, J = 245.58, 12.93 Hz), 136.39, 130.45, 123.20 (t, J = 10.07 Hz),
120.87 (d, J = 15.80 Hz), 120.69, 120.53, 118.63, 117.08 (d, J = 12.92 Hz), 114.26, 113.58,
55.72; HRMS Calcd for C20 H16 NO4F2 [M+H]+: 372.1047. Found 372.1058.
1
19c: Colorless prisms (chloroform); mp 168-169°C; H NMR (600 MHz, acetone-d6)  9.78
(br, 1 H), 8.22 (br, 1 H), 8.97 (d, J = 2.0 Hz, 2 H), 7.57 (d, J = 7.5 Hz, 1 H), 7.53 (t, J = 2.1
Hz, 1 H), 7.43 (t, J = 7.9 Hz, 1 H), 7.16 (dd, J = 8.3, 2.8 Hz, 1 H), 6.78 (dt, J = 9.0, 2.1 Hz, 2
13
H), 6.70 (dt, J = 9.6, 2.8 Hz, 2 H), 3.87 (s, 3 H); C NMR (150 MHz, CD3OD)  168.62,
161.37, 153.73, 151.73, 145.13, 138.16, 137.02, 130.82, 122.28, 120.79, 119.07, 116.89,
116.73, 113.98, 55.95; HRMS Calcd for C20H16NO4Cl2 [M+H]+:404.0456. Found 404.0459.
4.9. SC-3 Growth Inhibition Assay18
SC-3 cells were cultured in MEMα (Wako Co.) supplemented with 2% FBS (fetal bovine
serum) and 1 nM DHT at 37°C in an incubator under an atmosphere of 5% CO2 in humidified
air. All experiments were performed in triplicate or more. Cells were trypsinized and diluted
to 20,000 cells/mL with MEMα supplemented with 2% charcoal-stripped FBS. This cell
suspension was seeded in 96-well plates at a volume of 100 μL/well and the plates were
incubated for 24 h. Then, 10 μL of medium was removed from each well, and replaced with
10 μL of drug solution supplemented with serial dilutions of test compound or DMSO as a
dilution control in the presence of 1 nM DHT. The plates were incubated at 37°C under 5%

CO2 in air for 3 days, and the cell number was determined using a Cell Counting Kit-8
(Dojindo). A 10 μL aliquot of WST-8 [2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-
5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt] was added to each well, then the
cells were incubated for 2 h, and the absorbance at 450 nm was measured with a microplate
reader. This parameter is proportional to the number of living cells in the culture. Initially we
tested the activity of compounds at the concentration of 10 μM, and if the inhibition exceeded
50% at the concentration of 10 μM, the IC50 value was determined in the concentration range
of 0.01 to 10 μM. IC50 (Table 1) is the concentration of test compound that reduces
DHT-induced cell growth to 50% of the control.
4.10. LNCaP cell proliferation assay18
The human prostate adenocarcinoma cell line LNCaP was routinely cultivated in RPMI-1640
supplemented with 10% FBS at 37°C in an incubator under an atmosphere of 5% CO2 in
humidified air. All experiments were performed in triplicate or more. Cells were trypsinized
and diluted to 20,000 cells/mL with RPMI-1640 supplemented with 10% charcoal-stripped
FBS. This cell suspension was seeded in 96-well plates at a volume of 100 μL/well and the
plates were incubated at 24 h. Then, 10 μL of medium was removed from each well, and
replaced with 10 μL of the drug solution supplemented with serial dilutions of the test
compounds or DMSO as a dilution control in the presence or absence of 10 nM DHT. The
final concentrations of compounds were 10 nM to 10 μM. Cells were incubated for 6 days,
and half of the medium was removed and replaced once after 3 days with medium containing
test compound or DMSO as a dilution control in the presence or absence of 10 nM DHT. At
the end of the incubation, proliferation was evaluated by adding 10 μM WST-8 to
microcultures and incubating the cells for 2 h. The absorbance at 450 nm was measured. This
parameter is proportional to the number of living cells in the culture.
4.11. 22Rv1 growth inhibition assay18
The human prostate cancer cell line 22Rv1 was routinely cultivated in RPMI-1640
supplemented with 10% FBS at 37 °C in an incubator under an atmosphere of 5% CO2 in
humidified air. All experiments were performed in triplicate or more. Cells were trypsinized
and diluted to 20,000 cells/mL with RPMI-1640 supplemented with 10% charcoal-stripped
FBS. This cell suspension was seeded in 96-well plates at a volume of 100 μL/well and the
plates were incubated at 24 h. Then, 10 μL of medium was removed from each well, and
replaced with 10 μL of drug solution supplemented with serial dilutions of test compound or
DMSO as a dilution control in the presence of 1 nM DHT in triplicate microcultures. After
incubation for 3 days, cell proliferation was evaluated by adding 10 μM WST-8 to
microcultures and incubating the cells for 2 h. The absorbance at 450 nm was measured. This
parameter is proportional to the number of living cells in the culture.

4.12. PC-3 growth inhibition assay18
The human prostate cancer cell line PC-3 was routinely cultivated in RPMI-1640 medium
supplemented with 10% FBS at 37°C in an incubator under an atmosphere of 5% CO2 in
humidified air. Cells were trypsinized and diluted to 20,000 cells/mL with RPMI-1640
supplemented with 10% charcoal-stripped FBS. This cell suspension was seeded in 96-well
plates at a volume of 100 μL/well and incubated at 24 h. Then, 10 μL of medium was
removed from each well, and replaced with 10 μL of drug solution supplemented with serial
dilutions of test compound or DMSO as a dilution control in the presence of 1 nM DHT. Cells
were incubated for 3 days, and at the end of the incubation time, proliferation was evaluated
by adding 10 μM WST-8 to the microcultures and incubating the cells for 2 h. The absorbance
at 450 nm was measured. This parameter is proportional to the number of living cells in the
culture.
Acknowledgments
This work was partly supported by JSPS KAKENHI Grant No 16K08318 (to AT), 17H03887
and 16K15138 (to SF), JSPS Core-to-Core Program, A. Advanced Research Networks, and
MEXT Platform for Drug Discovery, Informatics, and Structural Life Science, Japan. A part
of this research is based on the Cooperative Research Project of Research Center for
Biomedical Engineering. A.T. thanks The Naito Foundation and the Tokyo Biochemical
Research Foundation for financial support.
References
1) Evans, R. M. The steroid and thyroid hormone receptor superfamily. Science 1988, 240,
889-894.
2) McEwan, I. J. Gene regulation through chromatin remodeling by members of the nuclear
receptor superfamily. Biochem. Soc. Trans. 2000, 28, 369-373.
3) Mooradian, A. D.; Morley, J. E.; Korenman, S. G. Biological actions of androgens. Endocr.
Rev. 1987, 8, 1-28.
4) Bagatell, C. J.; Bremner, W. J. Androgens in men; uses and abuses. N. Engl. J. Med. 1996,
334, 707-714.
5) Chen, Y.; Sawyers, C.L.; Scher, H.I. Targeting the androgen receptor pathway in prostate
cancer. Curr. Opin. Pharmacol. 2008, 8, 440-448.
6) Neri, R.; Florance, K.; Koziol, P.; Cleave, S. van. A biological profile of a nonsteroidal
antiandrogen, SCH 13521 (4’-nitro-3’-tri-fluoromethylisobutyranilide). Endocrinology
1972, 91, 427-437.
7) Schellhammer, P. An update on bicalutamide in the treatment of prostate cancer. Expert
Opin. Invest. Drugs 1999, 8, 849-860.

8) Taplin, M.-E. Androgen receptor: role and novel therapeutic prospects in prostate cancer.
Expert Rev. Anticancer Ther. 2008, 8, 1495-1508.
9) Heinlein, C. A.; Chang, C. Androgen receptor in prostate cancer. Endocr. Rev. 2004, 25,
276-308.
10) Scher, H. I.; Steineck, G.; Kelly, W. K. Hormone-refractory (D3) prostate cancer: refining
the concept. Urology 1995, 46, 142-148.
11) Miyamoto, H.; Rahman, M. M.; Chang, C. Molecular basis for the antiandrogen
withdrawal syndrome. J. Cell Biochem. 2003, 91, 3-12.
12) Bohl, C. E.; Gao, W.; Miller, D. D.; Bell, C. E.; Dalton, J. T. Structural basis for
antagonism and resistance of bicalutamide in prostate cancer. Proc. Natl. Acad. Sci. U. S.
A. 2005, 102, 6201-6206.
13) Steketee, K.; Timmerman, L.; Ziel-van der Made, A. C.; Doesburg, P.; Brinkmann, A. O.;
Trapman, J. Broadened ligand responsiveness of androgen receptor mutants obtained by
random amino acid substitution of H874 and mutation hot spot T877 in prostate cancer.
Int. J. Cancer 2002, 100, 309-317.
14) Jung, M. E.; Ouk, S.; Yoo, D.; Sawyers, C. L.; Chen, C.; Tran, C. Wongvipat, J.
Structure-activity relationship for thiohydantoin androgen receptor antagonists for
castration-resistant prostate cancer (CRPC). J. Med. Chem. 2010, 53, 2779-2796.
15) Yang, S. H.; Song, C.-H.; Van, H. T. M.; Park, E.; Khadka, D. B.; Gong, E.-Y.; Lee, K.;
Cho, W.-J. SAR based design of nicotinamides as a novel class of androgen receptor
antagonists for prostate cancer. J. Med. Chem. 2013, 56, 3414−3418.
16) Ishioka, T.; Tanatani, A.; Nagasawa, K.; Hashimoto, Y. Anti-androgens with full
antagonistic activety toward human prostate tumor LNCaP cells with mutated androgen
receptor. Bioorg. Med. Chem. Lett. 2003, 13, 2655-2658.
17) Inoue, K.; Urushibara, K.; Kanai, M.; Yura, K.; Fujii, S.; Hashimoto, Y.; Ishigami-Yuasa,
M.; Mori. S.; Kawachi, E.; Matsumura, M.; Hirano, T.; Kagechika, H.; Tanatani, A.
Design and synthesis of 4-benzyl-1-(2H)-phthalazinone derivatives as novel androgen
receptor antagonists. Eur. J. Med. Chem. 2015, 102, 310-319.
18) Yamada, A.; Fujii, S.; Mori, S.; Kagechika, H. Design and synthesis of
4-(4-benzoylaminophenoxy)phenol derivatives as androgen receptor antagonists. ACS
Med. Chem. Lett. 2013, 4, 937-941.
19) (a) Ohtsu, H.; Xiao, Z. Y.; Ishida, J.; Nagai, M.; Wang, H. K.; Itokawa, H.; Su, C. Y.; Shih,
C.; Chiang, T. Y.; Chang, E.; Lee, Y. F.; Tsai, M. Y.; Chang, C. S.; Lee, K. H. Antitumor
agents. 217. Curcumin analogues as novel androgen receptor antagonists with potential as
anti-prostate cancer agents. J. Med. Chem. 2002, 45, 5037−5042. (b) Zhou, J.; Geng, G.;
Shi, Q.; Sauriol, F.; Wu, J. H. Design and synthesis of androgen receptor antagonists with
bulky side chains for overcoming antiandrogen resistance. J. Med. Chem. 2009, 52, 5546−
5550.

20) (a) Kagechika, H.; Himi, T.; Kawachi, E.; Hashimoto, Y.; Shudo, K. Retinobenzoic acids.
4. Conformation of aromatic amides with retinoidal activity. Importance of trans-amide
structure for the activity. J. Med. Chem. 1989, 32, 2292−2296. (b) Itai, A.; Toriumi, Y.;
Tomioka, N.; Kagechika, H.; Azumaya, I.; Shudo, K. Stereochemistry of
N-methylbenzanilide and benzanilide. Tetrahedron Lett. 1989, 30, 6177−6180. (c)
Yamaguchi, K.; Matsumura, G.; Kagechika, H.; Azumaya, I.; Ito, Y.; Itai, A.; Shudo, K.
Aromatic architecture. Use of the N-methylamide structure as a molecular splint. J. Am.
Chem. Soc. 1991, 113, 5474−5475.
21) Hiraoka, D.; Nakamura, N.; Nishizawa, Y.; Uchida , N.; Noguchi, S.; Matsumoto, K.; Sato,
B. Inhibitory and stimulatory effects of glucocorticoid on androgen- induced growth of
murine shionogi carcinoma 115 in vivo and in cell cul- ture, Cancer Res. 1987, 47,
6560-6564.
22) Veldscholte, J.; Berrevoets, C. A.; Brinkmann, A. O.; Grootegoed, J. A.; Mulder, E.
Anti-androgens and the mutated androgen receptor LNCaP cells: Differential effects on
binding affinity, heat-shock protein interaction, and transcription activation. Biochemistry
1992, 31, 2393−2399.
23) Zhou, J.; Liu, B.; Geng, G.; Wu, J. H. Study of the impact of the T877A mutation on
ligand induced helix-12 positioning of the androgen receptor resulted in design and
synthesis of novel antiandrogens. Proteins 2010, 78, 623−637.
24) Sramkoski, R. M.; Pretlow, T. G., II; Giaconia, J. M.; Pretlow, T. P.; Schwartz, S.; Sy,
M.-S.; Marengo, S. R.; Rhim, J. S.; Zhang, D.; Jacobberger, J. W. A new human prostate
carcinoma cell line, 22Rv1. In Vitro Cell. Dev. Biol.-Anim. 1999, 35, 403−409.
Graphical Abstract
Structure-Activity Relationship of Novel (Benzoylaminophenoxy)phenol
Derivatives as Anti-prostate Cancer Agents
Yuko Kazui, Shinya Fujii, Ayumi Yamada, Mari Ishigami-Yuasa, Hiroyuki Kagechika, and
Aya Tanatani,