Design, Synthesis, and Biological Evaluation of 4-Phenoxyquinoline Derivatives Containing Benzo[d]thiazole-2-yl Urea as c-Met Kinase Inhibitors

Article abstract of DOI:10.1002/ardp.201600003

A series of novel 4-phenoxyquinoline derivatives containing the benzo[d]thiazole-2-yl urea moiety were synthesized and evaluated for their cytotoxicity against the HT-29, MKN-45, and H460 cell lines. The structures of the target compounds were confirmed by ¹H NMR and MS spectra. Most of them showed moderate to excellent potency against the three tested cell lines. Especially, compound 23 was identified a promising agent (c-Met IC₅₀ = 17.6 nM), showing the most potent anticancer activities with IC₅₀ values of 0.18, 0.06, and 0.01 μM against the HT-29, MKN-45, and H460 cell lines, respectively. The docking results of 23 with the c-Met kinase model 3LQ8 showed a specific binding mode between the ligand and the target protein.

Full text of DOI:10.1002/ardp.201600003

RCHH HARM
A P
Arch. Pharm. Chem. Life Sci. 2016, 349, 1–11
Archiv der Pharmazie
Full Paper
Design, Synthesis, and Biological Evaluation of
4-Phenoxyquinoline Derivatives Containing Benzo[d]thiazole-2-yl
Urea as c-Met Kinase Inhibitors
Hongrui Lei, Gang Hu, Yu Wang, Pei Han, Zijian Liu, Yanfang Zhaoꢀ, and Ping Gong
Key Laboratory of Structure-Based Drug Design and Discovery, Ministry of Education, Shenyang
Pharmaceutical University, Shenyang, P. R. China
A series of novel 4-phenoxyquinoline derivatives containing the benzo[d]thiazole-2-yl urea moiety
were synthesized and evaluated for their cytotoxicity against the HT-29, MKN-45, and H460 cell lines.
1
The structures of the target compounds were confirmed by H NMR and MS spectra. Most of them
showed moderate to excellent potency against the three tested cell lines. Especially, compound 23
was identified a promising agent (c-Met IC50 ¼ 17.6 nM), showing the most potent anticancer activities
with IC50 values of 0.18, 0.06, and 0.01 mM against the HT-29, MKN-45, and H460 cell lines,
respectively. The docking results of 23 with the c-Met kinase model 3LQ8 showed a specific binding
mode between the ligand and the target protein.
Keywords: 4-Phenoxyquinoline derivatives / Benzo[d]thiazole-2-yl urea / c-Met kinase inhibitor /
Cytotoxicity / Synthesis
Received: January 8, 2016; Revised: May 11, 2016; Accepted: May 20, 2016
DOI 10.1002/ardp.201600003
Additional supporting information may be found in the online version of this article at the publisher’s web-site.
:
tyrosine kinase expressed in both normal and malignant
cells [2, 3], is activated in physiological conditions by its
Introduction
Cancer has been a leading cause of death due to drug
resistance and adverse side-effects [1]. c-Met, a receptor
natural ligand, hepatocyte growth factor (HGF). It plays an
important role in establishing the normal tissue pattern and
protecting the normal architecture of organs [4, 5]. When the
HGF/c-Met signaling pathway gets disordered, the activation
of this pathway would result in tumor growth, division, and
transfer. The disordered pathway shows high expression
activity, high mutation rate, more tumor tissue changes in vast
majority of cancers, such as lung cancer, pancreatic cancer,
breast cancer, prostate cancer, and ovarian cancer [6].
Correspondence: Prof. Ping Gong, Key Laboratory of
Structure-Based Drug Design and Discovery, Ministry of
Education, Shenyang Pharmaceutical University, Shenyang
110016, P. R. China.
E-mail: gongpinggp@126.com
At present, small molecule c-Met kinase inhibitors can be
divided into two groups based on their different binding
mode with the active site: type I inhibitors and type II
inhibitors [7–10]. Type I inhibitors (Fig. 1) function as an
analog of ATP to competitively bind the hinge region of c-Met
kinase [11–16]. As the ATP binding region is a highly
conservative amino acid sequence, there remains little space
Fax: þ86-24-23986429
Abbreviations: ATP, adenosine triphosphate; BT2U, benzo[d]-
thiazole-2-yl urea; DTT, dithiothreitol; EDG, electron-donating
group; EDTA, ethylene diamine tetraacetic acid; ELISA, enzyme-
linked immunosorbent assay; ESI, electrospray ionization; EWG,
electron-withdrawing group; FBS, fetal bovine serum; HEPES, 4-2-
hydroxyethyl-1-piperazineethanesulfonic acid; HTRF, homoge-
neous time-resolved fluorescence; IC50, half maximal inhibitory
concentration; MEM, minimum essential medium; MTT, 3-(4,5-
dimethylthiazae-2-yl)-2,3-diphenyltetrazolium bromide; SAR,
structure–activity relationship; UV, ultraviolet.
ꢀAdditional correspondence: Dr. Yanfang Zhao,
E-mail: yanfangzhao@126.com
ß 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com
1

RCHH HARM
A P
Arch. Pharm. Chem. Life Sci. 2016, 349, 1–11
H. Lei et al.
Archiv der Pharmazie
Figure 1. Representative structures of type I inhibitors.
for further development of new drugs. However, type II
inhibitors (Fig. 2) can stretch into the ATP binding region and
the hinge region to form crucial hydrogen bonds. Meanwhile,
they also have lipophilic interactions with the hydrophobic
region of the “allosteric site” to make this type of inhibitors
more amendable. Consequently, study on type II inhibitors to
develop novel chemical entities is much more rational and of
great significance.
phenoxyquinoline derivatives as c-Met inhibitors (Fig. 3).
From the study of the structure–activity relationships
(SARs), the reported linkers had two common character-
istics between moiety A and moiety B. One was “5-atom
regulation”, which meant the six-chemical-bond distance
between moiety A and moiety B; the other was the linker
containing “hydrogen-bond donor and acceptor” [18].
Therefore, we had introduced quinoline, semicarbazone,
and 1,4-dihydrocinnoline fragments into the 5-atom
linker, and the resulting derivatives showed moderate-
to-excellent potency [19–22]. Additionally, R1 moiety
Foretinib is an oral multi-kinase type II inhibitor
targeting MET, VEGFR, RON, KIT, and AXL kinases [17].
Previously, we described some series of 6,7-disubstituted-4-
Figure 2. Representative structures of type II inhibitors.
ß 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com
2

RCHH HARM
A P
Arch. Pharm. Chem. Life Sci. 2016, 349, 1–11
4-Phenoxyquinoline Derivatives as c-Met Kinase Inhibitors
Archiv der Pharmazie
Figure 3. Representative structures of previously reported c-Met inhibitors.
usually existed in 7-position of quinoline, where a water-
soluble fatty-amino fragment was added to the three-
carbon moiety to improve its pharmacokinetic properties.
While, the R2 moiety varied from electron-donating groups
(EDGs) to electron-withdrawing groups (EWGs). The EDGs
included hydroxyl group, methoxyl group, while the EWGs
are often halogen atoms.
thiazole-2-yl urea (BT2U) moiety were designed and synthe-
sized (Fig. 5).
Results and discussion
Chemistry
Benzothiazole derivatives had
a
variety of biological
As shown in Scheme 1, the key intermediate 3-fluoro-4-(6-
methoxy-7-((3-substituted aminopropyl)oxy)quinolin-4-yloxy)-
aniline (20a–d) was prepared from acetovanillone which was
claimed in detail in our previous study [27].
activities such as antifungal [23], antihypoglycemic [24],
antitumor activity [25], and so on. In recent years, a series
of antitumor compounds with benzothiazole moiety had
been designed and synthesized, such as PMX610, NSC745689,
and triflorcas (Fig. 4) [26]. PMX610 exhibited a 0.1 nM GI50
value toward human breast carcinoma cells MCF-7 and
MDA-MB-468, while NSC745689 selectively inhibited human
non-small-cell lung cancer cells (NSCLC) with an IC50 value
of 2.5 mM. Triflorcas, containing a benzothiazole moiety,
exerted a 171 nM IC50 value toward c-Met kinase, which
accordingly drew our attention on it. According to the SARs of
quinoline derivatives as c-Met inhibitors, aminobenzothiazole
moiety was introduced into 4-phenoxy-6,7-disubstituted
quinolines on the basis of combination principles. As a result,
a series of novel quinoline derivatives bearing benzo[d]-
Compounds 23–43 were synthesized according to the
procedures outlined in Scheme 2. Addition reactions of the
commercially available anilines with NH4SCN were achieved in
acetic acid by adding Br2 dropwise at a temperature lower
than 10°C. The reaction was continued for 2 h after the adding
of Br2 and another 1 h was conducted at room temperature to
yield intermediates 21a–j [28]. Subsequently, intermediates
21a–j reacted with phenyl chloroformate in THF under basic
conditions (pyridine) to provide 22a–j. Finally, intermediates
20a–d refluxed with 22a–j in dioxane for 7 h to obtain target
compounds 23–43. Their spectral data were in full agreement
with the proposed structures.
Figure 4. Structures of antitumor compounds with benzothiazole moiety reported in recent years.
ß 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com
3

RCHH HARM
A P
Arch. Pharm. Chem. Life Sci. 2016, 349, 1–11
H. Lei et al.
Archiv der Pharmazie
Figure 5. Design strategy for the 4-phenoxyquinoline derivatives containing benzo[d]thiazole-2-yl urea moiety.
Biology
specific effect of the benzo[d]thiazole-2-yl urea moiety.
Secondly, introduction of different R1 groups (4-methyl
piperazinyl, morpholinyl, 4-methyl piperidinyl, piperidinyl)
at 7-position of quinoline only slightly changed cytotoxicity,
which suggested that these R1 groups had little influence on
the cytotoxicity. For instance, compounds 25, 31, 38, and 39
exhibited comparable anticancer activity with IC50 values
ranging from 1.4 to 1.8 mM against the HT-29 cell line. This
exactly achieves our goal to introduce R1 with an intention to
improve their pharmacokinetic properties (water solubility).
Thirdly, different biological properties were obtained when
various R2 groups were introduced into the benzo[d]thiazole
ring. For example, compounds 32, 33, and 34 with trifluoro-
methyl or trifluoromethoxy group showed lower anticancer
activity than compounds 30 and 31 with no substituent or
bromine substitution. Further analysis indicated that the
nature and the quantity of R2 group had a significant impact
on the activity. The lipophilicity affects the interaction with
the active site much more than the effect resulting from steric
In vitro cytotoxicity activities and SARs
As illustrated in Table 1, most of the prepared compounds
showed moderate-to-excellent potency against three tested
cell lines in the single-digit mM range. The pharmacological
data indicated that five compounds (23, 24, 29, 30, and 37)
showed significant anticancer activity against HT-29 cell line.
Among them, the anticancer activities of compounds 23
(IC50 ¼ 0.18 mM) and 29 (IC50 ¼ 0.21 mM) were comparable
with foretinib. Six compounds (23, 24, 26, 29, 30, and 37)
showed significant anticancer activity against MKN-45 cell
line, in which compound 24 (IC50 ¼ 0.02 mM) showed superior
activity to foretinib, especially. Most compounds showed
excellent activity against H460 and 11 (23, 24, 25, 26, 29, 30,
31, 37, 38, 39, and 42) of them showed superior activity to
foretinib.
From the above results, firstly, we could see that introduc-
tion of the benzo[d]thiazole-2-yl urea moiety could produce
similar anticancer activity with foretinib which indicates the
Scheme 1. Synthetic route to the 4-phenoxy quinoline moieties. Reagents and conditions: a. Br(CH2)3Cl, acetone, 0ºC, 30 min, rt,
12 h; b. 98% HNO3, CH2Cl2, 0ºC, 4 h; c. DMF-DMA, toluene, 110ºC, 10 h; d. Fe powder, AcOH, rt, 30 min, 80ºC, 2 h; e. secondary
amines, CH3CN, 85ºC, 10 h; f. POCl3, 85ºC, 6 h; g. 2-fluoro-4-nitrophenol, PhCl, 140ºC, 30 h; h. Fe powder, NH4Cl (cat.), EtOH/H2O,
reflux, 5 h.
ß 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com
4

RCHH HARM
A P
Arch. Pharm. Chem. Life Sci. 2016, 349, 1–11
4-Phenoxyquinoline Derivatives as c-Met Kinase Inhibitors
Archiv der Pharmazie
Scheme 2. Synthetic route to the target compounds 23–43. Reagents and conditions: (a) NH4SCN, Br2, AcOH, r.t; (b) phenyl
chloroformate, Py, THF, r.t; (c) dioxane, reflux.
hindrance. Judging from compounds 23 to 28, we could
realize that antiproliferative activity decreased significantly
when the benzo[d]thiazole ring was substituted with an
EWG. And the more EWGs existed, the more the activity
decreased, such as compound 23 (4,7-(CH3)2, IC50 ¼ 0.06 mM)
and 28 (4-F, 6-Br, IC50 ¼ 0.42 mM) against MKN-45 cell lines.
At the end, all the compounds tested showed a stronger
selectivity upon MKN-45 and H460 than HT-29, which may
suggest that c-Met kinase are higher expressed in the former
two kinds of cell lines.
Table 1. Cytotoxic activities of target compounds against HT-29, MKN-45, and H460 cancer cell lines in vitro.
IC50 (mM)
Compd.
23
R1
R2
HT-29
MKN-45
H460
N-Methyl-1-
piperazinyl
4,7-(CH3)2
0.18 ꢁ 0.02
0.06 ꢁ 0.01
0.01 ꢁ 0.003
24
25
26
27
28
N-Methyl-1-
piperazinyl
N-Methyl-1-
piperazinyl
N-Methyl-1-
piperazinyl
N-Methyl-1-
piperazinyl
N-Methyl-1-
piperazinyl
H
0.26 ꢁ 0.03
1.5 ꢁ 0.10
2.4 ꢁ 0.11
6.4 ꢁ 0.12
7.5 ꢁ 0.21
0.02 ꢁ 0.005
0.11 ꢁ 0.01
0.22 ꢁ 0.02
0.30 ꢁ 0.02
0.42 ꢁ 0.05
0.05 ꢁ 0.007
0.08 ꢁ 0.006
0.15 ꢁ 0.02
0.27 ꢁ 0.06
0.37 ꢁ 0.09
5-Br
4-OCF3
6-Cl
4-F,6-Br
29
30
31
32
33
34
35
36
37
4-Morpholinyl
4-Morpholinyl
4-Morpholinyl
4-Morpholinyl
4-Morpholinyl
4-Morpholinyl
4-Morpholinyl
4-Morpholinyl
4-Methyl-1-
piperidinyl
4,7-(CH3)2
H
0.21 ꢁ 0.03
0.70 ꢁ 0.05
1.6 ꢁ 0.09
3.4 ꢁ 0.13
6.5 ꢁ 0.15
5.3 ꢁ 0.15
4.5 ꢁ 0.11
7.2 ꢁ 0.16
0.46 ꢁ 0.04
0.03 ꢁ 0.005
0.04 ꢁ 0.005
0.20 ꢁ 0.01
0.44 ꢁ 0.03
0.56 ꢁ 0.05
0.55 ꢁ 0.08
0.42 ꢁ 0.06
0.43 ꢁ 0.05
0.04 ꢁ 0.004
0.09 ꢁ 0.02
0.14 ꢁ 0.03
0.09 ꢁ 0.02
0.86 ꢁ 0.08
0.91 ꢁ 0.06
0.23 ꢁ 0.06
0.24 ꢁ 0.06
0.18 ꢁ 0.06
0.06 ꢁ 0.02
5-Br
4-OCH3
4-CF3
6-Cl
5-CF3
4,6-(Cl)2
H
38
4-Methyl-1-
piperidinyl
5-Br
1.8 ꢁ 0.08
0.25 ꢁ 0.01
0.08 ꢁ 0.02
39
40
41
42
43
Foretinib
1-Piperidinyl
1-Piperidinyl
1-Piperidinyl
1-Piperidinyl
1-Piperidinyl
4-Morpholinyl
5-Br
6-Br
4-OCF3
6-Cl
5-CF3
–
1.4 ꢁ 0.09
2.1 ꢁ 0.15
3.2 ꢁ 0.13
4.2 ꢁ 0.18
3.2 ꢁ 0.11
0.18 ꢁ 0.02
0.23 ꢁ 0.04
0.30 ꢁ 0.04
0.49 ꢁ 0.06
0.52 ꢁ 0.05
0.34 ꢁ 0.04
0.031 ꢁ 0.004
0.12 ꢁ 0.03
0.24 ꢁ 0.05
0.45 ꢁ 0.07
0.10 ꢁ 0.02
0.49 ꢁ 0.06
0.18 ꢁ 0.04
ß 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com
5

RCHH HARM
A P
Arch. Pharm. Chem. Life Sci. 2016, 349, 1–11
H. Lei et al.
Archiv der Pharmazie
Table 3. Inhibition of tyrosine kinases by compound 23.
In vitro enzymatic assays
As shown in Table 2, compounds 23, 24, and 29 all exhibited
excellent c-Met enzymatic potency, suggesting that the
inhibition of c-Met is a candidate underlying mechanism
for the anticancer effect. Compound 23 showed the most
potent activity with an IC50 value of 17.6 nM, which was
comparable to foretinib (IC50 ¼ 11.7 nM). Thus, compound 23
is worth further studying as a new potential anticancer agent.
Kinase
Enzyme IC50 (nM)
Flt-3
VEGFR-2
EGFR
136.7
3361
>20000
Enzymatic selectivity assays
an important role in stabilizing the conformation of ligand–
To examine the selectivity of compound 23 on c-Met over
other kinases, it was screened against three other tyrosine
kinases (Table 3). Compared with its relatively high potency
against c-Met (IC50 ¼ 17.6 nM), 23 also exhibited inhibitory
effects against Flt-3 and VEGFR-2, although the potency was
8-, 190-fold lower than that against c-Met. Moreover, this
compound showed little or no kinase inhibition activity
against EGFR kinase (IC50 > 20000 nM). These data suggested
that compound 23 is a promising selective kinase inhibitor.
protein complex.
Conclusions
In summary, 21 novel 4-phenoxyquinoline derivatives con-
taining benzo[d]thiazole-2-yl urea moiety were synthesized
and evaluated for their cytotoxicity against three cell lines
with foretinib as the reference. Most compounds showed
moderate-to-excellent activity against H460 and MKN-45 cell
lines. Eleven (23, 24, 25, 26, 29, 30, 31, 37, 38, 39, and 42) of
them showed superior activity to foretinib. Compound 23
(c-Met IC50 ¼ 17.6 nM) showed the most potent anticancer
activities with IC50 values of 0.18, 0.06, and 0.01 mM against
HT-29, MKN-45, and H460 cell lines, respectively. The potency
against H460 was much higher than foretinib by 18-fold. The
preliminary SARs indicated that the nature and quantity of R2
groups on the benzothiazole moiety have the most important
influence on the anticancer activity. The docking results of 23
with c-Met kinase 3LQ8 showed a specific binding mode
between ligand and target protein.
Binding model analysis
To further elucidate the binding mode of compounds, a
docking analysis was performed. In our study, the co-crystal
structure of foretinib (GSK1363089) with c-Met kinase was
selected as the docking model (PDB ID: 3LQ8). The docking
simulation was conducted with Molegro Virtual Docker
(MVD), since MVD uses a hierarchical series of filters to
search for possible locations of the ligand in the active-site
region of the receptor. The shape and properties of the
receptor were represented on a grid by several different sets
of fields that provide progressively more accurate scoring of
the ligand poses. The image files were generated by Accelrys
DS visualizer 4.0 system. The binding model was exemplified
by the interaction of compound 23 with c-Met. As shown
in Fig. 6, the nitrogen atom of quinoline in compound 23
formed H-bond interactions with residue Met1160, while
the carbonyl oxygen atom of urea moiety in compound 23
formed a hydrogen bond with Asp1222. In the meantime,
there exists one p–p interaction between the phenyl ring
(moiety A) and Phe1223. The N-methylpiperazinyl moiety
stretches into the solvent region to improve its pharmaco-
kinetic property. Both p–p and H-bond interactions played
Experimental
Chemistry
All melting points were obtained on a Buchi Melting Point
B-540 apparatus (Buchi Labortechnik AG, Flawil, Switzerland)
€
€
Table 2. Inhibitory activity against c-Met kinase of
compounds 23, 24, and 29 in vitro.
a)
Compd.
IC50 on c-Met (nM)
23
24
17.6 ꢁ 1.17
22.4 ꢁ 2.31
23.4 ꢁ 1.73
11.7 ꢁ 0.95
29
Foretinibb)
a) Data presented is the mean ꢁ SD value of three independent
determinations. b)Used as positive control.
Figure 6. The c-Met active site in complex with compound 23.
ß 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com
6

RCHH HARM
A P
Arch. Pharm. Chem. Life Sci. 2016, 349, 1–11
4-Phenoxyquinoline Derivatives as c-Met Kinase Inhibitors
Archiv der Pharmazie
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-400, 400 MHz spectrometers
(Bruker Bioscience, Billerica, MA, USA) with TMS as an internal
standard.
Enzyme assays were performed at the Shanghai Chem-
partner Co., Ltd. Unless otherwise noted, all materials were
obtained from commercially available sources and were used
without further purification.
CH2CH2O), 3.96 (s, 3H, CH3O), 2.48 (m, 10H, CH2NCH2, NCH2),
2.44 (s, 3H, NCH3), 2.29 (s, 3H, benzothiazol-4-CH3), 2.25 (s, 3H,
benzothiazol-7-CH3), 1.96 (m, 2H, CH2CH2CH2); 13C NMR
(100 MHz, CDCl3) d 161.3, 155.9, 153.3, 152.9, 152.5, 150.0,
148.5, 146.8, 139.2, 139.0, 138.3, 134.8, 127.3, 124.0, 123.9,
116.3, 115.5, 115.1, 111.3, 108.5, 108.2, 102.2, 99.7, 67.5,
56.3, 55.2 (2C), 53.2, 46.2 (2C), 26.3, 20.6, 17.3, 10.5. MS (ESI),
m/z: 645.2 [MþH]þ.
1-(Benzothiazol-2-yl)-3-(3-fluoro-4-(6-methoxy-7-(3-(4-
methylpiperazin-1-yl)propoxy)quinolin-4-yloxy)phenyl)-
urea (24)
The InChI codes of the new compounds are provided in the
online Supporting Information.
Yellow solid; yield: 89.6%; mp: 187–188°C; 1H NMR (400 MHz,
CDCl3), d: 8.75 (s, 2H, NH), 8.44 (d, J ¼ 5.0 Hz, 1H, quinolin-2H),
7.67–7.51 (m, 4H, Ar-H), 7.48–7.42 (m, 2H, Ar-H), 7.38 (s, 1H,
quinolin-8H), 7.16 (m, 1H, Ar-H), 7.14 (t, J ¼ 8.7 Hz, 1H, Ar-H),
6.39 (d, J ¼ 5.0 Hz, 1H, quinolin-3H), 4.19 (t, J ¼ 5.2 Hz, 2H,
CH2CH2O), 4.01 (s, 3H, CH3O), 2.61 (m, 10H, CH2NCH2, NCH2),
2.38 (s, 3H, NCH3), 2.11 (m, 2H, CH2CH2CH2). MS (ESI), m/z:
617.2 [MþH]þ.
General procedure for the preparation of compounds
21a–j
Addition reactions of the commercial available anilines
(0.01 mol) with NH4SCN (0.05 mol) were achieved in acetic
acid by adding Br2 dropwise at a temperature lower than
10°C. The reaction was continued for 2 h after the adding of
Br2 (0.011 mol) and another 1 h was conducted at room
temperature to yield the intermediates 21a–j. Then, the
reaction mixture was filtered, and the cake was collected,
dissolved in hot water and filtrated. Finally, the filtrates were
combined and adjusted to pH 9 with aqueous ammonia to get
the yellow solids.
1-(5-Bromobenzothiazol-2-yl)-3-(3-fluoro-4-(6-methoxy-7-
(3-(4-methylpiperazin-1-yl)propoxy)quinolin-4-yloxy)-
phenyl)urea (25)
Yellow solid; yield: 85.5%; mp: 233–234°C; 1H NMR (400 MHz,
CDCl3), d: 9.54 (s, 1H, NH), 8.49 (d, J ¼ 5.0 Hz, 1H, quinolin-2H),
8.41 (dd, J ¼ 8.4 Hz, 2.0 Hz, 1H, benzothiazol-6H), 7.98 (d,
J ¼ 1.9 Hz, 1H, benzothiazol-4H), 7.76–7.61 (m, 2H, Ar-H), 7.59
(s, 1H, quinolin-5H), 7.47 (d, J ¼ 8.4 Hz, 1H, benzothiazol-7H),
7.42 (s, 1H, quinolin-8H), 7.30 (s, 1H, NH), 7.17 (t, J ¼ 8.7 Hz, 1H,
phenyl-6H), 6.42 (d, J ¼ 5.0 Hz, 1H, quinolin-3H), 4.23 (t,
J ¼ 5.6 Hz, 2H, CH2CH2O), 4.04 (s, 3H, CH3O), 2.78–2.33 (m, 10H,
CH2NCH2, NCH2), 2.30 (s, 3H, NCH3), 2.09 (m, 2H, CH2CH2CH2).
MS (ESI), m/z: 696.1 [MþH]þ.
General procedure for the preparation of compounds
22a–j
Amonolysis reaction between the sufficiently dried 21a–j
(0.008 mol) and the phenyl chloroformate (0.016 mol) was
achieved under basic conditions in THF by treating with 1
equiv of pyridine under ice bath conditions. The phenyl
chloroformate was dissolved in THF and added dropwise
during an interval of 10 min. The reaction mixture was
stirred for another 30 min at room temperature, and it was
evaporated to yield yellow solid after the reaction.
1-(4-Trifluoromethoxybenzothiazol-2-yl)-3-(3-fluoro-4-(6-
methoxy-7-(3-(4-methylpiperazin-1-yl)propoxy)quinolin-
4-yloxy)phenyl)urea (26)
General procedure for the preparation of compounds
23–43
Yellow solid; yield: 83.5%; mp: 214–215°C; 1H NMR (400 MHz,
CDCl3), d: 9.91 (s, 1H, NH), 8.62 (d, J ¼ 8.8 Hz, 1H, benzothiazol-
5H), 8.57 (d, J ¼ 8.7 Hz, 1H, quinolin-2H), 8.47 (s, 1H, NH), 7.70
(dd, J ¼ 12.4, 2.2 Hz, 1H, phenyl-2H), 7.58 (s, 1H, quinolin-5H),
7.49–7.43 (m, 2H, Ar-H), 7.42 (s, 1H, quinolin-8H), 7.36
(d, J ¼ 8.7 Hz, 1H, benzothiazol-7H), 7.16 (t, J ¼ 8.7 Hz, 1H,
benzothiazol-6H), 6.41 (d, J ¼ 8.7 Hz, 1H, quinolin-3H), 4.24 (t,
J ¼ 6.3 Hz, 2H, CH2CH2O), 4.03 (s, 3H, CH3O), 2.98–2.50 (m, 10H,
CH2NCH2, NCH2), 2.44 (s, 3H, NCH3), 2.13 (m, 2H, CH2CH2CH2).
MS (ESI), m/z: 701.1 [MþH]þ.
The key intermediate 3-fluoro-4-(6-methoxy-7-((3-substituted
aminopropyl)oxy)quinolin-4-yloxy)aniline (20a–d) (0.001 mol)
and 22a–j (0.003 mol) was dissolved in dioxane (20 mL)
and refluxed at 100°C for 7 h. The reaction mixture was
evaporated to obtain the crude product, which was purified
by silica gel chromatography with CH2Cl2/MeOH (30:1, v/v) to
afford the target compounds 23–43.
1-(4,7-Dimethylbenzothiazol-2-yl)-3-(3-fluoro-4-(6-
methoxy-7-(3-(4-methylpiperazin-1-yl)propoxy)quinolin-
4-yloxy)phenyl)urea (23)
1-(6-Chlorobenzothiazol-2-yl)-3-(3-fluoro-4-(6-methoxy-7-
(3-(4-methylpiperazin-1-yl)propoxy)quinolin-4-yloxy)-
phenyl)urea (27)
Yellow solid; yield: 83.1%; mp: 195–196°C; 1H NMR (400 MHz,
DMSO-d6), d: 9.94 (s, 1H, NH), 8.48 (d, J ¼ 5.1 Hz, 1H, quinolin-
2H), 8.44 (s, 1H, NH), 8.07 (s, 1H, quinolin-5H), 7.80 (dd,
J ¼ 12.9 Hz, 2.2 Hz, 1H, phenyl-2H), 7.58–7.50 (m, 2H, Ar-H),
7.47–7.36 (m, 2H, Ar-H), 7.27 (d, J ¼ 8.4 Hz, 1H, benzothiazol-
H), 6.45 (d, J ¼ 5.1 Hz, 1H, quinolin-3H), 4.19 (t, J ¼ 5.6 Hz, 2H,
Yellow solid; yield: 79.8%; mp: 203–205°C; 1H NMR (400 MHz,
CDCl3), d: 8.48 (d, J ¼ 5.1 Hz, 1H, quinolin-2H), 7.74 (dd,
J ¼ 12.1 Hz, 2.0 Hz, 1H, phenyl-2H), 7.69 (d, J ¼ 2.2 Hz, 1H,
benzothiazol-7H), 7.62 (d, J ¼ 8.8 Hz, 1H, phenyl-5H), 7.52
ß 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com
7

RCHH HARM
A P
Arch. Pharm. Chem. Life Sci. 2016, 349, 1–11
H. Lei et al.
Archiv der Pharmazie
(s, 1H, quinolin-5H), 7.41 (s, 1H, quinolin-8H), 7.38 (t,
J ¼ 8.8 Hz, 1H, phenyl-6H), 7.27 (m, 4H, NH, Ar-H), 6.40 (d,
J ¼ 5.1 Hz, 1H, quinolin-3H), 4.21 (t, J ¼ 6.5 Hz, 2H, CH2CH2O),
3.99 (s, 3H, CH3O), 2.70–2.40 (m, 10H, CH2NCH2, NCH2), 2.29 (s,
3H, NCH3), 2.10 (m, 2H, CH2CH2CH2). MS (ESI), m/z: 651.2
[MþH]þ.
(t, J ¼ 8.8 Hz, 1H, phenyl-6H), 7.40 (s, 1H, quinolin-8H), 7.28 (d,
J ¼ 8.8 Hz, 1H, benzothiazol-7H), 6.45 (d, J ¼ 5.2 Hz, 1H,
quinolin-3H), 4.20 (t, J ¼ 6.2 Hz, 2H, CH2CH2O), 3.96 (s, 3H,
CH3O), 3.59 (t, J ¼ 4.0 Hz, 4H, CH2OCH2), 2.46 (t, J ¼ 7.0 Hz, 2H,
NCH2), 2.39 (s, 4H, CH2NCH2), 1.98 (m, 2H, CH2CH2CH2).
MS (ESI), m/z: 682.1 [MþH]þ.
1-(6-Bromo-4-fluorobenzothiazol-2-yl)-3-(3-fluoro-4-(6-
methoxy-7-(3-(4-methyl-piperazin-1-yl)propoxy)quinolin-
4-yloxy)phenyl)urea (28)
1-(4-Trifluoromethoxybenzothiazol-2-yl)-3-(3-fluoro-4-(6-
methoxy-7-(3-(morpholin-4-yl)propoxy)quinolin-4-yloxy)-
phenyl)urea (32)
Yellow solid; yield: 87.6%; mp: 268–269°C; 1H NMR (400 MHz,
CDCl3), d: 10.16 (br, 1H, NH), 8.47 (d, J ¼ 5.2 Hz, 1H, quinolin-
2H), 7.78–7.65 (m, 2H, Ar-H), 7.57 (s, 1H, quinolin-5H), 7.42 (s,
1H, quinolin-8H), 7.33–7.15 (m, 4H, Ar-H, NH), 6.39 (d,
J ¼ 5.2 Hz, 1H, quinolin-3H), 4.24 (t, J ¼ 6.5 Hz, 2H, CH2CH2O),
4.02 (s, 3H, CH3O), 2.65–2.35 (m, 10H, CH2NCH2, NCH2), 2.31 (s,
3H, NCH3), 2.13 (m, 2H, CH2CH2CH2). MS (ESI), m/z: 713.9
[MþH]þ.
Yellow solid; yield: 89.5%; mp: 252–253°C; 1H NMR
(400 MHz, DMSO-d6), d: 9.74 (s, 1H, NH), 8.87 (s, 1H, NH),
8.48 (d, J ¼ 4.8 Hz, 1H, quinolin-2H), 8.45 (d, J ¼ 8.8 Hz, 1H,
benzothiazol-5H), 7.85–7.75 (m, 2H, Ar-H), 7.71 (d, J ¼ 8.7
Hz, 1H, phenyl-2H), 7.54 (s, 1H, quinolin-5H), 7.44 (t,
J ¼ 8.8 Hz, 1H, benzothiazol-6H), 7.41 (s, 1H, quinolin-8H),
7.28 (d, J ¼ 8.6 Hz, 1H, benzothiazol-7H), 6.46 (d, J ¼ 4.8 Hz,
1H, quinolin-3H), 4.21 (t, J ¼ 5.6 Hz, 2H, CH2CH2O), 3.96
(s, 3H, CH3O), 3.60 (s, 4H, CH2OCH2), 2.51 (s, 2H, NCH2),
2.42 (s, 4H, CH2NCH2), 2.00 (m, 2H, CH2CH2CH2). MS (ESI),
m/z: 688.1 [MþH]þ.
1-(4,7-Dimethylbenzothiazol-2-yl)-3-(3-fluoro-4-(6-
methoxy-7-(3-(morpholin-4-yl)propoxy)quinolin-4-yloxy)-
phenyl)urea (29)
Yellow solid; yield: 88.9%; mp: 193–194°C; 1H NMR (400 MHz,
CDCl3), d: 8.66 (s, 1H, NH), 8.35 (d, J ¼ 5.4 Hz, 1H, quinolin-2H),
7.91 (s, 1H, NH), 7.59 (s, 1H, quinolin-5H), 7.56 (dd, J ¼ 8.8 Hz,
2.2 Hz, 1H, phenyl-5H), 7.41 (m, 2H, Ar-H), 7.38 (s, 1H, quinolin-
8H), 7.20 (dd, J ¼ 10.5 Hz, 2.0 Hz, 1H, phenyl-2H), 7.14 (t,
J ¼ 8.8 Hz, 1H, phenyl-6H), 6.39 (d, J ¼ 5.4 Hz, 1H, quinolin-3H),
4.24 (t, J ¼ 6.3 Hz, 2H, CH2CH2O), 4.03 (s, 3H, CH3O), 3.74 (t,
J ¼ 4.4 Hz, 4H, CH2OCH2), 2.63 (t, J ¼ 7.0 Hz, 2H, NCH2), 2.56 (s,
4H, CH2NCH2), 2.45 (s, 3H, benzothiazol-4-CH3), 2.17 (s, 3H,
benzothiazol-7-CH3), 2.13 (m, 2H, CH2CH2CH2). MS (ESI), m/z:
632.2 [MþH]þ.
1-(4-Trifluoromethylbenzothiazol-2-yl)-3-(3-fluoro-4-(6-
methoxy-7-(3-(morpholin-4-yl)propoxy)quinolin-4-yloxy)-
phenyl)urea (33)
Yellow solid; yield: 82.1%; mp: 245–247°C; 1H NMR (400 MHz,
CDCl3), d: 9.83 (s, 1H, NH), 8.46 (d, J ¼ 5.0 Hz, 1H, quinolin-2H),
8.43 (s, 1H, NH), 7.80 (s, 1H, Ar-H), 7.73 (m, 2H, Ar-H), 7.68–7.49
(m, 2H, Ar-H), 7.40 (s, 1H, quinolin-8H), 7.17 (m, 2H, Ar-H), 6.43
(d, J ¼ 5.0 Hz, 1H, quinolin-3H), 4.22 (s, 2H, CH2CH2O), 4.05 (s,
3H, CH3O), 3.71 (s, 4H, CH2OCH2), 2.56 (t, J ¼ 6.6 Hz, 2H, NCH2),
2.48 (s, 4H, CH2NCH2), 2.12 (m, 2H, CH2CH2CH2). MS (ESI), m/z:
672.0 [MþH]þ.
1-(Benzothiazol-2-yl)-3-(3-fluoro-4-(6-methoxy-7-(3-
(morpholin-4-yl)propoxy)quinolin-4-yloxy)phenyl)urea
(30)
1-(6-Chlorobenzothiazol-2-yl)-3-(3-fluoro-4-(6-methoxy-7-
(3-(morpholin-4-yl)propoxy)quinolin-4-yloxy)phenyl)urea
(34)
Yellow solid; yield: 85.2%; mp: 187–189°C; 1H NMR (400 MHz,
DMSO-d6), d: 9.40 (s, 2H, NH), 8.48 (d, J ¼ 5.0 Hz, 1H, quinolin-
2H), 7.76 (d, J ¼ 13.1 Hz, 1H, phenyl-2H), 7.70–7.58 (m, 4H, Ar-
H), 7.54 (s, 1H, quinolin-5H), 7.42 (s, 1H, quinolin-8H), 7.40 (t,
J ¼ 9.2 Hz, 1H, Ar-H), 7.29 (d, J ¼ 8.4 Hz, 1H, Ar-H), 6.46 (d,
J ¼ 5.0 Hz, 1H, quinolin-3H), 4.22 (t, J ¼ 5.2 Hz, 2H, CH2CH2O),
3.96 (s, 3H, CH3O), 3.65 (s, 4H, CH2OCH2), 2.61 (s, 2H, NCH2),
2.51 (s, 4H, CH2NCH2), 2.04 (m, 2H, CH2CH2CH2). MS (ESI), m/z:
626.2 [MþNa]þ.
Yellow solid; yield: 80.5%; mp: 214–215°C; 1H NMR (400 MHz,
DMSO-d6), d: 11.89 (s, 1H, NH), 10.26 (s, 1H, NH), 8.48 (d,
J ¼ 4.6 Hz, 1H, quinolin-2H), 8.02 (s, 1H, quinolin-5H), 7.82 (d,
J ¼ 12.6 Hz, 1H, phenyl-2H), 7.61 (d, J ¼ 8.1 Hz, 1H, Ar-H), 7.54
(s, 1H, quinolin-8H), 7.40 (m, 4H, Ar-H), 6.46 (s, J ¼ 4.6 Hz, 1H,
quinolin-3H), 4.21 (s, 2H, CH2CH2O), 3.96 (s, 3H, CH3O), 3.59 (s,
4H, CH2OCH2), 2.48 (s, 2H, NCH2), 2.40 (s, 4H, CH2NCH2), 1.98 (s,
2H, CH2CH2CH2). MS (ESI), m/z: 638.6 [MþH]þ.
1-(5-Trifluoromethylbenzothiazol-2-yl)-3-(3-fluoro-4-(6-
methoxy-7-(3-(morpholin-4-yl)propoxy)quinolin-4-yloxy)-
phenyl)urea (35)
1-(5-Bromobenzothiazol-2-yl)-3-(3-fluoro-4-(6-methoxy-7-
(3-(morpholin-4-yl)propoxy)quinolin-4-yloxy)phenyl)urea
(31)
Yellow solid; yield: 84.1%; mp: 240–242°C; 1H NMR (400 MHz,
CDCl3), d: 9.27 (s, 1H, NH), 8.92 (s, 1H, NH), 8.46 (d, J ¼ 5.4 Hz,
1H, quinolin-2H), 7.99 (d, J ¼ 1.9 Hz, 1H, benzothiazol-4H),
7.82–7.70 (m, 2H, Ar-H), 7.63 (dd, J ¼ 8.6 Hz, 2.0 Hz, 1H, phenyl-
5H), 7.60 (s, 1H, quinolin-5H), 7.39 (s, 1H, quinolin-8H), 7.21
(dd, J ¼ 9.2 Hz, 2.0 Hz, 1H, phenyl-2H), 7.15 (t, J ¼ 8.6 Hz,
1H, phenyl-6), 6.42 (d, J ¼ 5.2 Hz, 1H, quinolin-3H), 4.22
Yellow solid; yield: 88.6%; mp: 272–273°C; 1H NMR (400 MHz,
DMSO-d6), d: 10.06 (s, 1H, NH), 8.50 (s, 1H, NH), 8.48 (d,
J ¼ 5.2 Hz, 1H, quinolin-2H), 8.26 (dd, J ¼ 8.8 Hz, 2.0 Hz, 1H,
benzothiazol-6H), 8.00 (d, J ¼ 1.9 Hz, 1H, benzothiazol-4H),
7.77 (dd, J ¼ 12.7 Hz, 2.0 Hz, 1H, phenyl-2H), 7.68 (dd, J ¼ 8.8
Hz, 1.9 Hz, 1H, phenyl-5H), 7.54 (s, 1H, quinolin-5H), 7.44
ß 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com
8

RCHH HARM
A P
Arch. Pharm. Chem. Life Sci. 2016, 349, 1–11
4-Phenoxyquinoline Derivatives as c-Met Kinase Inhibitors
Archiv der Pharmazie
(t, J ¼ 6.5 Hz, 2H, CH2CH2O), 4.04 (s, 3H, CH3O), 3.71 (t,
J ¼ 4.4 Hz, 4H, CH2OCH2), 2.56 (t, J ¼ 7.1 Hz, 2H, NCH2), 2.48 (s,
4H, CH2NCH2), 2.10 (m, 2H, CH2CH2CH2). MS (ESI), m/z: 672.3
[MþH]þ.
(d, J ¼ 1.9 Hz, 1H, benzothiazol-4H), 7.77(dd, J ¼ 10.8 Hz, 2.1 Hz,
1H, phenyl-2H), 7.68 (dd, J ¼ 9.2 Hz, 2.0 Hz, 1H, phenyl-5H), 7.55
(s, 1H, quinolin-5H), 7.48–7.37 (m, 2H, Ar-H), 7.29 (d, J ¼ 8.6 Hz,
1H, benzothiazol-7H), 6.47 (d, J ¼ 4.9 Hz, 1H, quinolin-3H),
4.23 (t, J ¼ 4.8 Hz, 2H, CH2CH2O), 3.96 (s, 3H, CH3O), 2.79 (s,
2H, NCH2), 2.50 (s, 4H, CH2NCH2), 2.12 (s, 2H, CH2CH2CH2),
1.64 (s, 4H, NCH2CH2), 1.47 (s, 2H, CH2CH2CH2). MS (ESI), m/z:
680.0 [MþH]þ.
1-(4,6-Dichlorobenzothiazol-2-yl)-3-(3-fluoro-4-(6-
methoxy-7-(3-(morpholin-4-yl)propoxy)quinolin-4-yloxy)-
phenyl)urea (36)
Yellow solid; yield: 80.9%; mp: 250–252°C; 1H NMR (400 MHz,
DMSO-d6), d: 11.31 (s, 1H, NH), 9.42 (s, 1H, NH), 8.49 (d,
J ¼ 5.2 Hz, 1H, quinolin-2H), 8.11 (d, J ¼ 1.9 Hz, 1H, benzo-
thiazol-5H), 7.77 (dd, J ¼ 12.8 Hz, 2.0 Hz, 1H, phenyl-2H), 7.63
(d, J ¼ 1.6 Hz, 1H, benzothiazol-7H), 7.55 (s, 1H, quinolin-5H),
7.50–7.35 (m, 3H, Ar-H), 6.47 (d, J ¼ 5.2 Hz, 1H, quinolin-3H),
4.23 (t, J ¼ 6.0 Hz, 2H, CH2CH2O), 3.96 (s, 3H, CH3O), 3.67 (s, 4H,
CH2OCH2), 2.73 (s, 2H, NCH2), 2.67 (s, 4H, CH2NCH2), 2.07 (m,
2H, CH2CH2CH2). MS (ESI), m/z: 672.9 [MþH]þ.
1-(6-Bromobenzothiazol-2-yl)-3-(3-fluoro-4-(6-methoxy-7-
(3-(piperidin-1-yl)propoxy)quinolin-4-yloxy)phenyl)urea
(40)
Yellow solid; yield: 82.4%; mp: 228–230°C; 1H NMR (400 MHz,
CDCl3), d: 10.47 (s, 1H, NH), 8.47 (d, J ¼ 5.0 Hz, 1H, quinolin-2H),
7.76 (dd, J ¼ 12.1 Hz, 2.1 Hz, 1H, phenyl-2H), 7.72 (d, J ¼ 2.0 Hz,
1H, benzothiazol-7H), 7.57 (s, 1H, quinolin-5H), 7.51 (d,
J ¼ 8.5 Hz, 1H, benzothiazol-5H), 7.42 (s, 1H, quinolin-8H),
7.34–7.27 (m, 2H, Ar-H), 7.18 (t, J ¼ 8.6 Hz, 1H, phenyl-6H), 6.40
(d, J ¼ 5.0 Hz, 1H, quinolin-3H), 4.24(t, J ¼ 6.5 Hz, 2H, CH2CH2O),
4.02 (s, 3H, CH3O), 2.56 (t, J ¼ 7.3 Hz, 2H, NCH2), 2.46 (s, 4H,
CH2NCH2), 2.14 (m, 2H, NCH2CH2CH2O), 1.60 (m, 4H, NCH2CH2),
1.45 (s, 2H, CH2CH2CH2). MS (ESI), m/z: 680.1 [MþH]þ.
1-(Benzothiazol-2-yl)-3-(3-fluoro-4-(6-methoxy-7-(3-(4-
methylpiperidin-1-yl)propoxy)quinolin-4-yloxy)phenyl)-
urea (37)
Yellow solid; yield: 86.1%; mp: 202–204°C; 1H NMR (400 MHz,
CDCl3), d: 9.01 (s, 2H, NH), 8.31 (d, J ¼ 5.2 Hz, 1H, quinolin-2H),
7.64 (dd, J ¼ 12.8 Hz, 2 Hz, 1H, phenyl-2H), 7.62–7.55 (m, 2H,
Ar-H), 7.51 (s, 1H, quinolin-5H), 7.44–7.37 (m, 2H, Ar-H), 7.29
(s, 1H, quinolin-8H), 7.12 (dd, J ¼ 8.4 Hz, 1.9 Hz, 1H, phenyl-
5H), 7.03 (t, J ¼ 8.4 Hz, 1H, phenyl-6H), 6.27 (d, J ¼ 5.2 Hz, 1H,
quinolin-3H), 4.19 (t, J ¼ 5.5 Hz, 2H, CH2CH2O), 3.94 (s, 3H,
CH3O), 3.37 (d, J ¼ 11.0 Hz, 2H, CH2NCH2), 3.02 (t, J ¼ 6.8 Hz,
2H, NCH2), 2.52 (s, 2H, CH2NCH2), 2.29 (m, 2H, CH2CH2CH2),
1.81 (d, J ¼ 9.1 Hz, 2H, NCH2CH2), 1.56 (m, 3H, NCH2CH2,
CHCH3), 0.98 (d, J ¼ 4.4 Hz, 3H, CHCH3). MS (ESI), m/z: 616.0
[MþH]þ.
1-(4-Trifluoromethoxybenzothiazol-2-yl)-3-(3-fluoro-4-(6-
methoxy-7-(3-(piperidin-1-yl)propoxy)quinolin-4-yloxy)-
phenyl)urea (41)
1
Yellow solid; yield: 83.9%; mp: 273–275°C; H NMR (400 MHz,
CDCl3), d: 9.71 (s, 1H, NH), 8.59 (d, J ¼ 8.8 Hz, 1H, benzothiazol-
5H), 8.45 (d, J ¼ 5.2 Hz, 1H, quinolin-2H), 8.38 (s, 1H, NH), 7.59
(s, 1H, quinolin-5H), 7.56 (dd, J¼ 9.0 Hz, 2.0 Hz, 1H, phenyl-5H),
7.48 (dd, J ¼ 8.8 Hz, 2.0 Hz, 1H, phenyl-2H), 7.39 (t, J ¼ 9.0 Hz, 1H,
phenyl-6H), 7.37 (s, 1H, quinolin-8H), 7.20–7.06 (m, 2H, Ar-H),
6.41 (d, J ¼ 5.2 Hz, 1H, quinolin-3H), 4.18 (t, J ¼ 6.5 Hz, 2H,
CH2CH2O), 4.02 (s, 3H, CH3O), 2.57 (t, J ¼ 7.2 Hz, 2H, NCH2), 2.46
(s, 4H, CH2NCH2), 2.12 (m, 2H, NCH2CH2CH2O), 1.59 (m, 4H,
NCH2CH2), 1.44 (m, 2H, CH2CH2CH2).MS(ESI),m/z:686.1[MþH]þ.
1-(5-Bromobenzothiazol-2-yl)-3-(3-fluoro-4-(6-methoxy-7-
(3-(4-methylpiperidin-1-yl)propoxy)quinolin-4-yloxy)-
phenyl)urea (38)
Yellow solid; yield: 79.8%; mp: 242–244°C; 1H NMR (400 MHz,
DMSO-d6),d:10.28(s,1H,NH),8.58(s,1H,NH),8.48(d,J ¼ 5.0 Hz,
1H, quinolin-2H), 8.26 (d, J ¼ 8.6 Hz, 1H, benzothiazol-6H), 8.00
(d, J ¼ 2.0 Hz, 1H, benzothiazol-4H), 7.78(dd, J ¼ 12.8 Hz, 2.1 Hz,
1H, phenyl-2H), 7.68 (dd, J ¼ 8.8 Hz, 2.0 Hz, 1H, phenyl-5H), 7.54
(s, 1H, quinolin-5H), 7.45 (t, J ¼ 8.8 Hz, 1H, phenyl-6H), 7.40
(s, 1H, quinolin-8H), 7.29 (d, J ¼ 8.4 Hz, 1H, benzothiazol-7H),
6.46 (d, J ¼ 4.9 Hz, 1H, quinolin-3H), 4.20 (s, 2H, CH2CH2O), 3.96
(s, 3H, CH3O), 2.98 (s, 2H, CH2NCH2), 2.61 (s, 2H, NCH2), 2.11 (s,
2H, CH2NCH2), 2.03 (s, 2H, CH2CH2CH2), 1.63 (d, J ¼ 11.9 Hz, 2H,
NCH2CH2),1.21(m, 3H, NCH2CH2, CHCH3), 0.90(d, J ¼ 6.2 Hz, 3H,
CHCH3). MS (ESI), m/z: 694.0 [MþH]þ.
1-(6-Chlorobenzothiazol-2-yl)-3-(3-fluoro-4-(6-methoxy-7-
(3-(piperidin-1-yl)propoxy)quinolin-4-yloxy)phenyl)urea
(42)
Yellow solid; yield: 88.4%; mp: 214–215°C; 1H NMR (400 MHz,
CDCl3), d: 10.46 (s, 1H, NH), 8.47 (d, J ¼ 5.1 Hz, 1H, quinolin-2H),
7.76 (dd, J ¼ 11.8 Hz, 2.2 Hz, 1H, phenyl-2H), 7.72 (d, J ¼ 1.8 Hz,
1H, benzothiazol-7H), 7.57 (s, 1H, quinolin-5H), 7.51 (d,
J ¼ 8.5 Hz, 1H, benzothiazol-5H), 7.42 (s, 1H, quinolin-8H),
7.35–7.28 (m, 2H, Ar-H), 7.18 (t, J ¼ 8.6 Hz, 1H, phenyl-6H), 6.40
(d, J ¼ 5.0 Hz, 1H, quinolin-3H), 4.24(t, J ¼ 6.3 Hz, 2H, CH2CH2O),
4.02 (s, 3H, CH3O), 2.56 (t, J ¼ 7.2 Hz, 2H, NCH2), 2.46 (s, 4H,
CH2NCH2), 2.14 (m, 2H, NCH2CH2CH2O), 1.60 (m, 4H, NCH2CH2),
1.45 (s, 2H, CH2CH2CH2). MS (ESI), m/z: 636.0 [MþH]þ.
1-(5-Bromobenzothiazol-2-yl)-3-(3-fluoro-4-(6-methoxy-7-
(3-(piperidin-1-yl)propoxy)quinolin-4-yloxy)phenyl)urea
(39)
1-(5-Trifluoromethylbenzothiazol-2-yl)-3-(3-fluoro-4-(6-
methoxy-7-(3-(piperidin-1-yl)propoxy)quinolin-4-yloxy)-
phenyl)urea (43)
Yellow solid; yield: 78.5%; mp: 219–221°C; 1H NMR (400 MHz,
DMSO-d6),d:10.24(s,1H,NH),8.56(s,1H,NH),8.48(d,J ¼ 5.0 Hz,
1H, quinolin-2H), 8.26 (d, J ¼ 8.6 Hz, 1H, benzothiazol-6H), 8.00
Yellow solid; yield: 85.2%; mp: 187–188°C; 1H NMR (400 MHz,
CDCl3), d: 9.64 (s, 1H, NH), 9.29 (s, 1H, NH), 8.41 (d, J ¼ 5.0 Hz,
ß 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com
9

RCHH HARM
A P
Arch. Pharm. Chem. Life Sci. 2016, 349, 1–11
H. Lei et al.
Archiv der Pharmazie
1H, quinolin-2H), 8.05 (d, J ¼ 2.1 Hz, 1H, benzothiazol-4H),
7.90 (dd, J ¼ 8.6 Hz, 2.4 Hz, 1H, phenyl-5H), 7.73 (d, J ¼ 8.7 Hz,
1H, benzothiazol-6H), 7.68 (dd, J ¼ 12.4 Hz, 2.3 Hz, 1H, phenyl-
2H), 7.55 (s, 1H, quinolin-5H), 7.37 (s, 1H, quinolin-8H), 7.27
(m, 1H, benzothiazol-7H), 7.11 (t, J ¼ 8.6 Hz, 1H, phenyl-6H),
6.35 (d, J ¼ 5.0 Hz, 1H, quinolin-3H), 4.26 (t, J ¼ 6.0 Hz, 2H,
CH2CH2O), 3.99 (s, 3H, CH3O), 2.94 (t, J ¼ 6.8 Hz, 2H, NCH2),
2.84 (s, 4H, CH2NCH2), 2.32 (m, 2H, NCH2CH2CH2O), 1.83
(m, 4H, NCH2CH2), 1.58 (s, 2H, CH2CH2CH2). MS (ESI), m/z: 670.0
[MþH]þ.
Financial support was provided by the Program for Innovative
Research Team of the Ministry of Education of China and the
Program for Liaoning Innovative Research Team in University,
China (no. IRT1073), National Basical Science Talent Training
Foundation of China (no. J1103606), and National Natural
Foundation of China (grant no. 81573295). Enzyme assays
were performed at the Shanghai Chempartner Co., Ltd.
The authors have declared no conflicts of interest.
Biology
References
Cytotoxic activities against tumor cells assay
All of the target compounds were evaluated against the
cancer cell lines HT-29 (human colon cancer), MKN-45 (human
gastric cancer), and H460 (human lung cancer) using standard
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
(MTT) assay.
[1] M. Heron, Natl. Vital Stat. Rep. 2010, 58(14), 1–100.
[2] C. M. Stellrecht, V. Gandhi, Cancer Lett. 2009, 280(1), 1.
[3] Y. C. Li, S. L Zhang, Z. G. Tang, J. Chen, W. J. Kong,
Tumour Biol. 2011, 32(6), 1217.
[4] S. Corso, P. M. Comoglio, S. Giordano, Trends Mol. Med.
2005, 11(6), 284.
[5] G. C. Chu, H. E. Zhau, R. X. Wang, A. Rogatko, X. Feng,
M. Zayzafoon, Y. H. Liu, M. C. Farach-Carson, S. Y. You,
J. Kim, M. R. Freeman, L. W. K. Chung, Endocr-relat.
Cancer 2014, 21(2), 311.
[6] I. T. Ocal, M. Dolled-Filhart, T. G. D’Aquila, Cancer 2003,
97(8), 1841.
[7] L. N. Johnson, E. D. Lowe, M. E. Noble, D. J. Owen, FEBS
Lett. 1998, 430(1–2), 1–11.
[8] H. Morgan, K. John, Cell 2002, 109(3), 275–282.
[9] G. Manning, D. B. Whyte, R. Martinez, Science, 2002,
298(5600), 1912–1916, 1933–1934.
[10] P. Traxler, P. Furet, Pharmacol. Therapeut. 1999, 82(2–3),
195–206.
[11] J. J. Cui, M. Tran-Dube, H. Shen, M. Nambu, P. Kung,
M. Pairish, L. Jia, J. Meng, L. Funk, I. Botrous, J. Med.
Chem. 2011, 54(18), 6342–6363.
The cancer cell lines were cultured in the minimum essential
medium (MEM), which was supplemented with 10% fetal
bovine serum (FBS). The cells were seeded into each well of a
96-well plate and incubated in 5% CO2 at 37°C for 24 h. About
4 ꢂ 103 cells were suspended in the MEM medium. At the
indicated final concentrations, the subject compounds were
added into the culture medium at a three-time concentration
gradient (23 for example, 10000, 3333.3, 1111.1, 370.4, 123.5,
41.2, 13.7, 4.6, 1.5, 0.5 nmol/L) and the cells were cultured
continuously for 72 h. At a terminal concentration of 5 mg/mL,
fresh MTT was added into each well and incubated with the
cells at 37°C for 4 h. The formazan crystals were dissolved in
100 mL DMSO and added into each well. Use the ELISA
(enzyme-linked immunosorbent assay) reader to measure the
absorbance at 492 nm (for absorbance of MTT formazan) and
630 nm (for the reference wavelength).
HTRF kinase assay
[12] B. K. Albrecht, J. C. Harmange, D. Bauer, L. Berry, C. Bode,
A. A. Boezio, A. Chen, D. Choquette, I. Dussault,
C. Fridrich, J. Med. Chem. 2008, 51(10), 2879–2882.
[13] J. P. Eder, G. F. Vande-Woude, S. A. Boerner, P. M. Lo
Russo, Clin. Cancer Res. 2009, 15(7), 2207–2214.
[14] J. Porter, S. Lumb, R. J. Franklin, J. M. Gascon-Simorte, M.
Calmiano, R. K. Le, B. Lallemand, J. Keyaerts, H. Edwards, A.
Maloney, Bioorg. Med. Chem. Lett. 2009, 19(3), 2780–2784.
[15] J. G. Christensen, R. Schreck, J. Burrows, P. Kuruganti,
E. Chan, P. Le, J. Chen, X. Wang, L. Ruslim, R. Blake,
Cancer Res. 2003, 63(21), 7345–7355.
[16] X. Wang, P. Le, C. Liang, J. Chan, D. Kiewlich, T. Miller,
D. Harris, L. Sun, A. Rice, S. Vasile, Mol. Cancer Ther.
2003, 2(11), 1085–1092.
[17] G. I. Shapiro, S. McCallum, L. M. Adams, L. Sherman,
S. Weller, S. Swann, H. Keer, D. Miles, Invest. New Drugs
2013, 31(3), 742–750.
The c-Met kinase assays of tested compounds were performed
by homogeneous time-resolved fluorescence (HTRF) assay as
per the previously reported protocol [29, 30]. To examine the
selectivity of the most promising compound 23 on c-Met over
other tyrosine kinases, it was screened against Flt-3, VEGFR-2,
and EGFR. Briefly, 20 mg/mL poly(Glu, Tyr) 4:1 (Sigma) was
preloaded as a substrate in 384-well plates. Then 50 mL of
10 mM ATP (Invitrogen) solution diluted in kinase reaction
buffer (50 mM HEPES, pH 7.0, 1 mM DTT, 1 mM MgCl2, 1 mM
MnCl2, 0.1% NaN3) was added to each well. Various
concentrations of compounds were diluted in 10 mL of 1%
DMSO (v/v), with blank DMSO solution as the negative
control. The kinase reaction was initiated by the addition of
purified tyrosine kinase proteins diluted in 39 mL of kinase
reaction buffer solution. Reactions were incubated for 30 min
at 25°C and stopped by the addition of 5 mL Streptavidin-
XL665 and 5 mL Tk Antibody Cryptate working solution to all
wells. The plate was read by Envision (Perkin-Elmer) at 320
and 615 nm. IC50 values were calculated from the inhibition
curves.
[18] S. Li, R. Jiang, M. Z. Qin, H. C. Liu, G. Y. Zhang, P. Gong,
Arch. Pharm. 2013, 346(7), 521–533.
[19] Q. D. Tang, Y. F. Zhao, X. M. Du, L. E. Chong, P. Gong,
C. Guo, Eur. J. Med. Chem. 2013, 69, 77.
ß 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com
10

RCHH HARM
A P
Arch. Pharm. Chem. Life Sci. 2016, 349, 1–11
4-Phenoxyquinoline Derivatives as c-Met Kinase Inhibitors
Archiv der Pharmazie
[20] B. H. Qi, B. Mi, X. Zhai, Z. Y. Xu, X. L. Zhang, Z. R. Tian,
P. Gong, Bioorg. Med. Chem., 2013, 21, 5246.
[21] S. Li, Q Huang, Y. J. Liu, X. L. Zhang, S. Liu, C. He, P. Gong,
Eur. J. Med. Chem. 2013, 64, 62.
[22] S. Li, Y. F. Zhao, K. W. Wang, Y. L. Gao, J. M. Han, B. B. Cui,
P. Gong, Bioorg.Med. Chem. 2013, 21, 2843.
[23] A. Catalano, A. Carocci, I. Defrenza, Eur. J. Med. Chem.
2013, 64, 357–364.
[24] E. Meltzer-Mats, G. Babai-Shani, L. Pasternak, N. Uritsky,
T. Getter, O. Viskind, J. Eckel, H. Senderowitz, S. Sasson, J.
Med. Chem. 2013, 56(13), 5335–5350.
[26] M. Singh, S. K. Singh, Med. Chem. 2014, 14, 127–146.
[27] B. H. Qi, H. Tao, D. Wu, J. Y. Bai, Y.D. Shi, P. Gong, Arch.
Pharm. 2013, 346(8), 596–609.
[28] H. H. Li, J. Li, H. X Chen, Y. Zhang, D. Huang, J. Chem.
Crystallogr. 2011, 41(12), 1844–1849.
[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. 2008,
51, 3688.
[25] A. Furlan, F. Colombo, A. Kover, N. Issaly, C. Tintori,
L. Angeli, V. Leroux, S. Letard, M. Amat, Y. Asses, Eur. J.
Med. Chem. 2012, 47(1), 239–254.
[30] M. L. Peach, N. Tan, N. Tan, S. J. Choyke, A. Giubellino,
G. Athauda, T. R. Burke, M. C. Nicklaus, D. P. Bottaro, J.
Med. Chem. 2009, 52, 943.
ß 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com
11