Anticancer Activity of 2-Amino-substituted-1,4-naphthoquinone Derivatives in Ovarian Cancer Cells

Article abstract of DOI:10.1002/bkcs.11315

2-Amino-substituted-1,4-naphthoquinone derivatives (10–17) were synthesized from coupling reaction between 1,4-dihydroxynaphthalene and amines in the presence of catalyst CeCl₃^.7H₂O. Their anticancer activity was evaluated by using three ovarian cancer cells (A2780, SKOV3, and OVCAR3). Among the eight compounds, compound 13 containing metal chelating moiety had a relatively potent cytotoxic activity (IC₅₀ < 10 μM) on all three ovarian cancer cells.

Full text of DOI:10.1002/bkcs.11315

Article
DOI: 10.1002/bkcs.11315
BULLETIN OF THE
S. Shin et al.
KOREAN CHEMICAL SOCIETY
Anticancer Activity of 2-Amino-substituted-1,4-naphthoquinone
Derivatives in Ovarian Cancer Cells
†
†
†
‡
‡
Sujeong Shin, Haneul Lee, Cheolmin Jeon, Umma Hafsa Preya, Jung-Hye Choi, and
†,
*
Jeong Ho Park
†Department of Chemical and Biological Engineering, Hanbat National University, Daejeon 34158,
South Korea. *E-mail: jhpark@hanbat.ac.kr
Department of Life and Nanopharmaceutical Sciences and College of Pharmacy, Kyung Hee University,
Seoul 02447, South Korea
‡
Received September 2, 2017, Accepted October 11, 2017
2
-Amino-substituted-1,4-naphthoquinone derivatives (10–17) were synthesized from coupling reaction
.
between 1,4-dihydroxynaphthalene and amines in the presence of catalyst CeCl 7H O. Their anticancer
3
2
activity was evaluated by using three ovarian cancer cells (A2780, SKOV3, and OVCAR3). Among the
eight compounds, compound 13 containing metal chelating moiety had a relatively potent cytotoxic activ-
ity (IC50 < 10 μM) on all three ovarian cancer cells.
Keywords: 2-Amino-substituted-1,4-naphthoquinone derivatives, Anticancer activity, Ovarian cancer
cells
Introduction
Experimental
1
13
Quinone moieties are widely distributed in nature and are
the key pharmcophore of many clinically important
drugs such as anthracycline, daunorubicin, doxorubicin,
General Methods. H NMR and C NMR spectra were
recorded on a Varian Mercury 400 (400 MHz) spectrometer
(Palo Alto, California, USA) and the mass spectra taken using
an Agilent G1956B mass spectrometer (Santa Clara, Califor-
nia, USA). Flash column chromatography was performed
using E. Merck silica gel (60, particle size 0.040–0.063 mm)
and analytical thin layer chromatography (TLC) performed
using pre-coated TLC plates with silica Gel 60 F254
(E. Merck). All chemicals were reagent grade unless otherwise
1
mitomycin, mitoxantrones, and saintopin. They are applied
2
3
4
5
as anticancer, antifungal, antiviral, antiplasmodial,
6
7
antibiotics, and endothelial vasodilation agent. Among
the quinone derivatives, 2-amino-substituted-1,4-naphtho-
quinone derivatives show many interesting biological activ-
8
9
10
ities such as antitumour, antimalarials, antibacterials,
.
3
1
1
12
specified. 1,4-dihydroxy naphthalene, CeCl 7H O, and amine
compounds were purchased from Sigma-Aldrich Chemical
Co. and used without purification.
Cytotoxicity Test. The cytotoxicity was analyzed using
MTT assay. Cells were seeded in 96-cell culture well at a
density of 1.0 × 10 cells/mL in each well. After 24 h, vari-
ous concentrations of 1,4-naphthoquinone derivatives were
added. After 48 h, 25 μL of MTT (3-(4,5-Dimethylthiazol-2-
yl)-2,5-Diphenyltetrazolium Bromide, 5 mg/mL stock con-
centration) was added, and the plates were incubated for 4 h.
The medium was discarded, and the formazan blue salt,
which was generated within the cells, was dissolved in
0 μL DMSO. The optical density was measured at 540 nm
using a micro plate spectrophotometer (Spectra Max; Molec-
ular Devices, Sunnyvale, CA, USA).
2
antituberculars, and fungicides.
There are many chemotherapy drugs for the treatment of
ovarian cancer such as platinum compounds, taxane com-
pounds, altretamine, capecitabine, cyclophosphamide, eto-
poside, gemcitabine, ifosfamide, irinotecan, liposomal
doxorubicin, melphalan, pemetrexed, topotecan, and vinor-
elbine. Alhough these chemotherapeutic drugs effectively
kill cancer cells, there are many common side effects
caused by chemotheraphy. There are still needs for devel-
opment of more effective but less toxic drug candidates.
Benites et al. reported that 4-methoxyphenyl-2-amino-
5
1
2
,4-naphthoquinone (compound 10) and 4-hydroxyphenyl-
-amino-1,4-naphthoquinone (compound 11) showed good
5
cytotoxic value against three human-derived tumor cell
lines such as DU145 (prostate, EC50 = 27 and 3.4 μM),
T24 (bladder, EC50 = 29 and 8.7 μM), and MCF7 (breast,
Synthesis
8
EC50 = 13 and 3.0 μM), respectively. Therefore, we have
synthesized new 2-amino-substituted-1,4-naphthoquinone
derivatives including compound 10 and 11 to evaluate their
biological activity in ovarian cancer cells.
General Procedure. The following procedure is the gener-
alized synthetic procedure used for the synthesis of 2-
amino-substituted-1,4-naphthoquinone derivatives. To the
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1
mixture
of
1,4-dihydroxy
naphthalene
(100 mg,
by the general procedure. H NMR (400 MHz, CDCl ) δ
3
0
0
.62 mmol), amine (0.75 mmol), and CeCl 7H O (20 mg,
.062 mmol) in 10 mL ethanol, TEA (1.25 mmol) was
0.88 (t, J = Hz, 3H), 1.26 (s, 22H), 1.70 (m, 2H), 2.02
(m,4H), 3.17 (q, J = 6.8 Hz, J = 13.2 Hz, 2H), 5.35 (t,
J = 5.6 Hz, 2H), 5.73 (s, 1H), 5.89 (s, 1H), 7.61 (t,
J = 7.6 Hz, 1H), 7.73 (t, J = 7.2 Hz, 1H), 8.04 (d,
J = 7.6 Hz, 1H), 8.10(d, J = 7.6 Hz, 1H).
3
2
added at room temperature. The reaction mixture was stir-
red for 4 h at room temperature. After the reaction was con-
firmed as completed by TLC (DCM: MeOH = 19: 1),
13
10 mL cold water was added to the reaction solution. The
C NMR (100 MHz, CDCl ) δ 14.11, 22.68, 27.03,
3
resulting precipitate was filtered and washed with 20 mL
EtOH. The desired 1,4-naphthoquinone derivatives were
obtained as a solid compound.
27.16, 27.22, 28.25, 29.17, 29.23, 29.32, 29.39, 29.52, 29.67,
29.70, 29.76, 31.90, 42.59, 100.73, 126.18, 126.24, 129.72,
130.02, 130.53, 131.89, 133.73, 134.73, 147.96, 181.97,
+
2-((4-methoxyphenyl)amino)naphthalene-1,4-dione
182.90. ESI-MS: m/z [M + H] 424.3 (Calcd 423.6).
0
(
10): Compound 10 was obtained as a solid in 30% yield
2,2 -(((methylazanediyl)bis(propane-3,1-diyl))bis(aza-
1
by the general procedure. H NMR (400 MHz, DMSO‑d )
nediyl))bis(naphthalene-1,4-dione) (15): Compound 15
6
δ 3.78 (s, 3H), 5.92 (s, 1H), 7.01 (d, J = 8.8 Hz, 2H), 7.29
was obtained as a solid in 18% yield by the general proce-
1
(
d, J = 8.8 Hz, 2H), 7.77 (t, J = 7.2 Hz, 1H), 7.85 (t,
dure. H NMR (400 MHz, DMSO‑d ) δ 1.92 (m, 4H),
6
J = 7.6 Hz, 1H), 7.94 (d, J = 7.6 Hz, 1H), 8.05 (d,
2.31 (s, 3H), 2.53 (t, J = 6 Hz, 2.8 Hz, 4H), 3.31 (m, 4H),
5.73 (s, 2H), 6.96 (bs, 2H), 7.58 (dt, J = 7.5 Hz, 1.0 Hz,
1
3
J = 7.2 Hz, 1H), 9.15 (s, 1H). C NMR (DMSO‑d6,
13
1
1
1
00 MHz) δ 55.8, 101.5, 115(2C), 125, 126.1(2C), 126.5,
2H), 7.69 (dt, J = 7.5 Hz, 1.0 Hz, 2H), 8.04 (m, 4H).
C
30.9, 131.1, 132.9, 133.2, 135.4, 147.4, 157.4, 182.2,
NMR (DMSO‑d , 100 MHz) δ 25.3(2C), 41.3(2C), 42.1,
6
+
825.7 ESI-MS: m/z [M + H] 280.1 (Calcd 279.29).
55.8(2C), 99.7(2C), 125.7(2C), 126.3(2C), 130.8(2C),
132.5(2C), 133.7(2C), 135.2(2C), 149.0(2C), 181.6(2C),
2-((4-hydroxyphenyl)amino)naphthalene-1,4-dione
+
(
11): Compound 11 was obtained as a solid in 45% yield
1812.0(2C) ESI-MS: m/z [M + H] 458.2 (Calcd 457.5).
1
by the general procedure. H NMR (400 MHz, DMSO‑d )
2-((2-(thiophen-3-yl)ethyl)amino)naphthalene-1,4-
6
δ 5.87 (s, 1H), 6.83 (d, J = 8.7 Hz, 2H), 7.16 (d,
J = 8.7 Hz, 2H), 7.76 (t, J = 7.6 Hz, 1H), 7.85 (t,
J = 7.6 Hz, 1H), 7.93 (d, J = 7.6 Hz, 1H), 8.04 (d,
dione (16): Compound 16 was obtained as a solid in 98%
yield by the general procedure. H NMR (400 MHz,
1
DMSO‑d ) δ 3.10 (t, J = 7.1 Hz, 2H), 3.44 (m, 2H), 5.72
6
1
3
J = 7.6 Hz, 1H), 9.07 (s, 1H), 9.56 (s, 1H). C NMR
(s, 1H), 6.95 (m, 2H), 7.34 (dd, J = 4.5 Hz & 1.7 Hz, 1H),
7.57 (t, J = 5.9 Hz, 1H), 7.72 (t, J = 7.5 Hz, 1H), 7.82 (t,
(
(
1
DMSO‑d , 100 MHz) δ 101.2, 116.28(2C), 125.7, 126.3
6
13
2C), 126.5, 129.4, 130.9, 132.9, 133.3, 135.3, 147.5,
J = 7.5 Hz, 1H), 7.95 (m, 2H). C NMR (100 MHz,
DMSO) δ 28.04, 43.80, 100.15, 124.76, 125.80, 126.04,
126.36, 127.49, 130.79, 132.67, 133.55, 135.33, 141.36,
+
55.8, 182.2, 182.6 ESI-MS: m/z [M + H] 266.1
(Calcd 265.3).
+
2-((2-(3a,7a–dihydro-1H-indol-3-yl)ethyl)amino)naph-
148.73, 181.83, 181.94 ESI-MS: m/z [M + H] 284.3
thalene-1,4-dione (12): Compound 12 was obtained as a
solid in 82% yield by the general procedure. H NMR
(Calcd 283.3).
1
(
2
400 MHz, DMSO‑d ) δ 3.01 (t, J = 7.2 Hz, 2H), 3.48 (m,
Results and Discussion
6
H), 5.74 (s, 1H), 6.98 (t, J = 7.2 Hz, 1H), 7.07 (t,
J = 7.6 Hz, 1H), 7.26 (d, J = 1.5 Hz, 1H), 7.34 (d,
J = 8 Hz, 1H), 7.52 (t, J = 5.8 Hz, 1H), 7.57 (d,
J = 7.8 Hz, 1H), 7.72 (t, J = 7 Hz, 1H), 7.82 (t, J = 7 Hz,
In general, 2-amino-substituted-1,4-naphthoquinone derivatives
were synthesized by three synthetic methods such as the nucle-
13
ophilic substitution of 2-halo-substituted benzoquinone, the
1,4-addition of amines to 1,4-naphthoquinone in the pres-
1
3
1H), 7.96 (m, 2H), 10.86 (s, 1H). C NMR (DMSO‑d6,
100 MHz) δ 23.8, 43.1, 99.8, 111.6, 111.9, 118.7, 118.8,
121.5, 123.6, 125.8, 126.4, 127.6, 130.8, 132.6, 133.7+,
135.3, 136.7, 148.8, 181.7, 182.0 ESI-MS: m/z [M + H]
317.3 (Calcd 316.3).
14
ence of Lewis acid, or the addition of amines to 1,4-
naphthoquinone in the presence of molecular iodine under
15
ultrasonic irradiation. In this present work, a series of 2-
amino-substituted-1,4-naphthoquinone derivatives were syn-
thesized by using 1,4-dihydroxy naphthalene as the starting
2-(bis(pyridin-2-ylmethyl)amino)naphthalene-1,4-
16
dione (13): Compound 13 was obtained as a solid in 92%
compound rather than 1,4-naphthoquinone. Coupling reac-
tion between 1,4-dihydroxy naphthalene and amine com-
1
yield by the general procedure. H NMR (400 MHz,
DMSO‑d ) δ 4.97 (s, 4H), 5.77 (s, 1H), 7.29 (m, 2H), 7.44
pounds in the presence of CeCl catalyst (Scheme 1) were
carried out at room temperature.
6
3
(
d, J = 7.9 Hz, 1H), 7.71 (m, 2H), 7.78 (m, 2H), 7.83 (m,
1
3
2
H), 8.52 (d, J = 5.2 Hz, 2H) C NMR (100 MHz,
The following amines (Figure 1) were used to synthesize
the 2-amino-substituted-naphthoquinone derivatives. Since
the antioxidant effect of natural polyphenols resulted from
phenolic hydroxy moiety, compounds 10 and 11 were syn-
thesized from coupling with 4-OH or 4-MeO substituted ani-
lines. Coupling with tryptamine known as a neuromodulator
or neurotransmitter gave rise to compound 12.
DMSO) δ 58.10(2C), 107.50, 121.92(2C), 122.87(2C),
1
1
25.06, 126.78, 132.35, 132.98(2C), 134.47, 137.38(2C),
49.67(2C), 152.86, 157.12(2C), 182.25, 183.37 ESI-MS:
+
m/z [M + H] 356.1 (Calcd 355.3).
(Z)-2-(octadec-9-en-1-ylamino)naphthalene-1,4-dione
(14): Compound 14 was obtained as a solid in 30% yield
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Anticancer Activity of 2-Amino-substituted-1,4-naphthoquinone
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Table 1. The IC50 value of naphthoquinone derivatives
synthesized.
IC50 (μM)
Compounds
A2780
SKOV3
OVCAR3
1
1
1
1
1
1
1
1
0
1
2
3
4
5
6
7
29.3
13.2
3.8
29.2
29.3
41.3
2.8
50.9
>100
40.3
8.4
7.8
19
78
Scheme 1. The general synthetic scheme for a series of 1,4-
naphthoquinone derivatives at the 2-position.
5.6
6.8
18.3
>100
44.7
17.6
15.4
>100
5.9
74
lipid solubility. A heteroaromatic thiophene compound was
coupled with compound 1 and resulted in compound 17.
The naphthoquinone derivatives (Figure 2) were synthe-
sized using 1,4-dihydroxy naphthalene and amine com-
pounds in the presence of CeCl catalyst. Without TEA the
3
yield was low and 3.0 equivalent of organic base TEA gave
rise to the optimal yield in the work.
The IC50 values of compound 10 for prostate (DU145
cells), bladder (T24 cells), and breast (MCF7 cells) cancer
cell lines are 27, 29, and 13 μM, respectively, and these of
8
compound 11 are 3.4, 8.7, and 3.0 μM, respectively. The
anti-cancer activity of naphthoquinone derivatives was
evaluated using A2780, SKOV3, and OVCAR3 ovarian
cancer cells (Table 1). The anti-ovarian cancer activity of
compound 10 shows the similar activity with prostate, blad-
der, and breast cell lines. But the anti-ovarian cancer activ-
ity of compound 11 is less effective than the anticancer
activity for prostate, bladder, and breast cell lines. There is
no big difference between compound 10 and 11 for ovarian
cancer cells. Effective antioxidant moiety –OH may not
give any influence for anticancer activity.
Figure 1. The structure of starting amine compounds.
Dipicolylamine well known as a metal chelator was coupled
with 1 and produced compound 13. To increase water solu-
bility of compounds, tertiary amine moiety was introduced
compounds 14 and 16). On the contrary with compounds
4 and 16, compound 15 was synthesized to increase its
(
1
Compound 12 containing an indole moiety showed the
best anticancer activity against A2780 cell line. Compound
1
3 having metal chelating moiety showed good anticancer
activity for A2780 and OVCAR3 cell line. Increasing hydro-
philicity (compound 14 and 16) gave rise to better anticancer
activity than increasing lipophilicity (compound 15). Com-
pound 17 containing a heteroaromatic thiophene moiety
showed the best anticancer activity against SKOV3 cell line.
Conclusion
A series of 2-amino-substituted-naphthoquinone derivatives
were synthesized and they generally showed anticancer
activity against ovarian cancer cell lines such as A2780,
SKOV3, and OVCAR3. Overall, among the eight com-
pounds, compound 13 containing metal chelating moiety
had a potent cytotoxic activity on all ovarian cancer cells
we used in this study. We now showed 2-amino naphtho-
quinone derivatives would be a good candidate for ovarian
cancer drug development. The further detail anticancer
mechanism study for each compound will be carried out.
Figure 2. The structure of naphthoquinone derivatives
synthesized.
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Anticancer Activity of 2-Amino-substituted-1,4-naphthoquinone
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