Design, synthesis, and structure-activity relationships of novel 6,7-disubstituted-4-phenoxyquinoline derivatives as potential antitumor agents

Article abstract of DOI:10.1016/j.ejmech.2013.08.019

Two series of quinoline derivatives bearing the pyridine/pyrimidine scaffold were synthesized, and evaluated for their c-Met kinase inhibitory activity and antiproliferative activity against 5 cancer cell lines (HT-29, H460, MKN-45, A549, and U87MG) were evaluated in vitro. Most compounds showed moderate to excellent potency, and compared to foretinib, the most promising analog 18b (c-Met half-maximal inhibitory concentration [IC₅₀] = 1.39 nM) showed a 7.3-fold increase in activity against HT-29 cell line in vitro. Structure-activity relationship studies indicated that regulation of the electron density on the pyridine/pyrimidine ring to a proper degree was a key factor in improving the antitumor activity.

Full text of DOI:10.1016/j.ejmech.2013.08.019

European Journal of Medicinal Chemistry 69 (2013) 77e89
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Design, synthesis, and structureeactivity relationships of novel 6,7-
disubstituted-4-phenoxyquinoline derivatives as potential antitumor
agents
*
**
Qidong Tang, Yanfang Zhao, Xinming Du, Lian’e Chong, Ping Gong , Chun Guo
Key Laboratory of Structure-Based Drug Design and Discovery (Shenyang Pharmaceutical University), Ministry of Education, 103 Wenhua Road,
Shenhe District, Shenyang 110016, PR China
a r t i c l e i n f o
a b s t r a c t
Article history:
Two series of quinoline derivatives bearing the pyridine/pyrimidine scaffold were synthesized, and
evaluated for their c-Met kinase inhibitory activity and antiproliferative activity against 5 cancer cell lines
(HT-29, H460, MKN-45, A549, and U87MG) were evaluated in vitro. Most compounds showed moderate
to excellent potency, and compared to foretinib, the most promising analog 18b (c-Met half-maximal
inhibitory concentration [IC50] ¼ 1.39 nM) showed a 7.3-fold increase in activity against HT-29 cell
line in vitro. Structureeactivity relationship studies indicated that regulation of the electron density on
the pyridine/pyrimidine ring to a proper degree was a key factor in improving the antitumor activity.
Ó 2013 Elsevier Masson SAS. All rights reserved.
Received 7 May 2013
Received in revised form
7 August 2013
Accepted 8 August 2013
Available online 19 August 2013
Keywords:
Synthesis
Quinoline derivatives
c-Met
Antiproliferative activity
1. Introduction
dihydrocinnoline and quinoline scaffolds as part of the 5-atom
linker on the basis of “5 atoms regulation”, and the resulting de-
Cancer is a widespread, complex, and lethal disease. In 2008, 7.6
million people died of cancer (around 13% of all deaths), and this
number is projected to increase with an estimated 13.1 million in
2030 [1]. Despite the efforts to discover and develop small molecule
anticancer drugs in the last decade [2e5], development of new
antitumor agents with improved tumor selectivity, efficiency, and
safety remains desirable.
Recently, a number of new quinoline derivatives with excellent
antitumor activity have been reported [6e18]. Among them, 6,7-
disubstituted-4-phenoxyquinoline derivatives, which could
inhibit c-Met kinase, have attracted our attention. Many of these
derivatives are already being marketed or are under clinical/pre-
clinical studies, such as cabozantinib, foretinib, MG10, Amgen, and
AM7 (1e5, Fig. 1). The main modification of these quinoline de-
rivatives was focused on the 5-atom linker between moiety A and
moiety B, which was characterized by the illustrated “5 atoms
regulation” [19]. In our previous study, we introduced 1,4-
rivatives N-(3-fluoro-4-(6,7-disubstitutedquinolin-4-yloxy)phenyl)
-4-oxo-1-(substitutedphenyl)-1,4-dihydrocinnoline-3-carboxami-
des (6, Fig. 1) and N-(3-fluoro-4-(6,7-disubstitutedquinolin-4-
yloxy)phenyl)-2-substitutedphenylquinoline-4-carboxamides (7,
Fig. 1) showed excellent potency [19,20].
Pyridine- and pyrimidine-based compounds are amongst the
most commonly occurring heterocycles in anticancer drugs, such
as imatinib, sorafenib, crizotinib, and abiraterone [5]. In this study,
pyridine/pyrimidine as a part of the 5-atom linker was attached to
the 6,7-disubstituted-4-phenoxyquinoline moiety via an amide
bond and then 2 series of antitumor agents were designed (Fig. 2).
According to our previous study, the 3-carbon tether at the 7
position of quinoline was reserved, while the morpholinyl group
was replaced by other water-soluble substituents, including
piperidinyl, pyrrolidinyl, and 4-methyl piperazinyl groups, to
observe the effects of the different cyclic tertiary amino groups on
activity of the new compounds. Furthermore, various substituents
were introduced at the phenyl ring (moiety B) to investigate their
effects on activity. Herein,
2 series of 6,7-disubstituted-4-
phenoxyquinoline derivatives were designed and synthesized to
study the structureeactivity relationships (SARs) and find prom-
ising antitumor agents.
* Corresponding author. Tel./fax: þ86 24 23986429.
** Corresponding author. Tel./fax: þ86 24 23986425.
E-mail addresses: gongpinggp@126.com (P. Gong), gc_66888@163.com (C. Guo).
0223-5234/$ e see front matter Ó 2013 Elsevier Masson SAS. All rights reserved.
http://dx.doi.org/10.1016/j.ejmech.2013.08.019

78
Q. Tang et al. / European Journal of Medicinal Chemistry 69 (2013) 77e89
Fig. 1. Structures of small-molecule c-Met inhibitors based on the 6,7-disubstituted-4-phenoxyquinoline scaffold.
2. Chemistry
cross-coupling reactions with substituted phenylboronic acids in
hot 1,4-dioxane:H2O (3:1, 85 ꢀC) [23,24] and then yielded the target
2.1. Synthesis of 6,7-disubstituted-4-phenoxyquinolines
compounds 18aead.
The synthesis of the key intermediates of 6,7-disubstituted-4-
phenoxyquinolines 15aed was achieved in 8 steps from the
commercially available 1-(4-hydroxy-3-methoxyphenyl)ethanone
as shown in Scheme 1, which has been illustrated in detail in our
previous study [19,20].
2.3. Synthesis of the target compounds of pyrimidine-based
quinolines
The synthesis of the target compounds 25aen is described in
Scheme 3. The commercially available substituted acetophenones
were condensed with N,N-dimethyl formamide dimethyl acetal
(DMF-DMA) at 80 ꢀC to afford intermediates 20aed as yellow
solids. Cyclization of 20aed with formamidine acetate in refluxing
ethanol (EtOH) under basic conditions yielded 4-phenyl
pyrimidines 21aed, which were subjected to free radical reaction
with formamide catalyzed with 98% sulfuric acid (H2SO4)/ferrous
sulfate (FeSO4$7H2O)/30% hydrogen peroxide (H2O2) [25e27] to
give the corresponding products 22aed in low to moderate yield
(32e48%). Simple procedures such as hydrolysis and acyl chlori-
nation were used to convert 4-phenylpyrimidine-2-carboxamides
2.2. Synthesis of the target compounds of pyridine-based quinolines
The target compounds 18aead were prepared as illustrated in
Scheme 2. Using sodium bromide (NaBr) as catalyst, chlorination of
the commercially available 2-picolinic acid with thionyl chloride
resulted in a high yield of the intermediate 16 [21,22]. Acylation of
amides 15aed with acyl chloride 16 in the presence of N,N-diiso-
propylethylamine (DIPEA) proceeded smoothly to yield 17aed,
which were subjected to palladium-catalyzed SuzukieMiyaura
Fig. 2. Design strategy for pyridine- and pyrimidine-based quinoline derivatives.

Q. Tang et al. / European Journal of Medicinal Chemistry 69 (2013) 77e89
79
Scheme 1. Reagents and conditions: (i) Br(CH2)3Cl, acetone, 0 ꢀC, 30 min, rt, 12 h; (ii) 98%HNO3, CH2Cl2, 0 ꢀC, 4 h; (iii) DMF-DMA, toluene, 110 ꢀC, 10 h; (iv) Fe powder, AcOH, rt,
30 min, 80 ꢀC, 2 h; (v) secondary amines, CH3CN, 85 ꢀC, 10 h; (vi) POCl3, 85 ꢀC, 6 h; (vii) 2-fluoro-4-nitrophenol, PhCl, 140 ꢀC, 30 h; (viii) Fe powder, NH4Cl(cat.), EtOH/H2O, reflux,
5 h.
22aed to the corresponding acyl chlorides 24aed; the reactions
proceeded with 20% H2SO4 and thionyl chloride, respectively. Re-
action of amides 15aed with acyl chlorides 24aed promoted by
DIPEA in dichloromethane at room temperature yielded the target
compounds 25aen.
replacement of N-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide
framework of foretinib with 4-substitutedphenylpyridine-2-
carboxamide or 4-substitutedphenylpyrimidine-2-carboxamide
moiety maintained the potent cytotoxic activity (Table 1). The IC50
values of the most promising compound 18b were 0.026
0.037 M, and 0.81 M against HT29, H460, and U87MG cell lines,
respectively; these values indicated that this compound was 7.3, 5.8,
and 1.2 times more active than foretinib (IC50 values: 0.19 M,
0.21 M, and 0.98 M, respectively).
mM,
m
m
3. Results and discussion
m
m
m
3.1. In vitro cytotoxic activities and SAR
The cell lines data revealed a clear preference for activity when
the R1 group was 4-methyl piperazinyl group, which indicated that
a more water-soluble cyclic tertiary amino group at the 7 position
of quinoline contributed to the potency of the target compounds.
The cytotoxic activities of the target compounds 18aead and
25aen have been evaluated in HT-29 (human colon cancer), H460
(human lung cancer), MKN-45 (human gastric cancer), A549 (hu-
man lung adenocarcinoma), and U87MG (human glioblastoma) cell
lines using the MTT assay. Foretinib was used as the positive con-
trol, and the results expressed as half-maximal inhibitory concen-
tration (IC50) values and are presented in Table 1, as mean values of
experiments performed in triplicate.
For example, the IC50 value of compound 18a, 0.082
mM, was clearly
lower than that of 18i, 18q, and 18y, 0.33
m
M, 0.12
m
M, and 0.27 M,
m
respectively, against HT29 cells.
Further investigations were performed to study the effect of
different substituents on the phenyl ring (moiety B) on the cyto-
toxic activity. Introduction of mono-electron-withdrawing groups
(mono-EWGs) on the phenyl ring showed totally different in-
fluences on the activity of the two series of compounds. Introduc-
tion of mono-EWGs to group R2 reduced the antitumor activity,
All the target compounds showed moderate to excellent cytotoxic
activity against the different cancer cells with potencies in the single-
digit
mM range, and 21 of these compounds were more potent than
foretinib against one or more cell lines, which suggested that
Scheme 2. Reagents and conditions: (i) SOCl2, NaBr, PhCl, 50 ꢀC, 1 h, reflux, 20 h; (ii) 4-chloropicolinoyl chloride, DIPEA, CH2Cl2, 0 ꢀC, 1 h, rt, 7 h; (iii) substituted phenylboronic acid,
PdCl2(PPh3)2, KI, NaCO3, N2, 1,4-dioxane/H2O, rt, 30 min, 85 ꢀC, 20e30 h.

80
Q. Tang et al. / European Journal of Medicinal Chemistry 69 (2013) 77e89
Scheme 3. Reagents and conditions: (i) DMF-DMA, 80 ꢀC, 8e12 h; (ii) formamidine acetate, EtONa, EtOH, 70 ꢀC, 1 h, reflux, 15e20 h; (iii) acetamide, 98%H2SO4, FeSO4$7H2O, 30%
H2O2, 0e10 ꢀC, 1 h, 10e15 ꢀC, 30 min; (iv) 20% H2SO4, 100 ꢀC, 5e10 h; (v) SOCl2, reflux, 6 h; (vi) appropriate aniline, carbonyl chloride, DIPEA, 0 ꢀC, 1 h, rt, 7e10 h.
while introduction of mono-EWGs to group R02 improved the ac-
M), with no
substituent on the phenyl ring, showed strong cytotoxicity against
HT29 cells. The introduction of mono-EWGs to R2 (18d, R2 ¼ 4-
Flt-3 (IC50 ¼ 1.08 nM) and PDGFR- (IC50 ¼ 1.42 nM). Moreover,
b
tivity. For example, compound 18a (IC50 ¼ 0.082
m
compound 18b showed inhibitory effects against EGFR, c-Kit and
VEGFR-2 although the potency was 65.1-, 323.5-, and 382.7-fold
lower than that of c-Met. These data suggested that compound
18b is a promising multitarget kinase inhibitor.
fluoro, IC50 ¼ 1.46
mM) reduced the activity by 18.3-fold, which
could be further confirmed by introduction of these groups in
compounds 18l, 18t, and 18aa. On the contrary, the activity of 25g
(R02 ¼ 4 ꢁ fluoro, IC50 ¼ 0.12
mM) against HT29 cells after intro-
4. Conclusions
duction of mono-EWGs to R02 was 2.2 times that of 25e (R02 ¼ H,
In summary, we designed and synthesized 44 quinoline de-
rivatives on the basis of the pyridine/pyrimidine scaffold. Five hu-
man cell lines were used to evaluate the potency of the synthesized
compounds, and 21 of these derivatives were more potent than
foretinib. Compound 18b (c-Met IC50 ¼ 1.39 nM, a multitarget
tyrosine kinase inhibitor) showed the strongest cytotoxic activities
against HT-29, H460, and U87MG cell lines; this compound was 7.3,
5.8, and 1.2 times more active than foretinib against the 3 cell lines,
respectively. The analysis of SARs indicated that a more water-
soluble cyclic tertiary amine (R1 ¼ 4-methyl piperazinyl) contrib-
uted to the potency. A proper degree of the electron density on
pyridine/pyrimidine ring was necessary, which could be achieved
by substituting substituents (fluoro or methyl) with different
electrical properties on the phenyl ring (moiety B). In addition, the
hydrophobic pocket of c-Met was probably not sufficiently large to
accommodate moiety B with bulky groups, while the fluoro and
methyl groups were well tolerated. Further studies on SARs and
mechanism of action of these compounds are in progress, and their
results will be reported in the future.
IC50 ¼ 0.66
mM), and the same trend was observed in compounds
25c and 25n. However, incorporation of double electron-
withdrawing groups (double-EWGs) or strong electron-donating
groups (EDGs) reduced the inhibitory activity in the 2 series of
compounds. For example, the activities of compounds 18o and 25d
reduced by 4.1-fold and 3.7-fold against MKN45 cells, respectively.
The pharmacological data suggested that a proper degree of elec-
tron density on pyridine/pyrimidine ring was probably necessary to
improve the antitumor activity. The pyridine ring, which is a part of
the 5-atom linker, requires weak EDGs (such as methyl) to increase
the lower electron density on it. EWGs (such as fluoro) are required
to reduce the higher electron density on the pyrimidine ring.
Incorporation of bulky groups such as methoxy or tri-
fluoromethyl group (18f, 18g, 18n, 18o, and 18v) on the phenyl ring
(moiety B) reduced the antitumor activity. Moreover, the potency
decreased with the number of substituents (18h, 18p, 18x, 25d, and
25h). The pharmacological data suggested that the hydrophobic
pocket probably was not sufficiently large to accommodate moiety
B with bulky groups, while the fluoro and methyl groups were well
tolerated.
5. Experimental
3.2. In vitro enzymatic assays
5.1. Chemistry
As shown in Table 2, the seven tested compounds all exhibited
excellent c-Met enzymatic potency, suggesting that the inhibition
of c-Met may be a mechanism for the antitumor effect of these
derivatives. Compound 18b showed the most potent activity with
an IC50 value of 1.39 nM, which was comparable to that of the
positive control foretinib (IC50 ¼ 1.14 nM), and this compound
should be studied further.
Unless otherwise noted, all materials were obtained from
commercial suppliers and were used without further purification.
Reactions’ time and purity of the products were monitored by TLC
on FLUKA silica gel aluminum cards (0.2 mm thickness) with
fluorescent indicator 254 nm. Column chromatography was run on
silica gel (200e300 mesh) from Qingdao Ocean Chemicals (Qing-
dao, Shandong, China). All melting points were obtained on a Büchi
Melting Point B-540 apparatus (Büchi Labortechnik, Flawil,
Switzerland) and were uncorrected. Mass spectra (MS) were taken
in ESI mode on Agilent 1100 LC-MS (Agilent, Palo Alto, CA, USA). 1H
NMR and 13C NMR spectra were recorded on Bruker ARX-300,
300 MHz or Bruker ARX-600, 600 MHz spectrometer (Bruker
Bioscience, Billerica, MA, USA) with TMS as an internal standard.
3.3. Enzymatic selectivity assays
In order to examine the selectivity of compound 18b on c-Met
over other kinases, it was screened against other 5 tyrosine kinases
(Table 3). Compared with its high potency against c-Met
(IC50 ¼ 1.39 nM), 18b also exhibited high inhibitory effects against

Q. Tang et al. / European Journal of Medicinal Chemistry 69 (2013) 77e89
81
Table 1
Structures and cytotoxic activities of compounds 18aead and 25aen against HT-29, H460, MKN-45, A549, and U87MG cancer cell lines in vitro.
Compd.
R1
R2=R02
IC50
(m
M) ꢂ SDa
HT-29
H460
MKN-45
A549
U87MG
18a
18b
18c
18d
18e
18f
18g
18h
18i
18j
18k
18l
18m
18n
18o
18p
18q
18r
4-Methyl piperazinyl
4-Methyl piperazinyl
4-Methyl piperazinyl
4-Methyl piperazinyl
4-Methyl piperazinyl
4-Methyl piperazinyl
4-Methyl piperazinyl
4-Methyl piperazinyl
Morpholinyl
Morpholinyl
Morpholinyl
Morpholinyl
Morpholinyl
Morpholinyl
Morpholinyl
Morpholinyl
Pyrrolidinyl
Pyrrolidinyl
Pyrrolidinyl
Pyrrolidinyl
Pyrrolidinyl
Pyrrolidinyl
Pyrrolidinyl
Pyrrolidinyl
Piperidinyl
Piperidinyl
Piperidinyl
Piperidinyl
Piperidinyl
Piperidinyl
Morpholinyl
Morpholinyl
Morpholinyl
Morpholinyl
Piperidinyl
Piperidinyl
Piperidinyl
H
0.082 ꢂ 0.01
0.026 ꢂ 0.003
0.18 ꢂ 0.02
1.46 ꢂ 0.27
0.22 ꢂ 0.01
0.20 ꢂ 0.05
0.39 ꢂ 0.06
0.42 ꢂ 0.02
0.33 ꢂ 0.02
0.29 ꢂ 0.03
1.05 ꢂ 0.16
1.00 ꢂ 0.12
0.40 ꢂ 0.02
5.07 ꢂ 0.24
1.34 ꢂ 0.12
1.53 ꢂ 0.24
0.12 ꢂ 0.02
0.35 ꢂ 0.04
1.15 ꢂ 0.003
0.43 ꢂ 0.01
0.23 ꢂ 0.02
0.72 ꢂ 0.02
0.35 ꢂ 0.05
0.63 ꢂ 0.02
0.27 ꢂ 0.01
0.34 ꢂ 0.02
1.37 ꢂ 0.24
0.32 ꢂ 0.01
0.23 ꢂ 0.01
0.69 ꢂ 0.03
0.15 ꢂ 0.02
0.24 ꢂ 0.17
0.11 ꢂ 0.01
2.19 ꢂ 0.12
0.26 ꢂ 0.01
0.62 ꢂ 0.02
0.12 ꢂ 0.02
0.40 ꢂ 0.01
0.21 ꢂ 0.01
0.042 ꢂ 0.01
0.48 ꢂ 0.03
0.39 ꢂ 0.02
0.67 ꢂ 0.17
0.16 ꢂ 0.05
0.19 ꢂ 0.01
0.091 ꢂ 0.02
0.037 ꢂ 0.01
0.15 ꢂ 0.02
0.43 ꢂ 0.05
0.14 ꢂ 0.02
0.16 ꢂ 0.03
0.37 ꢂ 0.03
0.43 ꢂ 0.03
0.35 ꢂ 0.01
0.57 ꢂ 0.05
2.13 ꢂ 0.23
1.87 ꢂ 0.14
0.68 ꢂ 0.03
52.72 ꢂ 0.24
3.12 ꢂ 0.25
12.69 ꢂ 0.26
0.13 ꢂ 0.03
0.16 ꢂ 0.02
0.42 ꢂ 0.03
0.37 ꢂ 0.02
0.14 ꢂ 0.01
0.25 ꢂ 0.01
0.20 ꢂ 0.04
0.52 ꢂ 0.01
0.12 ꢂ 0.02
0.16 ꢂ 0.04
0.25 ꢂ 0.01
0.13 ꢂ 0.03
0.36 ꢂ 0.02
0.20 ꢂ 0.01
0.24 ꢂ 0.05
0.30 ꢂ 0.15
0.22 ꢂ 0.02
1.03 ꢂ 0.11
0.33 ꢂ 0.02
0.23 ꢂ 0.05
0.24 ꢂ 0.01
0.55 ꢂ 0.02
0.16 ꢂ 0.02
0.063 ꢂ 0.02
0.45 ꢂ 0.02
1.12 ꢂ 0.15
0.44 ꢂ 0.05
0.30 ꢂ 0.02
0.21 ꢂ 0.03
0.15 ꢂ 0.01
0.073 ꢂ 0.01
0.24 ꢂ 0.04
0.51 ꢂ 0.04
0.15 ꢂ 0.002
0.33 ꢂ 0.01
0.25 ꢂ 0.02
0.33 ꢂ 0.03
0.28 ꢂ 0.02
0.44 ꢂ 0.06
1.14 ꢂ 0.12
0.87 ꢂ 0.01
1.31 ꢂ 0.12
78.75 ꢂ 036
1.15 ꢂ 0.11
6.68 ꢂ 035
0.21 ꢂ 0.01
0.44 ꢂ 0.05
0.44 ꢂ 0.02
0.35 ꢂ 0.05
0.23 ꢂ 0.01
0.26 ꢂ 0.04
0.15 ꢂ 0.03
0.74 ꢂ 0.05
0.27 ꢂ 0.15
1.67 ꢂ 0.26
1.24 ꢂ 0.27
0.18 ꢂ 0.03
0.87 ꢂ 0.01
0.57 ꢂ 0.04
0.26 ꢂ 0.01
0.31 ꢂ 0.11
0.36 ꢂ 0.02
0.96 ꢂ 0.05
1.03 ꢂ 0.12
0.58 ꢂ 0.06
0.93 ꢂ 0.03
1.12 ꢂ 0.11
0.34 ꢂ 0.03
0.09 ꢂ 0.01
0.44 ꢂ 0.01
1.36 ꢂ 0.13
0.24 ꢂ 0.03
0.49 ꢂ 0.03
0.032 ꢂ 0.005
0.34 ꢂ 0.02
0.10 ꢂ 0.02
1.33 ꢂ 0.23
ND
ND
ND
ND
ND
3.45 ꢂ 0.28
3.45 ꢂ 0.32
ND
1.01 ꢂ 0.15
0.81 ꢂ 0.19
2.03 ꢂ 0.31
ND
ND
ND
ND
ND
1.28 ꢂ 0.13
6.79 ꢂ 0.18
ND
4-Methyl
2-Methyl
4-Fluoro
2-Fluoro
4-Trifluoromethyl
4-Methoxyl
2,4-Dimethoxyl
H
4-Methyl
2-Methyl
4-Fluoro
ND
ND
2-Fluoro
3.99 ꢂ 0.13
3.03 ꢂ 0.27
4-Trifluoromethyl
4-Methoxyl
2,4-Dimethoxyl
H
4-Methyl
2-Methyl
4-Fluoro
ND
ND
ND
ND
ND
ND
1.58 ꢂ 0.17
1.06 ꢂ 0.15
ND
1.80 ꢂ 0.21
0.43 ꢂ 0.02
ND
1.01 ꢂ 0.12
0.56 ꢂ 0.06
ND
1.78 ꢂ 0.18
0.61 ꢂ 0.05
ND
18s
18t
18u
18v
18w
18x
18y
18z
18aa
18ab
18ac
18ad
25a
25b
25c
25d
25e
25f
25g
25h
25i
2-Fluoro
4-Trifluoromethyl
4-Methoxyl
2,4-Dimethoxyl
H
4-Methyl
4-Fluoro
ND
ND
ND
ND
1.80 ꢂ 0.15
0.79 ꢂ 0.04
ND
ND
10.09 ꢂ 0.24
ND
12.51 ꢂ 0.16
ND
2-Fluoro
4-Trifluoromethyl
4-Methoxyl
H
4-Methyl
4-Fluoro
2,4-Difluoro
H
4-Methyl
4-Fluoro
2,4-Difluoro
4-Methyl
4-Fluoro
2,4-Difluoro
H
1.64 ꢂ 0.15
1.63 ꢂ 0.11
ND
ND
0.51 ꢂ 0.02
2.24 ꢂ 0.21
ND
ND
ND
1.35 ꢂ 0.13
2.40 ꢂ 0.23
ND
ND
ND
0.90 ꢂ 0.05
1.14 ꢂ 0.22
ND
ND
Piperidinyl
0.47 ꢂ 0.03
ND
0.68 ꢂ 0.03
0.66 ꢂ 0.05
ND
0.78 ꢂ 0.02
ND
0.91 ꢂ 0.02
0.78 ꢂ 0.06
ND
4-Methyl piperazinyl
4-Methyl piperazinyl
4-Methyl piperazinyl
Pyrrolidinyl
Pyrrolidinyl
Pyrrolidinyl
25j
25k
25l
25m
25n
Foretinibb
4-Methyl
4-Fluoro
ND
ND
0.99 ꢂ 0.06
1.13 ꢂ 0.17
0.11 ꢂ 0.01
1.08 ꢂ 0.12
Bold values show the IC50 values of the target compounds lower than the values of the positive control. ND: Not determined.
a
IC50: concentration of the compound (mM) producing 50% cell growth inhibition after 72 h of drug exposure, as determined by the MTT assay. Each experiment was carried
out in triplicate.
b
Used as the positive control.
The IR spectra were recorded by means of the KBr pellet technique
on a Bruker FTS 135 spectrometer.
5.2.1. 3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperdine-1-yl)
propoxy)quinolin-4-yl-oxy)aniline (15a)
White solid; yield: 77%; m.p. 193e194 ꢀC; 1H NMR (300 MHz,
5.2. Preparation of 3-fluoro-4-(6,7-disubstituted quinolin-4-yloxy)
anilines (15aed)
CDCl3)
d
8.46 (d, J ¼ 5.3 Hz, 1H), 7.58 (s, 1H), 7.41 (s, 1H), 7.03 (t,
J ¼ 8.7 Hz,1H), 6.56 (dd, J ¼ 11.8, 2.6 Hz,1H), 6.50 (dd, J ¼ 9.0, 2.9 Hz,
1H), 6.39 (dd, J ¼ 5.3, 0.8 Hz,1H), 4.25 (t, J ¼ 6.7 Hz, 2H), 4.03 (s, 3H),
3.82 (s, 2H), 2.94 (d, J ¼ 11.5 Hz, 2H), 2.57 (m, 2H), 2.15 (m, 2H), 1.98
(t, J ¼ 10.9 Hz, 2H), 1.63 (d, J ¼ 10.4 Hz, 2H), 1.28 (m, 3H), 0.93 (d,
J ¼ 6.0 Hz, 3H); MS (ESI) m/z (%): 441.4 [M þ H]þ, 463.3 [M þ Na]þ.
The preparation of the key intermediates 15aed has been
illustrated in detail in our previous work [15,16], and so the syn-
thesis method would not be listed here.

82
Q. Tang et al. / European Journal of Medicinal Chemistry 69 (2013) 77e89
Table 2
5.3.2. General procedure for the preparation of N-(3-fluoro-4-(6,7-
disubstiteted quinolin-4-yloxy)phenyl)-4-chloropicolinamides
(17aed)
c-Met kinase activity of selected compounds 18a, 18b, 18q,
18y, 25a, 25j, 25n, and foretinib in vitro.
Compd.
IC50 on c-Met (nM)
A solution of an appropriate aniline 15aed (0.05 mol) and dii-
sopropylethylamine (10.5 mL, 0.06 mol) in dichloromethane
(400 mL) was added drop-wise to a solution of 4-chloropicolinoyl
chloride (19.4 g, obtained in the last step) in dichloromethane
(100 mL) in an ice bath. Upon completion of the addition, the re-
action mixture was removed from the ice bath and placed in room
temperature for 7 h and monitored by thin-layer chromatography
(TLC). The mixture was washed with 10% K2CO3 (150 mL ꢃ 3) fol-
lowed by brine (150 mL ꢃ 1), and the organic phase was separated,
dried, and evaporated to yield 17aed as light yellow solids.
18a
18b
18q
18y
25a
25j
25n
Foretinib
4.48
1.39
12.64
9.37
6.23
7.96
16.36
1.14
5.2.2. 3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)quinolin-
4-yloxy)aniline (15b)
5.3.2.1. 4-Chloro-N-(3-fluoro-4-(6-methoxy-7-(3-(4-methylpiperazin
-1-yl)propoxy)quinolin-4-yloxy)phenyl)picolinamide
(þ17a).
White solid; yield: 82%; m.p. 217e218 ꢀC; 1H NMR (300 MHz,
Yield: 82%; m.p. 168e169 ꢀC; MS (ESI) m/z (%): 580.4 [M þ H] ; 1H
CDCl3)
d
8.47 (d, J ¼ 5.2 Hz, 1H), 7.58 (s, 1H), 7.44 (s, 1H), 7.04 (t,
NMR (300 MHz, CDCl3)
d
10.02 (s, 1H), 8.51 (d, J ¼ 5.1 Hz, 1H), 8.47
J ¼ 8.7 Hz, 1H), 6.57 (dd, J ¼ 11.9, 2.6 Hz, 1H), 6.50 (m, 1H), 6.41 (d,
J ¼ 5.3 Hz, 1H), 4.27 (t, J ¼ 6.6 Hz, 2H), 4.04 (s, 3H), 3.82 (s, 2H), 3.74
(m, 4H), 2.60 (t, J ¼ 7.1 Hz, 2H), 2.51 (d, J ¼ 4.2 Hz, 4H), 2.13 (m, 2H);
MS (ESI) m/z (%): 428.3 [M þ H]þ.
(d, J ¼ 5.2 Hz, 1H), 8.29 (d, J ¼ 1.9 Hz, 1H), 7.95 (dd, J ¼ 14.7, 2.4 Hz,
1H), 7.55 (s, 1H), 7.50 (dd, J ¼ 5.2, 1.9 Hz, 1H), 7.46 (d, J ¼ 8.9 Hz, 1H),
7.41 (s, 1H), 7.27 (d, J ¼ 8.9 Hz, 1H), 6.40 (d, J ¼ 5.2 Hz, 1H), 4.24 (t,
J ¼ 6.5 Hz, 2H), 4.02 (s, 3H), 2.62e2.35 (m, 10H), 2.27 (s, 3H), 2.15e
2.07 (m, 2H).
5.2.3. 3-Fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)
quinolin-4-yloxy)aniline (15c)
5.3.2.2. 4-Chloro-N-(3-fluoro-4-(6-methoxy-7-(3-morpholino
propoxy)quinolin-4-yloxy)phenyl)picolinamide (17b). Yield: 84%;
m.p. 176e178 ꢀC; MS (ESI) m/z (%): 567.3 [M þ H]þ, 589.7
Light yellow solid; yield: 72%; m.p. 208e209 ꢀC; 1H NMR
(300 MHz, DMSO-d6)
d
8.49 (d, J ¼ 5.2 Hz, 1H), 7.55 (s, 1H), 7.44 (s,
[M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.02 (s, 1H), 8.51 (d,
1H), 7.08 (t, J ¼ 9.0 Hz,1H), 6.57 (d, J ¼ 14.1 Hz,1H), 6.46 (m, J ¼ 12.8,
7.1 Hz, 2H), 4.28 (t, J ¼ 5.7 Hz, 2H), 3.96 (s, 3H), 3.59 (s, 2H), 3.04 (m,
9H), 2.28 (m, 2H), 1.96 (d, J ¼ 28.0 Hz, 4H); MS (ESI) m/z (%): 412.2
[M þ H]þ.
J ¼ 5.3 Hz, 1H), 8.47 (d, J ¼ 5.2 Hz, 1H), 8.29 (d, J ¼ 1.9 Hz, 1H), 7.95
(dd, J ¼ 11.9, 2.4 Hz, 1H), 7.55 (s, 1H), 7.50 (dd, J ¼ 5.3, 1.9 Hz, 1H),
7.46 (d, J ¼ 8.6 Hz, 1H), 7.41 (s, 1H), 7.27 (d, J ¼ 8.6 Hz, 1H), 6.42 (d,
J ¼ 5.2 Hz, 1H), 4.26 (t, J ¼ 6.6 Hz, 2H), 4.02 (s, 3H), 3.69e3.62 (m,
4H), 2.55 (t, J ¼ 7.2 Hz, 2H), 2.48e2.42 (m, 4H), 2.11 (p, J ¼ 6.8, 2H).
5.2.4. 3-Fluoro-4-(6-methoxy-7-(3-(piperdine-1-yl)propoxy)
quinolin-4-yloxy)aniline (15d)
Gray solid; yield: 86%; m.p. 196e197 ꢀC; IR (KBr, cmꢁ1): 3482.2,
3387.0, 2946.5, 2835.1, 2788.1, 1621.1, 1587.8, 1512.8, 1483.4, 1252.9,
5.3.2.3. 4-Chloro-N-(3-fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)
propoxy)quinolin-4-yloxy)phenyl)picolinamide (17c). Yield: 79%;
m.p. 145e146 ꢀC; MS (ESI) m/z (%): 551.2 [M þ H]þ; 1H NMR
1215.4, 853.5; 1H NMR (300 MHz, CDCl3)
d
8.46 (d, J ¼ 5.3 Hz, 1H),
7.58 (s, 1H), 7.41 (s, 1H), 7.03 (t, J ¼ 8.7 Hz, 1H), 6.56 (dd, J ¼ 11.8,
2.6 Hz, 1H), 6.50 (m, 1H), 6.39 (dd, J ¼ 5.3, 1.1 Hz, 1H), 4.24 (t,
J ¼ 6.8 Hz, 2H), 4.04 (d, J ¼ 3.8 Hz, 3H), 3.81 (s, 2H), 2.54 (m, 2H),
2.43 (s, 4H), 2.14 (m, 2H), 1.60 (m, 4H), 1.45 (d, J ¼ 5.2 Hz, 2H);
MS (ESI) m/z (%): 426.4 [M þ H]þ, 448.4 [M þ Na]þ.
(300 MHz, CDCl3)
d
10.02 (s, 1H), 8.51 (d, J ¼ 5.2 Hz, 1H), 8.47 (d,
J ¼ 5.3 Hz, 1H), 8.29 (d, J ¼ 1.9 Hz, 1H), 7.95 (dd, J ¼ 12.0, 2.4 Hz, 1H),
7.55 (s, 1H), 7.50 (dd, J ¼ 5.2, 2.0 Hz, 1H), 7.46 (d, J ¼ 9.4 Hz, 1H), 7.41
(s, 1H), 7.27 (d, J ¼ 8.7 Hz, 1H), 6.43 (d, J ¼ 5.1 Hz, 1H), 4.25 (t,
J ¼ 6.7 Hz, 2H), 4.02 (s, 3H), 2.70e2.61 (m, 2H), 2.52 (br, 4H), 2.15 (p,
J ¼ 6.8, 2H), 1.77 (br, 4H).
5.3. Preparation of pyridine-based quinoline derivatives
5.3.2.4. 4-Chloro-N-(3-fluoro-4-(6-methoxy-7-(3-(piperidin-1-yl)
propoxy)quinolin-4-yloxy)phenyl)picolinamide (17d). Yield: 82%;
m.p. 153e154 ꢀC; MS (ESI) m/z (%): 565.2 [M þ H]þ, 587.4
5.3.1. Preparation of 4-chloropicolinoyl chloride (16)
Thionyl chloride (30 mL) was added drop-wise to a stirred
mixture of 2-picolinic acid (12.3 g, 0.1 mol) and anhydrous sodium
bromide (1.0 g, 0.01 mol) in phenyl chloride (20 mL) while main-
taining the temperature at 50 ꢀC. Upon completion of the addition,
the reaction mixture was heated at reflux for 20 h. The solvent was
concentrated in vacuum to give 4-chloropicolinoyl chloride as
yellow oil (19.4 g), and the mixture was used for the next step
immediately without further purification.
[M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.02 (s, 1H), 8.51 (d,
J ¼ 5.3 Hz, 1H), 8.47 (d, J ¼ 5.3 Hz, 1H), 8.29 (d, J ¼ 1.9 Hz, 1H), 7.95
(dd, J ¼ 12.4, 2.4 Hz, 1H), 7.55 (s, 1H), 7.50 (dd, J ¼ 5.3, 1.9 Hz, 1H),
7.46 (d, J ¼ 8.4 Hz, 1H), 7.41 (s, 1H), 7.27 (d, J ¼ 8.4 Hz, 1H), 6.43 (d,
J ¼ 5.0 Hz, 1H), 4.25 (t, J ¼ 6.7 Hz, 2H), 4.02 (s, 3H), 2.54 (t, J ¼ 7.2 Hz,
2H), 2.45e2.38 (m, 4H), 2.16e2.04 (m, 2H), 1.61e1.51 (m, 4H), 1.4e
1.29 (m, 2H).
5.3.3. General procedure for the preparation of the target
compounds (18aead)
Table 3
To the mixture of an appropriate amount of N-(3-fluoro-4-(6,7-
Inhibition of tyrosine kinases by compound 18b.
disubstituted
quinolin-4-yloxy)phenyl)-4-chloropicolinamides
17aed (0.9 mmol), substituted phenylboronic acid (0.18 mmol),
anhydrous sodium carbonate (0.29 g, 2.7 mmol), anhydrous po-
tassium iodide (0.15 g, 0.9 mmol),1,4-dioxane (30 mL), H2O (10 mL),
and bis(triphenylphosphine)palladium(II) dichloride (0.09 g,
0.135 mmol) were added under an atmosphere of nitrogen at room
temperature. The reaction mixture then was stirred at 85 ꢀC for 20e
Kinase
Enzyme IC50 (nM)
Flt-3
1.08
1.42
90.05
449.7
531.9
PDGFR-
EGFR
c-Kit
b
VEGFR-2

Q. Tang et al. / European Journal of Medicinal Chemistry 69 (2013) 77e89
83
30 h and monitored by TLC. The solvent was concentrated in vac-
NMR (600 MHz, CDCl3)
d
163.14, 162.35, 160.06, 152.24, 150.36,
uum and the residue was resolved with dichloromethane (100 mL),
washed with water (30 mL ꢃ 3), brine (30 mL), dried over anhy-
drous Na2SO4, and concentrated in vacuum to give a black solid. To
the mixture of the black solid, activated charcoal (3 g), silica gel
(1 g), dichloromethane (60 mL), and methanol (10 mL) were added,
and the mixture was stirred at reflux for 0.5 h. The mixture was
filtered immediately, and the filtrate was concentrated in vacuum
to give a yellow solid. The crude product was purified by chroma-
tography on silica gel using MeOH/CH2Cl2 to afford the target
compounds 18aead as white solids.
149.83, 149.68, 148.76, 148.54, 146.93, 137.28, 136.48, 134.14, 129.38
(2C), 124.24, 123.91, 120.08, 116.43 (2C), 115.88, 115.44, 109.26,
109.09, 108.76, 102.22, 99.52, 67.44, 56.18, 55.17 (2C), 54.99, 53.25
(2C), 46.08, 26.35. Anal. calcd. for C36H35F2N5O4 (%): C, 67.59; H,
5.51; N, 10.95. Found (%): C, 67.55; H, 5.53; N, 10.97.
5.3.3.5. N-(3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperazin-1-yl)
propoxy)quinolin-4-yloxy)phenyl)-4-(2-fluorophenyl)picolinamide
(18e). Yield: 62%; m.p. 142e143 ꢀC; MS (ESI) m/z (%): 640.4
[M þ H]þ; 1H NMR (300 MHz, CDCl3)
d 10.18 (s, 1H), 8.65 (d,
J ¼ 5.1 Hz, 1H), 8.50 (d, J ¼ 1.2 Hz, 1H), 8.48 (d, J ¼ 5.3 Hz, 1H), 8.00
(dd, J ¼ 12.2, 2.4 Hz, 1H), 7.73 (dd, J ¼ 5.3, 1.9 Hz, 1H), 7.56 (s, 1H),
7.52e7.31 (m, 4H), 7.30e7.26 (m, 3H), 6.43 (s, 1H), 4.25 (t, J ¼ 6.7 Hz,
2H), 4.02 (s, 3H), 2.62 (br,10H), 2.39 (s, 3H), 2.19e2.07 (m, 2H). Anal.
calcd. for C36H35F2N5O4 (%): C, 67.59; H, 5.51; N, 10.95. Found (%): C,
67.56; H, 5.48; N, 10.98.
5.3.3.1. N-(3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperazin-1-yl)
propoxy)quinolin-4-yloxy)phenyl)-4-phenylpicolinamide
(18a).
Yield: 64%; m.p. 127e128 ꢀC; IR (KBr, cmꢁ1): 3411.5, 3333.3, 2940.4,
2793.0, 1687.9, 1596.8, 1527.6, 1479.0, 1431.1, 1349.3, 1305.6, 1250.1,
1209.6, 1170.8, 1013.4, 853.4, 761.7, 694.5, 616.8; MS (ESI) m/z (%):
622.3 [M þ H]þ; 1H NMR (300 MHz, CDCl3)
d 10.20 (s, 1H), 8.65 (d,
J ¼ 5.0 Hz, 1H), 8.54 (d, J ¼ 1.2 Hz, 1H), 8.48 (d, J ¼ 5.2 Hz, 1H), 8.00
(dd, J ¼ 12.1, 2.5 Hz, 1H), 7.76e7.68 (m, 3H), 7.57 (s, 1H), 7.54e7.45
(m, 4H), 7.42 (s, 1H), 7.28 (d, J ¼ 8.6 Hz, 1H), 6.43 (d, J ¼ 4.3 Hz, 1H),
4.25 (t, J ¼ 6.7 Hz, 2H), 4.02 (s, 3H), 2.64e2.53 (m, 4H), 2.48 (br, 6H),
2.28 (s, 3H), 2.16e2.06 (m, 2H). Anal. calcd. for C36H36FN5O4 (%): C,
69.55; H, 5.84; N, 11.26. Found (%): C, 69.53; H, 5.86; N, 11.29.
5.3.3.6. N-(3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperazin-1-yl)
propoxy)quinolin-4-yloxy)phenyl)-4-(4-(trifluoromethyl)phenyl)
picolinamide (18f). Yield: 62%; m.p. 149e150 ꢀC; MS (ESI) m/z (%):
690.6 [M þ H]þ; 1H NMR (300 MHz, CDCl3)
d 10.17 (s, 1H), 8.71 (d,
J ¼ 5.1 Hz, 1H), 8.55 (d, J ¼ 1.1 Hz, 1H), 8.48 (d, J ¼ 5.3 Hz, 1H), 8.00
(dd, J ¼ 12.2, 2.4 Hz, 1H), 7.82 (d, J ¼ 8.4 Hz, 2H), 7.78 (d, J ¼ 8.4 Hz,
2H), 7.73 (dd, J ¼ 5.1, 1.9 Hz, 1H), 7.57 (s, 1H), 7.50 (d, J ¼ 8.5 Hz, 1H),
7.42 (s, 1H), 7.29 (d, J ¼ 8.6 Hz, 1H), 6.42 (d, J ¼ 5.3 Hz, 1H), 4.24 (t,
J ¼ 6.7 Hz, 2H), 4.03 (s, 3H), 2.65e2.52 (m, 4H), 2.47 (br, 6H), 2.28 (s,
3H), 2.15e2.06 (m, 2H). Anal. calcd. for C37H35F4N5O4 (%): C, 64.43;
H, 5.11; N, 10.15. Found (%): C, 64.39; H, 5.08; N, 10.17.
5.3.3.2. N-(3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperazin-1-yl)
propoxy)quinolin-4-yloxy)phenyl)-4-p-tolylpicolinamide
(18b).
Yield: 64%; m.p. 131e132 ꢀC; IR (KBr, cmꢁ1): 3408.2, 3331.3, 2938.3,
2792.5, 1688.7, 1598.0, 1526.4, 1479.1, 1431.3, 1349.1, 1305.7, 1249.2,
1209.5, 1167.2, 1113,6, 1013.6, 854.0, 814.4, 725.3, 686.6; MS (ESI) m/
z (%): 636.2 [M þ H]þ; 1H NMR (300 MHz, CDCl3)
d
10.20 (s, 1H),
5.3.3.7. N-(3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperazin-1-yl)
propoxy)quinolin-4-yloxy)phenyl)-4-(4-methoxyphenyl)picolinamide
(18g). Yield: 61%; m.p. 147e148 ꢀC; IR (KBr, cmꢁ1): 3419.1, 2934.0,
2834.1, 1681.2, 1597.9, 1514.5, 1479.6, 1432.0, 1349.7, 1252.6, 1210.9,
1177.2, 1023.7, 855.7, 830.6, 694.2, 573.5; MS (ESI) m/z (%): 674.7
8.62 (d, J ¼ 5.1 Hz, 1H), 8.52 (d, J ¼ 1.1 Hz, 1H), 8.48 (d, J ¼ 5.3 Hz,
1H), 8.00 (dd, J ¼ 12.1, 2.4 Hz, 1H), 7.70 (dd, J ¼ 5.2, 1.8 Hz, 1H), 7.63
(d, J ¼ 8.2 Hz, 2H), 7.57 (s, 1H), 7.49 (d, J ¼ 8.4 Hz, 1H), 7.42 (s, 1H),
7.32 (d, J ¼ 8.0 Hz, 2H), 7.27 (d, J ¼ 8.7 Hz, 1H), 6.43 (d, J ¼ 5.2 Hz,
1H), 4.25 (t, J ¼ 6.6 Hz, 2H), 4.02 (s, 3H), 2.65e2.53 (m, 4H), 2.48 (br,
6H), 2.42 (s, 3H), 2.28 (s, 3H), 2.16e2.06 (m, 2H); 13C NMR
[M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.20 (s, 1H), 8.60 (d,
J ¼ 5.2 Hz, 1H), 8.50 (d, J ¼ 1.1 Hz, 1H), 8.48 (d, J ¼ 4.9 Hz, 1H), 8.00
(dd, J ¼ 12.1, 2.5 Hz, 1H), 7.72e7.65 (m, 3H), 7.57 (s, 1H), 7.49 (d,
J ¼ 8.5 Hz, 1H), 7.42 (s, 1H), 7.26 (s, 1H), 7.03 (d, J ¼ 8.8 Hz, 2H), 6.43
(d, J ¼ 5.3 Hz, 1H), 4.25 (t, J ¼ 6.7 Hz, 2H), 4.02 (s, 3H), 3.87 (s, 3H),
2.64e2.52 (m, 4H), 2.47 (br, 6H), 2.27 (s, 3H), 2.15e2.07 (m, 2H).
Anal. calcd. for C37H38FN5O5 (%): C, 68.19; H, 5.88; N, 10.75. Found
(%): C, 68.16; H, 5.84; N, 10.79.
(600 MHz, CDCl3)
d 162.32, 160.06, 152.26, 150.32, 149.83, 149.70,
148.83, 148.51, 146.93, 140.05, 137.22, 136.55, 134.14, 130.06 (2C),
126.97 (2C), 124.18, 123.91, 120.09, 115.89, 115.44, 109.23, 109.07,
108.78, 102.21, 99.52, 67.40, 56.18, 55.15 (2C), 54.95, 53.14 (2C),
46.04, 26.34, 21.31. Anal. calcd. for C37H38FN5O4 (%): C, 69.90; H,
6.02; N, 11.02. Found (%): C, 69.87; H, 5.98; N, 11.06.
5.3.3.3. N-(3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperazin-1-yl)
5.3.3.8. N-(3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperazin-1-yl)
propoxy)quinolin-4-yloxy)phenyl)-4-(2,4-dimethoxyphenyl)picolina-
mide (18h). Yield: 56%; m.p. 151e152 ꢀC; MS (ESI) m/z (%): 682.4
propoxy)quinolin-4-yloxy)phenyl)-4-o-tolylpicolinamide
(18c).
Yield: 61%; m.p. 135e136 ꢀC; MS (ESI) m/z (%): 636.5 [M þ H]þ,
644.4 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d
10.20 (s, 1H), 8.63 (d,
[M þ H]þ, 704.2 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.21 (s,
J ¼ 5.1 Hz, 1H), 8.52 (d, J ¼ 1.2 Hz, 1H), 8.48 (d, J ¼ 5.3 Hz, 1H), 8.00
(dd, J ¼ 12.1, 2.4 Hz, 1H), 7.57 (s, 1H), 7.55e7.44 (m, 2H), 7.43 (s, 1H),
7.40e7.28 (m, 4H), 7.25 (d, J ¼ 8.4 Hz, 1H), 6.43 (d, J ¼ 4.8 Hz, 1H),
4.26 (t, J ¼ 6.7 Hz, 2H), 4.03 (s, 3H), 2.62e2.51 (m, 4H), 2.47 (br, 6H),
2.39 (s, 3H), 2.27 (s, 3H), 2.1e2.08 (m, 2H). Anal. calcd. for
1H), 8.56 (d, J ¼ 4.6 Hz, 1H), 8.47 (d, J ¼ 1.2 Hz, 1H), 8.46 (d,
J ¼ 5.3 Hz, 1H), 8.00 (dd, J ¼ 12.1, 2.4 Hz, 1H), 7.68 (dd, J ¼ 5.1, 1.9 Hz,
1H), 7.57 (s, 1H), 7.49 (d, J ¼ 7.7 Hz, 1H), 7.42 (s, 1H), 7.37 (d,
J ¼ 8.3 Hz, 1H), 7.25 (d, J ¼ 8.2 Hz, 1H), 6.64e6.55 (m, 2H), 6.42 (d,
J ¼ 4.8 Hz, 1H), 4.25 (t, J ¼ 6.7 Hz, 2H), 4.02 (s, 3H), 3.86 (s, 3H), 3.84
(s, 3H), 2.61e2.53 (m, 4H), 2.46 (br, 6H), 2.27 (s, 3H), 2.16e2.06 (m,
2H). Anal. calcd. for C38H40FN5O6 (%): C, 66.95; H, 5.91; N, 10.27.
Found (%): C, 66.94; H, 5.89; N, 10.25.
C
37H38FN5O4 (%): C, 69.90; H, 6.02; N, 11.02. Found (%): C, 69.89; H,
6.05; N, 10.96.
5.3.3.4. N-(3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperazin-1-yl)
propoxy)quinolin-4-yloxy)phenyl)-4-(4-fluorophenyl)picolinamide
(18d). Yield: 65%; m.p. 144e145 ꢀC; MS (ESI) m/z (%): 640.6
5.3.3.9. N-(3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)quino-
lin-4-yloxy)phenyl)-4-phenylpicolinamide (18i). Yield: 64%; m.p.
176e177 ꢀC; MS (ESI) m/z (%): 609.5 [M þ H]þ, 631.3 [M þ Na]þ; 1H
[M þ H]þ; 1H NMR (300 MHz, CDCl3)
d 10.18 (s, 1H), 8.65 (d,
J ¼ 5.1 Hz, 1H), 8.49 (d, J ¼ 1.2 Hz, 1H), 8.48 (d, J ¼ 5.3 Hz, 1H), 8.00
(dd, J ¼ 12.2, 2.4 Hz, 1H), 7.74e7.66 (m, 3H), 7.57 (s, 1H), 7.49 (d,
J ¼ 9.0 Hz,1H), 7.42 (s, 1H), 7.28 (d, J ¼ 8.7 Hz,1H), 7.19 (d, J ¼ 8.5 Hz,
2H), 6.42 (d, J ¼ 5.4 Hz, 1H), 4.25 (t, J ¼ 6.9 Hz, 2H), 4.02 (s, 3H),
NMR (300 MHz, CDCl3)
d
10.20 (s, 1H), 8.65 (d, J ¼ 4.9 Hz, 1H), 8.54
(d, J ¼ 1.2 Hz, 1H), 8.48 (d, J ¼ 5.2 Hz, 1H), 8.00 (dd, J ¼ 11.2, 2.4 Hz,
1H), 7.71 (br, 3H), 7.57 (s, 1H), 7.50 (d, J ¼ 6.9 Hz, 4H), 7.42 (s, 1H),
7.28 (d, J ¼ 8.2 Hz, 1H), 6.43 (d, J ¼ 5.0 Hz, 1H), 4.26 (t, J ¼ 6.4 Hz,
2H), 4.02 (s, 3H), 3.70 (br, 4H), 2.56 (t, J ¼ 6.8 Hz, 2H), 2.47 (br, 4H),
2.61e2.52 (m, 4H), 2.45 (br, 6H), 2.27 (s, 3H), 2.14e2.08 (m, 2H); 13
C

84
Q. Tang et al. / European Journal of Medicinal Chemistry 69 (2013) 77e89
2.16e2.05 (m, 2H). Anal. calcd. for C35H33FN4O5 (%): C, 69.07; H,
5.46; N, 9.20. Found (%): C, 69.05; H, 5.45; N, 9.23.
J ¼ 7.1 Hz, 2H), 2.50e2.40 (m, 4H), 2.11 (p, J ¼ 6.8 Hz, 2H); 13C NMR
(600 MHz, CDCl3) d 162.08, 160.06, 152.22, 149.80, 149.61, 148.83,
148.25, 146.88, 145.64, 137.27, 136.47, 131.36, 130.33, 126.72, 125.35,
124.96, 123.92, 122.25, 116.70, 116.56, 115.94, 115.44, 109.26, 109.11,
108.73, 102.22, 99.53, 67.24, 67.03 (2C), 56.18, 55.40, 53.71 (2C),
25.99. Anal. calcd. for C35H32F2N4O5 (%): C, 67.08; H, 5.15; N, 8.94.
Found (%): C, 67.06; H, 5.13; N, 8.96.
5.3.3.10. N-(3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)qui-
nolin-4-yloxy)phenyl)-4-p-tolylpicolinamide (18j). Yield: 59%; m.p.
178e179 ꢀC; IR (KBr, cmꢁ1): 3427.3, 3329.4, 2954.5, 1691.8, 1597.7,
1511.2, 1479.4, 1432.4, 1348.5, 1304.7, 1250.7, 1210.0, 1170.5, 1116.2,
855.1, 817.8, 724.2, 578.7; MS (ESI) m/z (%): 623.6 [M þ H]þ; 1H NMR
(300 MHz, CDCl3)
d
10.21 (s, 1H), 8.62 (d, J ¼ 5.2 Hz, 1H), 8.53e8.44
5.3.3.14. N-(3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)qui-
nolin-4-yloxy)phenyl)-4-(4-(trifluoromethyl)phenyl)picolinamide
(18n). Yield: 62%; m.p. 191e192 ꢀC; IR (KBr, cmꢁ1): 3428.5, 2935.7,
1688.7, 1601.0, 1506.8, 1484.7, 1433.0, 1350.5, 1326.0, 1260.8, 1214.3,
1171.0,1117.0,1071.5,1015.5, 837.0, 739.3, 694.3, 532.9; MS (ESI) m/z
(br, 2H), 8.01 (dd, J ¼ 12.1, 2.4 Hz, 1H), 7.69 (dd, J ¼ 5.1, 1.9 Hz, 1H),
7.64 (d, J ¼ 8.4 Hz, 2H), 7.58 (s, 1H), 7.54 (d, J ¼ 8.5 Hz, 1H), 7.49 (s,
1H), 7.32 (d, J ¼ 7.8 Hz, 2H), 7.27 (d, J ¼ 8.6 Hz, 1H), 6.47 (d,
J ¼ 5.1 Hz, 1H), 4.28 (t, J ¼ 6.8 Hz, 2H), 4.01 (s, 3H), 3.78 (br, 4H),
2.27e2.61 (m, 6H), 2.42 (s, 3H), 2.25e2.11 (m, 2H). Anal. calcd. for
(%): 677.2 [M þ H]þ; 1H NMR (300 MHz, CDCl3)
d 10.17 (s, 1H), 8.70
C
38H35FN4O5 (%): C, 69.44; H, 5.67; N, 9.00. Found (%): C, 69.43; H,
(d, J ¼ 4.9 Hz, 1H), 8.54 (d, J ¼ 1.2 Hz, 1H), 8.48 (d, J ¼ 5.1 Hz, 1H),
8.00 (dd, J ¼ 12.1, 2.4,1H), 7.87e7.75 (m, 4H), 7.72 (dd, J ¼ 5.1, 1.8 Hz,
1H), 7.57 (s, 1H), 7.50 (d, J ¼ 5.9 Hz, 1H), 7.42 (s, 1H), 7.28 (d,
J ¼ 8.5 Hz,1H), 6.42 (d, J ¼ 5.0 Hz,1H), 4.26 (t, J ¼ 6.5 Hz, 2H), 4.02 (s,
3H), 3.75e3.64 (m, 4H), 2.56 (t, J ¼ 7.2 Hz, 2H), 2.51e2.38 (m, 4H),
2.17e2.03 (m, 2H). Anal. calcd. for C36H32F4N4O5 (%): C, 63.90; H,
4.77; N, 8.28. Found (%): C, 63.88; H, 4.76; N, 8.30.
5.66; N, 9.02.
5.3.3.11. N-(3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)quino-
lin-4-yloxy)phenyl)-4-o-tolylpicolinamide (18kþ). Yield: 57%; m.þp.
181e182 ꢀC; MS (ESI) m/z (%): 623.5 [M þ H] , 645.3 [M þ Na] ;
1H NMR (300 MHz, CDCl3)
d
10.20 (s, 1H), 8.65 (d, J ¼ 5.1 Hz, 1H),
8.52 (d, J ¼ 1.2 Hz, 1H), 8.48 (d, J ¼ 5.3 Hz, 1H), 8.00 (dd, J ¼ 12.1,
2.4 Hz, 1H), 7.57 (s, 1H), 7.53e7.46 (m, 2H), 7.43 (s, 1H), 7.37e7.28
(m, 4H), 7.26 (d, J ¼ 8.1 Hz, 1H), 6.43 (d, J ¼ 4.9 Hz, 1H), 4.26 (t,
J ¼ 6.7 Hz, 2H), 4.03 (s, 3H), 3.74e3.69 (m, 4H), 2.57 (t, J ¼ 7.0 Hz,
2H), 2.52e2.44 (m, 4H), 2.31 (s, 3H), 2.17e2.09 (m, 2H). Anal. calcd.
for C38H35FN4O5 (%): C, 69.44; H, 5.67; N, 9.00. Found (%): C, 69.41;
H, 5.65; N, 9.04.
5.3.3.15. N-(3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)qui-
nolin-4-yloxy)phenyl)-4-(4-methoxyphenyl)picolinamide
(18o).
Yield: 62%; m.p. 189e190 ꢀC; MS (ESI) m/z (%): 639.4 [M þ H]þ,
661.6 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.20 (s, 1H), 8.59 (d,
J ¼ 5.0 Hz, 1H), 8.54e8.43 (m, 2H), 8.00 (dd, J ¼ 12.1, 2.3 Hz, 1H),
7.71e7.63 (m, 3H), 7.57 (s, 1H), 7.49 (d, J ¼ 8.7 Hz, 1H), 7.43 (s, 1H),
7.27 (d, J ¼ 8.4 Hz, 1H), 7.03 (d, J ¼ 8.6 Hz, 2H), 6.43 (d, J ¼ 4.8 Hz,
1H), 4.26 (t, J ¼ 6.4 Hz, 2H), 4.02 (s, 3H), 3.87 (s, 3H), 3.71 (br, 4H),
2.56 (d, J ¼ 7.3 Hz, 2H), 2.49 (br, 4H), 2.18e2.07 (m, 2H). Anal. calcd.
for C36H35FN4O6 (%): C, 67.70; H, 5.52; N, 8.77. Found (%): C, 67.68;
H, 5.50; N, 8.78.
5.3.3.12. N-(3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)qui-
nolin-4-yloxy)phenyl)-4-(4-fluorophenyl)picolinamide
(18l).
Yield: 63%; m.p. 186e187 ꢀC; IR (KBr, cmꢁ1): 3401.2, 3340.6, 2934.8,
1691.9, 1599.0, 1511.4, 1481.2, 1431.4, 1348.9, 1308.5, 1255.4, 1212.1,
1169.8,1115.8, 853.6, 834.7, 558.3; MS (ESI) m/z (%): 627.4 [M þ H]þ;
1H NMR (300 MHz, CDCl3)
d
10.18 (s, 1H), 8.64 (d, J ¼ 4.7 Hz, 1H),
5.3.3.16. N-(3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)qui-
nolin-4-yloxy)phenyl)-4-(2,4-dimethoxyphenyl)picolinamide (18p).
Yield: 56%; m.p. 196e197 ꢀC; IR (KBr, cmꢁ1): 3420.3, 3321.9, 2960.2,
2796.5, 1683.9, 1598.4, 1577.9, 1510.4, 1479.9, 1430.7, 1349.9, 1303.6,
1252.8, 1210.2, 1169.4, 1118.2, 1058.3, 1026.3, 854.4, 738.1, 571.5; MS
8.48 (d, J ¼ 5.3 Hz, 2H), 7.99 (dd, J ¼ 12.1, 2.3 Hz, 1H), 7.75e7.63 (m,
3H), 7.57 (s, 1H), 7.49 (d, J ¼ 8.7 Hz, 1H), 7.42 (s, 1H), 7.27 (d,
J ¼ 8.6 Hz,1H), 7.19 (d, J ¼ 8.7 Hz, 2H), 6.42 (d, J ¼ 4.7 Hz,1H), 4.26 (t,
J ¼ 6.6 Hz, 2H), 4.02 (s, 3H), 3.75e3.65 (m, 4H), 2.55 (t, J ¼ 7.1 Hz,
2H), 2.51e2.40 (m, 4H), 2.17e2.05 (m, 2H); 13C NMR (151 MHz,
(ESI) m/z (%): 669.7 [M þ H]þ; 1H NMR (300 MHz, CDCl3)
d 10.22 (s,
CDCl3)
d
163.02, 162.13, 160.05, 152.23, 149.85, 149.81, 149.38,
1H), 8.56 (d, J ¼ 4.6 Hz, 1H), 8.52 (d, J ¼ 1.2 Hz, 1H), 8.46 (d,
J ¼ 5.3 Hz, 1H), 8.01 (dd, J ¼ 11.9, 2.5 Hz, 1H), 7.69 (dd, J ¼ 5.3, 1.9 Hz,
1H), 7.58 (s, 1H), 7.49 (d, J ¼ 7.8 Hz, 1H), 7.42 (s, 1H), 7.37 (d,
J ¼ 8.4 Hz, 1H), 7.25 (d, J ¼ 8.3 Hz, 1H), 6.65e6.56 (m, 2H), 6.42 (d,
J ¼ 4.6 Hz, 1H), 4.26 (t, J ¼ 6.7 Hz, 2H), 4.02 (s, 3H), 3.76e3.63 (m,
4H), 2.56 (t, J ¼ 7.2 Hz, 2H), 2.50e2.37 (m, 4H), 2.15e2.03 (m, 2H).
Anal. calcd. for C37H37FN4O7 (%): C, 66.46; H, 5.58; N, 8.38. Found
(%): C, 66.45; H, 5.56; N, 8.39.
148.84, 148.65, 146.92, 137.28, 136.46, 133.25, 129.02 (2C), 124.26,
123.94, 120.17, 116.45 (2C), 115.93, 115.45, 109.26, 109.11, 108.77,
102.22, 99.52, 67.25, 67.05 (2C), 56.19, 55.40, 53.73 (2C), 26.00. Anal.
calcd. for C35H32F2N4O5 (%): C, 67.08; H, 5.15; N, 8.94. Found (%): C,
67.09; H, 5.16; N, 8.93.
13C NMR (600 MHz, CDCl3)
d 162.08, 160.06, 152.22, 149.80,
149.61, 148.83, 148.25, 146.88, 145.64, 137.27, 136.47, 131.36, 130.33,
126.72, 125.35, 124.96, 123.92, 122.25, 116.70, 116.56, 115.94, 115.44,
109.26, 109.11, 108.73, 102.22, 99.53, 67.24, 67.03 (2C), 56.18, 55.40,
53.71 (2C), 25.99.
5.3.3.17. N-(3-Fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-phenylpicolinamide (18q). Yield: 65%;
m.p. 121e122 ꢀC; IR (KBr, cmꢁ1): 3402.1, 3334.4, 2930.2, 1685.0,
1597.0, 1508.8, 1478.6, 1430.5, 1349.2, 1305.7, 1249.8, 1209.9, 1170.3,
854.5, 762.3, 616.2; MS (ESI) m/z (%): 615.6 [M þ Na]þ; 1H NMR
5.3.3.13. N-(3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)qui-
nolin-4-yloxy)phenyl)-4-(2-fluorophenyl)picolinamide
(18m).
Yield: 61%; m.p. 188e189 ꢀC; IR (KBr, cmꢁ1): 3415.3, 3322.5,
2940.8, 2750.4, 1691.1, 1597.6, 1527.2, 1479.2, 1432.0, 1349.0,
1304.9, 1248.9, 1206.8, 1173.2, 1115.2, 854.0, 695.3, 583.2; MS (ESI)
(300 MHz, CDCl3)
d
10.22 (s, 1H), 8.65 (d, J ¼ 5.1 Hz, 1H), 8.53 (d,
J ¼ 1.2 Hz, 1H), 8.48 (d, J ¼ 5.3 Hz, 1H), 8.02 (dd, J ¼ 12.2, 2.4 Hz, 1H),
7.72 (d, J ¼ 5.6 Hz, 3H), 7.58 (s, 1H), 7.54e7.45 (m, 5H), 7.28 (d,
J ¼ 8.7 Hz, 1H), 6.49 (d, J ¼ 5.1 Hz, 1H), 4.27 (t, J ¼ 7.1 Hz, 2H), 4.01 (s,
3H), 3.54e3.13 (m, 4H), 2.56 (t, J ¼ 7.2 Hz, 2H), 2.16 (br, 6H); 13C
m/z (%): 649.4 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.18 (s,
1H), 8.66 (dd, J ¼ 5.1, 1.7 Hz, 1H), 8.48 (d, J ¼ 1.6 Hz, 1H), 8.47 (d,
J ¼ 4.8 Hz, 1H), 8.00 (dd, J ¼ 12.1, 2.5 Hz, 1H), 7.72 (dt, J ¼ 5.1,
1.8 Hz, 1H), 7.58 (dd, J ¼ 3.8, 1.7 Hz, 1H), 7.57 (s, 1H), 7.52 (dd,
J ¼ 4.8, 1.6 Hz, 1H), 7.48 (dd, J ¼ 4.6, 3.3 Hz, 1H), 7.45 (dd, J ¼ 4.7,
2.8 Hz, 1H), 7.42 (s, 1H), 7.30 (dd, J ¼ 7.5, 1.2 Hz, 1H), 7.25 (d,
J ¼ 4.2 Hz, 1H), 7.21 (dd, J ¼ 2.6, 1.1 Hz, 1H), 6.43 (dd, J ¼ 5.3, 1.1 Hz,
1H), 4.26 (t, J ¼ 6.7 Hz, 2H), 4.02 (s, 3H), 3.75e3.65 (m, 4H), 2.56 (t,
NMR (600 MHz, CDCl3)
d 162.12, 160.71, 152.02, 149.92, 149.53,
148.27, 145.68, 136.97, 136.80, 131.41, 131.36, 130.32 (2C), 126.76
(2C), 125.34, 124.96, 123.84, 122.26, 116.71, 116.56, 116.01, 109.29,
109.14, 108.12, 102.41, 99.87, 65.92, 56.14, 53.74 (2C), 52.9, 25.4,
23.48 (2C). Anal. calcd. for C35H33FN4O4 (%): C, 70.93; H, 5.61; N,
9.45. Found (%): C, 70.91; H, 5.59; N, 9.47.

Q. Tang et al. / European Journal of Medicinal Chemistry 69 (2013) 77e89
85
5.3.3.18. N-(3-Fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)
1H NMR (300 MHz, CDCl3)
d
10.22 (s, 1H), 8.59 (d, J ¼ 5.2 Hz, 1H),
quinolin-4-yloxy)phenyl)-4-p-tolylpicolinamide (18r). Yielþd: 62%;
8.50 (s, 2H), 8.02 (dd, J ¼ 12.0, 2.7 Hz, 1H), 7.69 (d, J ¼ 8.7 Hz, 2H),
7.66 (s, 1H), 7.58 (s, 1H), 7.50 (d, J ¼ 7.7 Hz, 2H), 7.28 (d, J ¼ 8.6 Hz,
1H), 7.03 (d, J ¼ 8.8 Hz, 2H), 6.46 (d, J ¼ 5.3 Hz, 1H), 4.27 (t,
J ¼ 6.4 Hz, 2H), 4.01 (s, 3H), 3.86 (s, 3H), 3.48e3.28 (m, 4H), 2.56 (t,
J ¼ 7.2 Hz, 2H), 2.23e2.08 (m, 4H), 2.07e1.98 (m, 2H). Anal. calcd.
for C36H35FN4O5 (%): C, 69.44; H, 5.67; N, 9.00. Found (%): C, 69.42;
H, 5.66; N, 9.01.
m.p. 124e125 ꢀC; MS (ESI) m/z (%): 607.5 [M þ H] , 629.2
[M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.21 (s, 1H), 8.64 (d,
J ¼ 5.2 Hz, 1H), 8.52 (d, J ¼ 1.2 Hz, 1H), 8.48 (d, J ¼ 5.1 Hz, 1H), 8.00
(dd, J ¼ 12.2, 2.4 Hz, 1H), 7.70 (d, J ¼ 5.4 Hz, 1H), 7.64 (d, J ¼ 8.3 Hz,
2H), 7.57 (s, 1H), 7.51 (d, J ¼ 8.1 Hz, 1H), 7.42 (s, 1H), 7.32 (d,
J ¼ 7.8 Hz, 2H), 7.27 (d, J ¼ 8.4 Hz,1H), 6.42 (d, J ¼ 5.3 Hz,1H), 4.27 (t,
J ¼ 6.8 Hz, 2H), 4.01 (s, 3H), 3.42e3.27 (m, 4H), 2.56 (t, J ¼ 7.2 Hz,
2H), 2.16 (br, 6H). Anal. calcd. for C36H35FN4O4 (%): C, 71.27; H, 5.81;
N, 9.23. Found (%): C, 71.26; H, 5.82; N, 9.24.
5.3.3.24. N-(3-Fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-(2,4-dimethoxyphenyl)picolinamide
(18x). Yield: 56%; m.p. 143e144 ꢀC; MS (ESI) m/z (%): 653.8
5.3.3.19. N-(3-Fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-o-tolylpicolinamide (18s). Yield: 64%;
m.p. 127e128 ꢀC; MS (ESI) m/z (%): 607.7 [M þ H]þ; 1H NMR
[M þ H]þ, 665.5 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.23 (s,
1H), 8.56 (d, J ¼ 5.0 Hz, 1H), 8.52 (br, 1H), 8.45 (d, J ¼ 5.3 Hz, 1H),
8.02 (dd, J ¼ 12.4, 2.6 Hz, 1H), 7.69 (d, J ¼ 3.5 Hz, 1H), 7.58 (s, 1H),
7.49 (d, J ¼ 8.6 Hz, 2H), 7.37 (d, J ¼ 8.3 Hz, 1H), 7.28 (d, J ¼ 8.4 Hz,
1H), 6.66e6.53 (m, 2H), 6.46 (d, J ¼ 4.6 Hz, 1H), 4.28 (t, J ¼ 6.7 Hz,
2H), 4.01 (s, 3H), 3.86 (s, 3H), 3.84 (s, 3H), 3.48e3.27 (m, 4H), 2.55
(s, 2H), 2.15 (br, 4H), 2.01 (br, 2H). Anal. calcd. for C37H37FN4O6 (%):
C, 68.08; H, 5.71; N, 8.58. Found (%): C, 68.07; H, 5.70; N, 8.59.
(300 MHz, CDCl3)
d
10.20 (s, 1H), 8.64 (d, J ¼ 5.1 Hz, 1H), 8.52 (d,
J ¼ 1.2 Hz, 1H), 8.48 (d, J ¼ 5.3 Hz, 1H), 8.00 (dd, J ¼ 11.9, 2.5 Hz, 1H),
7.57 (s, 1H), 7.54e7.45 (m, 2H), 7.42 (s, 1H), 7.39e7.29 (m, 4H), 7.26
(d, J ¼ 8.4 Hz,1H), 6.43 (d, J ¼ 5.1 Hz,1H), 4.27 (t, J ¼ 6.7 Hz, 2H), 4.01
(s, 3H), 3.45e3.29 (m, 4H), 2.57 (t, J ¼ 7.1 Hz, 2H), 2.15 (br, 6H). Anal.
calcd. for C36H35FN4O4 (%): C, 71.27; H, 5.81; N, 9.23. Found (%): C,
71.25; H, 5.79; N, 9.24.
5.3.3.25. N-(3-Fluoro-4-(6-methoxy-7-(3-(piperidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-phenylpicolinamide (18y). Yield: 66%;
m.p. 136e137 ꢀC; IR (KBr, cmꢁ1): 3416.8, 3325.0, 2939.1, 2761.0,
1690.8, 1621.5, 1596.8, 1526.4, 1478.2, 1430.9, 1439.6, 1306.6, 1247.9,
1208.4, 1173.6, 853.6, 763.8, 696.4, 578.3; MS (ESI) m/z (%): 607.3
5.3.3.20. N-(3-Fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-(4-fluorophenyl)picolinamide
(18t).
Yield: 63%; m.p. 131e132 ꢀC; MS (ESI) m/z (%): 611.2 [M þ H]þ; 1H
NMR (300 MHz, CDCl3)
d
10.20 (s, 1H), 8.64 (d, J ¼ 5.1 Hz, 1H), 8.49
[M þ H]þ, 629.2 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.20 (s,
(br, 2H), 8.01 (dd, J ¼ 12.2, 2.4 Hz, 1H), 7.75e7.64 (m, 3H), 7.59 (s,
1H), 7.50 (d, J ¼ 8.0 Hz, 2H), 7.28 (d, J ¼ 8.8 Hz, 1H), 7.19 (d,
J ¼ 8.5 Hz, 2H), 6.48 (d, J ¼ 6.6 Hz,1H), 4.27 (t, J ¼ 6.7 Hz, 2H), 4.01 (s,
3H), 3.45e3.29 (m, 4H), 2.55 (t, J ¼ 7.2 Hz, 2H), 2.16 (br, 6H). Anal.
calcd. for C35H32F2N4O4 (%): C, 68.84; H, 5.28; N, 9.17. Found (%): C,
68.82; H, 5.26; N, 9.20.
1H), 8.65 (d, J ¼ 5.1 Hz, 1H), 8.54 (d, J ¼ 1.2 Hz, 1H), 8.48 (d,
J ¼ 5.3 Hz,1H), 8.00 (dd, J ¼ 11.8, 2.3 Hz,1H), 7.72 (s, 3H), 7.57 (s,1H),
7.51 (d, J ¼ 7.5 Hz, 4H), 7.42 (s, 1H), 7.28 (d, J ¼ 9.0 Hz, 1H), 6.43 (d,
J ¼ 5.4 Hz,1H), 4.24 (t, J ¼ 6.7 Hz, 2H), 4.02 (s, 3H), 2.55 (t, J ¼ 7.0 Hz,
2H), 2.45 (s, 4H), 2.14 (br, 2H), 1.60 (br, 4H), 1.43 (br, 2H). Anal. calcd.
for C36H35FN4O4 (%): C, 71.27; H, 5.81; N, 9.23. Found (%): C, 71.26;
H, 5.79; N, 9.25.
5.3.3.21. N-(3-Fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-(2-fluorophenyl)picolinamide
(þ18u).
5.3.3.26. N-(3-Fluoro-4-(6-methoxy-7-(3-(piperidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-p-tolylpicolinamide (18z). Yield: 65%;
m.p. 143e144 ꢀC; IR (KBr, cmꢁ1): 3420.9, 2937.6, 1682.2, 1597.3,
1509.1, 1479.6, 1432.2, 1349.9, 1305.8, 1252.3, 1211.0, 1172.7, 854.1,
817.7, 694.5, 532.7; MS (ESI) m/z (%): 643.3 [M þ Na]þ; 1H NMR
Yield: 59%; m.p. 129e130 ꢀC; MS (ESI) m/z (%): 611.5 [M þ H] ; 1H
NMR (300 MHz, CDCl3)
d
10.19 (s, 1H), 8.66 (d, J ¼ 5.1 Hz, 1H), 8.47
(s, 2H), 8.01 (dd, J ¼ 12.6, 2.4 Hz, 1H), 7.73 (s, 1H), 7.58 (s, 1H), 7.52e
7.36 (m, 4H), 7.35e7.25 (m, 2H), 7.18 (d, J ¼ 8.4 Hz, 1H), 6.46 (d,
J ¼ 6.4 Hz, 1H), 4.27 (t, J ¼ 6.7 Hz, 2H), 4.00 (s, 3H), 3.46e3.28 (m,
4H), 2.55 (t, J ¼ 7.2 Hz, 2H), 2.15 (br, 6H); 13C NMR (600 MHz, CDCl3)
(300 MHz, CDCl3)
d
10.20 (s, 1H), 8.62 (d, J ¼ 5.2 Hz, 1H), 8.53 (d,
J ¼ 1.2 Hz, 1H), 8.48 (d, J ¼ 5.2 Hz, 1H), 8.00 (dd, J ¼ 12.2, 2.4 Hz, 1H),
7.70 (dd, J ¼ 5.1, 1.8 Hz, 1H), 7.64 (d, J ¼ 8.2 Hz, 2H), 7.57 (s, 1H), 7.51
(d, J ¼ 8.3 Hz, 1H), 7.42 (s, 1H), 7.32 (d, J ¼ 7.9 Hz, 2H), 7.27 (d,
J ¼ 8.6 Hz,1H), 6.42 (d, J ¼ 5.3 Hz,1H), 4.23 (t, J ¼ 6.8 Hz, 2H), 4.02 (s,
3H), 2.54 (t, J ¼ 7.2 Hz, 2H), 2.42 (s, 3H), 2.41e2.34 (m, 4H), 2.14e
2.09 (m, 2H), 1.60e1.55 (m, 4H), 1.45e1.39 (m, 2H); 13C NMR
d
162.10, 160.12, 153.71, 151.64, 149.62, 148.94, 148.26, 146.57,
145.67, 137.25, 136.51, 131.40, 130.34, 130.06, 126.85, 125.40, 124.97,
123.89, 122.25, 116.71, 116.56, 115.94, 109.27, 109.12, 109.05, 102.43,
99.79, 66.10, 56.10, 53.83 (2C), 53.09, 26.05, 23.45 (2C). Anal. calcd.
for C35H32F2N4O4 (%): C, 68.84; H, 5.28; N, 9.17. Found (%): C, 68.83;
H, 5.26; N, 9.21.
(600 MHz, CDCl3)
d 162.36, 160.06, 155.39, 153.73, 152.28, 149.84,
149.66, 148.78, 148.50, 146.93, 140.12, 137.22, 136.54, 130.12 (2C),
129.23, 126.89 (2C), 123.91, 123.72, 119.72, 115.88, 115.44, 109.21,
108.76, 102.20, 99.50, 67.62, 56.18, 55.82, 54.56 (2C), 26.35, 25.95
(2C), 24.46, 21.35. Anal. calcd. for C37H37FN4O4 (%): C, 71.59; H, 6.01;
N, 9.03. Found (%): C, 71.57; H, 5.99; N, 9.02.
5.3.3.22. N-(3-Fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-(4-(trifluoromethyl)phenyl)picolinamide
(18v). Yield: 60%; m.p. 140e141 ꢀC; IR (KBr, cmꢁ1): 3421.3, 2936.4,
2605.1, 1691.7, 1598.0, 1532.6, 1508.6, 1480.3, 1432.2, 1350.9, 1328.9,
1253.2, 1212.0, 1170.9, 1117.5, 1071.3, 1014.8, 856.4, 734.1, 572.0; MS
(ESI) m/z (%): 661.5 [M þ H]þ; 1H NMR (300 MHz, CDCl3)
d
10.19 (s,
5.3.3.27. N-(3-Fluoro-4-(6-methoxy-7-(3-(piperidin-1-yl)propoxy)
1H), 8.70 (d, J ¼ 4.7 Hz, 1H), 8.53 (s, 2H), 8.02 (dd, J ¼ 12.8, 2.5 Hz,
1H), 7.81 (s, 2H), 7.77 (d, J ¼ 8.2 Hz, 2H), 7.73 (s,1H), 7.59 (s, 1H), 7.51
(d, J ¼ 7.8 Hz, 2H), 7.29 (d, J ¼ 8.4 Hz, 1H), 6.50 (s, 1H), 4.29 (t,
J ¼ 6.4 Hz, 2H), 4.01 (s, 3H), 3.47e3.27 (m,4H), 2.56 (br, 2H), 2.16 (br,
6H). Anal. calcd. for C36H32F4N4O4 (%): C, 65.45; H, 4.88; N, 11.50.
Found (%): C, 65.43; H, 4.86; N, 11.52.
quinolin-4-yloxy)phenyl)-4-(4-fluorophenyl)picolinamide
(18aa).
Yield: 65%; m.p. 148e149 ꢀC; MS (ESI) m/z (%): 625.1 [M þ H]þ,
645.3 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.19 (s, 1H), 8.64 (d,
J ¼ 5.1 Hz, 1H), 8.49 (br, 2H), 8.01 (dd, J ¼ 11.1, 2.3 Hz, 1H), 7.75e7.65
(m, 3H), 7.58 (s, 1H), 7.50 (d, J ¼ 8.2 Hz, 1H), 7.42 (s, 1H), 7.28 (d,
J ¼ 8.7 Hz,1H), 7.19 (d, J ¼ 8.6 Hz, 2H), 6.46 (d, J ¼ 5.1 Hz,1H), 4.27 (t,
J ¼ 6.7 Hz, 2H), 4.01 (s, 3H), 2.54 (t, J ¼ 7.1 Hz, 2H), 2.43 (s, 4H), 2.11e
2.01 (m, 2H), 1.62e1.57 (m, 4H), 1.41 (br, 2H). Anal. calcd. for
5.3.3.23. N-(3-Fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-(4-methoxyphenyl)picolinamide (18w).
Yield: 58%; m.p. 138e139 ꢀC; MS (ESI) m/z (%): 645.4 [M þ Na]þ;
C36H34F2N4O4 (%): C, 69.22; H, 5.49; N, 8.97. Found (%): C, 66.23; H,
5.49; N, 9.01.

86
Q. Tang et al. / European Journal of Medicinal Chemistry 69 (2013) 77e89
5.3.3.28. N-(3-Fluoro-4-(6-methoxy-7-(3-(piperidin-1-yl)propoxy)
[M þ Na]þ; 1H NMR (300 MHz, DMSO)
d 7.98e7.91 (m, 2H), 7.75 (d,
J ¼ 12.5 Hz,1H), 7.18e7.09 (m, 2H), 5.67 (d, J ¼ 12.4 Hz, 1H), 3.04 (br,
quinolin-4-yloxy)phenyl)-4-(2-fluorophenyl)picolinamide
(1þ8ab).
Yield: 64%; m.p. 146e147 ꢀC; MS (ESI) m/z (%): 625.3 [M þ H] ; 1H
6H).
NMR (300 MHz, CDCl3)
d
10.18 (s, 1H), 8.67 (d, J ¼ 5.1 Hz, 1H), 8.48
(br, 2H), 8.00 (dd, J ¼ 10.2, 2.2 Hz, 1H), 7.73 (dd, J ¼ 4.7, 1.8 Hz, 1H),
7.56 (s, 1H), 7.53e7.43 (m, 3H), 7.42 (s, 1H), 7.34e7.25 (m, 3H), 6.43
(d, J ¼ 5.1 Hz, 1H), 4.23 (t, J ¼ 7.0 Hz, 2H), 4.02 (s, 3H), 2.54 (t,
J ¼ 7.3 Hz, 2H), 2.43 (br, 4H), 2.16e2.10 (m, 2H), 1.63e1.55 (m, 4H),
1.47e1.40 (m, 2H). Anal. calcd. for C36H34F2N4O4 (%): C, 69.22; H,
5.49; N, 8.97. Found (%): C, 66.21; H, 5.47; N, 8.99.
5.4.1.4. (E)- 3-(Dimethylamino)-1-(2,4-difluorophenyl)prop-2-en-1-
one (20d). Yield: 81%; m.p. 109e110 ꢀC; MS (ESI) m/z (%): 212.1
[M þ H]þ; 1H NMR (300 MHz, DMSO)
d 8.01e7.92 (m, 1H), 7.75 (d,
J ¼ 12.6 Hz, 1H), 6.94e6.81 (m, 2H), 5.63 (d, J ¼ 12.5 Hz, 1H), 3.06
(br, 6H).
5.4.2. General procedure for the preparation of 4-
5.3.3.29. N-(3-Fluoro-4-(6-methoxy-7-(3-(piperidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-(4-(trifluoromethyl)phenyl)picolinamide
(18ac). Yield: 63%; m.p. 149e150 ꢀC; MS (ESI) m/z (%): 695.4
substititedphenylpyrimidines (21aed)
To the mixture of formamidine acetate (9.9 g, 0.096 mol), so-
dium ethoxide (10.9 g, 0.16 mol) and anhydrous EtOH (300 mL), an
appropriate (E)-3-(dimethylamino)-1-substititedphenylprop-2-en-
1-one (0.08 mol) was added slowly at 70 ꢀC. Upon the completion of
addition, the reaction mixture then was stirred at reflux for 15e
20 h and monitored by TLC. The solvent was concentrated in vac-
uum and the residue was resolved with dichloromethane (300 mL),
washed with water (30 mL ꢃ 3), brine (60 mL), dried over anhy-
drous Na2SO4, and concentrated in vacuum to give the corre-
sponding 4-substititedphenylpyrimidines as light yellow solids.
[M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.17 (s, 1H), 8.71 (d,
J ¼ 5.8 Hz, 1H), 8.55 (s, 1H), 8.48 (d, J ¼ 5.3 Hz, 1H), 7.99 (dd, J ¼ 11.9,
2.5 Hz, 1H), 7.84 (d, J ¼ 8.4 Hz, 2H), 7.78 (d, J ¼ 8.3 Hz, 2H), 7.73 (dd,
J ¼ 5.0, 1.9 Hz, 1H), 7.56 (s, 1H), 7.50 (d, J ¼ 11.1 Hz, 1H), 7.42 (s, 1H),
7.29 (d, J ¼ 8.6 Hz, 1H), 6.43 (d, J ¼ 6.3 Hz, 1H), 4.24 (t, J ¼ 6.7 Hz,
2H), 4.02 (s, 3H), 2.55 (t, J ¼ 7.7 Hz, 2H), 2.44 (br, 4H), 2.17e2.04 (m,
2H), 1.60 (br, 4H), 1.45 (br, 2H). Anal. calcd. for C37H34F4N4O4 (%): C,
65.87; H, 5.08; N, 8.30. Found (%): C, 65.84; H, 5.07; N, 8.32.
5.3.3.30. N-(3-Fluoro-4-(6-methoxy-7-(3-(piperidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-(4-methoxyphenyl)picolinamide (18ad).
Yield: 60%; m.p. 143e144 ꢀC; MS (ESI) m/z (%): 637.3 [M þ H]þ,
5.4.2.1. 4-Phenylpyrimidine (21a). Yield: 66%; m.p. 60e61 ꢀC; MS
(ESI) m/z (%): 157.5 [M þ H]þ; 1H NMR (300 MHz, CDCl3)
d 9.29 (s,
1H), 8.79 (d, J ¼ 5.4 Hz, 1H), 8.11 (d, J ¼ 8.6 Hz, 2H), 7.74 (d,
J ¼ 5.6 Hz, 1H), 7.54 (m, 3H).
659.2 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.20 (s, 1H), 8.59 (d,
J ¼ 5.1 Hz, 1H), 8.49 (d, J ¼ 1.2 Hz, 1H), 8.47 (d, J ¼ 5.2 Hz, 1H), 7.99
(dd, J ¼ 12.1, 2.4 Hz, 1H), 7.68 (t, J ¼ 7.5 Hz, 3H), 7.56 (s, 1H), 7.49 (d,
J ¼ 8.2 Hz,1H), 7.41 (s,1H), 7.27 (d, J ¼ 8.5 Hz,1H), 7.02 (d, J ¼ 8.7 Hz,
2H), 6.42 (d, J ¼ 5.1 Hz, 1H), 4.23 (t, J ¼ 6.6 Hz, 2H), 4.02 (s, 3H), 3.86
(s, 3H), 2.54 (t, J ¼ 7.3 Hz, 2H), 2.40 (br, 4H), 2.15e2.08 (m, 2H),
1.65e1.54 (m, 4H), 1.48e1.40 (m, 2H); 13C NMR (600 MHz, CDCl3)
5.4.2.2. 4-p-Tolylpyrimidine (21b). Yield: 63%; m.p. 63e65 ꢀC; MS
(ESI) m/z (%): 193.5 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 9.27 (s,
1H), 8.78 (d, J ¼ 5.6 Hz, 1H), 8.12 (d, J ¼ 8.8 Hz, 2H), 7.75 (d,
J ¼ 5.6 Hz, 1H), 7.36 (d, J ¼ 8.6 Hz, 2H), 2.35 (s, 3H).
d
162.37, 161.06, 160.06, 155.41, 153.75, 152.30, 149.86, 149.67,
5.4.2.3. 4-(4-Fluorophenyl)pyrimidine (þ21c). Yield: 66%;þm.p. 82e
83 ꢀC; MS (ESI) m/z (%): 175.1 [M þ H] , 197.3 [M þ Na] ; 1H NMR
148.78, 148.49, 146.93, 137.23, 136.52, 129.25, 128.38 (2C), 123.90,
123.74, 119.68, 115.88, 115.43, 114.74 (2C), 109.14, 108.75, 102.20,
99.51, 67.65, 56.18, 55.79, 55.46, 54.57 (2C), 26.35, 25.97 (2C), 24.44.
Anal. calcd. for C37H37FN4O5 (%): C, 69.80; H, 5.86; N, 8.80. Found
(%): C, 69.78; H, 5.84; N, 8.83.
(300 MHz, CDCl3)
d
9.28 (s, 1H), 8.76 (d, J ¼ 5.6 Hz, 1H), 8.22 (d,
J ¼ 8.6 Hz, 2H), 7.74 (d, J ¼ 5.6 Hz, 1H), 7.24 (d, J ¼ 8.6 Hz, 2H).
5.4.2.4. 4-(2,4-Difluorophenyl)pyrimidine (21d). Yield: 65%; m.p.
88e90 ꢀC; MS (ESI) m/z (%): 215.4 [M þ Na]þ; 1H NMR (300 MHz,
5.4. Preparation of pyrimidine-based quinoline derivatives
CDCl3)
d
9.29 (s, 1H), 8.78 (d, J ¼ 5.4 Hz, 1H), 8.31e8.24 (m, 1H), 7.78
(d, J ¼ 5.6 Hz, 1H), 7.59e7.51 (m, 2H).
5.4.1. General procedure for the preparation of (E)-3-
(dimethylamino)-1-substititedphenylprop-2-en-1-ones (20aed)
A mixture of an appropriate substituted acetophenone (0.1 mol)
and DMF-DMA (39.8 mL, 0.3 mol) was heated at 80 ꢀC for 8e12 h
and monitored by TLC. Upon cooling to room temperature of the
reaction mixture, Et2O (40 mL) and petroleum ether (120 mL) were
added and stirred for 0.5 h, and then the precipitate was collected
by filtration and dried to give the corresponding (E)-3-(dimethy-
lamino)-1-substititedphenylprop-2-en-1-one as yellow solids.
5.4.3. General procedure for the preparation of 4-
substititedphenylpyrimidine-2-carboxamides (22aed)
To the mixture of an appropriate 4-substititedphenylpyrimidine
(0.06 mol) and acetamide (500 mL), concentrated sulfuric acid
(20 mL) was added drop-wise at the temperature of 0e10 ꢀC. Upon
the completion of addition, 30% H2O2 (50 mL) and saturated FeS-
O4$7H2O (aq.100 mL) were added drop-wise at the same time at the
temperature of 0e10 ꢀC. The reaction mixture the was stirred at 10e
15 ꢀC for 0.5 h and then poured into water (2000 mL), basified with
potassium hydroxide to pH 9, and extracted with dichloromethane
(200 mL ꢃ 3). The combined extracts were washed with brine
(200 mL ꢃ 2), dried over anhydrous Na2SO4, and concentrated in
vacuum to give the corresponding 4-substititedphenylpyrimidine-
2-carboxamides as white solids in a moderate yield.
5.4.1.1. (E)-3-(Dimethylamino)-1-phenylprop-2-en-1-one
þ (20a).
Yield: 89%; m.p. 90e91 ꢀC; MS (ESI) m/z (%): 176.3 [M þ H] , 198.6
[M þ Na]þ; 1H NMR (300 MHz, DMSO)
d 7.89e7.41 (m, 5H), 7.75 (d,
J ¼ 12.4 Hz, 1H), 5.71 (d, J ¼ 12.5 Hz, 1H), 3.05 (br, 6H).
5.4.1.2. (E)-3-(Dimethylamino)-1-p-tolylprop-2-en-1-one
(þ20b).
Yield: 82%; m.p. 94e96 ꢀC; MS (ESI) m/z (%): 190.4 [M þ H] ; 1H
5.4.3.1. 4-Phenylpyrimidine-2-carboxamide (22a). Yield: 47%;þm.p.
152e153 ꢀC; MS (ESI) m/z (%): 200.4 [M þ H]þ, 222.5 [M þ Na] ; 1H
NMR (300 MHz, DMSO)
1H), 7.39 (d, J ¼ 8.0 Hz, 2H), 5.71 (d, J ¼ 12.5 Hz, 1H), 3.03 (br, 6H),
d
7.94 (d, J ¼ 8.1 Hz, 2H), 7.75 (d, J ¼ 12.4 Hz,
NMR (300 MHz, CDCl3)
1H), 8.24 (d, J ¼ 8.4 Hz, 2H), 7.84 (s, 2H), 7.61e7.46 (m, 3H).
d
9.28 (d, J ¼ 1.3 Hz, 1H), 8.38 (d, J ¼ 1.2 Hz,
2.35 (s, 3H).
5.4.1.3. (E)-3-(Dimethylamino)-1-(4-fluorophenyl)prop-2-en-1-one
(20c). Yield: 84%; m.p. 102e103 ꢀC; MS (ESI) m/z (%): 216.6
5.4.3.2. 4-p-Tolylpyrimidine-2-carboxamide (þ22b). Yield: 37%;þm.p.
155e156 ꢀC; MS (ESI) m/z (%): 214.3 [M þ H] , 236.2 [M þ Na] ; 1H

Q. Tang et al. / European Journal of Medicinal Chemistry 69 (2013) 77e89
87
NMR (300 MHz, CDCl3)
d
9.27 (d, J ¼ 1.3 Hz, 1H), 8.35 (d, J ¼ 1.3 Hz,
concentrated in vacuum. The crude product was purified by chro-
matography on silica gel using MeOH/CH2Cl2 to afford the target
compounds 25aen as white solids.
1H), 8.10 (d, J ¼ 8.2 Hz, 2H), 7.82 (s, 2H), 7.28 (d, J ¼ 8.2 Hz, 2H), 2.35
(s, 3H).
5.4.3.3. 4-(4-Fluorophenyl)pyrimidine-2-carboxamide
(22c).
5.4.6.1. N-(3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)quino-
Yield: 45%; m.p.161e162 ꢀC; MS (ESI) m/z (%): 218.7 [M þ H]þ, 240.5
lin-4-yloxy)phenyl)-4-phenylpyrimidine-2-carboxamide
(25a).
[M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d
9.29 (d, J ¼ 1.3 Hz, 1H), 8.38
Yield: 61%; m.p. 183e184 ꢀC; IR (KBr, cmꢁ1): 3321.8, 2941.5, 1694.2,
1587.0, 1528.1, 1480.1, 1432.0, 1350.2, 1248.3, 1209.3, 1174.4, 1114.8,
850.2, 582.6; MS (ESI) m/z (%): 610.2 [M þ H]þ; 1H NMR (300 MHz,
(d, J ¼ 1.3 Hz, 1H), 8.22 (d, J ¼ 8.2 Hz, 2H), 7.81 (s, 2H), 7.16 (d,
J ¼ 8.4 Hz, 2H).
CDCl3)
d
10.02 (s, 1H), 9.30 (d, J ¼ 1.3 Hz, 1H), 8.62 (d, J ¼ 1.3 Hz, 1H),
5.4.3.4. 4-(2,4-Difluorophenyl)pyrimidine-2-carboxamide
(22d).
8.48 (d, J ¼ 5.2 Hz, 1H), 8.22 (dd, J ¼ 6.5, 3.3 Hz, 2H), 7.97 (dd,
J ¼ 11.9, 2.5 Hz, 1H), 7.57 (s, 1H), 7.55 (d, J ¼ 8.2 Hz, 2H), 7.54 (d,
J ¼ 1.3 Hz, 1H), 7.52e7.47 (m, 1H), 7.44 (s, 1H), 7.29 (d, J ¼ 8.6 Hz,
1H), 6.43 (d, J ¼ 5.2 Hz, 1H), 4.27 (t, J ¼ 6.6 Hz, 2H), 4.02 (s, 3H),
3.72e3.67 (m, 4H), 2.56 (t, J ¼ 7.1 Hz, 2H), 2.49e2.44 (m, 4H), 2.10
Yield: 32%; m.p. 168e169 ꢀC; MS (ESI) m/z (%): 236.6 [M þ H]þ,
258.5 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d
9.27 (d, J ¼ 1.2 Hz,
1H), 8.36 (d, J ¼ 1.3 Hz, 1H), 8.30e8.24 (m, 1H), 7.82 (s, 2H), 7.18e
7.04 (m, 2H).
(p, J ¼ 6.8 Hz, 2H); 13C NMR (600 MHz, CDCl3)
d 166.89, 160.82,
5.4.4. General procedure for the preparation of 4-substitited
phenylpyrimidine-2-carboxylic acids (23aed)
159.93, 157.85, 156.32, 155.42, 152.27, 149.86, 148.81, 146.96, 137.84,
135.76, 131.97, 129.26 (2C), 127.53 (2C), 124.05, 116.18, 115.44,
114.09,109.55,109.40,108.79,102.25, 99.48, 67.26, 67.05 (2C), 56.19,
55.40, 53.73 (2C), 26.01. Anal. calcd. for C34H32FN5O5 (%): C, 66.98;
H, 5.29; N, 11.49. Found (%): C, 66.96; H, 5.28; N, 11.51.
A mixture of an appropriate 4-substititedphenylpyrimidine-2-
carboxamide (0.025 mol) and 20% H2SO4 (100 mL) was heated at
100 ꢀC for 5e10 h and monitored by TLC. The reaction mixture was
poured onto ice/water with vigorously stirring and then the
precipitate was collected by filtration and dried to give the corre-
sponding 4-substititedphenylpyrimidine-2-carboxylic acids as
light yellow solids.
5.4.6.2. N-(3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)quino-
lin-4-yloxy)phenyl)-4-p-tolylpyrimidine-2-carboxamide
(25b).
Yield: 56%; m.p.188e189 ꢀC; IR (KBr, cmꢁ1): 3425.5, 3343.3, 2953.5,
1696.8, 1588.6, 1533.7, 1480.7, 1432.6, 1347.9, 1208.2, 1115.7, 853.9,
740.8, 618.3; MS (ESI) m/z (%): 624.5 [M þ H]þ, 646.2 [M þ Na]þ; 1H
5.4.4.1. 4-Phenylpyrimidine-2-carboxylic acid (23a). Yield: 87%;
m.p. 155e157 ꢀC; MS (ESI) m/z (%): 199.5 [M ꢁ H]ꢁ; 1H NMR
NMR (300 MHz, CDCl3)
d
10.07 (s, 1H), 9.28 (d, J ¼ 1.3 Hz, 1H), 8.59
(300 MHz, DMSO)
8.24 (dd, J ¼ 7.3, 2.5 Hz, 2H), 7.63e7.48 (m, 3H).
d
9.36 (d, J ¼ 1.2 Hz, 1H), 8.44 (d, J ¼ 1.3 Hz, 1H),
(d, J ¼ 1.3 Hz, 1H), 8.50 (d, J ¼ 5.2 Hz, 1H), 8.13 (d, J ¼ 8.2 Hz, 2H),
8.02 (dd, J ¼ 12.1, 2.4 Hz, 1H), 7.59 (s, 1H), 7.52 (d, J ¼ 10.6 Hz, 2H),
7.35 (d, J ¼ 8.5 Hz, 2H), 7.29 (d, J ¼ 8.8 Hz, 1H), 6.47 (d, J ¼ 5.3 Hz,
1H), 4.29 (t, J ¼ 6.6 Hz, 2H), 4.03 (s, 3H), 3.92e3.81 (m, 4H), 2.87 (br,
2H), 2.79 (br, 4H), 2.44 (s, 3H), 2.34e2.23 (m, 2H). Anal. calcd. for
5.4.4.2. 4-p-Tolylpyrimidine-2-carboxylic acid (23b). Yield: 81%;
m.p. 159e161 ꢀC; MS (ESI) m/z (%): 212.9 [M ꢁ H]ꢁ; 1H NMR
(300 MHz, DMSO)
8.14 (d, J ¼ 8.2 Hz, 2H), 7.34 (d, J ¼ 8.0 Hz, 2H), 2.35 (s, 3H).
d
9.31 (d, J ¼ 1.3 Hz, 1H), 8.40 (d, J ¼ 1.3 Hz, 1H),
C35H34FN5O5 (%): C, 67.40; H, 5.49; N, 11.23. Found (%): C, 67.42; H,
5.70; N, 11.24.
5.4.4.3. 4-(4-Fluorophenyl)pyrimidine-2-carboxylic
acid
(23c).
5.4.6.3. N-(3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)quino-
lin-4-yloxy)phenyl)-4-(4-fluorophenyl)pyrimidine-2-carboxamide
(25c). Yield: 58%; m.p. 201e202 ꢀC; IR (KBr, cmꢁ1): 3326.2, 2940.4,
1693.4, 1587.9, 1509.7, 1480.7, 1430.9, 1352.1, 1211.4, 1170.8, 1114.7,
990.6, 844.1, 572.2; MS (ESI) m/z (%): 628.7 [M þ H]þ, 650.4
Yield: 76%; m.p. 162e163 ꢀC; MS (ESI) m/z (%): 217.6 [M ꢁ H]ꢁ; 1H
NMR (300 MHz, DMSO)
1H), 8.26 (d, J ¼ 8.1 Hz, 2H), 7.22 (d, J ¼ 8.6 Hz, 2H).
d
9.32 (d, J ¼ 1.3 Hz, 1H), 8.42 (d, J ¼ 1.3 Hz,
5.4.4.4. 4-(2,4-Difluorophenyl)pyrimidine-2-carboxylic acid (ꢁ23d).
Yield: 77%; m.p. 166e167 ꢀC; MS (ESI) m/z (%): 235.4 [M ꢁ H] ; 1H
[M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.04 (s, 1H), 9.29 (d,
J ¼ 1.3 Hz, 1H), 8.58 (d, J ¼ 1.3 Hz, 1H), 8.49 (d, J ¼ 5.3 Hz, 1H), 8.25
(dd, J ¼ 8.9, 5.3 Hz, 2H), 7.99 (dd, J ¼ 11.8, 2.3 Hz, 1H), 7.56 (s, 1H),
7.50 (d, J ¼ 8.9 Hz, 1H), 7.43 (s, 1H), 7.30 (d, J ¼ 8.6 Hz, 1H), 7.26 (s,
1H), 7.20 (s, 1H), 6.42 (d, J ¼ 5.0 Hz, 1H), 4.27 (t, J ¼ 6.6 Hz, 2H), 4.03
(s, 3H), 3.76e3.66 (m, 4H), 2.58 (t, J ¼ 7.0 Hz, 2H), 2.48 (br, 4H),
2.17e2.06 (m, 2H). Anal. calcd. for C34H31F2N5O5 (%): C, 65.06; H,
4.98; N, 11.16. Found (%): C, 65.07; H, 5.01; N, 11.15.
NMR (300 MHz, DMSO)
1H), 8.30e8.24 (m, 1H), 7.13e6.94 (m, 2H).
d
9.37 (d, J ¼ 1.3 Hz, 1H), 8.48 (d, J ¼ 1.3 Hz,
5.4.5. General procedure for the preparation of 4-
substititedphenylpyrimidine-2-carbonyl chlorides (24aed)
An appropriate 4-substititedphenylpyrimidine-2-carboxylic
acid was added to thionyl chloride (0.02 mol) and refluxed for
6 h. The reaction mixture was evaporated to yield the corre-
sponding 4-substititedphenylpyrimidine-2-carbonyl chlorides
24aed, which were used for the next step immediately without
further purification.
5.4.6.4. N-(3-Fluoro-4-(6-methoxy-7-(3-morpholinopropoxy)quino-
lin-4-yloxy)phenyl)-4-(2,4-difluorophenyl)pyrimidine-2-
carboxamide (25d). Yield: 53%; m.p. 208e209 ꢀC; IR (KBr, cmꢁ1):
3423.3, 2939.4, 1685.2, 1587.1, 1510.5, 1431.9, 1347.8, 1300.8, 1248.6,
1214.9, 1116.6, 856.4, 572.1; MS (ESI) m/z (%): 646.6 [M þ H]þ, 668.3
5.4.6. General procedure for the preparation of the target
compounds 25aen
[M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 9.97 (s, 1H), 9.33 (s, 1H),
8.68 (s,1H), 8.48 (d, J ¼ 5.0 Hz,1H), 8.27 (dd, J ¼ 8.7, 5.3 Hz,1H), 7.96
(d, J ¼ 11.6 Hz, 1H), 7.56 (s, 1H), 7.50 (d, J ¼ 8.6 Hz, 1H), 7.43 (s, 1H),
7.28 (d, J ¼ 8.5 Hz, 1H), 7.12e6.91 (m, 2H), 6.43 (d, J ¼ 5.3 Hz, 1H),
4.27 (t, J ¼ 6.6 Hz, 2H), 4.01 (s, 3H), 3.79e3.62 (m, 4H), 2.56 (t,
J ¼ 7.1 Hz, 2H), 2.51e2.42 (m, 4H), 2.17e2.04 (m, 2H); 13C NMR
To a solution of an appropriate aniline 15aed (2 mmol) and
diisopropylethylamine (2.4 mmol) in dichloromethane (40 mL), an
appropriate carbonyl chloride 24aed (4 mmol) dissolved in dried
dichloromethane (20 mL) was added drop-wise in an ice bath.
Upon completion of the addition, the reaction mixture was
removed to room temperature for 7e10 h and monitored by TLC.
The mixture was washed with 10% K2CO3 (20 mL ꢃ 3) followed by
brine (20 mL ꢃ 1), and the organic phase was separated, dried, and
(600 MHz, CDCl3)
d 162.39, 160.55, 159.92, 157.67, 156.37, 152.27,
149.86, 148.80, 146.96, 137.96, 135.69, 132.39, 132.30, 124.06, 117.91,
117.83, 116.15, 115.44, 112.70, 112.56, 109.55, 109.39, 108.78, 105.16,
102.25, 99.48, 67.26, 67.04 (2C), 56.19, 55.40, 53.73 (2C), 26.01. Anal.

88
Q. Tang et al. / European Journal of Medicinal Chemistry 69 (2013) 77e89
calcd. for C34H30F3N5O5 (%): C, 63.25; H, 4.68; N, 10.85. Found (%): C,
63.24; H, 4.67; N, 10.86.
(d, J ¼ 5.3 Hz, 1H), 8.13 (d, J ¼ 8.3 Hz, 2H), 7.97 (dd, J ¼ 11.8, 2.4 Hz,
1H), 7.56 (s, 1H), 7.49 (d, J ¼ 7.9 Hz, 1H), 7.43 (s, 1H), 7.35 (d,
J ¼ 7.9 Hz, 2H), 7.29 (d, J ¼ 8.6 Hz, 1H), 6.43 (d, J ¼ 4.6 Hz, 1H), 4.25
(t, J ¼ 6.6 Hz, 2H), 4.02 (s, 3H), 2.57 (t, J ¼ 7.1 Hz, 2H), 2.49 (d,
J ¼ 12.7 Hz, 8H), 2.44 (s, 3H), 2.28 (s, 3H), 2.10 (dt, J ¼ 13.4, 6.6 Hz,
5.4.6.5. N-(3-Fluoro-4-(6-methoxy-7-(3-(piperidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-phenylpyrimidine-2-carboxamide (25e).
Yield: 61%; m.p. 151e152 ꢀC; MS (ESI) m/z (%): 608.6 [M þ H]þ,
2H); 13C NMR (600 MHz, CDCl3)
d 166.82, 160.93, 159.95, 157.80,
630.2 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d
10.03 (s, 1H), 9.28 (d,
156.48, 156.13, 152.32, 149.89, 148.76, 146.92, 142.66, 135.74,
132.98, 130.01 (2C), 127.46 (2C), 124.04, 116.16, 115.42, 113.72,
109.52, 109.40, 108.75, 102.24, 99.48, 67.36, 56.18, 55.03 (2C),
54.90, 53.00 (2C), 45.90, 26.30, 21.57. Anal. calcd. for C36H37FN6O4
(%): C, 67.91; H, 5.86; N, 13.22. Found (%): C, 67. 90; H, 5.84; N,
13.24.
J ¼ 1.3 Hz, 1H), 8.61 (d, J ¼ 1.3 Hz, 1H), 8.48 (d, J ¼ 5.2 Hz, 1H), 8.22
(dd, J ¼ 8.2, 5.3 Hz, 2H), 7.96 (dd, J ¼ 11.9, 2.5 Hz, 1H), 7.57 (s, 1H),
7.55 (d, J ¼ 2.3 Hz, 2H), 7.54 (d, J ¼ 1.2 Hz,1H), 7.49 (d, J ¼ 7.8 Hz,1H),
7.44 (s, 1H), 7.29 (d, J ¼ 8.6 Hz, 1H), 6.43 (d, J ¼ 5.2 Hz, 1H), 4.25 (t,
J ¼ 6.7 Hz, 2H), 4.01 (s, 3H), 2.54 (t, J ¼ 7.2 Hz, 2H), 2.42e2.33 (m,
4H), 2.15e2.05 (m, 2H), 1.62e1.52 (m, 4H), 1.46e1.38 (m, 2H). Anal.
calcd. for C35H34FN5O4 (%): C, 69.18; H, 5.64; N, 11.52. Found (%): C,
69.17; H, 5.64; N, 11.66.
5.4.6.10. N-(3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperazin-1-yl)
propoxy)quinolin-4-yloxy)phenyl)-4-(4-fluorophenyl)pyrimidine-2-
carboxamide (25j). Yield: 62%; m.p. 175e176 ꢀC; IR (KBr, cmꢁ1):
3329.8, 2936.3, 2794.5, 1693.6, 1509.4, 1480.1, 1431.7, 1351.2, 1251.8,
1211.5, 1169.5, 1014.6, 992.1, 843.0, 572.5; MS (ESI) m/z (%): 641.6
5.4.6.6. N-(3-Fluoro-4-(6-methoxy-7-(3-(piperidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-p-tolylpyrimidine-2-carboxamide (25f).
Yield: 66%; m.p. 157e158 ꢀC; IR (KBr, cmꢁ1): 3343.3, 2931.8, 1697.4,
1587.8, 1528.6, 1479.6, 1431.9, 1348.2, 1248.4, 1209.6, 854.8, 581.5;
MS (ESI) m/z (%): 621.3 [M þ H]þ, 643.4 [M þ Na]þ; 1H NMR
[M þ H]þ, 663.5 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.00 (s,
1H), 9.29 (d, J ¼ 1.3 Hz, 1H), 8.58 (d, J ¼ 1.3 Hz, 1H), 8.48 (d,
J ¼ 5.2 Hz, 1H), 8.25 (dt, J ¼ 8.9, 5.3 Hz, 2H), 7.96 (dd, J ¼ 11.9, 2.5 Hz,
1H), 7.56 (s, 1H), 7.49 (d, J ¼ 9.0 Hz, 1H), 7.43 (s, 1H), 7.29 (d,
J ¼ 8.6 Hz,1H), 7.21 (d, J ¼ 8.6 Hz, 2H), 6.43 (d, J ¼ 5.3 Hz,1H), 4.25 (t,
J ¼ 6.6 Hz, 2H), 4.02 (s, 3H), 2.57 (t, J ¼ 7.1 Hz, 2H), 2.47 (d,
J ¼ 23.5 Hz, 8H), 2.27 (s, 3H), 2.16e2.04 (m, 2H); 13C NMR (600 MHz,
(300 MHz, CDCl3)
d
10.00 (s, 1H), 9.27 (d, J ¼ 1.3 Hz, 1H), 8.59 (d,
J ¼ 1.3 Hz, 1H), 8.48 (s, 1H), 8.13 (d, J ¼ 8.2 Hz, 2H), 7.96 (dd, J ¼ 11.8,
2.2 Hz, 1H), 7.57 (s, 1H), 7.49 (d, J ¼ 7.8 Hz, 1H), 7.44 (s, 1H), 7.35 (d,
J ¼ 8.0 Hz, 2H), 7.28 (d, J ¼ 8.6 Hz, 1H), 6.43 (d, J ¼ 5.9 Hz, 1H), 4.25
(t, J ¼ 6.7 Hz, 2H), 4.01 (s, 3H), 2.54 (t, J ¼ 7.2 Hz, 2H), 2.44 (s, 3H),
2.43e2.36 (m, 4H), 2.15e2.05 (m, 2H), 1.62e1.51 (m, 4H), 1.41e1.10
(m, 2H). Anal. calcd. for C36H36FN5O4 (%): C, 69.55; H, 5.84; N, 11.26.
Found (%): C, 69.54; H, 5.82; N, 11.38.
CDCl3)
d 165.70, 164.42, 160.71, 159.90, 157.84, 156.40, 155.42,
152.34, 149.90, 148.76, 146.99, 135.63, 131.92, 129.74 (2C), 124.05,
116.41 (2C), 116.16, 115.42, 113.69, 109.56, 109.40, 108.79, 102.23,
99.46, 67.43, 56.18, 55.19 (2C), 54.97, 53.22 (2C), 46.09, 26.35. Anal.
calcd. for C35H34F2N6O4 (%): C, 65.61; H, 5.35; N, 13.12. Found (%): C,
65.59; H, 5.34; N, 13.13.
5.4.6.7. N-(3-Fluoro-4-(6-methoxy-7-(3-(piperidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-(4-fluorophenyl)pyrimidine-2-
carboxamide (25g). Yield: 58%; m.p. 161e162 ꢀC; IR (KBr, cmꢁ1):
3327.8, 2935.4, 2769.5, 1693.7, 1587.7, 1509.5, 1480.0, 1431.5, 1351.5,
1306.8, 1251.3, 1211.4, 1173.0, 843.5, 756.2, 572.4; MS (ESI) m/z (%):
5.4.6.11. N-(3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperazin-1-yl)
propoxy)quinolin-4-yloxy)phenyl)-4-(2,4-difluorophenyl)pyrimidine-
2-carboxamide (25k). Yield: 59%; m.p.179e180 ꢀC; MS (ESI) m/z (%):
625.6 [M þ H]þ, 647.2 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d
10.00
659.3 [M þ H]þ, 681.6 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 9.97 (s,
(s, 1H), 9.28 (d, J ¼ 1.3 Hz, 1H), 8.57 (d, J ¼ 1.3 Hz, 1H), 8.48 (d,
J ¼ 5.2 Hz, 1H), 8.13 (d, J ¼ 8.3 Hz, 2H), 7.97 (dd, J ¼ 11.9, 2.4 Hz, 1H),
7.56 (s, 1H), 7.49 (d, J ¼ 8.7 Hz, 1H), 7.43 (s, 1H), 7.29 (d, J ¼ 8.6 Hz,
1H), 7.25 (d, J ¼ 1.9 Hz, 1H), 7.21 (d, J ¼ 8.6 Hz, 1H), 6.42 (d,
J ¼ 4.5 Hz,1H), 4.24 (t, J ¼ 6.7 Hz, 2H), 4.02 (s, 3H), 2.52 (t, J ¼ 6.9 Hz,
2H), 2.44e2.37 (m, 4H), 2.16e2.05 (m, 2H), 1.62e1.53 (m, 4H), 1.47e
1.39 (m, 2H). Anal. calcd. for C35H33F2N5O4 (%): C, 67.19; H, 5.32; N,
11.19. Found (%): C, 67.18; H, 5.30; N, 11.21.
1H), 9.33 (d, J ¼ 1.4 Hz,1H), 8.68 (d, J ¼ 1.5 Hz,1H), 8.48 (d, J ¼ 5.2 Hz,
1H), 8.28 (td, J ¼ 8.8, 6.5 Hz,1H), 7.96 (dd, J ¼ 11.9, 2.5 Hz,1H), 7.56 (s,
1H), 7.49 (d, J ¼ 7.5 Hz,1H), 7.43 (s, 1H), 7.28 (d, J ¼ 8.6 Hz, 1H), 7.11e
6.93 (m, 2H), 6.42 (d, J ¼ 5.2 Hz, 1H), 4.25 (t, J ¼ 6.7 Hz, 2H), 4.01 (s,
3H), 2.57 (t, J ¼ 7.2 Hz, 2H), 2.47 (d, J ¼ 23.4 Hz, 8H), 2.27 (s, 3H), 2.17e
2.04 (m, 2H). Anal. calcd. for C35H33F3N6O4 (%): C, 63.82; H, 5.05; N,
12.76. Found (%): C, 63.80; H, 5.04; N, 12.75.
5.4.6.12. N-(3-Fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-phenylpyrimidine-2-carboxamide (25l).
Yield: 61%; m.p. 128e129 ꢀC; MS (ESI) m/z (%): 594.2 [M þ H]þ,
5.4.6.8. N-(3-Fluoro-4-(6-methoxy-7-(3-(piperidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-(2,4-difluorophenyl)pyrimidine-2-
carboxamide (25h). Yield: 56%; m.p. 166e167 ꢀC; MS (ESI) m/z (%):
616.7 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.04 (s, 1H), 9.27
644.8 [M þ H]þ, 666.7 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d
9.96
(d, J ¼ 1.3 Hz, 1H), 8.59 (d, J ¼ 1.3 Hz, 1H), 8.48 (d, J ¼ 5.2 Hz, 1H),
8.22 (dd, J ¼ 8.4, 5.3 Hz, 2H), 7.96 (dd, J ¼ 11.9, 2.2 Hz, 1H), 7.57 (s,
1H), 7.55 (d, J ¼ 2.3 Hz, 2H), 7.54 (d, J ¼ 1.2 Hz, 1H), 7.49 (d,
J ¼ 7.8 Hz, 1H), 7.44 (s, 1H), 7.29 (d, J ¼ 8.6 Hz, 1H), 6.43 (d,
J ¼ 5.2 Hz, 1H), 4.28 (t, J ¼ 6.7 Hz, 2H), 4.02 (s, 3H), 3.45e3.21 (m,
4H), 2.56 (t, J ¼ 7.2 Hz, 2H), 2.16 (br, 6H). Anal. calcd. for
(s, 1H), 9.33 (d, J ¼ 1.3 Hz, 1H), 8.68 (d, J ¼ 1.3 Hz, 1H), 8.48 (d,
J ¼ 5.2 Hz, 1H), 8.25 (dd, J ¼ 15.3, 6.4 Hz, 2H), 7.96 (dd, J ¼ 11.9,
2.4 Hz, 1H), 7.56 (s, 1H), 7.49 (d, J ¼ 8.9 Hz, 1H), 7.43 (s, 1H), 7.28 (d,
J ¼ 8.7 Hz, 1H), 7.10e6.94 (m, 2H), 6.42 (d, J ¼ 5.3 Hz, 1H), 4.25 (t,
J ¼ 6.6 Hz, 2H), 4.01 (s, 3H), 2.51 (t, J ¼ 7.2 Hz, 2H), 2.44e2.37 (m,
4H), 2.16e2.05 (m, 2H), 1.63e1.53 (m, 4H), 1.47e1.38 (m, 2H). Anal.
calcd. for C35H32F3N5O4 (%): C, 65.31; H, 5.01; N, 10.88. Found (%): C,
65. 30; H, 5.00; N, 10.89.
C
34H32FN5O4 (%): C, 68.79; H, 5.43; N, 11.80. Found (%): C, 68.78;
H, 5.44; N, 11.79.
5.4.6.13. N-(3-Fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-p-tolylpyrimidine-2-carboxamide (25m).
Yield: 53%; m.p. 133e134 ꢀC; IR (KBr, cmꢁ1): 3424.0, 2961.1, 2809.2,
1684.4, 1586.6, 1527.4, 1480.0, 1431.7, 1350.5, 1252.6, 1212.9, 1173.5,
855.2, 749,7, 574.9; MS (ESI) m/z (%): 608.7 [M þ H]þ, 630.5
5.4.6.9. N-(3-Fluoro-4-(6-methoxy-7-(3-(4-methylpiperazin-1-yl)
propoxy)quinolin-4-yloxy)phenyl)-4-p-tolylpyrimidine-2-
carboxamide (25i). Yield: 63%; m.p. 167e168 ꢀC; IR (KBr, cmꢁ1):
3326.1, 2938.0, 2795.0, 1690.0, 1588.8, 1528.8, 1479.8, 1432.2,
1349.2, 1248.6, 1209.9, 1173.0, 1013.4, 855.6, 574.7; MS (ESI) m/z
(%): 637.3 [M þ H]þ, 659.5 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
[M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 10.07 (s, 1H), 9.28 (d,
J ¼ 1.2 Hz,1H), 8.59 (d, J ¼ 1.3 Hz,1H), 8.50 (d, J ¼ 6.4 Hz,1H), 8.12 (d,
d
10.02 (s, 1H), 9.27 (d, J ¼ 1.3 Hz, 1H), 8.59 (d, J ¼ 1.3 Hz, 1H), 8.48
J ¼ 8.2 Hz, 2H), 8.02 (dd, J ¼ 11.8, 2.2 Hz, 1H), 7.58 (s, 1H), 7.51 (d,

Q. Tang et al. / European Journal of Medicinal Chemistry 69 (2013) 77e89
89
J ¼ 5.9 Hz, 2H), 7.35 (d, J ¼ 8.0 Hz, 2H), 7.28 (d, J ¼ 8.8 Hz, 1H), 6.49
Acknowledgments
(d, J ¼ 4.5 Hz,1H), 4.29 (t, J ¼ 6.7 Hz, 2H), 4.02 (s, 3H), 3.43e3.22 (m,
4H), 2.54 (t, J ¼ 7.2 Hz, 2H), 2.44 (s, 3H), 2.16 (br, 6H). Anal. calcd. for
The work was supported by Program for Innovative Research
Team of the Ministry of Education of the People’s Republic of China
and Program for Liaoning Innovative Research Team in University.
C
35H34FN5O4 (%): C, 69.18; H, 5.64; N, 11.52. Found (%): C, 69.17; H,
5.63; N, 11.53.
5.4.6.14. N-(3-Fluoro-4-(6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)
quinolin-4-yloxy)phenyl)-4-(4-fluorophenyl)pyrimidine-2-
carboxamide (25n). Yield: 59%; m.p. 138e139 ꢀC; MS (ESI) m/z (%):
Appendix A. Supplementary data
Supplementary data associated with this article can be found in
the online version, at http://dx.doi.org/10.1016/j.ejmech.2013.08.
019.
612.3 [M þ H]þ, 634.6 [M þ Na]þ; 1H NMR (300 MHz, CDCl3)
d 9.99
(s, 1H), 9.28 (s, 1H), 8.57 (s, 1H), 8.48 (d, J ¼ 5.3 Hz, 1H), 8.24 (dd,
J ¼ 8.7, 5.3 Hz, 2H), 7.96 (dd, J ¼ 11.9, 2.2 Hz,1H), 7.56 (s, 1H), 7.49 (d,
J ¼ 8.9 Hz,1H), 7.43 (s, 1H), 7.28 (d, J ¼ 8.5 Hz,1H), 7.21 (d, J ¼ 8.8 Hz,
2H), 6.42 (d, J ¼ 5.3 Hz, 1H), 4.27 (t, J ¼ 6.6 Hz, 2H), 4.01 (s, 3H), 2.67
(t, J ¼ 7.2 Hz, 2H), 2.54 (br, 4H), 2.20e2.08 (m, 2H), 1.77 (br, 4H).
Anal. calcd. for C34H31F2N5O4 (%): C, 66.77; H, 5.11; N, 11.45. Found
(%): C, 66.76; H, 5.09; N, 11.46.
References
[1] Cancer e Fact sheet Nꢀ297, reviewed January 2013. WHO website: http://
www.who.int/mediacentre/factsheets/fs297/en/.
[2] A.K. Samadi, J. Bazzill, X. Zhang, R. Gallagher, H.P. Zhang, R. Gollapudi,
K. Kindscher, B. Timmermann, M.S. Cohen, Surgery 152 (2012) 1238e1247.
[3] W.K. You, B. Sennino, C.W. Williamson, B. Falcón, H. Hashizume, L.C. Yao,
D.T. Aftab, D.M. McDonald, Cancer Res. 71 (2011) 4758e4768.
[4] F. Dayyani, G.E. Gallick, C.L. Christopher, P.G. Corn, J. Natl. Cancer Inst. 103
(2011) 1665e1675.
[5] S. Hoelder, P.A. Clarke, P. Workman, Mol. Oncol. 6 (2012) 155e176.
[6] M. Zillhardt, S.M. Park, I.L. Romero, K. Sawada, A. Montag, T. Krausz,
S.D. Yamada, M.E. Peter, E. Lengyel, Clin. Cancer Res. 17 (12) (2011) 4042e
4051.
[7] F.M. Yakes, J. Chen, J. Tan, K. Yamaguchi, Y.C. Shi, P.W. Yu, F. Qian, F. Chu,
F. Bentzien, B. Cancilla, J. Orf, A. You, A.D. Laird, S. Engst, L. Lee, J. Lesch,
Y.C. Chou, A.H. Joly, Mol. Cancer Ther. 10 (2011) 2298e2308.
[8] M. Mannion, S. Raeppel, S. Claridge, N. Zhou, O. Saavedra, L. Isakovic, L.J. Zhan,
F. Gaudette, F. Raeppel, R. Déziel, N. Beaulieu, H. Nguyen, I. Chute, C. Beaulieu,
I. Dupont, M.F. Robert, S. Lefebvre, M. Dubay, J. Rahil, J. Wang, H.S. Croix,
A.R. Macleod, J.M. Besterman, A. Vaisburg, Bioorg. Med. Chem. Lett. 19 (2009)
6552e6556.
[9] M.H. Norman, L.B. Liu, M. Lee, N. Xi, I. Fellows, N.D. D’Angelo, C. Dominguez,
K. Rex, S.F. Bellon, T.S. Kim, I. Dussault, J. Med. Chem. 55 (2012) 1858e1867.
[10] N.D. D’Angelo, S.F. Bellon, S.K. Booker, Y. Cheng, A. Coxon, C. Dominguez,
I. Fellows, D. Hoffman, R. Hungate, P.K. Lefko, M.R. Lee, C. Li, L.B. Liu,
E. Rainbeau, P.J. Reider, K. Rex, A. Siegmund, Y.X. Sun, A.S. Tasker, N. Xi,
S.M. Xu, Y.J. Yang, Y.H. Zhang, T.L. Burgess, I. Dussault, T.S. Kim, J. Med. Chem.
51 (2008) 5766e5779.
[11] P.P. Kung, L. Funk, J. Meng, G. Alton, E. Padrique, B. Mroczkowski, Eur. J. Med.
Chem. 43 (2008) 1321e1329.
[12] X.D. Liu, R.C. Newton, P.A. Scherle, Trends Mol. Med. 16 (2009) 1471e1491.
[13] Y. Kataoka, T. Mukohara, H. Tomioka, Y. Funakoshi, N. Kiyota, Y. Fujiwara,
M. Yashiro, K. Hirakawa, M. Hirai, H. Minami, Invest. New Drugs 30 (2012)
1352e1360.
[14] F. Cecchi, D.C. Rabe, D.P. Bottaro, Eur. J. Cancer 4 6 (2010) 1260e1270.
[15] F. Qian, S. Engst, K. Yamaguchi, P. Yu, K.A. Won, L. Mock, T. Lou, J. Tan, C. Li,
D. Tam, J. Lougheed, F.M. Yakes, F. Bentzien, W. Xu, T. Zaks, R. Wooster,
J. Greshock, A.H. Joly, Cancer Res. 69 (2009) 8009e8016.
[16] I. González-Sánchez, J.D. Solano, M.A. Loza-Mejía, S. Olvera-Vázquez,
R. Rodríguez-Sotres, J. Morán, A. Lira-Rocha, M.A. Cerbón, Eur. J. Med. Chem.
46 (2011) 2102e2108.
[17] R. Abonia, D. Insuasty, J. Castillo, B. Insuasty, J. Quiroga, M. Nogueras, J. Cobo,
Eur. J. Med. Chem. 57 (2012) 29e40.
[18] Y. Ai, Y.J. Liang, J.C. Liu, H.W. He, Y. Chen, C. Tang, G.Z. Yang, L.W. Fu, Eur. J.
Med. Chem. 47 (2012) 206e213.
[19] S. Li, Y.F. Zhao, K.W. Wang, Y.L. Gao, J.M. Han, B.B. Cui, P. Gong, Bioorg. Med.
Chem. 21 (2013) 2843e2855.
[20] P. Gong, Y.J. Liu, Y.F. Zhao, X. Zhai, S. Li, CN Patent ZL201310045375.5.
[21] J.W. Yao, J. Chen, Z.P. He, W. Sun, W.F. Xu, Bioorg. Med. Chem. 20 (2012)
2923e2929.
[22] R.K. Anchoori, M.S.Q. Kortenhorst, M. Hidalgo, T. Sarkar, G. Hallur, R.L. Bai,
P.J.V. Diest, E. Hamel, S.R. Khan, J. Med. Chem. 51 (2008) 5953e5957.
[23] S. Kotha, K. Lahiri, D. Kashinath, Tetrahedron 58 (2002) 9633e9695.
[24] A.F. Littke, G.C. Fu, Angew. Chem., Int. Ed. Engl. 37 (1999) 3387e3388.
[25] F. Minisci, A. Citterio, E. Vismara, Tetrahedron 41 (1985) 4151e4170.
[26] I. Martin, J. Anvelt, L. Vares, I. Kühn, A. Claesson, Act. Chem. Scand. 49 (1995)
230e232.
5.5. Pharmacology
5.5.1. MTT assay in vitro
The anti-proliferative activities of compounds 18aead and 25ae
n were evaluated against HT-29, H460, MKN-45, A549 and U87MG
cell lines using the standard MTT assay in vitro, with foretinib as the
positive control. The cancer cell lines were cultured in minimum
essential medium (MEM) supplemented with 10% fetal bovine
serum (FBS). Approximate 4 ꢃ 103 cells, suspended in MEM me-
dium, were plated onto each well of a 96-well plate and incubated
in 5% CO2 at 37 ꢀC for 24 h. The compounds tested 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 of
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
three times in each cell line. The results expressed as IC50 (inhibi-
tory concentration 50%) were the averages of three determinations
and calculated by using the Bacus Laboratories Incorporated Slide
Scanner (Bliss) software.
5.5.2. Tyrosine kinases assay
The tyrosine kinases activities were evaluated using homoge-
neous time-resolved fluorescence (HTRF) assays, as previously
reported protocol [28,29]. Briefly, 20
(Sigma) was preloaded as a substrate in 384-well plates. Then
50 L of 10 mM ATP (Invitrogen) solution diluted in kinase reac-
mg/mL poly (Glu, Tyr) 4:1
m
tion buffer (50 mM HEPES, pH 7.0, 1 M DTT, 1 M MgCl2, 1 M MnCl2,
and 0.1% NaN3) was added to each well. Various concentrations of
compounds diluted in 10
negative control. The kinase reaction was initiated by the addition
of purified tyrosine kinase proteins diluted in 39 L of kinase re-
mL of 1% DMSO (v/v) were used as the
m
action buffer solution. The incubation time for the reactions was
30 min at 25 ꢀC, and the reactions were stopped by the addition of
5 mL of Streptavidin-XL665 and 5 mL Tk Antibody Cryptate working
solution to all of wells. The plates were read using Envision (Per-
kinElmer) at 320 nm and 615 nm. The inhibition rate (%) was
calculated using the following equation: % inhibition ¼ 100
[27] H. Bernard, G. Bülow, U.E.W. Lange, H. Mack, T. Pfeiffer, B. Schäfer, W. Seitz,
T. Zierkea, Synthesis 14 (2004) 2367e2375.
ꢁ
[(activity of enzyme with tested compounds
ꢁ
Min)/
[28] B.K. Albrecht, J.C. Harmange, D. Bauer, L. Berry, C. Bode, A.A. Boezio, A. Chen,
D. Choquette, I. Dussault, C. Fridrich, R. Shimanovich, S.K. Springer, Y. Teffera,
Y. Yang, Y. Zhang, S.F. Bellon, J. Med. Chem. 51 (2008) 2879e2882.
[29] L.B. Liu, A. Siegmund, N. Xi, P. Kaplan-Lefko, K. Rex, A. Chen, J. Lin, J. Moriguchi,
L. Berry, L.Y. Huang, Y. Teffera, Y.J. Yang, Y.H. Zhang, S.F. Bellon, M. Lee,
R. Shimanovich, A. Bak, C. Dominguez, M.H. Norman, J.C. Harmange,
I. Dussault, T.S. Kim, J. Med. Chem. 51 (2008) 3688e3691.
(Max
ꢁ
Min)]
ꢃ
100 (Max: the observed enzyme activity
measured in the presence of enzyme, substrates, and cofactors;
Min: the observed enzyme activity in the presence of substrates,
cofactors and in the absence of enzyme). IC50 values were calcu-
lated from the inhibition curves.