Synthesis and antiproliferative evaluation of certain 4-anilino-8-methoxy- 2-phenylquinoline and 4-anilino-8-hydroxy-2-phenylquinoline derivatives

Article abstract of DOI:10.1016/j.bmc.2005.12.017

The present report describes the synthesis and antiproliferative evaluation of certain 4-anilino-8-methoxy-2-phenylquinoline and 4-anilino-8-hydroxy-2- phenylquinoline derivatives. The antiproliferative activity of 4′-COMe-substituted derivatives decrease

Full text of DOI:10.1016/j.bmc.2005.12.017

Bioorganic & Medicinal Chemistry 14 (2006) 3098–3105
Synthesis and antiproliferative evaluation of certain
4-anilino-8-methoxy-2-phenylquinoline and 4-anilino-
8-hydroxy-2-phenylquinoline derivatives
Yeh-Long Chen, Chao-Jhieh Huang, Zun-Yuan Huang, Chih-Hua Tseng,
Feng-Shuo Chang, Sheng-Huei Yang, Shinne-Ren Lin and Cherng-Chyi Tzeng*
Faculty of Medicinal and Applied Chemistry, College of Life Science, Kaohsiung Medical University, Kaohsiung City 807, Taiwan
Received 18 November 2005; accepted 15 December 2005
Available online 18 January 2006
Abstract—The present report describes the synthesis and antiproliferative evaluation of certain 4-anilino-8-methoxy-2-phenylquin-
oline and 4-anilino-8-hydroxy-2-phenylquinoline derivatives. The antiproliferative activity of 40-COMe-substituted derivatives
decreased in an order of 6-OMe (1, 3.89 lM) > 8-OMe (8, 10.47 lM) > 8-OH (9, 14.45 lM), indicating that the position of substi-
tution at the quinoline ring is crucial. For 30-COMe derivatives, the antiproliferative activity of 8-OH (11, 1.20 lM) is more potent
than its 8-OMe counterpart (10, 8.91 lM), indicating that a H-bonding donating substituent is more favorable than that of a H-
bonding accepting group. Comparison of 8-OH derivatives, the antiproliferative effect of COMe (11) is more potent than its oxime
derivative (15a, 2.88 lM), which in turn is more potent than the methyloxime counterpart (15b, 5.50 lM). Compound 11 is espe-
cially active against the growth of certain solid cancer cells such as HCT-116 (colon cancer), MCF7, and MDA-MB-435 (breast
cancer) with GI50 values of 0.07, <0.01, and <0.01 lM, respectively. Flow cytometric analyses revealed that growth inhibition by
11 and 15a was due to accumulation in S-phase. This result is interesting because 2-phenylquinolone derivatives have been reported
to be antimitotic agents which induced cell cycle arrest in G2/M phase.
ꢀ 2005 Elsevier Ltd. All rights reserved.
1. Introduction
number of anticancer agents.13–17 Among them, 4-(4-
acetylanilino)-6-methoxy-2-phenylquinoline (1) and its
oxime (2a) and methyloxime (2b) derivatives exhibited
potent antiproliferative activity with a mean GI50 value
of 3.89, 3.02, and 3.89 lM, respectively.13 Structures of
these antiproliferative agents can also be considered as
the 4-anilino-substituted derivatives of antimitotic 2-
phenylquinolin-4(1H)-ones. In continuation of our
study to explore more potent anticancer drug candi-
dates, we described herein the preparation and antipro-
liferative evaluation on 8-substituted isomers of 1, 2a,
and 2b (Chart 1). A number of carboxamide derivatives
comprising polycyclic chromophores and a flexible cat-
ionic side chain are known to show antiproliferative ef-
fects18–21 prompted us to prepare certain 4-(8-methoxy-
2-phenylquinolin-4-ylamino)benzamides for evaluation.
Selected compounds were further evaluated on their
effect of cell cycle distribution.
Quinolin-4(1H)-one moiety is a characteristic compo-
nent of a large number of antibacterial and/or antican-
cer agents.1–5 The biological activity of these
quinolone derivatives depends not only on the bicyclic
heteroaromatic pharmacophore but also on the nature
of the peripheral substituents and their spatial relation-
ship. With a phenyl group appended on C-2 position of
quinolin-4(1H)-one, a number of 2-phenylquinolone
derivatives have been discovered to possess antimitotic
activity.6–8 Recently, we have synthesized certain AMSA
analogs in which the tricyclic acridine was replaced with
its isosteric furoquinoline for evaluation of their anti-
cancer activity.9–12 Further modification of AMSA has
been explored by the replacement of acridine with its
isomeric 2-phenylquinoline, an aza-analog of antitumor
2-phenylnaphthalene skeleton which consists of a large
2. Chemistry
Keywords: 4-Anilino-2-phenylquinoline derivatives; Antiproliferative
activity; Cytotoxicity; Anticancer agents.
*
The preparation of 4-anilino-2-phenylquinolines is illus-
trated in Schemes 1–3. Reaction of ethyl 3-chloro-2-
Corresponding author. Tel.: +886 7 3121101x6985; fax: +886 7
3125339; e-mail: tzengch@kmu.edu.tw
0968-0896/$ - see front matter ꢀ 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2005.12.017

Y.-L. Chen et al. / Bioorg. Med. Chem. 14 (2006) 3098–3105
3099
O
O
OR
N
O
OH
ii
R'
Cl
Me
Me
HN
HN
N
i
HN
HN
N
N
MeO
MeO
N
OMe
OMe
OMe
6
N
16
17 R' = NHCH2CH2N(CH3)2
18 R' = NHCH2CH2NCH2CH2OH
19 R' = NHCH2CH2N(CH2CH2)2NH
1
2a R = H (Mean GI50 = 3.02 μM)
2b R = Me (Mean GI50 = 3.89 μM)
Scheme 3. Reagents and conditions: (i) 4-aminobenzoic acid, pyridine,
EtOH, reflux (49–52%); (ii) a—CDI, DMF, rt, 18 h; b—1ꢁ-amine, rt,
5 h (21–84%).
(Mean GI50 = 3.89 μM)
Chart 1. Structures of 4-anilino-6-methoxy-2-phenylquinoline
derivatives.
EtOOC
methoxy-2-phenylquinoline (6), which was reacted with
48% HBr to give 4-bromo-8-hydroxy-2-phenylquinoline
(7) in a fairly good overall yield (Scheme 1). Treatment
of 6 with 4-aminoacetophenone and 3-aminoacetophe-
none, respectively, afforded 4-(4-acetylanilino)-8-meth-
oxy-2-phenylquinoline (8) and its 3-substituted isomer
10, respectively, which were reacted with NH2OH to
give exclusively (E)-oximes 12a and 14a, respectively.
The configuration of the oxime moiety was determined
by through-space nuclear Overhauser effect spectrosco-
py (NOESY) which revealed coupling connectivity to
CH3 protons. Accordingly, (E)-oximes 13a and 15a were
obtained from 9 and 11, respectively, which in turn were
prepared from 7 by the same reaction sequences. Reac-
tion of 8 with NH2OMe provided exclusively (E)-meth-
yloxime 12b in 72% yield. (E)-Methyloximes 13b, 14b,
and 15b were obtained, respectively, from 9, 10, and
11 by the same reaction sequences (Scheme 2). The con-
figuration of the oxime and methyloxime moieties was
further confirmed by the 13C NMR spectra. The carbon
of CH3, which is syn to the OH moiety, shifted upfield (d
value of CH3 is approximately 11.50 ppm for (E)-iso-
mers), while that of the anti-isomer shifted downfield
(d value of CH3 is approximately 18.75 ppm for (Z)-
isomers)23.
Cl
COOEt
OMe
COOEt
COOEt
H2N
i
+
N
H
OMe
3
4
O
Cl
N
Br
ii
iii
iv
N
H
N
OMe
OMe
OH
5
6
7
Scheme 1. Reagents and conditions: (i) K2CO3, DMF, 140 ꢁC, 3 h
(78%); (ii) a—Ph2O, 260–280 ꢁC, 0.5 h; b—4 N NaOH, reflux for 18 h;
(c) 20% HCl, reflux for 5 h (83%); (iii) POCl3, 80–90 ꢁC, 1 h (95%); (iv)
48% HBr, reflux for 24 h (90%).
OR'
O
N
Me
Me
HN
N
HN
N
i
ii
R
R
X
N
8 R = OMe
9 R = OH
12a R = OMe, R'= H
12b R = OMe, R' = Me
13a R = OH, R' = H
13b R = OH, R' = Me
Reaction of 6 with 4-aminobenzoic acid afforded 4-(8-
methoxy-2-phenylquinolin-4-ylamino)benzoic acid (16),
which was converted to 4-(8-methoxy-2-phenylquino-
lin-4-ylamino)benzamides 17–19 in an overall yield of
21–84% (Scheme 3).
R
6 X = Cl, R = OMe
7 X = Br, R = OH
Me
Me
HN
N
HN
N
O
N
OR'
i
ii
3. Pharmacological results and discussion
R
R
10 R = OMe
11 R = OH
14a R = OMe, R'= H
All compounds were evaluated in vitro against the full
panel of 60 human tumor cell lines derived from nine
cancer cell types (leukemia, non-small cell lung cancer,
colon cancer, CNS cancer, melanoma, ovarian cancer,
renal cancer, prostate cancer, and breast cancer). For
each compound, dose–response curves for each cell line
were measured with five different drug concentrations,
the concentration causing 50% cell growth inhibition
(GI50) compared with the control was calculated24 and
the results are summarized in Tables 1 and 2. The anti-
proliferative activity of 40-COMe substituted derivatives
decreasing in an order of 6-OMe (1, 3.89 lM) > 8-OMe
(8, 10.47 lM) > 8-OH (9, 14.45 lM) indicating that the
14b R = OMe, R' = Me
15a R = OH, R' = H
15b R = OH, R' = Me
Scheme 2. Reagents and conditions: (i) 3- or 4-aminoacetophenone,
pyridine, EtOH, reflux (49–86%); (ii) NH2OR, K2CO3, EtOH, reflux
for 0.5 h (70–94%).
(ethoxycarbonyl)-3-phenylpropenoate (3)22 with o-anisi-
dine gave 2-[(2-methoxyphenylamino)phenylmethyl-
ene]malonic acid diethyl ester (4), which was thermally
cyclized to afford 8-methoxy-2-phenyl-1H-quinolin-4-
one (5). Chlorination of 5 with POCl3 gave 4-chloro-8-

3100
Y.-L. Chen et al. / Bioorg. Med. Chem. 14 (2006) 3098–3105
Table 1. Antiproliferative assay of 4-substituted 4-anilino-2-phenylquinoline derivatives [GI50 (lM)a,b
]
Cell lines
Compound
1
2a
2.16
2b
2.79
8
9
12a
12b
3.62
13a
4.50
13b
RPMI-8226
NCI-H226
HCT-116
1.72
0.94
1.54
4.33
5.88
1.23
8.80
28.2
5.79
37.2
13.0
57.1
14.0
21.8
Ndd
5.53
24.0
23.6
0.86
14.4
1.79
15.8
22.4
22.2
12.7
24.0
18.8
19.6
14.1
Ndd
14.5
15.0
15.5
2.05
1.52
1.53
1.62
2.32
2.25
1.76
2.02
1.77
3.59
1.54
1.97
2.62
14.7
13.2
SF-295
<0.01
1.60
16.1
6.20
14.1
2.47
2.78
0.04
0.04
22.9
54.1
38.3
28.5
>100
54.1
2.69
2.52
5.18
2.89
3.18
5.24
SK-MEL-5
SK-OV-3
UO-31
Ndd
27.4
14.8
13.5
Ndd
15.2
14.9
1.74
10.0
20.3
41.3
5.19
5.36
5.01
2.61
1.23
1.76
2.45
8.96
1.94
3.43
2.87
2.13
DU-145
MCF7
1.57
1.47
2.25
0.73
1.79
6.93
5.79
NCI/ADR-RES
MDA-MB-231/ATCC
MDA-MB-435
8.55
12.4
2.46
6.72
7.46
Meanc
3.89
3.02
3.89
10.47
14.45
6.02
4.26
10.71
16.59
a GI50: drug molar concentration causing 50% cell growth inhibition.
b Data obtained from NCI’s in vitro disease-oriented tumor cell screen.
c Mean values over all 60 cell lines tested. These cell lines are: leukemia (CCRF-CEM, HL-60 (TB), K-562, MOLT-4, PRMI-8226, and SR); non-
small cell lung cancer (A549/ATCC, EKVX, HOP-62, HOP-92, NCI-H226, NCI-H23, NCI-H322M, and NCI-H522); colon cancer (COLC 205,
HCC-2998, HCT-116, HCT-15, HT29, KM12, and SW-620); CNS cancer (SF-268, SF-295, SF-539, SNB-19, SNB-75, and U251); melanoma (LOX
IMVI, MALME-3M, M14, SK-MEL-2, SK-MEL-28, SK-MEL-5, and UACC-257); ovarian cancer (IGR0V1, OVCAR-3, OVCAR-4, OVCAR-5,
OVCAR-8, and SK-OV-3); renal cancer (786-0, A498, ACHN, CAKI-1, RXF 393, SN12C, TK-10, and UO-31); prostate cancer (PC-3 and DU-
145); and breast cancer (MCF7, MCF7/ADR-RES, MDA-MB-231/ATCC, HS 578T, MDA-MB-435, MDA-N, and T-47D).
d Not determined.
Table 2. Antiproliferative assay of 3-substituted 4-anilino-2-phenylquinoline and 4-carboxamide derivatives
Cell lines
Compound
10
4.90
11.9
2.25
11
2.02
14.8
0.07
14a
7.10
14b
2.25
15a
15b
16
17
8.11
19.3
3.88
18
19
RPMI-8226
NCI-H226
HCT-116
2.34
4.60
1.48
1.31
2.49
0.63
8.83
3.36
0.35
0.98
2.02
0.25
2.91
19.8
>100
>100
>100
>100
>100
>100
>100
>100
Nd
24.5
58.1
34.6
40.9
38.6
86.7
18.0
>100
30.9
33.1
31.0
>100
39.4
71.0
23.7
>100
19.9
35.9
2.65
1.48
7.57
2.09
2.24
3.39
>100
6.23
18.6
>100
3.43
2.75
2.07
3.21
2.17
SF-295
3.66
7.61
1.58
0.46
10.8
22.9
17.3
29.4
13.9
16.4
Nd
SK-MEL-5
SK-OV-3
UO-31
10.7
14.5
16.6
32.4
4.95
32.1
1.13
1.36
8.62
4.58
9.20
3.92
38.0
0.38
>100
58.7
Nd
DU-145
MCF7
1.32
1.85
7.01
5.35
2.30
<0.01
0.10
1.64
Nd
NCI/ADR-RES
MDA-MB-231/ATCC
MDA-MB-435
4.62
4.71
3.19
2.02
2.74
1.97
>100
>100
>100
18.3
>100
4.15
25.5
27.8
<0.01
22.0
Mean
8.91
1.20
6.31
6.02
2.88
5.50
>100
14.45
28.84
41.69
position of substitution at quinoline ring is crucial. The
same antiproliferative SAR was observed for oxime (2a,
3.02 lM > 12a, 6.02 lM > 13a, 10.71 lM) and meth-
yloxime derivatives (2b, 3.89 lM > 12b, 4.26 lM > 13b,
16.59 lM) in which 6-OMe derivatives are preferred.
For the substituent at C(4) position of anilino moiety,
antiproliferative activity for oxime (2a, 3.02 lM), meth-
yloxime (2b, 3.89 lM), and the ketone precursor (1,
3.89 lM) is comparably indicated, a H-bonding accept-
ing group at C(4) position of 4-anilino-moiety is crucial.
The same antiproliferative SAR was observed for 8-OH
derivatives in which antiproliferative activity of 13a
(10.71 lM), 13b (16.59 lM), and 9 (14.45 lM) is compa-
rable (Table 1).
its 8-OMe counterpart (10, 8.91 lM) indicating that a
H-bonding donating substituent is more favorable
than that of a H-bonding accepting group. The same
antiproliferative SAR was observed for oxime (15a,
2.88 lM > 14a, 6.31 lM) and methyloxime derivatives
(15b, 5.50 lM > 14b, 6.02 lM). For 8-OH derivatives,
the antiproliferative effect of 30-COMe (11) is more
potent than its oxime derivative (15a), which in turn
is more potent than the methyloxime counterpart
(15b). This antiproliferative SAR is not obvious for
8-OMe derivatives in which the cytotoxicity of oxime
(14a), methyloxime (14b), and their ketone precursor
(10) is comparable. Compound 11 is especially active
against the growth of certain solid cancer cells such
as HCT-116 (colon cancer), MCF7, and MDA-MB-
435 (breast cancer) with GI50 values of 0.07, <0.01,
and <0.01 lM, respectively. Among those cancer
cells tested, breast cancer cells (MCF7, NCI/ADR-
RES, MDA-MB-231/ATCC, and MDA-MB-435)
The antiproliferative activity of 3-substituted 4-anili-
no-2-phenylquinoline derivatives is summarized in Ta-
ble 2. For 30-COMe derivatives, the antiproliferative
activity of 8-OH (11, 1.20 lM) is more potent than

Y.-L. Chen et al. / Bioorg. Med. Chem. 14 (2006) 3098–3105
Table 3. Effects of selected 4-anilino-2-phenylquinolines on cell cycle
progression
3101
5. Experimental
5.1. General
Compound
Sub-G1 (%) G0/G1 (%) S (%) G2/M (%)
Control
4.57
6.5
29.6
26.7
22.3
28.9
25.6
29.7
30.0
52.0
67.9
70.1
65.4
69.6
53.5
48.7
18.4
5.4
TLC: precoated (0.2 mm) silica gel 60 F254 plates from
EM Laboratories, Inc.; detection by UV light (254 nm).
All chromatographic separations were performed using
silica gel (Merck 60 230–400 mesh). Mp: Electrothermal
11 (3 lg/mL)
11 (10 lg/mL)
15a (3 lg/mL)
15a (10 lg/mL)
16 (3 lg/mL)
16 (10 lg/mL)
11.2
7.7
4.8
6.0
8.6
3.2
4.1
4.9
1
16.9
21.3
IA9100 melting point apparatus; uncorrected. H and
13C NMR spectra: Varian-Unity-400 spectrometer at
400 and 100 MHz, chemical shifts d in ppm with SiMe4
as an internal standard (=0 ppm), coupling constants J
in hertz. Elemental analyses were carried out on a
Heraeus CHN-O-Rapid elemental analyzer, and results
were within 0.4% of calculated values.
were found to be the most sensitive to 11 and 15a
with GI50 values less than 2.02 lM in each case. 4-
(8-Methoxy-2-phenylquinolin-4-ylamino)benzoic acid
(16) was devoid of antiproliferative activity, while its
carboxamide derivatives 17–19 were only weakly
active.
5.1.1. 2-[(2-Methoxyphenylamino)phenylmethylene]malon-
ic acid diethyl ester (4). A mixture of ethyl 3-chloro-2-
(ethoxycarbonyl)-3-phenylpropenoate
(3)
(2.84 g,
10 mmol), p-anisidine (1.48 g, 12 mmol), K2CO3 (1.95 g,
14 mmol), and dry DMF (50 mL) was heated in 140 ꢁC
for 3 h (TLC monitoring). The mixture was evaporated
under reduced pressure and then H2O (100 mL) was add-
ed. This aqueous mixture was extracted with CH2Cl2 (3·
100 mL), dried (MgSO4), and concentrated. The crude oil
thus obtained was purified by flash column chromatogra-
phy (FC, silica gel; using CH2Cl2 as the eluent) to give 4
(2.88 g, 78%). 1H NMR (CDCl3) d: 0.82 (t, 3H,
J = 7.2 Hz, OCH2CH3), 1.33 (t, 3H, J = 7.2 Hz, OCH2
CH3), 3.76 (s, 3H, OCH3), 3.74 (q, 2H, J = 7.2 Hz,
OCH2CH3), 4.26 (q, 2H, J = 7.2 Hz, OCH2CH3), 6.56–
6.72 (m, 4H, Ar-H), 7.26 (m, 5H, Ar-H), 11.12 (br s,
1H, NH). 13C NMR (CDCl3) d: 13.41, 14.36, 55.12,
60.08, 61.16, 97.05, 110.66, 115.41, 119.32, 121.09,
128.12 (2C), 128.68 (2C), 129.20, 131.82, 136.85, 148.16,
161.65, 167.27, 168.44. Anal. Calcd for C21H23NO5:
C, 68.27; H, 6.29; N, 3.79. Found: C, 68.19; H, 6.34; N,
3.68.
For 8-OH derivatives, the 30-COMe substituent is more
active than their respective 40-substituted couterparts
(11, mean GI50 = 1.20 lM > 9, 14.45 lM). The same
antiproliferative SAR was observed for oxime (15a,
2.88 lM > 13a, 10.71 lM) and methyloxime derivatives
(15b, 5.50 lM > 13b, 16.59 lM) in which 30-substituted
derivatives are preferred. This antiproliferative SAR is
not obvious for 8-OMe derivatives in which 40-substitut-
ed derivatives (8, 10.47 lM; 12a, 6.02 lM; 12b, 4.26 lM)
and their respective 30-substituted counterparts (10,
8.91 lM; 14a, 6.31 lM; 14b, 6.02 lM) are comparable.
Two of the most active compounds 11 and 15a along
with an inactive analog 16 were selected for evaluation
of cell cycle progression (Table 3). Flow cytometric anal-
yses indicated that 11 and 15a induced cell cycle arrest in
S phase. This result is interesting because 2-phenylqui-
nolone derivatives have been reported to be antimitotic
agents which induced cell cycle arrest in G2/M phase.
Thus, a substituent such as anilino group at C(4) posi-
tion of 2-phenylquinoline altered the mode of pharma-
cological mechanism of antimitotic 2-phenylquinolone
derivatives.
5.1.2. 8-Methoxy-2-phenyl-quinolin-4(1H)-one (5).
A
solution of 4 (3.68 g, 9.8 mmol) in Ph2O (10 mL) was
heated at 260–280 ꢁC for 0.5 h (TLC monitoring). The
reaction mixture was cooled and then n-hexane
(150 mL) was added and stirred at room temperature
for 6 h. The resulting precipitate was collected, dissolved
in EtOH (20 mL), added NaOH (4 N, 40 mL), and the
mixture was heated at reflux for 18 h. The solvent was
evaporated under reduced pressure and then H2O
(100 mL) was added and neutralized with 20% HCl solu-
tion. The resulting precipitate was collected and refluxed
with 20% HCl solution (200 mL) for 5 h (TLC monitor-
ing). The mixture was cooled to room temperature and
the precipitate was collected, washed with H2O, and
then crystallized from EtOH to give 5 (2.04 g, 83%).
Mp 170–171 ꢁC. (lit.,25 168–170 ꢁC). 1H NMR (CDCl3):
4.03 (3H, s, 8-OCH3), 6.59 (1H, s, 3-H), 7.06 (1H, d,
J = 7.6 Hz, 7-H), 7.27 (1H, m, Ar-H), 7.54 (3H, m,
Ar-H), 7.69 (2H, m, Ar-H), 7.93 (1H, d, J = 8.0 Hz, 5-
H), 8.84 (1H, br s, NH). 13C NMR (CDCl3): 56.07,
108.90, 110.56, 117.38, 123.18, 125.87, 126.44 (2C),
129.44 (2C), 130.65, 130.77, 134.55, 147.75, 148.40,
178.81. Anal. Calcd for C16H13NO2: C, 76.47; H, 5.22;
N, 5.58. Found: C, 76.39; H, 5.57; N, 5.31.
4. Conclusion
A number of 8-substituted 4-anilino-2-phenylquinoline
derivatives were synthesized and evaluated for their
antiproliferative activities. The results are: (1) for 40-
COMe-substituted derivatives, the antiproliferative
activity decreased in an order of 6-OMe (1,
3.89 lM) > 8-OMe
(8,
10.47 lM) > 8-OH
(9,
14.45 lM); (2) for 30-COMe derivatives, the antiprolifer-
ative activity of 8-OH (11, 1.20 lM) is more potent than
its 8-OMe counterpart (10, 8.91 lM); (3) for 8-OH
derivatives, the 30-COMe derivative (11, 1.20 lM) is
more active than its 40-substituted isomer (9,
14.45 lM). Flow cytometric analyses indicated that 11
and 15a induced cell cycle arrest in S-phase. This result
is interesting because 2-phenylquinolone derivatives
have been reported to be antimitotic agents which in-
duced cell cycle arrest in G2/M phase.

3102
Y.-L. Chen et al. / Bioorg. Med. Chem. 14 (2006) 3098–3105
5.1.3. 4-Chloro-8-methoxy-2-phenylquinoline (6). A mix-
ture of 5 (2.26 g, 9 mmol) and POCI3 (27 mL) was heat-
ed at 80–90 ꢁC for 1 h (TLC monitoring). After cooling,
the mixture was slowly poured into ice water (150 mL)
and neutralized with NH4OH. The precipitate was col-
lected, washed with H2O, and then crystallized from
EtOH to give 6 (2.30 g, 95%). Mp 100–101 ꢁC. (lit.,26
5.1.7.
4-(3-Acetylanilino)-8-methoxy-2-phenylquinoline
(10). A mixture of 6 (0.54 g, 2 mmol), 3-aminoacetophe-
none (0.27 g, 2 mmol), and pyridine (0.5 mL) in EtOH
(20 mL) was refluxed for 6 h (TLC monitoring). The mix-
ture was then cooled and evaporated in vacuo to yield a
yellow residue, treated with H2O (50 mL), and the result-
ing precipitate was filtered and washed with H2O. The
crude product was crystallized from EtOH to give 10
1
101–102 ꢁC). H NMR (CDCl3): 4.10 (3H, s, 8-OCH3),
1
7.12 (1H, d, J = 8.0 Hz, 7-H), 7.48–7.57 (4H, m, 6-H
and Ar-H), 7.81 (1H, d, J = 8.4 Hz, 5-H), 8.01 (1H, s,
3-H), 8.15–8.17 (2H, m, Ar-H). 13C NMR (CDCl3):
56.32, 108.90, 115.64, 119.68, 126.41, 127.36, 127.58
(2C), 128.85 (2C), 129.62, 138.68, 140.97, 143.10,
155.69, 155.99. Anal. Calcd for C16H12ClNO: C, 71.24;
H, 4.49; N, 5.19. Found: C, 71.11; H, 4.47; N, 5.20.
(0.51 g, 70%). Mp 200–201 ꢁC. H NMR (DMSO-d6):
2.62 (3H, s, COCH3), 4.02 (3H, s, 8-OCH3), 7.29 (1H,
d, J = 7.6 Hz, 7-H), 7.45–7.65 (6H, m, Ar-H), 7.78 (2H,
m, Ar-H), 7.96–8.03 (4H, m, Ar-H), 9.48 (1H, br s,
NH). 13C NMR (DMSO-d6): 26.75, 56.08, 100.74,
108.98, 113.10, 119.73, 121.90, 124.28, 125.67, 126.89,
127.38 (2C), 128.76 (2C), 129.57, 129.84, 130.24, 138.42,
138.54, 140.46, 149.12, 154.21, 155.48, 197.71. Anal.
Calcd for C24H20N2O2ÆH2O: C, 74.58; H, 5.75; N, 7.25.
Found: C, 74.60; H, 5.58; N, 7.09.
5.1.4. 4-Bromo-8-hydroxy-2-phenylquinoline (7). A mix-
ture of 6 (0.26 g, 1 mmol) and 48% HBr (20 mL) was re-
fluxed for 24 h (TLC monitoring). After cooling, the
mixture was neutralized with saturated NaOH aqueous.
The precipitate was collected, washed with H2O, and then
crystallized from EtOH to give 7 (0.23 g, 90%). Mp 121–
122 ꢁC. 1H NMR (CDCl3): 7.23 (1H, dd, J = 7.6,
1.2 Hz, 7-H), 7.47–7.56 (4H, m, 6-H and Ar-H), 7.62
(1H, dd, J = 8.4, 1.2 Hz, 5-H), 8.09-8.12 (2H, m, Ar-H),
8.18 (1H, s, 3-H), 8.38 (1H, br s, 8-OH). 13C NMR
(CDCl3): 111.12, 116.84, 123.48, 127.02, 127.36 (2C),
128.54, 128.96 (2C), 130.02, 134.97, 137.52, 138.39,
152.36, 154.72. Anal. Calcd for C15H10BrNO: C, 60.02;
H, 3.36; N, 4.67. Found: C, 60.11; H, 3.40; N, 4.68.
5.1.8. 4-(3-Acetylanilino)-8-hydroxy-2-phenylquinoline (11).
This compound was obtained from 7 and 3-aminoacetoph-
enone as described for 10 and was crystallized from EtOH
in 82% yield. Mp 201–202 ꢁC. 1H NMR (DMSO-d6): 2.65
(3H, s, COCH3), 6.95 (1H, s, 3-H), 7.45 (1H, d, J = 7.6 Hz,
7-H), 7.55–7.79 (7H, m, Ar-H), 7.89 (1H, m, Ar-H), 7.99
(1H, m, Ar-H), 8.10 (1H, m, Ar-H), 8.17 (1H, d,
J = 8.4 Hz, 5-H) 10.80 (1H, br s, OH), 11.60 (1H, br s,
NH). 13C NMR (DMSO-d6): 28.85, 99.82, 112.57,
116.25, 117.87, 124.50, 126.88, 127.60, 128.70 (2C),
128.82 (2C), 129.58, 130.36, 131.35, 132.68, 137.77,
138.28, 139.02, 148.37,152.97, 154.11, 197.30. Anal. Calcd
for C23H18N2O2Æ0.1H2O: C, 77.54; H, 5.16; N, 7.86.
Found: C, 77.20; H, 5.14; N, 7.79.
5.1.5. 4-(4-Acetylanilino)-8-methoxy-2-phenylquinoline (8).
A mixture of 6 (0.40 g, 1.5 mmol), 4-aminoacetophenone
(0.41 g, 3 mmol), and pyridine (0.5 mL) in EtOH (20 mL)
was added to a flask, which was placed in a sealed steel
bomb. The bomb was heated at 200 ꢁC for 18 h (TLC
monitoring). The resulting mixture was evaporated under
reduced pressure and then H2O (100 mL) was added. The
precipitate was collected, washed with H2O, and then
crystallized from EtOH to give 8 (0.48 g, 86%). Mp
5.1.9. (E)-4-[4-(1-Hydroxyiminoethyl)anilino]-8-methoxy-
2-phenylquinoline (12a). A mixture of 8 (0.52 g, 1.4 mmol),
NH2OHÆHCl (0.49 g, 7.0 mmol), and K2CO3 (0.48 g,
3.5 mmol) in EtOH (10 mL) was refluxed for 0.5 h (TLC
monitoring). The mixture was evaporated under reduced
pressure and then H2O (80 mL) was added. The resulting
precipitate was collected, washed with H2O, and crystal-
lized from EtOH to give 12a (0.51 g, 94%). Mp 235–
236 ꢁC. 1H NMR (DMSO-d6): 2.19 (3H, s, (C@N)CH3),
4.05 (3H, s, OCH3), 7.26 (1H, s, 3-H), 7.43 (1H, d,
J = 7.6, 7-H), 7.53–7.64 (6H, m, Ar-H), 7.78 (2H, m,
Ar-H), 7.88 (2H, m, Ar-H), 8.21 (1H, d, J = 8.4 Hz, 5-
H), 10.20 (1H, br s, NH), 11.25 (1H, s, NOH). 13C
NMR (DMSO-d6): 11.30, 56.21, 100.18, 111.31, 114.06,
118.74, 123.09 (2C), 126.12, 126.71 (2C), 127.98 (2C),
128.62 (2C), 130.21, 133.71, 135.90, 138.59, 139.20,
147.68, 151.22, 152.24, 153.94. Anal. Calcd for
C24H21N3O2Æ0.4H2O: C, 73.78, H, 5.64; N, 10.75. Found:
C, 73.77; H, 5.63; N, 10.57.
1
147–148 ꢁC. H NMR (CDCl3): 2.60 (3H, s, COCH3),
4.03 (3H, s, 8-OCH3), 7.06 (1H, d, J = 7.2 Hz, 7-H),
7.40–7.51 (6H, m, Ar-H), 7.57 (1H, s, 3-H), 7.91 (1H,
br s, NH), 7.95–8.01 (5H, m, Ar-H). 13C NMR (CDCl3):
26.39, 56.20, 103.08, 109.25, 113.38, 119.97 (2C), 120.51,
126.11, 127.40 (2C), 128.94 (2C), 129.70, 129.86, 130.31
(2C), 132.42, 139.29, 145.01, 147.52, 153.82, 155.34,
196.71. Anal. Calcd for C24H20N2O2Æ1.8H2O: C, 71.90;
H, 5.94; N, 6.99. Found: C, 72.11; H, 5.54; N, 6.83.
5.1.6. 4-(4-Acetylanilino)-8-hydroxy-2-phenylquinoline (9).
This compound was obtained from 7 and 4-aminoace-
tophenone as described for 8 and was crystallized from
1
EtOH in 77% yield. Mp 175–176 ꢁC. H NMR (CDCl3):
2.61 (3H, s, COCH3), 6.91 (1H, br s, NH), 7.20 (1H, dd,
J = 7.6, 2.0 Hz, 7-H), 7.35–7.52 (7H, m, Ar-H), 7.76 (1H,
s, 3-H), 8.00–8.05 (4H, m, Ar-H), 10.36 (1H, br s, OH).
13C NMR (CDCl3): 26.43, 102.83, 109.92, 110.44,
119.31 (2C), 119.54, 126.69, 127.27 (2C), 128.81 (2C),
129.59, 130.53 (2C), 132.16, 139.05, 139.24, 145.05,
146.35, 152.99, 155.54, 196.67. Anal. Calcd for
C23H18N2O2Æ0.1H2O: C, 77.54; H, 5.16; N, 7.86. Found:
C, 77.27; H, 5.31; N, 7.89.
The same procedure was used to convert each com-
pound 9, 10, and 11 to 13a, 14a, and 15a, respectively.
5.1.10. (E)-8-Hydroxy-4-[4-(1-hydroxyiminoethyl)anili-
no]-2-phenylquinoline (13a). Yield 75%. Mp 107–108 ꢁC
(EtOH). 1H NMR (DMSO-d6): 2.19 (3H, s, (C@N)CH3),
7.12 (1H, d, J = 7.6, 7-H), 7.437–7.50 (6H, m, Ar-H), 7.58
(1H, s, 3-H), 7.75 (2H, m, Ar-H), 7.83 (1H, d, J = 8.4 Hz,
5-H), 8.22 (2H, m, Ar-H), 9.15 (1H, br s, NH), 10.62 (1H,

Y.-L. Chen et al. / Bioorg. Med. Chem. 14 (2006) 3098–3105
3103
br s, OH), 11.14 (1H, s, NOH). 13C NMR (DMSO-d6):
11.38, 99.34, 111.13, 111.97, 119.48, 121.57 (2C), 125.47,
126.68 (2C), 127.26 (2C), 128.59 (2C), 129.26, 132.01,
138.52, 138.80, 141.04, 148.50, 152.43, 152.95, 154.30.
Anal. Calcd for C23H19N3O2Æ1.7H2O: C, 69.04, H, 5.65;
N, 10.50. Found: C, 69.01; H, 5.51; N, 10.35.
(1H, br s, NH), 7.45–7.54 (7H, m, Ar-H), 7.77 (2H, m,
Ar-H), 7.92 (1H, d, J = 8.4 Hz, 5-H), 8.08 (2H, m, Ar-
H), 9.85 (1H, br s, NH). 13C NMR (DMSO-d6): 12.13,
61.57, 99.78, 112.22 (2C), 112.88, 119.06, 122.43, 126.23,
127.13 (2C), 127.83 (2C), 128.66 (2C), 129.77, 129.99,
131.85, 140.53, 150.14, 151.47, 151.95, 153.47, 153.87.
Anal. Calcd for C24H21N3O2Æ1.4H2O: C, 70.52; H, 5.88;
N, 10.28. Found: C, 70.34; H, 5.62; N, 10.09.
5.1.11. (E)-4-[3-(1-Hydroxyiminoethyl)anilino]-8-methoxy-
2-phenylquinoline (14a). Purified by flash column chroma-
tography (FC, silica gel; using MeOH/CH2Cl2 = 1/30 as
the eluent) to give 14a in 93% yield. Mp 201–202 ꢁC.
1H NMR (DMSO-d6): 2.19 (3H, s, (C@N)CH3), 4.00
(3H, s, OCH3), 7.20 (1H, d, J = 7.6 Hz, 7-H), 7.41–7.56
(8H, m, Ar-H), 7.74 (1H, s, 3-H), 7.93 (1H, d,
J = 8.0 Hz, Ar-H), 8.00–8.03 (2H, m, Ar-H), 9.02 (1H,
br s, NH), 11.27 (1H, s, NOH). 13C NMR (DMSO-d6):
11.55, 55.80, 99.40, 109.11, 113.51, 119.03, 119.99,
120.86, 122.19, 124.84, 126.95 (2C), 128.64 (2C), 128.97,
129.48, 138.20, 139.82, 140.80, 140.87, 148.40, 152.67,
154.94, 155.59. Anal. Calcd for C24H21N3O2Æ0.2H2O: C,
74.46; H, 5.58; N, 10.86. Found: C, 74.42; H, 5.82; N,
10.49.
5.1.15. (E)-8-Methoxy-4-[3-(1-methoxyiminoethyl)anili-
no]-2-phenylquinoline (14b). Yield 80%. Mp 163–164 ꢁC
1
(EtOH). H NMR (CDCl3): 2.24 (3H, s, (C@N)CH3),
4.01 (3H, s, OCH3), 4.07 (3H, s, OCH3), 7.06 (1H, d,
J = 7.6 Hz, 7-H), 7.34–7.44 (7H, m, Ar-H), 7.54 (2H, m,
Ar-H), 7.67 (1H, m, Ar-H), 8.01–8.04 (2H, m, Ar-H),
9.19 (1H, br s, NH). 13C NMR (CDCl3): 12.65, 56.15,
62.05, 101.05, 108.29, 111.48, 119.71, 119.96, 122.02,
122.76, 125.33, 127.59 (2C), 128.56 (2C), 128.99, 129.71,
138.35, 139.24, 140.21, 140.34, 147.89, 154.10, 155.96,
156.76. Anal. Calcd for C25H23N3O2Æ0.2H2O: C, 74.85;
H, 5.89; N, 10.48. Found: C, 74.90; H, 6.03; N, 10.15.
5.1.16. (E)-8-Hydroxy-4-[3-(1-methoxyiminoethyl)anili-
no]-2-phenylquinoline (15b). Yield 82%. Mp 107–108 ꢁC
5.1.12. (E)-8-Hydroxy-4-[3-(1-hydroxyiminoethyl)anilino]-
2-phenylquinoline (15a). Yield 75%. Mp 148–150 ꢁC
(EtOH). 1H NMR (DMSO-d6): 2.19 (3H, s, (C@N)CH3),
6.90 (1H, d, J = 6.0 Hz, Ar-H), 7.46–7.81 (12H, m, Ar-
H), 8.18 (1H, d, J = 8.0 Hz, 5-H), 10.91 (1H, br s, OH),
11.38 (1H, br s, NOH), 13.20 (1H, br s, NH). 13C
NMR (DMSO-d6): 11.45, 99.22, 112.38, 114.35, 118.36,
123.03, 124.09, 126.60, 128.10 (2C), 128.64 (2C), 128.92,
129.70, 130.46, 138.42, 138.57, 139.37, 140.52, 140.69,
150.18, 152.32, 153.29. Anal. Calcd for C23H19N3O2Æ1.2-
H2O: C, 70.63, H, 5.53; N, 10.75. Found: C, 70.55; H,
5.36; N, 10.61.
1
(EtOH). H NMR (CDCl3): 2.20 (3H, s, (C@N)CH3),
3.97 (3H, s, OCH3), 7.17 (1H, s, 3-H), 7.21–7.49 (9H, m,
Ar-H), 7.81 (2H, m, Ar-H), 8.08 (1H, d, J = 8.0 Hz, 5-
H), 9.40 (1H, br s, NH), 10. 79 (1H, br s, OH). 13C
NMR (CDCl3): 12.58, 62.03, 99.13, 112.84, 115.28,
118.03, 121.60, 123.89, 124.76, 126.87, 127.36 (2C),
129.29 (2C), 129.85, 131.08, 133.20, 134.75, 138.25,
138.52, 149.35, 152.62, 152.73, 153.82. Anal. Calcd for
C24H21N3O2Æ1.7H2O: C, 70.21; H, 5.90; N, 10.24. Found:
C, 69.95; H, 5.58; N, 0.03.
5.1.17. 4-(8-Methoxy-2-phenylquinolin-4-ylamino)benzoic
acid (16). Obtained from 6 and 4-aminobenzoic acid as
5.1.13. (E)-8-Methoxy-4-[4-(1-methoxyiminoethyl)anilino]-
2-phenylquinoline (12b). A mixture of 8 (0.31 g, 0.8 mmol),
O-methylhydroxylamineÆHCl (0.35 g, 4.2 mmol), and
K2CO3 (0.29 g, 2.1 mmol) in EtOH (10 mL) was refluxed
for 0.5 h (TLC monitoring). The mixture was evaporated
under reduced pressure and then H2O (80 mL) was added.
The resulting precipitate was collected, washed with H2O,
and then crystallized from EtOH to give 12b (0.23 g, 72%).
Mp 181–182 ꢁC. 1H NMR (DMSO-d6): 2.20 (3H, s,
(C@N)CH3), 3.92 (3H, s, OCH3), 4.00 (3H, s, OCH3),
7.21 (1H, d, J = 7.2 Hz, 7-H), 7.44–7.52 (6H, m, Ar-H),
7.62 (1H, s, 3-H), 7.73 (2H, m, Ar-H), 7.91 (1H, m, Ar-
H), 8.05 (2H, m, Ar-H), 9.08 (1H, br s, NH). 13C NMR
(DMSO-d6): 12.12, 55.82, 61.51, 100.39, 109.12, 113.57,
119.86, 120.83 (2C), 124.96, 127.03 (2C), 127.69 (2C),
128.63 (2C), 129.80, 130.23, 139.80, 140.92, 142.10,
147.71, 153.57, 155.02, 155.65. Anal. Calcd for
C25H23N3O2Æ0.2H2O: C, 74.85; H, 5.89; N, 10.47. Found:
C, 74.82; H, 6.07; N, 10.26.
described for
9
and purified by FC (MeOH/
CH2Cl2 = 1/10) to give 16 in 52% yield. Mp 236–
237 ꢁC. 1H NMR (DMSO-d6): 4.01 (3H, s, OCH3),
7.23 (1H, d, J = 7.6 Hz, 7-H), 7.43–7.53 (6H, m, Ar-
H), 7.79 (1H, s, 3-H), 7.89 (1H, d, J = 8.4 Hz, 5-H),
7.97 (2H, m, Ar-H), 8.10 (2H, m, Ar-H), 9.27 (1H, br
s, NH) 12.60 (1H, br s, COOH). 13C NMR (100 MHz,
DMSO-d6): 55.86, 102.55, 109.21, 113.78, 118.92 (2C),
120.95, 124.03, 125.30, 127.15 (2C), 128.69 (2C),
129.14, 131.11 (2C), 139.62, 141.04, 146.00, 146.85,
155.01, 155.70, 167.18. Anal. Calcd for C23H18N2O3Æ0.6-
H2O: C, 72.46; H, 5.09; N, 7.35. Found: C, 72.33; H,
5.18; N, 7.28.
5.1.18. N-(2-Dimethylaminoethyl)-4-(8-methoxy-2-phenyl-
quinolin-4-ylamino)benzamide (17). A mixture of 16
(0.37 g, 1.0 mmol) and CDI (0.97 g, 6.0 mmol) was stirred
in dry DMF (10 mL) at room temperature for 2 h. To
this mixture was added N,N-dimethylethylenediamine
(0.61 g, 6.0 mmol), and the reaction mixture was stirred
at room temperature for a further 24 h (TLC monitor-
ing). The volatile components were then removed in vac-
uo, the residue was dissolved in CH2Cl2 (80 mL), washed
with H2O (2· 80 mL), and dried (MgSO4), and the sol-
vent was removed in vacuo to provide the crude product,
which was purified by FC (MeOH/CH2Cl2 = 1/5) to give
The same procedure was used to convert each com-
pounds 9, 10, and 11, to 13b, 14b, and 15b, respectively.
5.1.14. (E)-8-Hydroxy-4-[4-(1-methoxyiminoethyl)anili-
no]-2-phenylquinoline (13b). Yield 70%. Mp 154–155 ꢁC
(EtOH). 1H NMR (DMSO-d6): 2.21 (3H, s, (C@N)CH3),
3.93 (3H, s, OCH3), 7.21 (1H, d, J = 7.2 Hz, 7-H), 7.40

3104
Y.-L. Chen et al. / Bioorg. Med. Chem. 14 (2006) 3098–3105
17 (0.39 g, 84%). Mp 213–214 ꢁC. 1H NMR (DMSO-d6):
2.20 (6H, s, N(CH3)2), 2.43 (2H, t, J = 6.8 Hz, CH2N),
3.38 (2H, dd, J = 12.0, 6.8 Hz, NHCFb), 4.00 (3H, s, 8-
OCH3), 7.21 (1H, d, J = 8.0 Hz, 7-H), 7.42–7.53 (6H,
m, Ar-H), 7.69 (1H, s, 3-H), 7.90 (3H, m, Ar-H), 8.07
(2H, m, Ar-H), 8.33 (1H, t, J = 5.6 Hz, CONH), 9.14
(1H, br s, NH). 13C NMR (DMSO-d6): 37.32, 45.24
(2C), 55.82, 58.28, 101.13, 109.14, 113.62, 119.78 (2C),
120.52, 125.08, 127.04 (2C), 128.39, 128.63 (2C), 128.68
(2C), 129.04, 139.71, 140.97, 144.01, 147.36, 154.98,
155.67, 165.64. Anal. Calcd for C27H28N4O2Æ0.2H2O: C,
73.00; H, 6.46; N, 12.62. Found: C, 73.27; H, 6.67; N,
12.36.
RNase A solution at 37 ꢁC for 30 min. Cells were har-
vested by centrifugation at 1000 rpm for 5 min and fur-
ther stained with 250 lL DNA staining solution (10 mg
propidium iodide [PI], 0.1 mg trisodium citrate, and
0.03 mL Triton X-100 dissolved in 100 mL H2O) at
room temperature for 30 min in the dark. After loading
500 lL PBS, the DNA contents of 10,000 events were
measured by FACScan (Elite ESP, Beckman Coulter,
Brea, CA) and the cell cycle profile was analyzed from
the DNA content histograms by using WinCycle soft-
ware. When cells were apoptotic, the containing DNA
was digested by endonuclease and then the sub G1 pick
appeared. The percentage in sub G1 was analyzed by
gating on cell cycle dot blots using Windows Multiple
Document Interface software (WinMDI).
5.1.19. N-[2-(2-Hydroxyethylamino)ethyl]-4-(8-methoxy-
2-phenylquinolin-4-ylamino)benzamide (18). Obtained
from 16 and N-(2-hydroxyethyl)ethylenediamine as de-
scribed for 17. The crude product was purified by FC
(MeOH/CH2Cl2 = 1/5) to give 18 in 51% yield. Mp
117–119 ꢁC. 1H NMR (DMSO-d6): 2.62 (2H, t,
J = 6.0 Hz, NCH2CH2OH), 2.71 (2H, t, J = 6.4 Hz,
CH2NHCH2CH2OH), 3.36 (2H, dd, J = 11.6, 6.0 Hz,
NHCH2CH2), 3.47 (2H, t, J = 5.6 Hz, NCH2CH2OH),
4.00 (3H, s, 8-OCH3), 4.53 (2H, br s, OH and NH),
7.23 (1H, d, J = 7.6 Hz, 7-H), 7.42–7.53 (6H, m, Ar-
H), 7.69 (1H, s, 3-H), 7.91 (3H, m, Ar-H), 8.06 (2H,
m, Ar-H), 8.38 (1H, t, J = 5.2 Hz, CONH), 9.15 (1H,
br s, NH). 13C NMR (DMSO-d6): 40.76, 48.14, 50.96,
55.27, 59.91, 101.10, 109.15, 113.62, 119.77 (2C),
120.51, 125.05, 127.02 (2C), 128.51, 128.61 (2C),
128.70 (2C), 129.02, 139.70, 140.98, 143.98, 147.37,
154.97, 155.65, 165.82. Anal. Calc. for C27H28N4O3Æ1.3-
H2O: C, 67.55; H, 6.44; N, 11.67. Found: C, 67.31; H,
6.44; N, 11.56.
Acknowledgments
Financial support of this work by the National Science
Council of the Republic of China is gratefully acknowl-
edged. We also thank National Cancer Institute (NCI)
of the United States for the anticancer screenings and
the National Center for High-Performance Computing
for providing computer resources and chemical database
services.
References and notes
1. Fang, K. C.; Chen, Y. L.; Sheu, J. Y.; Wang, T. C.; Tzeng,
C. C. J. Med. Chem. 2000, 43, 3809.
2. Chen, Y. L.; Fang, K. C.; Sheu, J. Y.; Hsu, S. L.; Tzeng,
C. C. J. Med. Chem. 2001, 44, 2374.
3. Tzeng, C. C.; Chen, Y. L. Chin. Pharm. J. 2002, 54, 229.
4. Sheu, J. Y.; Chen, Y. L.; Tzeng, C. C.; Hsu, S. L.; Fang,
K. C.; Wang, T. C. Helv. Chim. Acta 2003, 86, 2481.
5. Zhao, Y. L.; Chen, Y. L.; Sheu, J. Y.; Chen, I. L.; Wang,
T. C.; Tzeng, C. C. Bioorg. Med. Chem. 2005, 13, 3921.
6. Chen, K.; Kuo, S. C.; Hsieh, M. C.; Mauger, A.; Lin, C.
M.; Hamel, E.; Lee, K. H. J. Med. Chem. 1997, 40, 3049.
7. Nakamura, S.; Kozuka, M.; Bastow, K. F.; Tokuda, H.;
Nishino, H.; Suzuki, M.; Tatsuzaki, J.; Natschke, S. L.
M.; Kuo, S. C.; Lee, K. H. Bioorg. Med. Chem. 2005, 13,
4396.
8. Lai, Y. Y.; Huang, L. J.; Lee, K. H.; Xiao, Z.; Bastow, K. F.;
Yamori, T.; Kuo, S. C. Bioorg. Med. Chem. 2005, 13, 265.
9. Chen, I. L.; Chen, Y. L.; Tzeng, C. C.; Chen, I. S. Helv.
Chim. Acta 2002, 85, 2214.
10. Chen, I. L.; Chen, Y. L.; Tzeng, C. C. Chin. Pharm. J.
2003, 55, 49.
5.1.20. 4-(8-Methoxy-2-phenylquinolin-4-ylamino)-N-(2-
piperazin-l-ylethyl)benzamide (19). Obtained from 16
and l-(2-aminoethyl)piperazine described for 17. The
crude product was purified by FC (MeOH/CH2Cl2 = 1/
1
5) to give 19 in 52% yield. Mp 125–126 ꢁC. H NMR
(DMSO-d6): 2.36 (4H, m, piperazinyl-H), 2.44 (2H, t,
J = 6.8 Hz, NHCH2CH2N), 2.70 (4H, m, piperazinyl-
H), 3.38 (2H, dd, J = 12.8, 6.4 Hz, NHCH2CH2N),
4.00 (3H, s, 8-OCH3), 4.09 (2H, br s, NH), 7.21 (1H,
d, J = 8.0 Hz, 7-H), 7.42–7.52 (6H, m, Ar-H), 7.69
(1H, s, 3-H), 7.89 (3H, m, Ar-H), 8.07 (2H, m, Ar-H),
8.33 (1H, t, J = 5.6 Hz, CONH), 9.15 (1H, br s, NH).
13C NMR (DMSO-d6): 37.12 (2C), 46.03, 54.71, 56.26
(2C), 58.33, 101.58, 109.58, 114.07, 120.22 (2C),
120.97, 125.52, 127.48 (2C), 128.88, 129.07 (2C),
129.11 (2C), 129.49, 140.16, 141.41, 144.45, 147.80,
155.42, 156.11, 166.08. Anal. Calcd for C29H31N5O2Æ1.1-
H2O: C, 69.45; H, 6.69; N, 13.96. Found: C, 69.25; H,
6.77; N, 13.60.
11. Huang, Y. T.; Huang, D. M.; Guh, J. H.; Chen, I. L.;
Tzeng, C. C.; Teng, C. M. J. Biol. Chem. 2005, 280, 2771.
12. Chen, Y. L.; Chen, I. L.; Wang, T. C.; Han, C. H.; Tzeng,
C. C. Eur. J. Med. Chem. 2005, 40, 928.
13. Zhao, Y. L.; Chen, Y. L.; Chang, F. S.; Tzeng, C. C. Eur.
J. Med. Chem. 2005, 40, 792.
14. Cheng, C. C. Med. Hypothesis 1986, 20, 157.
15. Atwell, G. J.; Baguley, B. C.; Denny, W. A. J. Med. Chem.
1989, 32, 396.
16. Kuo, S. C.; Lee, H. Z.; Juang, J. P.; Lin, Y. T.; Wu, T. S.;
Chang, J. J.; Lednicer, D.; Paull, K. D.; Lin, C. M.;
Hamel, E.; Lee, K. H. J. Med. Chem. 1993, 36, 1146.
17. Xia, Y.; Yang, Z. Y.; Xia, P.; Bastow, K. F.; Tachibana,
Y.; Kuo, S. C.; Hamel, E.; Hackl, T.; Lee, K. H. J. Med.
Chem. 1998, 41, 1155.
5.2. Cell cycle analysis
Flow cytometry was used to measure cell cycle profile
and apoptosis. For cell cycle analysis, K-562 cell treated
with compounds (3 and 10 lM) for 24 h was harvested
by centrifugation. After being washed with PBS, the cell
was fixed with ice-cold 70% ethanol for 30 min, washed
with PBS, and then treated with 1 mL of 1 mg/mL of

Y.-L. Chen et al. / Bioorg. Med. Chem. 14 (2006) 3098–3105
3105
18. Deady, L. W.; Desneves, J.; Kaye, A. J.; Finlay, G. J.;
Baguley, B. C.; Denny, W. A. Bioorg. Med. Chem. 2001, 9,
445.
19. Deady, L. W.; Rodemann, T.; Zhuang, L.; Baguley, B. C.;
Denny, W. A. J. Med. Chem. 2003, 46, 1049.
20. Deady, L. W.; Roger, M. L.; Zhuang, L.; Baguley, B. C.;
Denny, W. A. Bioorg. Med. Chem. 2005, 13, 1341.
21. Bu, X.; Chen, J.; Deady, L. W.; Smith, C. L.; Baguley, B.
C.; Greenhalgh, D.; Yang, S.; Denny, W. A. Bioorg. Med
Chem. 2005, 13, 3657.
23. Silverstein, R. M.; Webster, F. X. 13C NMR Spectrom-
etry. In Spectrometric Identification of Organic Com-
pounds; Rose, N., Ed., 6th ed.; John Wiley and Sons:
New York, 1998; pp 217–249.
24. Monks, A.; Scuderio, D.; Skehan, P.; Shoemaker, R.;
Paull, K.; Vistica, D.; Hose, C.; Langlay, J.; Cronise, P.;
Vaigro-Wolff, A.; Gray-Goodrich, M.; Campbell, H.;
Mayo, J.; Boyd, M. J. Natl. Cancer Inst. 1991, 83, 757.
25. Kuo, S. C.; Lee, H. Z.; Juang, J. P.; Lin, Y. T.; Wu, T. S.;
Chang, J. J.; Lednicer, D.; Paull, K. D.; Lin, C. M.;
Hamel, E.; Lee, K. H. J. Med. Chem. 1993, 63, 1146.
26. Venturella, P.; Bellino, A. J. Heterocycl. Chem. 1975, 12, 669.
22. Hormi, O. E. O.; Peltonen, C.; Heikkila, L. J. Org. Chem.
1990, 55, 2513.