Synthesis and antitumor activity of novel 4-(2-fluorophenoxy)quinoline derivatives bearing the 4-Oxo-1,4-dihydroquinoline-3-carboxamide moiety

Article abstract of DOI:10.1002/ardp.201300029

A series of 4-(2-fluorophenoxy)quinoline derivatives bearing the 4-oxo-1,4-dihydroquinoline-3-carboxamide moiety were designed, synthesized, and evaluated for their in vitro antitumor activity against the H460, HT-29, MKN-45, U87MG, and SMMC-7721 cancer cell lines. Most of the tested compounds showed potent activity and high selectivity toward the HT-29 and MKN-45 cell lines. Furthermore, compounds 21b, 21c, and 21i were further examined for their c-Met kinase activity and exhibited strong efficacy with IC₅₀ values in the single-digit nanomolar range, which was comparable with the positive control foretinib. The most promising compound 21c showed excellent cytostatic activity with IC₅₀ values from 0.01 to 0.53 μM against all tested cell lines, thus being 1.7-2.2 times more active than foretinib. Novel 4-(2-fluorophenoxy)quinoline derivatives bearing the 4-oxo-1,4-dihydroquinoline- 3-carboxamide moiety were evaluated for their in vitro antitumor activity against several cancer cell lines. Compounds 21b, 21c, and 21i were also examined for their c-Met kinase activity in comparison to foretinib. Compound 21c showed excellent cytostatic activity with IC₅₀ values from 0.01 to 0.53 μM against all tested cell lines and higher activity than foretinib.

Full text of DOI:10.1002/ardp.201300029

Arch. Pharm. Chem. Life Sci. 2013, 346, 521–533
521
Full Paper
Synthesis and Antitumor Activity of Novel 4-(2-Fluorophenoxy)-
quinoline Derivatives Bearing the 4-Oxo-1,4-dihydroquinoline-
3-carboxamide Moiety
Sai Li, Rui Jiang, Mingze Qin, Haicheng Liu, Guangyan Zhang, and Ping Gong
Key Laboratary of Original New Drugs Design and Discovery of Ministry of Education, Shenyang Pharmaceutical
University, Shenyang, P. R. China
A series of 4-(2-fluorophenoxy)quinoline derivatives bearing the 4-oxo-1,4-dihydroquinoline-3-
carboxamide moiety were designed, synthesized, and evaluated for their in vitro antitumor activity
against the H460, HT-29, MKN-45, U87MG, and SMMC-7721 cancer cell lines. Most of the tested
compounds showed potent activity and high selectivity toward the HT-29 and MKN-45 cell lines.
Furthermore, compounds 21b, 21c, and 21i were further examined for their c-Met kinase activity and
exhibited strong efficacy with IC50 values in the single-digit nanomolar range, which was comparable
with the positive control foretinib. The most promising compound 21c showed excellent cytostatic
activity with IC50 values from 0.01 to 0.53 mM against all tested cell lines, thus being 1.7–2.2 times more
active than foretinib.
Keywords: Antitumor activity / 4-Oxo-1,4-dihydroquinoline-3-carboxamide moiety / Synthesis
Received: February 1, 2013; Revised: April 20, 2013; Accepted: April 26, 2013
DOI 10.1002/ardp.201300029
Introduction
by some water-soluble fragments, such as the 3-morpholino-
propoxy group, while there is little change to the B moiety
except phenyl ring or substituted phenyl ring. In contrast, the
linkers between A moiety and B moiety seem remarkably
different, but have two common features. First, there is six
chemical bonds distance existing between A moiety and B
moiety, which is called as “5 atoms regulation”. Second, the
linkers contain both hydrogen-bond donor or acceptor and
amide or thioamide group. These structural characteristics
suggest that exploring a satisfactory linker will be a practicable
way of designing 4-(2-fluorophenoxy)quinoline derivatives.
To our knowledge, compounds bearing 4-oxo-1,4-dihydro-
quinoline-3-carboxam-ide fragment have been reported to
display a large field of biological activities, including
antitumor [13], antibacterial [14], and anti-inflammatory
activities [15], etc.
Interestingly, this fragment conformed to the two common
characteristics of linker mentioned above. Inspired by these
observations, we designed a series of novel 4-(2-fluorophe-
noxy)quinoline derivatives (I, Fig. 2) based on combination
principles, in which the A moiety of foretinib was preserved
and the 4-oxo-1-phenyl-1,4-dihydro-quinoline-3-carboxamide
framework was attached to the C-40 positon of A moiety as the
linker and B moiety. Meanwhile, the morpholinyl group at
The search for new antitumor chemotherapeutic agents
continues to be an active area of research in many companies.
During the last decade, many synthetic quinoline derivatives
have been reported with significant antitumor activity [1–5].
Among them, foretinib (1, Fig. 1), an orally available multi-
kinase inhibitor of c-Met and VEGFR-2, is currently undergoing
phase II clinical trials for head and neck cancer, gastric cancer
and papillary renal cell carcinoma [6]. Due to its virtue of
multi-targets effects and broad-spectrum anticancer potency,
researchers drew attentions to the optimization of foretinib.
In recent years, a variety of 4-(2-fluorophenoxy)quinoline
derivatives have been developed as potent multiple target c-Met
kinase inhibitors [7–12]. As shown in Fig. 1, these quinoline
derivatives have some similar structural characteristics. The
modifications of the A moiety focus on the 7-position of
the quinoline ring, where the methoxy group could be replaced
Correspondence: Prof. Ping Gong, Key Laboratary of Original New Drugs
Design and Discovery of Ministry of Education, Shenyang Pharmaceutical
University, 103 Wenhua Road, Shenhe District, Shenyang 110016,
P. R. China.
E-mail: gongpinggp@126.com
Fax: þ86 24 23882925
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Figure 1. Structures of foretinib and other 4-(2-fluorophenoxy)quinoline derivatives.
7-position of quinoline ring was replaced by other water-
soluble substituents, including piperidinyl, pyrrolidinyl, 4-
methyl piperazinyl, and 4-methyl piperidinyl group. Addi-
antitumor activities against five human cancer cell lines and
c-Met kinase were evaluated, respectively.
tionally, various R3 groups especially mono-electron-with- Results and discussion
drawing groups (mono-EWGs), especially halogen (F, Cl), and
trifluoromethyl group were introduced to the phenyl ring
(B moiety) to investigate their effect on activity, as well as a
7-fluoro group was added to the linker. In this paper, the
synthesis of target compounds was reported and their in vitro
Chemistry
The key intermediates 8a–8e were synthesized using a
convenient eight-step procedure starting from 1-(4-hydroxy-
3-methoxyphenyl)ethanone depicted in Scheme 1. The
commercially available acetophenone was alkylated with 1-
bromo-3-chloro-propane in acetone under basic condition to
provide compound 1 [16], which was converted to nitro
compound 2 using fuming nitric acid as nitration reagent in
dichloromethane at ꢀ20°C for 6 h with 70% yield [17]. Then,
condensation of 2 with dimethyl formamide dimethyl acetal
(DMF-DMA) in refluxing toluene gave yellow solid 3 [18],
which was reduced and cyclized using glacial acetic acid and
iron powder to afford hydroxy-quinoline 4 in high yield and
purity in a single step [19]. The reaction of 4 with excessive
secondary amine (piperidine, 4-methyl piperidine, morpho-
line, pyrrolidine, 4-methyl piperizine) in acetonitrile at reflux
provided intermediates 5a–5e, respectively [20], which were
converted to chloro-quinoline 6a–6e by refluxing in the mixed
solvent of phosphorus oxychloride and acetonitrile in
sequence [21]. Next, the etherification reaction of 6a–6e
and 2-fluoro-4-nitrophenol afforded purified 7a–7e, which
were reduced by iron powder and catalytic amounts of
ammonium chloride in ethanol to obtain amides 8a–8e [22].
Figure 2. General structure of the target compounds.
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4-(2-Fluorophenoxy)quinoline Derivatives
523
Scheme 1. Synthesis of intermediates 8a–e. Reagents and conditions: (i) Br(CH2)3Cl, (CH3)2CO, K2CO3, r.t.; (ii) fuming HNO3, CH2Cl2,
ꢀ20°C; (iii) DMF-DMA, PhMe, 110°C; (iv) Fe, HOAc, 85°C; (v) amine, CH3CN, reflux; (vi) POCl3, CH3CN, 85°C; (vii) 2-fluoro-4-nitrophenol,
PhCl, reflux; (viii) Fe, NH4Cl, EtOH, reflux.
The syntheses of target compounds 21a–21l and 22a–22g
were outlined in Scheme 2. 2-Fluorobenzoic acid or 2,4-
difluorobenzoic acid were refluxed in dichlorosulfoxide to
yield benzoyl chlorides 9a and 9b, which were separately
converted to ketones 10a and 10b by the acylation with 2,2-
dimethyl-1,3-dioxane-4,6-dione (meldrum’s acid) and 4-
dimethyl aminopyridine (DMAP) in dichloromethane at
0°C for 5–6 h [23]. Refluxing in ethanol, ketones 10a and 10b
yielded intermediates 11a and 11b [23], which via conden-
sation with DMF-DMA afforded light yellow solid 12a and
12b [24]. Reaction of 12a and 12b with various substituted
anilines afforded the intermediates 13a–13f and 14a–14d,
respectively [24], which were stirred in dimethyl formamide
in the presence of potassium carbonate at 80°C to provide
esters 15a–15f and 16a–16d. The esters were hydrolyzed
with 10% NaOH solution to obtain corresponding acids 17a–
17f and 18a–18d, which were further treated with
dichlorosulfoxide to provide acyl chlorides 19a–19f and
20a–20d in sequence. Finally, condensation of intermedi-
ates 8a–8d with 19a–19f or 20a–20d in the presence of
triethylamine in dichloromethane at room temperature for
a whole night provided compounds 21a–21l and 22a–22g,
respectively [25].
First screening for biological activities
The antitumor activities of target compounds were evaluated
against H460 (human lung cancer), HT-29 (human colon
cancer), MKN-45 (human gastric cancer cancer), U87MG
(human glioblastoma) and SMMC-7721 (human liver cancer)
cancer cell lines by a MTT assay. The results, expressed as
averaged IC50 values from at least three independent experi-
ments, were summarized in Table 1.
As illustrated in Table 1, all target compounds showed
moderate to excellent cytostatic potency against the different
cancer cells in the single-digit mM range. In general, most of
them displayed high selectivity toward HT-29 and MKN-45
cancer cells. It was worth noting that compounds 21b, 21c,
and 21i exhibited more potent antitumor activities against all
tested cell lines than foretinib with IC50 values ranging from
0.01 to 0.71 mM.
The analysis of Table 1 clearly reveals that the antitumor
activities of compounds 22a–22g (R2 ¼ F) was lower than
those of the corresponding compounds 21a–21l (R2 ¼ H). For
example, the activity of compound 21b (R2 ¼ H, IC50 ¼ 0.086,
0.054, 0.015, 0.67, and 0.15 mM) was 15.2, 16.6, 39, 1.8, and 9.1
times higher than that of 22a (R2 ¼ F) against H460, HT-29,
MKN-45, U87MG, and SMMC-7721, respectively, which
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Scheme 2. Synthesis of target compounds. Reagents and conditions: (i) SOCl2, PhMe, reflux; (ii) DMAP, Meldrum’s acid, CH2Cl2, ꢀ10°C;
(iii) EtOH, reflux; (iv) DMF-DMA, CH2Cl2, reflux; (v) phenylamine, EtOH, concentrated HCl, reflux; (vi) DMF, K2CO3, 50°C; (vii) EtOH/5%
NaOH, r.t.; (viii) SOCl2/PhMe, reflux; (ix) Et3N, CH2Cl2, r.t.
suggested that the introduction of a 7-fluoro group to the
4-oxo-1,4-dihydroquinoline scaffold led to an obvious decline
in activity.
Replacement of the 4-fluoro group with double-EWGs (3,4-
difluoro, 21g, IC50 ¼ 0.58 mM or 2,4-dichloro, 21k, IC50
¼ 0.49 mM) resulted in a slight improvement in activity. On
In addition, it was worth mentioning that compounds
with the same R3 groups but different R1 groups showed
minor differences in cytostatic activity, such as compounds
21b, 21f, and 21i, showing comparable potency with IC50
values ranging from 0.015 to 0.073 mM against MKN-45. In
contrast, the activities of compounds with the same R1
groups but different R3 groups were markedly different.
Compound 21l, in which R3 is a 4-fluoro group, exhibited
moderate potency with IC50 values of 1.0 mM agaist MKN-45.
the other hand, when the 4-fluoro group (21l, IC50
¼
1.0 mM) was shifted to 2-position (21a, IC50 ¼ 0.083 mM), the
inhibitory efficacy was increased to 12-fold. From com-
pounds 22a–22g, it was found that compounds with mono-
EWGs (2-fluoro, 2-chloro, and 2-trifluoromethyl group) were
all more active than that with electron-donating groups
(EGDs; 4-methyl).
As shown in Table 2, the three tested compounds exhibited
excellent c-Met inhibitory potency with IC50 values in the
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Arch. Pharm. Chem. Life Sci. 2013, 346, 521–533
4-(2-Fluorophenoxy)quinoline Derivatives
525
Table 1. Cytotoxicity of compounds 21a–l and 22a–g against H460, HT-29, MKN-45, U87MG, and SMMC-7721 cell lines.
Compd.
R1
R2
R3
IC50 (mM)
MKN-45
H460
HT-29
U87MG
SMMC-7721
21a
21b
21c
21d
21e
21f
21g
21h
21i
21j
21k
21l
22a
22b
22c
22d
22e
22f
Piperidinyl
Piperidinyl
Piperidinyl
H
H
H
H
H
H
H
H
H
H
H
H
F
F
F
F
F
F
F
2-F
2-Cl
2-CF3
2,4-(Cl)2
2-F
0.14
0.086
0.075
0.44
0.16
0.11
0.93
0.13
0.12
0.31
0.91
1.4
1.0
3.2
1.6
1.1
0.73
0.80
1.5
0.10
0.054
0.051
0.33
0.10
0.10
0.67
0.077
0.082
0.21
0.83
2.1
0.86
1.7
1.1
0.52
0.67
0.84
1.2
0.083
0.015
0.010
0.25
0.056
0.073
0.58
0.055
0.027
0.11
0.49
1.0
1.0
0.93
0.15
0.20
0.67
1.1
1.0
1.3
1.1
0.24
1.2
1.1
2.6
1.4
ND
0.73
1.1
1.2
0.92
3.0
0.67
0.53
1.2
1.1
1.1
Piperidinyl
4-Methyl piperidinyl
4-Methyl piperidinyl
4-Methyl piperidinyl
Morpholinyl
Morpholinyl
Pyrrolidinyl
Pyrrolidinyl
4-Methyl piperizinyl
Piperidinyl
Piperidinyl
Pyrrolidinyl
4-Methyl piperidinyl
4-Methyl piperidinyl
4-Methyl piperidinyl
Morpholinyl
2-Cl
3,4-(F)2
2-CF3
2-Cl
2-F
2,4-(Cl)2
4-F
2-Cl
4-CH3
2-F
2-Cl
2-CF3
2-F
1.1
0.75
0.71
1.0
1.3
1.5
1.2
2.4
1.5
1.8
0.59
1.4
0.82
0.59
0.86
0.77
1.2
1.7
0.84
1.7
22g
Foretinib
2-F
0.18
0.15
0.030
0.90
0.44
single-digit nM range, suggesting that their mechanism of
action may act via inhibiting c-Met kinase. Compound 21c,
showing the most potent activity with an IC50 value of
1.35 nM, which was comparable to the positive control
foretinib (IC50 ¼ 1.16 nM), deserves further investigation.
The results of flow cytometric analysis shown in Fig. 3A
indicated that compound 21c reduced cellular proliferation
through a G2-M cell-cycle arrest. These data suggested that the
mode of action of compound 21c seemed to be cell cycle arrest,
but not apoptosis induction. In addition, the data of Western
blot assay outlined in Fig. 3B showed that compound 21c
produced constitutive inhibition of c-Met phosphorylation in
MKN-45 cells, with an obvious abolishment at 1.5 mg/mL.
In summary, a series of 4-(2-fluorophenoxy)quinoline
derivatives bearing 4-oxo-1,4-dihydroquinoline-3-carboxa-
mide moiety were synthesized and evaluated for their
biological activity. Through anti-tumor activity screening
the following conclusion can be reached: (a) all the target
compounds showed moderate to excellent anti-tumor activity
and high selectivity for inhibition of HT-29 and MKN-45 cell
lines; (b) the substituents on the phenyl ring (R3) contribute a
lot to the antitumor activity. The introduction of mono-EWGs
at the 2-position of phenyl ring was favorable to cytotoxicity,
while double-EWGs or EDGs lowered the potency; (c) the
introduction of a 7-fluoro group to the 4-oxo-1,4-dihydroqui-
noline-3-carboxamide moiety led to an obvious decline in
activity; (d) the results of c-Met kinase evaluation suggested
that this series of compounds may act through inhibiting
c-Met. Further studies on structural optimization and
biological activities about these derivatives are still underway
in our laboratory.
Experimental
Chemistry
All melting points were obtained on a Büchi Melting Point B-540
apparatus (Büchi Labortechnik AG, Flawil, Switzerland) and are
uncorrected. Mass spectra (MS) were taken in ESI mode on Agilent
1100 LC–MS (Agilent, Palo Alto, CA, USA). Proton (1H) nuclear
magnetic resonance spectroscopy was performed using Bruker
ARX-300, 300 MHz spectrometers (Bruker Bioscience, Billerica,
MA, USA) with TMS as an internal standard. Column chromatog-
raphy was run on silica gel (200–300 mesh) from Qingdao Ocean
Chemicals (Qingdao, Shandong, China). Unless otherwise noted,
all materials were obtained from commercially available sources
and were used without further purification.
Table 2. c-Met kinase activity of selected compounds 21b, 21c,
21i and positive control foretinib.
Compd.
IC50 on c-Met (nM)
21b
21c
21i
Foretinib
2.76
1.35
2.64
1.16
1-(4-(3-Chloropropoxy)-3-methoxyphenyl)ethanone (1)
A stirred solution of 1-(4-hydroxy-3-methoxyphenyl) ethanone
(24.9 g, 0.15 mol) and anhydrous potassium carbonate (57.9 g,
0.21 mol) in acetone was added dropwise with an acetone
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Figure 3. Both apoptosis and senescence were fates of the cells undergoing mitotic catastrophe in MKN-45 cells. (A) SubG0/G1 ratio was
determined by flow cytometry after staining with PI. (B) Effects of compound 21c on c-Met phosphorylation in MKN-45 cells.
solution of 1-bromo-3-chloropropane (66.1 g, 4.2 mol, in 200 mL
acetone) while maintaining the temperature below 25°C. Upon
completion of the addition, the reaction mixture was stirred at
room temperature for a whole night. The solid was removed by
filtration, and the filtrate was poured slowly into 1 L ice water,
which was stirred vigorously. A lot of precipitated white solid was
filtered and dried in vacuo at 40°C for 24 h to afford the target
compound (69.5 g, yield: 92.5%). m.p. 71–73°C; MS m/z (ESI): 264.8
[MþNa]þ.
the cooled reaction mixture was filterd to obtain the target
compound as a yellow powder (31 g, yield 72.4%). m.p. 117–119°
C; MS m/z (ESI): 343.4 [MþH]þ, 355.3 [MþNa]þ.
7-(3-Chloropropoxy)-6-methoxyquinolin-4-ol (4)
(E)-1-(4-(3-Chloropropoxy)-5-methoxy-2-nitrophenyl)-3-(dimethyl-
amino)prop-2-en-1-one 3 (14.5 g, 0.05 mol) was dissolved in glacial
acetic acid (110 mL), and stirred at room temperature. Iron
powder was added slowly to keep the temperature below 60°C.
Upon the completion of addition, the reaction mixture was heated
to 80°C for 2 h, and then filtered immediately to remove iron
powder. The precipitate was filtered and the filter cake was
washed to be colorless with water, and dried at room temperature
to obtain a white solid (10.5 g, yield 78.4%). m.p. 232–233°C;
MS m/z (ESI): 268.1 [MþH]þ, 290.2 [MþNa]þ, 557.3 [2MþNa]þ.
1-(4-(3-Chloropropoxy)-5-methoxy-2-nitrophenyl)-
ethanone (2)
To a solution of 1-(4-(3-chloropropoxy)-3-methoxyphenyl)etha-
none 1 (35 g, 0.14 mol) in dichloromethane (150 mL), fuming
nitric acid (10 mL, 0.28 mol) was added dropwise at a rate to
maintain the reaction temperature below 0°C. Upon the
completion of addition, the mixture was stirred at 0°C for
another 4 h. The reaction mixture was poured slowly into the
mixed solution of cold water (200 mL) and dichloromethane
(100 mL). The organic layer was separated and washed with brine
(600 mL ꢁ 5), dried with anhydrous sodium sulfate and evapo-
rated. The residue was cooled and dried at room temperature
to give a light yellow solid (360 g, yield 86.7%). m.p. 60–61°C;
MS m/z (ESI): 288.0 [MþH]þ, 310.0 [MþNa]þ.
General procedure for the preparation of 4-hydroxy-
quinolines (5a–5e)
To a stirring solution of 7-(3-chloropropoxy)-6-methoxyquinolin-
4-ol (4) (10.15 g, 0.032 mol) in acetonitrile (80 mL) at 25°C was
added the appropriate secondary amine (0.3 mol), and the
resulting reaction mixture was heated to 85°C for 10 h, then
cooled to room temperature and filtered. The light yellow filter
cake was kept, and the filtrate was concentrated in vacuum. Ethyl
acetate (100 mL) was added to the residue and stirred fully. The
turbid fluid was filtered and the cake was combined with the
earlier one and recrystallized with ethanol and acetone (2:5) to
give the corresponding 4-hydroxyquinolines 5a–5e.
(E)-1-(4-(3-Chloropropoxy)-5-methoxy-2-nitrophenyl)-3-
(dimethylamino)prop-2-en-1-one (3)
1-(4-(3-Chloropropoxy)-5-methoxy-2-nitrophenyl)ethanone 2 (36 g,
0.125 mol) was added to toluene (100 mL) and heated to 110°C.
When the mixture became clear, N,N-dimethylformamide
dimethyl acetal (90 mL, 6.25 mol) was poured into the solution
and kept reacting for more 10 h. The resulting precipitate from
6-Methoxy-7-(3-(piperidin-1-yl)propoxy)quinolin-4-ol (5a)
White solid; yield: 85.5%; m.p. 175–176°C; MS m/z (ESI): 317.4
[MþH]þ.
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6-Methoxy-7-(3-(4-methylpiperidin-1-yl)propoxy)quinolin-
General procedure for the preparation of nitro compounds
4-ol (5b)
(7a–7e)
Sallow solid; yield: 93.4%; m.p. 182–183°C; MS m/z (ESI): 331.5
A stirring mixture of an appropriate 4-chloroquinoline (0.2 mol)
and 2-fluoro-4-nitrophenol (0.4 mol) in chlorobenzene (450 mL, 5
v/w) was heated to 140°C for 30 h. The resulting mixture was
cooled and filtered. The filter cake was washed with saturated
K2CO3 solution and recrystallized with ethanol to give the
corresponding nitro compounds 7a–7e.
[MþH]þ.
6-Methoxy-7-(3-(pyrrolidin-1-yl)propoxy)quinolin-4-ol (5c)
Light yellow solid; yield: 85.5%; m.p. 171–173°C; MS m/z (ESI):
303.2 [MþH]þ.
4-(2-Fluoro-4-nitrophenoxy)-6-methoxy-7-(3-(piperidin-1-
6-Methoxy-7-(3-morpholinopropoxy)quinolin-4-ol (5d)
White solid; yield: 97.6%; m.p. 187–188°C; MS m/z (ESI): 319.1
[MþH]þ, 659.3 [2MþNa]þ.
yl)propoxy)quinoline (7a)
Yellowish-white solid; yield: 90.4%; m.p. 137–138°C; MS m/z (ESI):
456.4 [MþH]þ.
6-Methoxy-7-(3-(4-methylpiperazin-1-yl)propoxy)-
4-(2-Fluoro-4-nitrophenoxy)-6-methoxy-7-(3-(4-
quinolin-4-ol (5e)
methylpiperidin-1-yl)propoxy)quinoline (7b)
Yellow solid; yield: 88.7%; m.p. 133–134°C; MS m/z (ESI): 470.6
[MþH]þ.
Sallow solid; yield: 91.2%; m.p. 193–195°C; MS m/z (ESI): 332.4
[MþH]þ.
General procedure for the preparation of 4-chloro-
quinolines (6a–6e)
4-(2-Fluoro-4-nitrophenoxy)-6-methoxy-7-(3-(pyrrolidin-1-
yl)propoxy)quinoline (7c)
To a solution of an appropriate 4-hydroxyquinoline (0.05 mol) in
acetonitrile (80 mL) at 85°C was dropped POCl3 (80 mL), and the
resulting mixture was kept at this temperature for 6 h. The solvent
was concentrated in vacuum and the residue was poured into
stirring ice-water (200 mL), basified with potassium hydroxide to
pH 12, and extracted with CH2Cl2 (3 ꢁ 500 mL). The combined
extracts were washed with brine, dried over anhydrous Na2SO4,
and concentrated in vacuum to give the corresponding 4-
chloroquinolines 6a–6e.
Yellow solid; yield: 86.3%; m.p. 140–141°C; MS m/z (ESI): 442.2
[MþH]þ.
4-(3-(4-(2-Fluoro-4-nitrophenoxy)-6-methoxyquinolin-
7-yloxy)propyl)morpholine (7d)
Light yellow solid; yield: 82.5%; m.p. 135–136°C; MS m/z (ESI):
458.4 [Mþ1]þ, 480.4 [Mþ23]þ.
4-(2-Fluoro-4-nitrophenoxy)-6-methoxy-7-(3-(4-
4-Chloro-6-methoxy-7-(3-(piperidin-1-yl)propoxy)-
methylpiperazin-1-yl)propoxy)quinoline (7e)
Yellow solid; yield: 85.0%; m.p. 152–153°C; MS m/z (ESI): 471.4
[MþH]þ.
quinoline (6a)
White solid; yield: 85.4%; m.p. 187–188°C; MS m/z (ESI): 335.3
[MþH]þ.
General procedure for the preparation of anilines (8a–8e)
To a refluxing solution of an appropriate nitro compound
(0.1 mol) in EtOH (450 mL, 10 v/w) was added saturated NH4Cl
solution (450 mL) and Fe powder in batches. The mixture was
kept at this temperature for more 5 h. After completion of the
reaction as indicated by TLC, the mixture was filtered immedi-
ately, and the filtrate was cooled and filtered to obtain the
corresponding anilines 8a–8e.
4-Chloro-6-methoxy-7-(3-(4-methylpiperidin-1-yl)propoxy)-
quinoline (6b)
White solid; yield: 79.7%; m.p. 179–180°C; MS m/z (ESI): 349.4
[MþH]þ.
4-Chloro-6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)-
quinoline (6c)
Yellow solid; yield: 78.2%; m.p. 177–178°C; MS m/z (ESI): 321.2
3-Fluoro-4-(6-methoxy-7-(3-(piperdine-1-yl)propoxy)-
[MþH]þ.
quinolin-4-yloxy)aniline (8a)
Gray solid; yield: 85.5%; m.p. 196–197°C; IR (KBr) cmꢀ1: 3482.2,
3387.0, 2946.5, 2835.1, 2788.1, 1621.1, 1587.8, 1512.8, 1483.4,
4-Chloro-6-methoxy-7-(3-morpholinopropoxy)quinoline
1252.9, 1215.4, 853.5; 1H NMR (300 MHz, CDCl3)
d 1.45
(6d)
White solid; yield: 82.4%; m.p. 174–175°C; 1H NMR (300 MHz,
CDCl3) d 2.12 (m, 2H), 2.48 (m, 4H), 2.57 (t, J ¼ 7.1 Hz, 2H), 3.72
(m, 4H), 4.04 (s, 3H), 4.27 (t, J ¼ 6.7 Hz, 2H), 7.34 (d, J ¼ 4.8 Hz,
1H), 7.39 (s, 1H), 7.44 (s, 1H), 8.57 (d, J ¼ 4.8 Hz, 1H); MS m/z (ESI):
337.4 [MþH]þ, 359.4 [MþNa]þ.
(d, J ¼ 5.2 Hz, 2H), 1.60 (m, 4H), 2.14 (m, 2H), 2.43 (s, 4H), 2.54
(m, 2H), 3.81 (s, 2H), 4.04 (d, J ¼ 3.8 Hz, 3H), 4.24 (t, J ¼ 6.8 Hz,
2H), 6.39 (dd, J ¼ 5.3, 1.1 Hz, 1H), 6.50 (m, 1H), 6.56 (dd, J ¼ 11.8,
2.6 Hz, 1H), 7.03 (t, J ¼ 8.7 Hz, 1H), 7.41 (s, 1H), 7.58 (s, 1H),
8.46 (d, J ¼ 5.3 Hz, 1H); MS m/z (ESI): 426.3 [MþH]þ.
4-Chloro-6-methoxy-7-(3-(4-methylpiperazin-1-yl)-
3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperdine-1-yl)-
propoxy)quinoline (6e)
propoxy)quinolin-4-yloxy)aniline (8b)
White solid; yield: 75.6%; m.p. 195–196°C; MS m/z (ESI): 350.3
White solid; yield: 77.4%; m.p. 193–194°C; 1H NMR (300 MHz,
CDCl3) d 0.93 (d, J ¼ 6.0 Hz, 3H), 1.28 (m, 3H), 1.63 (d, J ¼ 10.4 Hz,
[MþH]þ.
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Arch. Pharm. Chem. Life Sci. 2013, 346, 521–533
2H), 1.98 (t, J ¼ 10.9 Hz, 2H), 2.15 (m, 2H), 2.57 (m, 2H),
2.94 (d, J ¼ 11.5 Hz, 2H), 3.82 (s, 2H), 4.03 (s, 3H), 4.25
(t, J ¼ 6.7 Hz, 2H), 6.39 (dd, J ¼ 5.3, 0.8 Hz, 1H), 6.50
(dd, J ¼ 9.0, 2.9 Hz, 1H), 6.56 (dd, J ¼ 11.8, 2.6 Hz, 1H), 7.03
(t, J ¼ 8.7 Hz, 1H), 7.41 (s, 1H), 7.58 (s, 1H), 8.46 (d, J ¼ 5.3 Hz, 1H);
MS m/z (ESI): 440.3 [MþH]þ.
General procedure for the preparation of esters (11a–b)
A mixture of an appropriate dione (10a or 10b) (0.3 mol) and EtOH
(500 mL) was heated to 80°C refluxing for 5 h. Upon cooling to
room temperature, the solvent was evaporated to give the esters
11a or 11b as red oil.
Ethyl 3-(2-fluorophenyl)-3-oxopropanoate (11a)
Red oil; yield: 85%; MS m/z (ESI): 233.1 [MþNa]þ, 443.1 [2MþNa]þ.
3-Fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)-
quinolin-4-yloxy)aniline (8c)
Light yellow solid; yield: 72.3%; m.p. 208–209°C; 1H NMR
(300 MHz, DMSO-d6) d 1.96 (d, J ¼ 14.0 Hz, 4H), 2.28 (s, 2H),
3.04 (s, 2H), 3.24 (s, 4H), 3.59 (s, 2H), 3.97 (s, 3H), 4.28 (t, J ¼ 5.7 Hz,
2H), 6.46 (m, 2H), 6.57 (d, J ¼ 14.1 Hz, 1H), 7.08 (t, J ¼ 9.0 Hz, 1H),
7.44 (s, 1H), 7.55 (s, 1H), 8.49 (d, J ¼ 5.2 Hz, 1H); MS m/z (ESI): 412.5
[MþH]þ.
Ethyl 3-(2,4-difluorophenyl)-3-oxopropanoate (11b)
Red oil; yield: 90%; MS m/z (ESI): 251.1 [MþNa]þ.
General procedure for the preparation of compounds
12a–12b
A mixture of an appropriate intermediate 11a or 11b (0.04 mol),
and DMF-DMA (0.32 mol), in CH2Cl2 (100 mL) was heated to 40°C
for 4 h and monitored by TLC. The reaction mixture was
concentrated under reduced pressure and hexane (80 mL) was
added to the residue with vigorous stirring yielding a precipitate
in an ice bath. The mixture was then filtered to get the yellow
compounds 12a and 12b.
3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)-
quinolin-4-yloxy)aniline (8d)
White solid; yield: 81.8%; m.p. 217–218°C; 1H NMR (300 MHz,
CDCl3) d 2.13 (m, 2H), 2.51 (d, J ¼ 4.2 Hz, 4H), 2.60 (t, J ¼ 7.1 Hz,
2H), 3.74 (m, 4H), 3.82 (s, 2H), 4.04 (s, 3H), 4.27 (t, J ¼ 6.6 Hz, 2H),
6.41 (d, J ¼ 5.3 Hz, 1H), 6.50 (m, 1H), 6.57 (dd, J ¼ 11.9, 2.6 Hz,
1H), 7.04 (t, J ¼ 8.7 Hz, 1H), 7.44 (s, 1H), 7.58 (s, 1H), 8.47
(d, J ¼ 5.2 Hz, 1H); MS m/z (ESI): 428.2 [MþH]þ, 450.1 [MþNa]þ.
(Z)-Ethyl-3-(dimethylamino)-2-(2-fluorobenzoyl)-
acrylate (12a)
Light yellow solid; yield: 72%; m.p. 112–113°C; MS m/z (ESI): 266.1
3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperazin-1-yl)-
[MþH]þ, 288.1 [MþNa]þ, 553.1 [2MþNa]þ.
propoxy)quinolin-4-yl-oxy)aniline (8e)
Yellow white solid (71.3%); m.p. 223–224°C; 1H NMR (300 MHz,
CDCl3) d 1.88 (s, 2H), 2.16 (m, 2H), 2.33 (s, 3H), 2.59 (m, 8H), 3.84
(s, 2H), 4.06 (s, 3H), 4.28 (t, J ¼ 6.7 Hz, 2H), 6.42 (d, J ¼ 5.3 Hz, 1H),
6.53 (dd, J ¼ 8.7, 2.6 Hz, 1H), 6.59 (dd, J ¼ 11.8, 2.6 Hz, 1H), 7.06
(t, J ¼ 8.7 Hz, 1H), 7.44 (s, 1H), 7.61 (s, 1H), 8.49 (d, J ¼ 5.3 Hz, 1H);
MS m/z (ESI): 441.6 [MþH]þ.
(Z)-Ethyl 2-(2,4-difluorobenzoyl)-3-(dimethylamino)
acrylate (12b)
Light yellow solid; yield: 70%; m.p. 116–117°C; MS m/z (ESI): 284.1
[MþH]þ, 306.1 [MþNa]þ, 589.1 [2MþNa]þ.
General procedure for preparation of the intermediates
13a–13f and 14a–14d
General procedure for the preparation of diones 10a–10b
2-Fluorobenzoic acid and 2,4-difluorobenzoic acid (0.7 mol) were
added to the mixed solvent of toluene (200 mL) and SOCl2
(400 mL) and heated to 80°C for 8 h, respectively. The solvent was
evaporated in vacuum and the residue (9a and 9b) was used in the
next reaction step directly.
A mixture of 12a or 12b (0.05 mol), a substituted phenyl amine
(0.06 mol) and catalytic amount HCl (1 mL) in EtOH (50 mL) was
heated at reflux for 5 h and the reaction was monitored by TLC.
Part of solvent was evaporated and the residue was filtered to
yield white solids 13a–13f and 14a–14d.
To a mixture of 2,2-dimethyl-1,3-dioxane-4,6-dione (82.6 g,
0.574 mol), DMAP (87.5 g, 0.717 mol) and CH2Cl2 (600 mL), the
appropriate benzoyl chloride (9a or 9b) in CH2Cl2 was added drop-
wise at 0°C. Upon the completion of addition, the reaction
mixture was kept stirring at this temperature for more 6 h, and
then poured into HCl (1 mol/L). The organic phase was separated,
washed with brine, dried (MgSO4), filtered and concentrated to
give substituted diones 10a–10b.
General procedure for preparation of the intermediates
15a–15f and 16a–16d
An appropriate compound (13a–13f or 14a–14d) (0.03 mol) and
anhydrous K2CO3 (4.9 g, 0.036 mol) were added to DMF (30 mL)
and the mixture was heated to 50°C for 8 h. Upon the TLC
showing that the starting material disappeared, the mixture was
poured into ice/water (150 mL) with vigorous stirring for 30 min.
The precipitate was filtered, washed with water, and dried to
obtain white solids 15a–15f and 16a–16d. Yielded 90–95%.
5-(2-Fluorobenzoyl)-2,2-dimethyl-1,3-dioxane-4,6-
dione (10a)
Light yellow solid; yield: 50%; m.p. 232–233°C; MS m/z (ESI): 264.9
General procedure for the preparation of acids 17a–17f
and 18a–18d
[MꢀH]ꢀ.
To a solution of an appropriate ester (15a–15f or 16a–16d)
(0.025 mol) dissolved in EtOH (80 mL), was added drop-wise 10%
NaOH (50 mL) at room temperature. After the completion of
addition, the mixture was kept at this temperature for further
5 h, the organic solvent was evaporated, the residue was poured
5-(2,4-Difluorobenzoyl)-2,2-dimethyl-1,3-dioxane-4,6-
dione (10b)
White solid; yield: 53%; m.p. 236–238°C; MS m/z (ESI): 283.1
[MꢀH]ꢀ.
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Arch. Pharm. Chem. Life Sci. 2013, 346, 521–533
4-(2-Fluorophenoxy)quinoline Derivatives
529
into ice/water (150 mL) and acidified (HCl, 6 N) to pH 5. The
precipitate was collected by filtration and dried to give
substituted acids 17a–17f and 18a–18d.
7-Fluoro-4-oxo-1-p-tolyl-1,4-dihydroquinoline-3-carboxylic
acid (18d)
White solid; yield: 65%; m.p. 244–245°C; MS m/z (ESI): 320.4
[MþNa]þ, 617.8 [2MþNa]þ.
1-(2-Fluorophenyl)-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid (17a)
General procedure for preparation of the target compounds
White solid; yield: 73%; m.p. 268–269°C; 1H NMR (300 MHz, DMSO-
d6) d 7.30 (d, J ¼ 8.7 Hz, 1H), 7.72 (d, J ¼ 8.8 Hz, 6H), 7.90
(t, J ¼ 7.3 Hz, 1H), 8.34 (d, J ¼ 7.9 Hz, 1H), 14.18 (s, 1H); MS m/z
(ESI): 284.0 [MþH]þ, 306.1 [MþNa]þ, 589.0 [2MþNa]þ, 281.9 [MꢀH]ꢀ.
21a–21l and 22a–22g
A
mixture of an appropriate acid (17a–17f or 18a–18d)
(0.58 mmol), toluene (5 mL), and SOCl2 (10 mL) was heated to
80°C for 5 h. Upon cooling to room temperature, the solvent was
evaporated in vacuum and the residue was dissolved in dried
CH2Cl2 (10 mL) to add dropwise to a mixture of an appropriate
aniline (8a–8e) (0.48 mmol), Et3N (0.58 mol), and CH2Cl2 (10 mL)
in an ice bath, which was removed to room temperature for 10 h
and monitored by TLC. The mixture was washed with 10% K2CO3
(20 mL ꢁ 3), brine (20 mL ꢁ 3) in sequence, and the organic
phase was separated, dried, and evaporated. The crude product
was purified by chromatography on silica gel using CH2Cl2/MeOH
(40:1) to afford the solids 21a–21l and 22a–22g.
1-(2-Chlorophenyl)-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid (17b)
White solid; yield: 75%; m.p. 275–277°C; 1H NMR (300 MHz,
DMSO-d6) d 7.16 (d, J ¼ 8.5 Hz, 1H), 7.56 (t, J ¼ 8.7 Hz, 2H), 7.68
(t, J ¼ 7.5 Hz, 1H), 7.83 (m, 3H), 8.44 (d, J ¼ 8.0 Hz, 1H), 8.76
(s, 1H), 15.03 (s, 1H); MS m/z (ESI): 302.2 [MþH]þ, 324.3 [MþNa]þ,
625.3 [2MþNa]þ.
1-(2-(Trifluoromethyl)phenyl)-4-oxo-1,4-dihydroquinoline-
3-carboxylic acid (17c)
N-(3-Fluoro-4-(6-methoxy-7-(3-(piperidin-1-yl)propoxy)-
quinolin-4-yloxy)phenyl)-1-(2-fluorophenyl)-4-oxo-1,4-
dihydroquinoline-3-carboxamide (21a)
White solid; yield: 64%; m.p. 284–286°C; MS m/z (ESI): 334.4
[MþH]þ, 356.4 [MþNa]þ, 689.6 [2MþNa]þ.
1
Yield: 61.3%; m.p. 148–150°C; H NMR (300 MHz, DMSO-d6) d 1.49
1-(4-Fluorophenyl)-4-oxo-1,4-dihydroquinoline-3-
(s, 2H), 1.68 (s, 4H), 2.16 (s, 2H), 2.87 (s, 6H), 3.97 (s, 3H), 4.24 (s, 2H),
6.52 (d, J ¼ 5.1 Hz, 1H), 7.11 (d, J ¼ 8.6 Hz, 1H), 7.45 (d, J ¼ 6.2 Hz,
1H), 7.50 (d, J ¼ 8.7 Hz, 1H), 7.63 (m, 5H), 7.85 (m, 3H), 8.09
(d, J ¼ 12.7 Hz, 1H), 8.50 (d, J ¼ 5.1 Hz, 2H), 8.86 (s, 1H), 12.47
(s, 1H); MS m/z (ESI): 691.3 [MþH]þ; Anal. calcd. for C40H36F2N4O5 (%):
C, 69.55; H, 5.25; N, 8.11. Found (%): C, 69.67; H, 5.24; N, 8.10.
carboxylic acid (17d)
White solid; yield: 74%; m.p. 264–266°C; MS m/z (ESI): 284.1
[MþH]þ, 589.1 [2MþNa]þ.
1-(2,4-Dichlorophenyl)-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid (17e)
1-(2-Chlorophenyl)-N-(3-fluoro-4-(6-methoxy-7-(3-
(piperidin-1-yl)propoxy)quinolin-4-yloxy)phenyl)-4-oxo-
1,4-dihydroquinoline-3-carboxamide (21b)
White solid; yield: 61%; m.p. 292–293°C; MS m/z (ESI): 334.2
[MþH]þ, 356.2 [MþNa]þ, 691.2 [2MþNa]þ.
1-(3,4-Difluorophenyl)-4-oxo-1,4-dihydroquinoline-3-
Yield: 65.0%; m.p. 172–174°C; 1H NMR (300 MHz, DMSO-d6) d 1.56
(s, 2H), 1.79 (s, 4H), 2.29 (s, 2H), 3.17 (s, 6H), 3.98 (s, 3H), 4.28 (s, 2H),
6.53 (d, J ¼ 4.7 Hz, 1H), 6.97 (d, J ¼ 8.2 Hz, 1H), 7.47 (s, 1H), 7.52
(d, J ¼ 8.9 Hz, 1H), 7.58 (s, 2H), 7.70 (m, 2H), 7.75 (s, 1H), 7.81
(d, J ¼ 6.5 Hz, 1H), 7.93 (m, 2H), 8.10 (d, J ¼ 10.8 Hz, 1H), 8.51
(d, J ¼ 8.0 Hz, 2H), 8.81 (s, 1H), 12.49 (s, 1H); 13C NMR (CDCl3)
d 22.78, 24.01 (2C), 24.64, 53.29 (2C), 54.58, 56.27, 66.76, 99.54,
102.64, 109.15, 111.63, 115.07, 117.14, 117.71, 117.83, 118.27,
124.78, 126.58, 126.65, 126.80, 127.68, 130.32, 133.32, 134.61,
136.41, 137.81, 140.57, 146.74, 149.19, 149.41, 149.93, 152.11,
153.22, 154.85, 156.53, 158.19, 159.74, 162.78; MS m/z (ESI): 707.7
[MþH]þ; Anal. calcd. for C40H36ClFN4O5 (%): C, 67.93; H, 5.13;
N, 7.92. Found (%): C, 67.95; H, 5.22; N, 8.13.
carboxylic acid (17f)
White solid; yield: 78%; m.p. 283–284°C; MS m/z (ESI): 302.1 [MþH]þ.
7-Fluoro-1-(2-fluorophenyl)-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid (18a)
White solid; yield: 57%; m.p. 262–263°C; 1H NMR (300 MHz,
DMSO-d6) d 6.51 (d, J ¼ 10.1 Hz, 1H), 7.31 (t, J ¼ 7.9 Hz, 1H),
7.69 m, 2H), 7.82 (t, J ¼ 6.9 Hz, 2H), 8.37 (m, 1H), 8.49 (s, 1H); MS
m/z (ESI): 307.0 [MþNa]þ, 313.0 [MþK]þ.
1-(2-Chlorophenyl)-7-fluoro-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid (18b)
White solid; yield: 63%; m.p. 266–267°C; 1H NMR (300 MHz,
DMSO-d6) d 7.05 (d, J ¼ 8.6 Hz, 1H), 7.69 (t, J ¼ 7.6 Hz, 1H), 7.87
(m, 1H), 7.98 (m, 2H), 8.07 (m, 1H), 8.15 (d, J ¼ 7.4 Hz, 1H), 8.32
(m, 1H), 13.87 (s, 1H); MS m/z (ESI): 318.4 [MþH]þ, 340.3 [MþNa]þ,
657.5 [2MþNa]þ, 315.8 [MꢀH]ꢀ.
N-(3-Fluoro-4-(6-methoxy-7-(3-(piperidin-1-yl)propoxy)-
quinolin-4-yloxy)phenyl)-1-(2-(trifluoromethyl)phenyl)-4-
oxo-1,4-dihydroquinoline-3-carboxamide (21c)
Yield: 67.0%; m.p. 198–199°C; 1H NMR (300 MHz, DMSO-d6) d 1.58 (s,
2H), 1.88 (s, 4H), 2.39 (s, 2H), 3.02 (m, 6H), 4.03 (s, 3H), 4.27 (s, 2H),
6.54 (d, J ¼ 5.1 Hz, 1H), 7.12 (d, J ¼ 8.6 Hz, 1H), 7.48 (d, J ¼ 6.2 Hz,
1H), 7.51 (d, J ¼ 8.7 Hz, 1H), 7.66 (m, 5H), 7.87 (m, 3H), 8.11
(d, J ¼ 12.7 Hz, 1H), 8.52 (d, J ¼ 5.1 Hz, 2H), 8.88 (s, 1H), 12.50 (s, 1H);
13C NMR (CDCl3) d 22.91, 23.89 (2C), 24.64, 53.81 (2C), 55.34, 56.05,
66.55, 99.71, 102.36, 108.83, 109.81, 109.97, 115.67, 116.62, 117.46,
7-Fluoro-4-oxo-1-(2-(trifluoromethyl)phenyl)-1,4-
dihydroquinoline-3-carboxylic acid (18c)
White solid; yield: 58%; m.p. 273–274°C; MS m/z (ESI): 352.3
[MþH]þ, 374.2 [MþNa]þ, 725.3 [2MþNa]þ.
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121.62, 123.65, 125.60, 125.94, 127.33, 128.23, 130.05, 131.25,
133.90, 134.76, 136.80, 137.02, 137.18, 138.49, 142.36, 146.56,
148.77, 149.63, 151.78, 153.47, 155.12, 159.51, 160.07, 170.91;
MS m/z (ESI): 742.5 [MþH]þ; Anal. calcd. for C41H36F4N4O5 (%):
C, 66.48; H, 4.90; N, 7.56. Found (%): C, 66.51; H, 4.55; N, 7.55.
6.52 (d, J ¼ 4.8 Hz, 1H), 6.92 (d, J ¼ 8.4 Hz, 1H), 7.44 (s, 1H),
7.51 (m, 1H), 7.63 (m, 3H), 7.80 (t, J ¼ 7.7 Hz, 1H), 8.01
(d, J ¼ 7.8 Hz, 2H), 8.12 (m, 3H), 8.50 (d, J ¼ 7.2 Hz, 2H), 8.88
(s, 1H), 12.48 (s, 1H); MS m/z (ESI): 743.1 [MþH]þ; Anal. calcd. for
C
40H34F4N4O5 (%): C, 64.69; H, 4.61; N, 7.54. Found (%): C, 64.72;
H, 4.59; N, 7.54.
1-(2,4-Dichlorophenyl)-N-(3-fluoro-4(6-methoxy-7-(3-
(piperidin-1-yl)propoxy)quinolin-4-yloxy)phenyl)-4-oxo-
1-(2-Chlorophenyl)-N-(3-fluoro-4-(6-methoxy-7-(3-
1,4-dihydroquinoline-3-carboxamide (21d)
morpholinopropoxy)quinolin-4-yloxy)phenyl)-4-oxo-1,4-
dihydroquinoline-3-carboxamide (21i)
Yield: 61.5%; m.p. 185–187°C; 1H NMR (300 MHz, DMSO-d6) d 1.79
(s, 6H), 2.29 (s, 2H), 3.22 (s, 6H), 3.97 (s, 3H), 4.27 (s, 2H), 6.53
(d, J ¼ 5.0 Hz, 1H), 7.05 (d, J ¼ 8.5 Hz, 1H), 7.49 (m, 2H), 7.63
(m, 3H), 7.80 (m, 2H), 7.97 (d, J ¼ 8.6 Hz, 1H), 8.09 (m, 2H), 8.49
(t, J ¼ 6.7 Hz, 2H), 8.86 (s, 1H), 12.46 (s, 1H); MS m/z (ESI): 741.6
[MþH]þ; Anal. calcd. for C40H35Cl2FN4O5 (%): C, 64.75; H, 4.76;
N, 7.55. Found (%): C, 64.77; H, 4.73; N, 8.11.
Yield: 61.5%; m.p. 204–205°C; 1H NMR (300 MHz, DMSO-d6)
d 2.01 (s, 2H), 2.49 (s, 6H), 3.61 (s, 4H), 3.94 (s, 3H), 4.20 (s, 2H),
6.48 (d, J ¼ 4.8 Hz, 1H), 6.88 (d, J ¼ 8.4 Hz, 1H), 7.40 (s, 1H),
7.46 (m, 1H), 7.54 (t, J ¼ 7.2 Hz, 2H), 7.61 (m, 1H), 7.76
(t, J ¼ 7.7 Hz, 1H), 7.95 (s, 1H), 7.97 (s, 1H), 8.03 (d, J ¼ 6.9 Hz,
1H), 8.11 (m, 2H), 8.45 (d, J ¼ 7.2 Hz, 2H), 8.84 (s, 1H), 12.44
(s, 1H); 13C NMR (CDCl3) d 26.14, 53.83 (2C), 55.26, 56.23,
66.67 (2C), 67.13, 99.48, 102.55, 108.98, 109.12, 109.27, 114.93,
117.19, 117.19, 118.17, 124.74, 125.26, 125.51, 127.65, 128.73,
131.00, 132.22, 135.41, 135.70, 136.71, 137.59, 138.63, 139.49,
142.56, 146.85, 149.28, 150.04, 152.40, 153.15, 154.78, 159.65,
160.28, 169.82; MS m/z (ESI): 710.7 [MþH]þ; Anal. calcd. for
C39H34ClFN4O5 (%): C, 66.05; H, 4.83; N, 7.90. Found (%): C, 66.03;
H, 4.86; N, 7.92.
N-(3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperidin-1-yl)-
propoxy)quinolin-4-yloxy)phenyl)-1-(2-fluorophenyl)-4-
oxo-1,4-dihydroquinoline-3-carboxamide (21e)
Yield: 60.8%; m.p. 163–164°C; 1H NMR (300 MHz, DMSO-d6)
d 12.49 (s, 1H), 8.88 (s, 1H), 8.52 (d, J ¼ 5.2 Hz, 2H), 8.12
(d, J ¼ 12.9 Hz, 1H), 7.93 (t, J ¼ 7.4 Hz, 1H), 7.83 (m, 2H), 7.69
(t, J ¼ 7.9 Hz, 2H), 7.61 (m, 3H), 7.51 (t, J ¼ 8.9 Hz, 1H), 7.45
(s, 1H), 7.14 (d, J ¼ 8.6 Hz, 1H), 6.54 (d, J ¼ 5.0 Hz, 1H), 4.25 (s, 2H),
3.99 (s, 3H), 3.20 (s, 4H), 2.87 (s, 2H), 2.16 (s, 2H), 1.72 (d,
J ¼ 10.4 Hz, 2H), 1.52 (s, 1H), 1.35 (s, 2H), 0.94 (d, J ¼ 6.3 Hz, 3H);
MS m/z (ESI): 705.2 [MþH]þ; Anal. calcd. for C41H38F2N4O5 (%): C,
69.87; H, 5.43; N, 7.95. Found (%): C, 69.88; H, 5.41; N, 7.95.
N-(3-Fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)-
quinolin-4-yloxy)phenyl)-1-(2-fluorophenyl)-4-oxo-1,4-
dihydroquinoline-3-carboxamide (21j)
Yield: 66.2%; m.p. 171–173°C; 1H NMR (300 MHz, DMSO-d6)
d 1.91 (s, 4H), 2.22 (s, 2H), 3.14 (s, 6H), 3.98 (s, 3H), 4.28 (s, 2H),
6.53 (d, J ¼ 5.1 Hz, 1H), 7.12 (d, J ¼ 8.3 Hz, 1H), 7.45 (s, 1H),
7.51 (d, J ¼ 8.9 Hz, 1H), 7.64 (m, 5H), 7.86 (m, 3H), 8.10
(d, J ¼ 11.5 Hz, 1H), 8.51 (d, J ¼ 4.9 Hz, 2H), 8.87 (s, 1H),
12.47 (s, 1H); MS m/z (ESI): 677.6 [MþH]þ; Anal. calcd. for
C39H34F2N4O5 (%): C, 69.22; H, 5.06; N, 8.28. Found (%): C, 69.25;
H, 5.04; N, 8.27.
1-(2-Chlorophenyl)-N-(3-fluoro-4-(6-methoxy-7-(3-(4-
methylpiperidin-1-yl)propoxy)quinolin-4-yloxy)phenyl)- 4-
oxo-1,4-dihydroquinoline-3-carboxamide (21f)
Yield: 64.2%; m.p. 177–179°C; 1H NMR (300 MHz, DMSO-d6) d 0.94
(d, J ¼ 5.0 Hz, 3H), 1.59 (s, 3H), 1.79 (d, J ¼ 11.9 Hz, 2H), 2.39 (s, 2H),
2.94 (d, J ¼ 9.7 Hz, 2H), 3.23 (s, 4H), 4.06 (s, 3H), 4.36 (s, 2H), 6.95
(s, 1H), 7.01 (d, J ¼ 6.4 Hz, 1H), 7.64 (s, 1H), 7.67 (d, J ¼ 5.4 Hz, 2H),
7.77 (m, 5H), 7.92 (m, 2H), 8.17 (d, J ¼ 13.1 Hz, 1H), 8.50
(d, J ¼ 8.0 Hz, 1H), 8.83 (d, J ¼ 9.9 Hz, 2H), 12.55 (s, 1H); MS m/z
(ESI): 721.8 [MþH]þ; Anal. calcd. for C41H38ClFN4O5 (%): C, 68.28;
H, 5.31; N, 7.77. Found (%): C, 68.30; H, 5.31; N, 7.74.
1-(2,4-Dichlorophenyl)-N-(3-fluoro-4-(6-methoxy-7-(3-
(pyrrolidin-1-yl)propoxy)quinolin-4-yloxy)phenyl)-4-oxo-
1,4-dihydroquinoline-3-carboxamide (21k)
Yield: 62.7%; m.p. 197–199°C; 1H NMR (300 MHz, DMSO-d6)
d 1.94 (s, 4H), 2.25 (s, 2H), 3.18 (m, 6H), 3.97 (s, 3H), 4.28 (s, 2H),
6.52 (d, J ¼ 4.9 Hz, 1H), 7.05 (d, J ¼ 8.3 Hz, 1H), 7.48 (m, 2H),
7.63 (m, 3H), 7.81 (d, J ¼ 7.9 Hz, 2H), 7.97 (d, J ¼ 8.3 Hz, 1H),
8.08 (d, J ¼ 11.5 Hz, 2H), 8.49 (t, J ¼ 6.1 Hz, 2H), 8.86 (s, 1H),
12.46 (s, 1H); MS m/z (ESI): 727.5 [MþH]þ; Anal. calcd. for
C39H33Cl2FN4O5 (%): C, 64.38; H, 4.57; N, 7.70. Found (%): C, 64.41;
H, 4.56; N, 7.72.
N-(3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperidin-1-yl)-
propoxy)quinolin-4-yloxy)phenyl)-1-(3,4-difluorophenyl)-4-
oxo-1,4-dihydroquinoline-3-carboxamide (21g)
Yield: 67.3%; m.p. 164–166°C; 1H NMR (300 MHz, DMSO-d6) d 0.93
(d, J ¼ 5.6 Hz, 3H), 1.41 (s, 2H), 1.53 (s, 1H), 1.73 (d, J ¼ 11.6 Hz,
2H), 2.20 (s, 2H), 2.56 (s, 2H), 3.03 (m, 4H), 3.98 (s, 3H), 4.25 (s, 2H),
6.65 (s, 1H), 7.27 (d, J ¼ 15.5 Hz, 2H), 7.44 (s, 3H), 7.58 (s, 3H), 7.77
(m, 3H), 8.01 (s, 1H), 8.48 (d, J ¼ 14.1 Hz, 1H), 8.81 (d, J ¼ 9.8 Hz,
1H); MS m/z (ESI): 723.2 [MþH]þ; Anal. calcd. for C41H37F3N4O5 (%):
C, 68.13; H, 5.16; N, 7.75. Found (%): C, 68.15; H, 5.15; N, 7.72.
N-(3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperazin-1-yl)-
propoxy)quinolin-4-yloxy)phenyl)-1-(4-fluorophenyl)-4-
oxo-1,4-dihydroquinoline-3-carboxamide (21l)
Yield: 60.5%; m.p. 195–197°C; 1H NMR (300 MHz, DMSO-d6) d 2.03
(s, 2H), 2.55 (s, 3H), 2.80 (m, 10H), 3.96 (s, 3H), 4.21 (s, 2H),
6.64 (d, J ¼ 4.2 Hz, 1H), 7.22 (s, 2H), 7.47 (m, 7H), 7.76 (m, 4H),
8.52 (d, J ¼ 5.2 Hz, 1H), 8.76 (s, 1H); MS m/z (ESI): 706.3 [MþH]þ;
Anal. calcd. for C40H37F2N5O5 (%): C, 68.07; H, 5.28; N, 9.92.
Found (%): C, 68.10; H, 5.27; N, 9.95.
N-(3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)-
quinolin-4-yloxy)phenyl)-1-(2-(trifluoromethyl)phenyl)-4-
oxo-1,4-dihydroquinoline-3-carboxamide (21h)
Yield: 61.5%; m.p. 201–202°C; 1H NMR (300 MHz, DMSO-d6)
d 2.05 (s, 2H), 2.53 (s, 6H), 3.65 (s, 4H), 3.99 (s, 3H), 4.24 (s, 2H),
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
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Arch. Pharm. Chem. Life Sci. 2013, 346, 521–533
4-(2-Fluorophenoxy)quinoline Derivatives
531
1-(2-Chlorophenyl)-7-fluoro-N-(3-fluoro-4-(6-methoxy-7-
(3-piperidin-1-yl)propoxy)quinolin-4-yloxy)phenyl)-4-oxo-
773.2 [MþH]þ; Anal. calcd. for C42H37F5N4O5 (%): C, 65.28; H, 4.83;
N, 7.25. Found (%): C, 65.31; H, 4.81; N, 7.24.
1,4-dihydroquinoline-3-carboxamide (22a)
Yield: 62.4%; m.p. 212–213°C; 1H NMR (300 MHz, DMSO-d6) d 1.59
(m, 2H), 1.79 (s, 4H), 2.29 (s, 2H), 3.25 (m, 6H), 3.97 (s, 3H),
4.27 (s, 2H), 6.52 (d, J ¼ 5.6 Hz, 1H), 6.72 (d, J ¼ 9.0 Hz, 1H),
7.46 (s, 1H), 7.49 (s, 1H), 7.56 (m, 3H), 7.74 (m, 2H), 7.91 (m, 2H),
8.08 (d, J ¼ 12.3 Hz, 1H), 8.52 (m, 2H), 8.80 (s, 1H), 12.32
(d, J ¼ 16.8 Hz, 1H); MS m/z (ESI): 725.1 [MþH]þ; Anal. calcd. for
C40H35ClF2N4O5 (%): C, 66.35; H, 4.86; N, 7.73. Found (%): C, 66.37;
H, 4.85; N, 7.74.
7-Fluoro-N-(3-fluoro-4-(6-methoxy-7-(3-(4-
methylpiperidin-1-yl)propoxy)quinolin-4-yloxy)phenyl)-1-
(2-fluorophenyl)-4-oxo-1,4-dihydroquinoline-3-
carboxamide (22f)
Yield: 64.5%; m.p. 193–195°C; 1H NMR (300 MHz, DMSO-d6) d 0.92
(d, J ¼ 5.9 Hz, 3H), 1.43 (m, 3H), 1.72 (d, J ¼ 11.6 Hz, 2H), 2.19
(s, 2H), 2.51 (s, 2H), 2.93 (s, 2H), 3.45 (s, 2H), 3.97 (s, 3H), 4.24 (s, 2H),
6.51 (d, J ¼ 5.0 Hz, 1H), 7.24 (d, J ¼ 8.5 Hz, 1H), 7.43 (s, 1H), 7.61
(m, 6H), 7.78 (s, 1H), 7.90 (d, J ¼ 15.5, 2H), 8.04 (d, J ¼ 13.0 Hz, 1H),
8.38 (d, J ¼ 7.8 Hz, 1H), 8.49 (d, J ¼ 5.0 Hz, 1H), 11.71 (s, 1H);
MS m/z (ESI): 723.2 [MþH]þ; Anal. calcd. for C41H37F3N4O5 (%):
C, 68.13; H, 5,16; N, 7.75. Found (%): C, 68.15; H, 5.15; N, 7.74.
7-Fluoro-N-(3-fluoro-4-(6-methoxy-7-(3-(piperidin-1-yl)-
propoxy)quinolin-4-yloxy)phenyl)-4-oxo-1-p-tolyl-1,4-
dihydroquinoline-3-carboxamide (22b)
Yield: 66.5%; m.p. 166–188°C; 1H NMR (300 MHz, DMSO-d6)
d 1.41 (s, 2H), 1.82 (s, 4H), 2.33 (s, 3H), 2.42 (m, 2H), 2.82 (m, 6H),
3.99 (s, 3H), 4.29 (s, 2H), 6.55 (s, 1H), 6.82 (d, J ¼ 11.9 Hz,
1H), 7.27 (s, 1H), 7.40 (m, 1H), 7.52 (m, 4H), 7.60 (d, J ¼ 4.9 Hz,
3H), 8.10 (d, J ¼ 10.3 Hz, 1H), 8.54 (m, 2H), 8.73 (s, 1H),
12.47 (s, 1H); MS m/z (ESI): 705.0 [MþH]þ; Anal. calcd. for
7-Fluoro-N-(3-fluoro-4-(6-methoxy-7-(3-
morpholinopropoxy)quinolin-4-yloxy)phenyl)-1-(2-
fluorophenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide
(22g)
Yield: 62.3%; m.p. 214–215°C; 1H NMR (300 MHz, DMSO-d6) d 2.02
(m, J ¼ 6.4 Hz, 2H), 2.48 (m, J ¼ 15.4 Hz, 6H), 3.62 (s, 4H), 3.97
(s, 3H), 4.22 (t, J ¼ 6.3 Hz, 2H), 6.51 (d, J ¼ 5.2 Hz, 1H), 7.10
(d, J ¼ 8.6 Hz, 1H), 7.42 (s, 1H), 7.50 (d, J ¼ 8.9 Hz, 1H), 7.55
(d, J ¼ 5.0 Hz, 1H), 7.63 (d, J ¼ 8.8 Hz, 1H), 7.76 (m, 3H), 7.91
(m, 3H), 8.05 (dd, J ¼ 12.8, 2.3 Hz, 1H), 8.41 (d, J ¼ 8.0 Hz, 1H),
8.50 (d, J ¼ 5.2 Hz, 1H), 11.75 (s, 1H); MS m/z (ESI): 712.7 [MþH]þ;
Anal. calcd. for C39H33F3N4O6 (%): C, 65.91; H, 4.68; N, 7.88. Found
(%): C, 65.94; H, 4.65; N, 7.90.
C41H38F2N4O5 (%): C, 69.87; H, 5.43; N, 7.95. Found (%): C, 69.90;
H, 5.44; N, 7.93.
7-Fluoro-N-(3-fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)-
propoxy)quinolin-4-yloxy)phenyl)-1-(2-fluorophenyl)-4-
oxo-1,4-dihydroquinoline-3-carboxamide (22c)
1
Yield: 52%; m.p. 199–200°C; H NMR (300 MHz, DMSO-d6) d 1.97
(s, 4H), 2.28 (s, 2H), 3.29 (s, 6H), 4.00 (s, 3H), 4.31 (s, 2H), 6.51
(d, J ¼ 5.0 Hz, 1H), 7.24 (d, J ¼ 8.5 Hz, 1H), 7.43 (s, 1H), 7.50
(t, J ¼ 6.4 Hz, 1H), 7.66 (m, 5H), 7.78 (s, 1H), 7.90 (dd, J ¼ 15.5,
7.6 Hz, 2H), 8.04 (d, J ¼ 13.0 Hz, 1H), 8.38 (d, J ¼ 7.8 Hz, 1H), 8.49
(d, J ¼ 5.0 Hz, 1H), 11.71 (s, 1H); MS m/z (ESI): 696.1 [MþH]þ; Anal.
calcd. for C39H33F3N4O5 (%): C, 67.43; H, 4.39; N, 8.06. Found (%):
C, 67.45; H, 4.38; N, 8.04.
Pharmacology
MTT assay in vitro
The antiproliferative activities of compounds 21a–21l and
22a–22g were evaluated against four c-Met-dependent cancer
cell lines (HT-29, MKN-45, U87MG, and SMMC-7721) and one c-
Met-independent cancer cell line (H460) by the standard MTT
assay in vitro, with foretinib as the positive control [26–28]. The
cancer cell lines were cultured in minimum essential medium
(MEM) supplement with 10% fetal bovine serum (FBS).
Approximate 4 ꢁ 103 cells, suspended in MEM medium,
were plated onto each well of a 96-well plate and incubated in
5% CO2 at 37°C for 24 h. The tested compounds at the
indicated final concentrations were added to the culture
medium and the cell cultures were continued for 72 h. Fresh
MTT was added to each well at a terminal concentration of
5 mg/mL, and incubated with cells at 37°C for 4 h. The
formazan crystals were dissolved in 100 mL DMSO each well,
and the absorbency at 492 nm (for absorbance of MTT
formazan) and 630 nm (for the reference wavelength) was
measured with an ELISA reader. All of the compounds were
tested two times in each of the cell lines. The results expressed
as IC50 (inhibitory concentration 50%) were the averages of
three determinations and calculated by using the Bacus
Laboratories Incorporated Slide Scanner (Bliss) sofeware.
1-(2-Chlorophenyl)-7-fluoro-N-(3-fluoro-4-(6-methoxy-7-
(3-(4-methylpiperidin-1-yl)propoxy)quinolin-4-yloxy)-
phenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (22d)
Yield: 60.3%; m.p. 209–210°C; 1H NMR (300 MHz, DMSO-d6) d 0.91
(d, J ¼ 6.0 Hz, 3H), 1.34 (d, J ¼ 12.0 Hz, 2H), 1.50 (s, 1H), 1.71
(d, J ¼ 12.3 Hz, 2H), 2.17 (s, 2H), 2.91 (s, 2H), 3.24 (s, 4H), 3.96
(s, 3H), 4.23 (s, 2H), 6.50 (d, J ¼ 5.0 Hz, 1H), 6.71 (d, J ¼ 9.1 Hz, 1H),
7.55 (m, 5H), 7.98 (d, J ¼ 6.9 Hz, 2H), 8.10 (d, J ¼ 17.6 Hz, 3H), 8.50
(m, 2H), 8.85 (s, 1H); MS m/z (ESI): 739.2 [MþH]þ; Anal. calcd. for
C40H35ClF2N4O5 (%): C, 66.25; H, 4.86; N, 7.73. Found (%): C, 66.27;
H, 4.86; N, 7.74.
7-Fluoro-N-(3-fluoro-4-(6-methoxy-7-(3-(4-
methylpiperidin-1-yl)propoxy)quinolin-4-yloxy)phenyl)-1-
(2-(trifluoromethyl)phenyl)-4-oxo-1,4-dihydroquinoline-3-
carboxamide (22e)
Yield: 60.3%; m.p. 196–197°C; 1H NMR (300 MHz, DMSO-d6) d 0.95
(d, J ¼ 5.7 Hz, 3H), 1.46 (s, 2H), 1.58 (s, 1H), 1.78 (d, J ¼ 11.1 Hz, 2H),
2.27 (s, 2H), 2.84 (s, 2H), 3.14 (s, 4H), 3.98 (s, 3H), 4.27 (s, 2H), 6.53
(d, J ¼ 5.5 Hz, 1H), 6.73 (d, J ¼ 8.1 Hz, 1H), 7.49 (m, 2H), 7.59
(m, 3H), 7.75 (m, 2H), 7.92 (m, 2H), 8.08 (d, J ¼ 13.3 Hz, 1H),
8.54 (m, 2H), 8.81 (s, 1H), 12.33 (d, J ¼ 16.6 Hz, 1H); MS m/z (ESI):
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

532
S. Li et al.
Arch. Pharm. Chem. Life Sci. 2013, 346, 521–533
Flow cytometric analysis
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The treated MKN-45 cells were harvested, rinsed with cold PBS,
and fixed in 70% ethanol at 4°C for at least 18 h. Then the cell
pellets were stained with the fluorescent probe solution
containing 50 mg/mL PI and 1 mg/mL DNase-free RNase A in
PBS on ice in the dark for 1 h. The DNA fluorescence of PI-
stained cells was evaluated with the FACScan flow cytometer.
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and cofactors; min: the observed enzyme activity in the
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Western blot analysis
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