Design, synthesis and pharmacological evaluation of new 2-oxo-quinoline derivatives containing α-aminophosphonates as potential antitumor agents

Article abstract of DOI:10.1039/c7md00098g

A series of novel 2-oxo-quinoline derivatives containing α-aminophosphonates were designed and synthesized as antitumor agents. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay results demonstrated that some compounds exhibited moderate to high inhibitory activity against HepG2, SK-OV-3 and NCI-H460 tumor cell lines, and most compounds showed much lower cytotoxicity against HL-7702 normal cells than 5-FU and cisplatin. The action mechanism of representative compound 5b was investigated by fluorescence staining assay, flow cytometric analysis and western blot (WB) assay, which indicated that this compound induced apoptosis and G₂/M phase arrest accompanied by an increase in the production of intracellular Ca²⁺ and reactive oxygen species (ROS) and affecting associated enzymes and genes.

Full text of DOI:10.1039/c7md00098g

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Design, synthesis and pharmacological evaluation of new 2-oxo-
quinoline derivatives containing α-aminophosphonates as
potential antitumor agents‡
Received 00th January 20xx,
Accepted 00th January 20xx
Yan-Cheng Yua,b,†, Wen-Bin Kuangb,†, Ri-Zhen Huangb, Yi-Lin Fangb, Ye Zhanga,b,c,*, Zhen-Feng
Chenb,*, Xian-Li Maa,*
DOI: 10.1039/x0xx00000x
www.rsc.org/
A series of novel 2-oxo quinoline derivatives containing α-aminophosphonate were designed and synthesized as antitumor
agents. MTT(3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide) assay results demonstrated that some
compounds exhibited moderate to high inhibitory activity against HepG2, SK-OV-3 and NCI-H460 cells tumor cell lines, and
most compounds showed much lower cytotoxicy against HL-7702 normal cell than 5-FU and cisplatin. The action
mechanism of representative compound 5b was investigated by fluorescence staining assay, flow cytometric analysis and
western blot (WB) assay, which indicated that this compound induced apoptosis and involved G2/M phase arrest
accompying by increasing the production of intracellular Ca2+ and reactive oxygen species (ROS) and affecting associated
enzymes and genes.
1. Introduction
Quinoline alkaloids exhibit a variety of outstanding
biological activities, including inhibition of cellular proliferation
and development changes. So the design and synthesis of new
quinoline derivatives with better antitumor activity has
attracted many chemists’ interest. As the family of quinolines,
2-oxo-quinoline derivatives display potent proliferation
inhibitory activity on tumor cells.1,2 Our previous work has also
demonstrated that 2-oxo-quinoline schiff base derivatives
exhibited good antioxidant activity and obvious nontoxicity on
bald mice.3,4 In addition, antioxidants are fabricated as the
drug candidates to counter these multifarious chronic
diseases, containing carcinogenesis, atherogenesis and aging,5
and many antioxidants exhibit efficient antitumor activity.6-7
It is well known that cancer is a global health problem and
the main causes of mortality worldwide. In order to prevent
and combat this disease, a great deal of effort has been
devoted to the design and synthesis of new antitumor drugs
with high efficiency and low toxicity.
R2
OH
R1
HO
N
N
O
H
OH
2-oxo-quinoline schiff base derivatives
resveratrol
Fig. 1. The chemical structures of 2-oxo-quinoline schiff base For instance, the powerful antioxidant resveratrol is also
derivatives and resveratrol.
deemed as antitumor agents.7 Moreover, 2-oxo-quinoline
schiff base derivatives (Fig. 1) show similar structure with
resveratrol and can be considered as its bioisostere
derivatives. So it is considered that 2-oxo-quinoline schiff base
derivatives may exhibit potent proliferation inhibitory.
Furthermore, our previous work has demonstrated that the
introduction of aminophosphonate group to pharmacy core is
able to increase the antitumor activity and many
aminophosphonate derivatives have exhibited potent
inhibition activities against human tumors.8,9 It is thus to
expect that the combination of 2-oxo-quinoline schiff base
derivatives and aminophosphonate (APA) groups may lead to
good antitumor activity and low toxicity. In our previous work,
we have described the synthesis, antitumor activity,
preliminary apoptosis-inducing and cycle arresting effects of
some 2-oxo-quinoline APA derivatives.10 However, to the best
of our knowledge, the scale and mode of the synthesis is still
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limited. Particularly, the cytotoxicity on normal cells(i.e., obtained in good yields by the hydrolytic reaction of
toxicity), the further apoptosis-inducing and cycle arresting in the presence of 70% acetic acid aqueous solution.3 The
mechanism of 2-oxo-quinoline schiff base derivatives with APA target compounds 4a1-4d7 were synthesized by the
moieties has not been reported. Therefore, in the present Kabachnik-Fields reaction, which executed by the
work, as a continuation of our previous work, we introduced simultaneous condensation of compounds diethyl
2(2a-2d)
4(
)
3,
some APA moiety to different 2-oxo-quinoline schiff base phosphate and many kinds of amines, through a one-pot
skeleton and evaluated the in vitro cytotoxicity of the target three-component synthesis method. The other target
compounds. Moreover, the mechanism of apoptosis and cycle compounds
5
(
5a-5d
condensation of compounds
aminobenzylphosphonate. The structures of target compounds
4a1-4d7 and 5a 5d), were confirmed using various
spectroscopic methods, including H NMR, 13C NMR and high-
resolution mass spectrometry (HR-MS)(Part of
)
were also synthesized by the
arrest was further deeply and systematically investigated.
3
with diethyl 4-
4
(
)
5(
-
1
2. Results and discussion
3
2.1. Chemistry
supplementary data). In order to better understand the
structures of these target compounds, the crystal structures of
2-Oxo-quinoline
(compounds and
Firstly, 2-chloro-quinoline-3-carbaldehyde derivatives
derivatives
) were synthesized as shown in Scheme 1.
2a-2d)
bearing
APA
moiety
4
5
compounds 4c5, 4d2 and 5c were also determined by X-ray
2
(
diffraction (Fig. 2, selected relevant parameters see in part 1 of
supplementary data). As shown in Fig. 2, the crystals structure
were obtained via Vilsmeier-Haack-Arnold reaction according
to our previous work,3 which included the condensation of
of compounds 4c5
characterization data of NMR and HR-MS, indirectly confirming
the structures of target compounds
, 4d2 and 5c were well consistent with the
acetanilide derivatives
1(1a-1d) with N,N-dimethylformamide
(DMF) in the presence of phosphorusoxychloride. 2-Oxo-
4-5.
quinoline 3-carbaldehyde derivatives
3
(
3a-3d) were then
O
EtO
P
OEt
R2
R1
R2
NHCOCH3
(a)
R1
R2
CHO
Cl
R1
CHO
O
R1
R2
CH
N
(c)
(b)
H
N
H
R2
N
H
N
O
H
3(3a-3d)
1(1a-1d)
2(2a-2d)
4(4a1-4d7)
4a: R1 = R2 =H
(d)
4b: R1 = CH3, R2 =H
4c: R1 = OCH3, R2 =H
4d: R1,R2 = CH2-O-CH2
O
P
OEt
H
C
R1
R2
OEt
N
N
O
H
5
5a: R1 = R2 =H
5b: R1 = CH3, R2 =H
5c: R1 = OCH3, R2 =H
5d: R1,R2 = CH2-O-CH2
Scheme 1. General synthetic route for compound 3−5. Reagents and conditions: (a) POCl3/DMF; (b) 70% acetic acid aqueous
solution; (c) diethyl phosphite, amines, CH3CN; (d) diethyl 4-aminobenzylphosphonate, CH3CN.
4d2
5c
4c5
Fig. 2. The crystal structures of compounds 4c5, 4d2 and 5c.
2.2 Biological Activity
2.2.1 Cytotoxicity Test
The in vitro cytoxicity of the compounds 4(4a1-4d7) and
5
(
5a-5d) were evaluated by methylthiazoltetrazolium (MTT)
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assay against HepG2 (Human liver cancer cell line), SK-OV-3 also concluded that the schiff base bond had potent effect on
(human ovarian cancer cell line), NCI-H460 (Human large cell the antitumor activity.
lung cancer cell line) and HL-7702 (Human liver normal cell
In HepG2 assay, compounds 4c5, 5a, 5b, 5c and 5d exhibited
line) cell lines. Two commercial anticancer drugs 5-fluorouracil better cytotoxicity than the commercial anticancer drug 5-FU
(5-FU) and cisplatin were used as positive controls. The results (IC50=31.98 μM), with IC50 of 20.80 μM, 17.85 μM, 9.99 μM,
were shown in Table 1.
17.31 μM and 28.92 μM, respectively. It was worth noting that
As shown in Table 1, most of target compounds 4−5 compound 5b even exhibited better cytotoxic inhibition than
displayed much higher inhibitory activity than their cisplatin (IC50=10.12 μM), implying its favourable inhibition
corresponding 2-oxo-quinoline 3-carbaldehyde derivatives activities of on HepG2 cell line. So compound 5b was then
3
(
3a−3d) against the HepG2, SKOV3 and NCI-H460 cell lines, selected as representative compound to assay the action
indicating the introduction of α-aminophosphonates on 2-oxo- mechanism of these 2-oxo-quinoline APA derivatives 4-5
quinoline may improve the antitumor activity. It was important In SKOV-3 assay, compounds 4a7 4c5 4d6 and 4d7 displayed
.
,
,
to note that some target compounds showed better cytotoxic better cytotoxicity than 5-FU (IC50=26.34 μM), with IC50 of
inhibition against these three cell lines than 5-FU, while they 25.81μM, 25.13μM, 26.25μM and 22.36 μM, respectively. It
displayed lower cytotoxicity on HL-7702 normal cell line than was obvious that compound 4d7 showed the best cytotoxic
that of commercial anticancer drugs 5-fluorouracil (5-FU) and inhibition among these compounds. Pity of it was that no
cisplatin, indicating that they may be good candidate for anti- compound exhibited better cytotoxic inhibition than cisplatin
tumour drugs. Table 1 also demonstrated that, for compound in this cell line.
4, both of the substituents (R1) in 2-oxo-quinoline group and
In NCI-H460 assay, compounds 4c5 and 5b exhibited better
the substituents (R2) in α-aminophosphonates moiety had cytotoxic inhibition than 5-FU (IC50=45.44 μM), with IC50 of
important influence on the cytotoxic inhibition, though the 44.56 and 38.96μM, respectively, while these was no
influence was irregular. In addition, by the comparasion of compound demonstrated better cytotoxic inhibition than
cytotoxic inhibition activity of compounds
4 and 5, it could be cisplatin in this assay.
Table 1. αIC50 values of 2-oxo-quinoline derivatives bearing APA moiety (
4-
5) towards three selected tumor cell lines and normal
cell line for 48 h.
IC50(μM)
Compounds
HepG2
SKOV-3
NCI-H460
>100
HL-7702
>100
OEt
O
P
OEt
HN
61.43±2.14
38.30±1.74
N
O
H
4a1
OEt
O
P
OEt
HN
67.27±2.35
65.46±2.64
66.75±2.48
55.95±1.11
>100
>100
>100
>100
O
N
H
4a2
EtO
OEt
P
O
HN
O
N
H
4a3
EtO
OEt
P
O
HN
46.33±2.02
75.21±3.74
37.21±2.23
54.11±1.18
29.98±0.91
40.95±0.85
>100
>100
>100
>100
>100
>100
O
N
H
4a4
EtO
OEt
P
O
HN
O
N
H
NO2
4a5
EtO
OEt
P
O
HN
O
N
H
F
4a6
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EtO
OEt
HN
P
O
39.12±2.11
39.17±3.12
37.57±1.45
25.81±0.54
130.26±4.59
37.22±1.34
>100
>100
>100
>100
>100
>100
O
N
H
CF3
4a7
EtO
OEt
P
O
HN
O
N
H
4b1
EtO
OEt
P
O
HN
O
N
H
4b2
EtO
OEt
HN
P
O
45.95±1.78
342.40±4.26
>100
>100
O
N
H
4b3
EtO
OEt
HN
P
O
39.56±1.54
35.61±2.73
69.37±1.08
>200
>100
>100
>100
>100
O
N
H
4b4
EtO
OEt
P
O
HN
O
N
H
NO2
4b5
EtO
OEt
P
O
HN
46.21±2.11
68.11±2.64
43.25±3.35
57.63±2.75
30.33±0.66
191.26±4.02
58.21±1.75
59.96±2.77
>100
>100
>100
>100
>100
>100
>100
>100
O
N
H
F
4b6
EtO
OEt
P
O
HN
O
N
H
CF3
4b7
EtO
OEt
HN
P
O
O
O
N
H
4c1
EtO
OEt
HN
P
O
O
O
N
H
4c2
EtO
OEt
HN
P
O
O
45.54±2.85
60.77±3.01
73.011±2.29
54.66±1.95
>100
>100
>100
>100
O
N
H
4c3
EtO
P
OEt
HN
O
O
O
N
H
4c4
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EtO
OEt
HN
P
O
O
O
O
20.80±1.65
58.92±2.27
40.15±1.88
25.13±1.04
44.56±2.56
>100
O
N
H
NO2
4c5
EtO
OEt
HN
P
O
269.696±5.12
102.11±4.39
>100
>100
>100
O
N
H
F
4c6
EtO
OEt
P
O
HN
>100
O
N
H
CF3
4c7
EtO
OEt
HN
P
O
O
O
46.27±2.66
43.11±1.17
>100
37.23±1.95
48.89±1.53
>100
47.86±2.05
>100
>100
>100
>100
>100
>100
O
N
H
4d1
EtO
P
OEt
HN
O
O
O
O
N
H
4d2
EtO
OEt
HN
P
O
O
O
>100
O
N
H
4d3
EtO
P
OEt
HN
O
O
O
45.21±3.14
44.01±1.31
64.95±2.43
28.23±0.75
>100
O
N
H
4d4
OEt
OEt
O
P
O
O
HN
>100
O
N
H
NO2
4d5
OEt
O
P
OEt
O
HN
37.95±1.98
78.95±4.45
26.25±0.87
22.36±0.45
59.56±2.75
>100
>100
O
N
O
H
F
4d6
OEt
OEt
O
P
O
O
HN
>100
O
N
H
CF3
4d7
N
OEt
P
O
OEt
17.85±0.68
9.99±0.25
35.97±0.98
34.76±1.14
57.88±2.24
38.96±1.88
>100
>100
N
O
H
5a
N
OEt
P
OEt
O
N
O
H
5b
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OEt
P
OEt
O
O
N
17.31±1.13
28.92±1.47
52.991±1.61
47.92±1.14
>100
>100
>100
>100
N
O
H
5c
OEt
P
O
OEt
O
O
N
N
O
H
5d
O
N
H
O
>200
>200
>200
>200
>200
>200
>200
>200
>200
>200
>200
>200
>100
>100
>100
>100
3a
O
N
H
O
3b
O
O
N
O
H
3c
O
O
O
N
O
H
3d
5-FU
Cis-platin
31.98±0.56
10.12±0.71
26.34±0.57
15.60± 1.70
45.44±0.94
20.36±0.50
58.74±2.31
15.67±0.32
αIC50 values are presented as the mean ± SD (standard error of the mean) from three separated experiments.
2.2.2. Investigation of Cell Cycle Distribution
promoting accumulation of the inactive cyclin B-CDK complex,
by which lead to the regulation to G2/M transition.15. So the
expression of these regulatory proteins including cyclin B1,
CDK1 and CDC25C, as well as p53, p27 and p21 anti-
oncogenes, were further evaluated by western blot assay,
using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as
control. As shown in Fig.4, the HepG2 cell lysates
demonstrated that compound 5b dramatically decreased the
Cell cycle checkpoints are significant control mechanisms
that ensure the nonreversible performance of cell cycle
events.11 Understanding of cell cycle distribution may provide
important information to the regulation of the cell cycle.12 To
determine the possible role of cell cycle arrest in 2-oxo-
quinoline APA derivatives induced growth inhibition, HepG2
cells were treated with the representative compound 5b at
different concentrations (0, 8, 10 and 12 μM) for 48 h. As
shown in Fig. 3, treating of HepG2 cells with compound 5b
increased cell cycle arrest at the G2 phase, leading to a
significant increase in the G2-phase population (34.68%(8μM),
36.92%(10μM) and 46.77%(12μM)) compared with the control
cells (19.04%). The G1-phase and S-phase population of HepG2
cells reduced by 35.31%(8μM), 33.91%(10μM), 25.75%(12μM)
expression of cyclin B1, CDK1 and CDC25C in
a dose-
dependent manner, accompanied by a dose-dependent
increase in the level of p53, p21 and p27 protein compared
with controls. These results clearly displayed that compounds
5b potentially induced G2/M phase cell cycle arrest in HepG2
cells, which might be one of the reasons for its
antiproliferative effect.
2.2.3. Apoptosis Assay study by mitochondrial membrane
potential staining
and
30.01%(8μM),
29.16%(10μM),
27.49%(12μM),
respectively, compared with that of control (50.47% and
30.49%). The results evidently demonstrated that compound
Apoptosis is a key pathway leading to cell death and has
5b potentially arrested the cell cycle of HepG2 cells in the been considered as another effective approach in cancer
G2/M stage in a concentration-dependent manner.
treatment.16 Its assays may provide important information for
G2/M transition is basically mediated by timed activation of preliminary investigation of the mode of action.16-19 So it may
distinct cyclin B1/CDK1(CDC2) complexes which is mainly be interested to investigate the apoptosis-inducing effect of
regulated by the positive regulator CDC25C phosphatase.12,13 compound 5b. In order to investigate the apoptosis-inducing
CDC25C dephosphorylates CDK1 resulting to the activation of effect of target compound 5b, mitochondrial membrane
cyclin B1/CDK1 complex and performance of G2/M potential changes were designed and detected, using the
transition.12,13 In response to DNA damage or other alteration fluorescent probe JC-1. HepG2 cells treated with compound 5b
causing to G2/M arrest, phosphorylation of CDC25C results to at 10 μM for 12 h were stained with JC-1 and not treated with
its degradation and sequestration in the cytoplasm, which the compound 5b were used as control. The results were
eventually leads to the destabilization of cyclin-CDK shown in Fig. 5. For fluorescence microscopy, Fig. 5 showed
complexes.14 Moreover, the anti-oncogene p53, p27 and p21 that cells not treated with the compound 5b were normally
protein also plays a significant role in G2/M arrest through red, while for compound 5b treatment, cells showed strong
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green fluorescence and indicated typical apoptotic 2.2.5. Apoptosis Assay study by Acridine Orange/Ethidium
morphology after 12 h. The phenomenon suggested that Bromide (AO/EB) Staining
compound 5b was able to induce apoptotic cell morphology in
HepG2 cell line.17-19
To further characterize the cell apoptosis induced by
compound 5b, AO/EB staining was performed to evaluate the
2.2.4. Apoptosis Assay study by Hoechst 33258 Staining
accompanying changes in morphology. The cytotoxicity of
The apoptosis-inducing effect of compound 5b was also compound 5b was evaluated in HepG2 cells following
determined by Hoechst 33258 staining of HepG2 cancer cells treatment with 10 μM for 24 h. HepG2 cells not treated with
and the result were shown in Fig. 6. As shown in Fig. 6, HepG2 5b were used as control. The results (Fig. 7) showed that the
cells not treated with compound 5b were normally stained as morphology of 5b-treated HepG2 cells had changed
blue, while treated with 5b for 12 h showed strong blue significantly. The cell nuclei were stained yellow green or
fluorescence and had characteristic apoptotic morphologies. orange, and the morphology showed pycnosis, membrane
The observations demonstrated that compound 5b induced blebbing and cell budding characteristic of apoptosis. These
apoptosis of HepG2 cells, consistent with that of previous phenomena were associated with cell apoptosis, indicating
experiment of mitochondrial membrane potential staining.
that compound 5b was able to induce the apoptosis of HepG2
cells.
Fig. 3. Cell cycle analysis of compound 5b treated with HepG2 cells. HepG2 cells were treated with different concentrations
compound 5d ((a) 0μM, (b) 8μM, (c) 10μM and (d) 12μM) for 48 h to determine cell cycle phase distribution.
Fig. 4. Effect of compound 5b on the expression of cyclins and associated proteins. HepG2 cells were treated with compound 5b
for 48 h. Western blot analysis was carried out with antibodies against cyclin B1, CDK1, CDC25C and GAPDH was used as loading
control.
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2.2.6. Apoptosis Assay study by Flow Cytometry
19.08% late apoptotic cells) (8 μM), 43.62%(i.e., 9.24% early
apoptotic cells and 34.38% late apoptotic cells) (10 μM) and
54.90%(i.e., 15.49% early apoptotic cells and 19.08% late
apoptotic cells) (12 μM) in treated cells (i.e., 11.83% early
apoptotic cells and 43.07% late apoptotic cells). The results
demonstrated that the apoptosis of HepG2 cells treated with
compound 5b increased gradually with concentration and
compound 5b may suppress cell proliferation by inducing
apoptosis.
The apoptosis ratios induced by compound 5b in HepG2
tumor cells were quantitatively assayed by flow cytometry and
the result was shown in Fig. 8. The result contained the
differentiation of live cells (annexin V–/PI–), early apoptotic
cells (annexin V+/PI–), late apoptotic cells (annexin V+/PI+), and
necrotic cells (annexin V–/PI+). Treating of HepG2 cells with
different concentration (8, 10 and 12 μM) compound 5b led to
the increment of apoptotic cells population, from 8.41% in
controls to 34.57%(i.e., 15.49% early apoptotic cells and
Fig. 5. Mitochondrial membrane potential staining of compound 5b on HepG2 cells. (a) The cells not treated with 5b were
used as control, (b) compound 5b treated HepG2 cells at concentration of 10 μM, respectively.
Fig. 6. Effects of compound 5b on morphological changes of HepG2 cells after staining with Hoechst 33258 dye. (a) The cells
not treated with 5b were used as control, (b) compound 5b treated HepG2 cells at concentrations of 10 μM, respectively.
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Fig. 7. Compound 5b induced apoptosis in HepG2 cells were determined by AO/EB staining and were photographed via
fluorescence microscopy. (a) Not dealt with compound 5b was used as control at for 24 h, (b) dealt with compound 5b for 24 h at
concentrations of 10 μM, respectively.
Figure 8. Apoptosis ratio detection of compound 5b (b) 8μM, (c) 10μM, (d) 12μM) against HepG2 cells by Annexin V/PI assay
and the control (a) was used as comparation.
2.2.7. ROS generation assay
was synchronously detected by fluorescence microscopy with
a calcium indicator dye DCFH-DA (Fig. 10). As shown in Fig. 10,
treatment with 5b led to an increment of Ca2+ in the HepG2
cells. The results implied that 5b-induced apoptosis might be
associated with its induction of Ca2+ increment.
Previous work has proved that the generation of
intracellular reactive oxygen species (ROS) may lead to the
induction of apoptosis.20,21 To investigate the role of ROS in 5b
-
induced apoptosis in HepG2 cells, the ROS levels was
investigated by fluorescence microscopy using 4',6-diamidino-
2.2.9. Caspase-dependent Apoptosis in HepG2 Cells
2-phenylindole
dihydrochloride
(
DAPI
)
and
2,7-
As a physiological program of cellular death, apoptosis plays
dichlorofluorescein diacetate (DCFH-DA) as fluorescent probe.
a
vital role during normal development and in cellular
As shown in Fig. 9, for DAPI staining, HepG2 cells treated with homeostasis. It is well-known that Both Fas-dependent and
compound 5b displayed more strong fluorescence in the mitochondria-dependent apoptotic pathways are considered
cytoplasm, while the control that untreated control cells was as major pathways directly causing neuronal apoptosis, which
weak and spread over the cells. For DCFH-DA staining, HepG2 are associated with expression of Bax, Bcl-2 and cytochrome
cells treated with compound 5b exhibited stronger green proteins.24,25 To test the mechanism of 5b-induced apoptosis
fluorescence indicating that 5b significantly up-regulated in HepG2 cells, the expression of Bax, Bcl-2 and cytochrome
c
,
c
generation of ROS and induced apoptosis of HepG2 cells. The was also investigated by western blotting assay. As shown in
phenomenon indicated that compounds 5b significantly Fig. 11, treatment of HepG2 cells with 5b resulted to an
increased the intracellular level of ROS and was generally elevation in the expression of Bax and reduction in expression
considered as cues for the induction of apoptosis.17-19
2.2.8. Intracellular Ca2+ Release
of Bcl-2 accompanied by the release of mitochondrial
cytochrome into the cytosol. The results suggested that 5b
c
induced apoptosis by regulating the levels of Bcl-2 family
proteins, Bax and Bcl-2.
Intracellular calcium played important role in inducing cell
apoptosis and the overload of intracellular calcium can induce
cell apoptosis.22,23 Our above results have demonstrated that
compound 5b can induce apoptosis of HepG2 cells. To
determine the role of calcium signaling in 5b-induced
apoptosis, HepG2 cells were treated with 5b for 24 h and Ca2+
Caspases are
a family of cysteinyl aspartate specific
proteases contained in apoptosis and are dichotomized to
groups of initiators (caspases 8, 9 and 10) and executioners
(caspases 3, 6 and 7).19 The caspase cascade, which is initiated
by the proteolysis of inactive procaspases, is propagated by
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the cleavage of downstream caspases and substrates such as treated with compound 5b led to a significant increment in the
poly (ADP-ribose) polymerase cleavage (PARP). To determine expression of caspase-9, -3 and PARP compared to control.
whether caspases were activated in 5b-induced apoptosis, the These results indicated that compound 5b might induce
expression of caspase-9, -3 and PARP was investigated by apoptosis through a mitochondrial mediated pathway and
western blot assay (Fig. 11). As shown in Fig. 11, HepG2 cells caspase cascade.
Fig. 9. Compound 5b (10μM) affected the levels of intracellular ROS in HepG2 cells.
Fig. 10. Compound 5b (10μM) caused the levels of intracellular Ca2+ elevation in HepG2 cells.
Fig. 11. Effects of compound 5b on the level of cytochrome c, Bcl-2, Bax, Caspase-9, Caspase-3 and p53. HepG2 cells were
treated with of compound 5b for 24 h at different concentrations. Equal amount of protein was loaded on SDS-PAGE gel for
western blot analysis as described in experimental section. Glyceraldehyde-3-phosphate dehydrogenase(GAPDH) was used as an
internal control.
3.1. Chemistry
All chemicals (reagent grade) were commercially available
3. Experimental Section
and used without further purification. NMR spectra were
10 | J. Name., 2012, 00, 1-3
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assayed on a BRUKER AVANCE AV500 spectrometer using (ddd, J = 25.7, 13.6, 7.0 Hz, 2H), 1.36 (dt, J = 14.0, 7.0 Hz, 2H),
tetramethyl silane, (TMS) as the internal standard. The mass 1.26 (ddd, J = 16.7, 10.7, 5.3 Hz, 5H), 1.08 (t, J = 7.0 Hz, 3H),
spectra were determined on
spectrometer. GelRed nucleic acid stain was purchased from 161.62, 147.44, 147.33, 138.48, 137.76, 137.71, 130.87,
a
BRUKER ESQUIRE HCT 0.82 (t, J = 7.3 Hz, 3H).13C NMR (126 MHz, DMSO) δ 161.66,
Biotium.
129.99, 129.36, 128.16, 122.60, 119.39, 119.36, 117.79,
115.53, 113.65, 63.26, 63.21, 62.99, 62.93, 47.97, 46.73, 40.47,
40.30, 40.14, 39.97, 39.80, 39.63, 39.47, 16.76, 16.72, 16.58,
16.53.ESI-HRMS m/z Calc for C20H23N2O4P [M+Na]+ :409.1293;
found: 409.1293.
3.1.1 General procedure for compound 4(4a1-4d7)
2-oxo-quinoline 3-carbaldehyde derivatives
3 (1 mmol),
primarily amines (1.5 mmol), diethyl phosphate (1mmol) and 3
mL acetonitrile were mixed in pressure tube and reacted at
105oC for 4h. After the reaction, the mixture was cooled to
4a5: Yield73.01%, 1H NMR (400 MHz, DMSO) δ 12.13 (s, 1H),
8.13 (d, J = 3.4 Hz, 1H), 8.03 (d, J = 9.3 Hz, 2H), 7.95 (dd, J = 9.4,
5.6 Hz, 1H), 7.66 (d, J = 7.7 Hz, 1H), 7.54 (t, J = 7.5 Hz, 1H), 7.35
(d, J = 8.2 Hz, 1H), 7.25 – 7.17 (m, 1H), 6.86 (d, J = 9.3 Hz, 2H),
5.47 (dd, J = 22.6, 9.4 Hz, 1H), 4.21 – 3.88 (m, 4H), 1.21 (t, J =
7.0 Hz, 3H), 1.11 (t, J = 7.0 Hz, 3H)13C NMR (101 MHz, DMSO) δ
161.37, 161.31, 153.69, 153.58, 138.63, 138.45, 138.39,
137.66, 131.28, 128.59, 128.41, 126.45, 122.77, 119.18,
119.15, 115.65, 112.43, 63.46, 63.39, 63.36, 63.29, 47.73,
46.17, 40.60, 40.39, 40.18, 39.98, 39.77, 39.56, 39.35, 16.78,
16.72, 16.59, 16.54. ESI-HRMS m/z Calc for C20H22N3O6P
[M+Na]+ :454.1144; found: 454.1163.
room temperature and filtered to yield compound
or yellow powder.
4a1: Yield54.29%, 1H NMR (500 MHz, DMSO) δ 11.87 (s, 1H),
4 as white
8.05 (d,
J
= 3.7 Hz, 1H), 7.65 (d,
= 8.2 Hz, 1H), 7.19 (t,
= 21.9 Hz, 1H), 4.14 – 4.05 (m, 2H), 3.96 – 3.84 (m, 2H), 2.40
J
= 7.7 Hz, 1H), 7.49 (t,
J = 7.7
Hz, 1H), 7.31 (d,
J
J
= 7.5 Hz, 1H), 4.48 (d,
J
(ddd,
J
= 25.7, 13.6, 7.0 Hz, 2H), 1.36 (dt,
= 16.7, 10.7, 5.3 Hz, 5H), 1.08 (t,
= 7.3 Hz, 3H).13C NMR (126 MHz, DMSO) δ 161.98,
J
= 14.0, 7.0 Hz, 2H),
1.26 (ddd,
0.82 (t,
J
J
= 7.0 Hz, 3H),
J
161.93, 138.40, 137.70, 137.64, 130.60, 129.88, 128.18,
122.42, 119.55, 119.53, 115.37, 62.92, 62.87, 62.42, 62.36,
53.20, 51.96, 47.67, 47.55, 40.48, 40.31, 40.23, 40.14, 39.98,
39.81, 39.64, 39.48, 31.86, 20.17, 16.79, 16.74, 16.63, 16.58,
14.26.ESI-HRMS m/z Calc for C18H27N2O4P [M+H]+ :367.1787;
found: 367.1782.
4a6: Yield79.13%, 1H NMR (400 MHz, DMSO) δ 12.00 (s, 1H),
8.10 (d,
Hz, 1H), 7.32 (d,
(m, 2H), 6.71 (ddd,
6.3 Hz, 1H), 5.25 (dd,
7.1 Hz, 2H), 4.05 – 3.84 (m, 2H), 1.23 (t,
= 7.0 Hz, 3H).13C NMR (101 MHz, DMSO) δ 161.68, 161.62,
J
= 3.7 Hz, 1H), 7.59 (d,
J = 7.7 Hz, 1H), 7.50 (t, J = 7.7
J
= 8.2 Hz, 1H), 7.22 – 7.14 (m, 1H), 6.96 – 6.85
J
= 6.8, 5.2, 2.8 Hz, 2H), 6.37 (dd,
= 24.4, 10.3 Hz, 1H), 4.13 (dq,
J
J
= 10.3,
= 14.2,
4a2: Yield75.82%, 1H NMR (500 MHz, DMSO) δ 11.96 (s, 1H),
J
J
= 7.0 Hz, 3H), 1.09 (t,
8.08 (d,
Hz, 1H), 7.31 (d,
8.3 Hz, 2H), 6.62 (d,
1H), 5.26 (dd, = 24.5, 10.4 Hz, 1H), 4.11 (dq,
2H), 4.02 – 3.87 (m, 2H), 2.09 (s, 3H), 1.22 (t,
1.08 (t,
J
= 3.7 Hz, 1H), 7.57 (d,
J = 7.7 Hz, 1H), 7.48 (t, J = 7.6
J
J
= 8.2 Hz, 1H), 7.19 – 7.14 (m, 1H), 6.85 (d, J =
156.67, 154.36, 144.10, 143.95, 138.49, 137.78, 137.71,
130.92, 129.82, 128.19, 122.62, 119.35, 119.31, 115.88,
115.66, 115.54, 114.60, 114.53, 63.27, 63.20, 62.99, 62.92,
48.53, 46.98, 40.60, 40.39, 40.19, 39.98, 39.77, 39.56, 39.35,
16.79, 16.74, 16.59, 16.54.ESI-HRMS m/z Calc for C20H22FN2O4P
[M+Na]+ :427.1199; found: 427.1204.
J
= 8.5 Hz, 2H), 6.11 (dd,
J
J
= 10.4, 6.2 Hz,
J
= 14.2, 7.1 Hz,
= 7.1 Hz, 3H),
= 7.0 Hz, 3H).13C NMR (126 MHz, DMSO) δ 161.68,
J
J
161.64, 145.11, 144.99, 138.47, 137.65, 137.60, 130.80,
130.13, 129.78, 128.12, 126.32, 122.56, 119.40, 119.38,
115.50, 113.86, 63.22, 63.16, 62.92, 62.86, 48.28, 47.05, 40.50,
40.43, 40.34, 40.26, 40.17, 40.00, 39.84, 39.67, 39.50, 20.45,
16.77, 16.73, 16.58, 16.54.ESI-HRMS m/z Calc for C21H25N2O4P
[M+Na]+ :423.1450; found: 423.1447.
4a7: Yield80.64%, 1H NMR (400 MHz, DMSO) δ 12.13 (s, 1H),
8.13 (d,
5.6 Hz, 1H), 7.66 (d,
(d, = 8.2 Hz, 1H), 7.25 – 7.17 (m, 1H), 6.86 (d,
5.47 (dd, = 22.6, 9.4 Hz, 1H), 4.21 – 3.88 (m, 4H), 1.21 (t,
7.0 Hz, 3H), 1.11 (t,
= 7.0 Hz, 3H).13C NMR (101 MHz, DMSO)
J
= 3.4 Hz, 1H), 8.03 (d,
J
= 9.3 Hz, 2H), 7.95 (dd,
= 7.5 Hz, 1H), 7.35
= 9.3 Hz, 2H),
J = 9.4,
J
= 7.7 Hz, 1H), 7.54 (t,
J
J
J
4a3: Yield70.24%, 1H NMR (400 MHz, DMSO) δ 12.00 (s, 1H),
J
J =
J
8.10 (d,
Hz, 1H), 7.31 (d,
= 7.8 Hz, 1H), 6.57 (s, 1H), 6.50 (d,
J
= 3.7 Hz, 1H), 7.59 (d,
= 8.2 Hz, 1H), 7.17 (t,
= 8.1 Hz, 1H), 6.39 (d,
= 10.2, 6.2 Hz, 1H), 5.30 (dd,
= 14.2, 7.1 Hz, 2H), 4.04 – 3.85 (m, 2H),
= 7.0 Hz, 3H).13C
J
= 7.7 Hz, 1H), 7.48 (t,
J = 7.7
δ 161.52, 161.47, 150.77, 150.64, 138.56, 138.03, 137.97,
131.07, 129.29, 128.28, 126.88, 126.75, 126.71, 124.19,
122.68, 119.27, 119.24, 117.62, 117.30, 115.59, 112.99, 99.99,
63.35, 63.29, 63.16, 63.09, 47.66, 46.10, 40.60, 40.39, 40.18,
39.97, 39.76, 39.56, 39.35, 16.76, 16.71, 16.59, 16.53.ESI-
HRMS m/z Calc for C21H22F3N2O4P [M+Na]+: 477.1167; found:
477.1165.
J
J
= 7.5 Hz, 1H), 6.93 (t,
J
J
J
=
7.4 Hz, 1H), 6.25 (dd,
10.3 Hz, 1H), 4.12 (dq,
J
J
J = 24.4,
2.13 (s, 3H), 1.22 (t, J = 7.1 Hz, 3H), 1.09 (t, J
NMR (101 MHz, DMSO) δ 161.66, 161.61, 147.46, 147.32,
138.46, 138.35, 137.73, 137.67, 130.86, 130.14, 129.24,
128.16, 122.60, 119.39, 119.36, 118.69, 115.51, 114.43,
110.74, 63.26, 63.19, 62.97, 62.90, 48.02, 46.47, 40.57, 40.36,
40.15, 39.95, 39.74, 39.53, 39.32, 21.78, 16.79, 16.73, 16.60,
16.54. ESI-HRMS m/z Calc for C21H25N2O4P [M-H]- :399.1474;
found: 399.1493.
4a4: Yield63.62%, 1H NMR (500 MHz, DMSO) δ 11.87 (s, 1H),
8.05 (d, J = 3.7 Hz, 1H), 7.65 (d, J = 7.7 Hz, 1H), 7.49 (t, J = 7.7
Hz, 1H), 7.31 (d, J = 8.2 Hz, 1H), 7.19 (t, J = 7.5 Hz, 1H), 4.48 (d,
J = 21.9 Hz, 1H), 4.14 – 4.05 (m, 2H), 3.96 – 3.84 (m, 2H), 2.40
4b1: Yield38.61%, 1H NMR (500 MHz, DMSO) δ 11.87 (s, 1H),
8.05 (d,
J
= 3.7 Hz, 1H), 7.65 (d,
= 8.2 Hz, 1H), 7.19 (t,
= 21.9 Hz, 1H), 4.14 – 4.05 (m, 2H), 3.96 – 3.84 (m, 2H), 2.40
J
= 7.7 Hz, 1H), 7.49 (t,
J = 7.7
Hz, 1H), 7.31 (d,
J
J
= 7.5 Hz, 1H), 4.48 (d,
J
(ddd,
J
= 25.7, 13.6, 7.0 Hz, 2H), 1.36 (dt,
= 16.7, 10.7, 5.3 Hz, 5H), 1.08 (t,
= 7.3 Hz, 3H).13C NMR (126 MHz, DMSO) δ 161.86,
J
= 14.0, 7.0 Hz, 2H),
1.26 (ddd,
0.82 (t,
J
J
= 7.0 Hz, 3H),
J
161.82, 137.44, 137.39, 136.41, 131.83, 131.39, 129.77,
127.65, 119.50, 119.47, 115.29, 62.87, 62.82, 62.39, 62.34,
53.21, 51.97, 47.65, 47.53, 40.52, 40.35, 40.18, 40.02, 39.85,
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39.68, 39.52, 31.85, 20.83, 20.18, 16.80, 16.76, 16.64, 16.60, (d,
14.28.ESI-HRMS m/z Calc for C19H29N2O4P [M+H]+: 381.1943; (m, 4H), 2.33 (s, 3H), 1.22 (t,
found: 381.1953.
4b2: Yield75.87%, 1H NMR (400 MHz, DMSO) δ 11.88 (s, 1H), 150.66, 137.80, 137.74, 136.55, 132.32, 131.74, 129.16,
8.01 (d, = 3.6 Hz, 1H), 7.38 – 7.27 (m, 2H), 7.21 (d, = 8.3 Hz, 127.70, 126.88, 126.73, 126.69, 124.20, 119.23, 119.20,
1H), 6.85 (d, = 8.2 Hz, 2H), 6.61 (d, = 8.3 Hz, 2H), 6.12 (dd,
= 10.4, 6.2 Hz, 1H), 5.26 (dd, = 24.5, 10.5 Hz, 1H), 4.11 (p,
J
= 8.7 Hz, 2H), 5.37 (dd,
J
= 23.7, 9.7 Hz, 1H), 4.16 – 3.88
J
= 7.0 Hz, 3H), 1.10 (t, J = 7.0 Hz,
3H).13C NMR (101 MHz, DMSO) δ 161.42, 161.37, 150.79,
J
J
J
J
J
117.58, 117.27, 116.95, 115.49, 113.00, 63.31, 63.25, 63.16,
63.09, 47.66, 46.11, 40.60, 40.39, 40.18, 39.98, 39.77, 39.56,
J
J
=
7.2 Hz, 2H), 4.02 – 3.82 (m, 2H), 2.31 (s, 3H), 2.08 (s, 3H), 1.22 39.35, 20.77, 16.77, 16.72, 16.59, 16.53.ESI-HRMS m/z Calc for
(t, = 7.0 Hz, 3H), 1.07 (t,
= 7.0 Hz, 3H). 13C NMR (101 MHz, C21H24FN2O4P [M+Na]+ :441.1355; found: 441.1368.
DMSO) δ 161.58, 161.53, 145.14, 144.99, 137.44, 137.37,
4b7: Yield59.59%, 1H NMR (500 MHz, DMSO) δ 11.76 (s, 1H),
136.46, 132.05, 131.60, 129.99, 129.76, 127.57, 126.27, 8.00 (d, = 3.6 Hz, 1H), 7.25 (d, = 8.9 Hz, 1H), 7.18 (d, = 2.5
119.35, 119.32, 115.39, 113.87, 63.20, 63.13, 62.93, 62.86, Hz, 1H), 7.14 (dd, = 8.9, 2.6 Hz, 1H), 4.48 (d, = 21.8 Hz, 1H),
48.37, 46.82, 40.59, 40.38, 40.17, 39.96, 39.75, 39.54, 39.34, 4.13 – 4.05 (m, 2H), 3.94 – 3.85 (m, 2H), 3.79 (s, 3H), 2.49 –
20.78, 20.46, 16.79, 16.74, 16.60, 16.54.ESI-HRMS m/z Calc for 2.31 (m, 2H), 1.35 (dd, = 14.0, 7.0 Hz, 2H), 1.30 – 1.22 (m,
C22H27N2O4P [M+Na]+ :437.1606; found: 437.1616. = 7.2 Hz, 3H).13C NMR
8H), 1.08 (t, = 7.0 Hz, 3H), 0.83 (t,
4b3: Yield71.80%, 1H NMR (500 MHz, DMSO) δ 11.89 (s, 1H), (126 MHz, DMSO) δ 161.52, 161.47, 154.73, 137.33, 137.28,
J
J
J
J
J
J
J
J
J
J
8.02 (d,
J
= 3.7 Hz, 1H), 7.36 (s, 1H), 7.31 (d,
= 8.4 Hz, 1H), 6.92 (t, = 7.8 Hz, 1H), 6.56 (s, 1H), 6.49 62.44, 62.38, 55.93, 53.21, 51.97, 47.66, 47.54, 40.48, 40.41,
= 7.4 Hz, 1H), 6.21 (dd, 40.31, 40.24, 40.15, 39.98, 39.81, 39.65, 39.48, 31.85, 20.17,
= 24.4, 10.3 Hz, 1H), 4.12 – 3.86 16.79, 16.75, 16.64, 16.60, 14.27, 8.41.ESI-HRMS m/z Calc for
(m, 4H), 2.31 (s, 3H), 2.13 (s, 3H), 1.25 (t,
= 7.1 Hz, 1H), 1.21 C22H24F3N2O4P [M+Na]+ :491.1323; found: 491.1328.
(d, = 7.0 Hz, 3H), 1.08 (t,
= 7.0 Hz, 3H).13C NMR (126 MHz, 4c1: Yield59.59%, 1H NMR (500 MHz, DMSO) δ 11.76 (s, 1H),
DMSO) δ 161.55, 161.51, 147.46, 147.35, 138.32, 137.49, 8.00 (d, = 3.6 Hz, 1H), 7.25 (d, = 8.9 Hz, 1H), 7.18 (d, = 2.5
137.44, 136.47, 132.06, 131.62, 130.00, 129.20, 127.60, Hz, 1H), 7.14 (dd, = 8.9, 2.6 Hz, 1H), 4.48 (d, = 21.8 Hz, 1H),
119.36, 119.34, 118.66, 115.41, 114.48, 110.79, 63.19, 63.14, 4.13 – 4.05 (m, 2H), 3.94 – 3.85 (m, 2H), 3.79 (s, 3H), 2.49 –
62.94, 62.89, 48.00, 46.76, 40.49, 40.42, 40.32, 40.25, 40.16, 2.31 (m, 2H), 1.35 (dd, = 14.0, 7.0 Hz, 2H), 1.30 – 1.22 (m,
39.99, 39.82, 39.66, 39.49, 21.77, 20.77, 16.77, 16.73, 16.59, 8H), 1.08 (t, = 7.0 Hz, 3H), 0.83 (t,
= 7.2 Hz, 3H).13C NMR
16.54.ESI-HRMS m/z Calc for C22H27N2O4P [M-H]-:413.1681; (126 MHz, DMSO) δ 161.52, 161.47, 154.73, 137.33, 137.28,
found: 413.1681. 132.91, 130.20, 120.11, 119.90, 116.69, 109.52, 62.91, 62.85,
4b4: Yield71.72%, 1H NMR (500 MHz, DMSO) δ 11.89 (s, 1H), 62.44, 62.38, 55.93, 53.21, 51.97, 47.66, 47.54, 40.48, 40.41,
J = 8.4 Hz, 1H), 132.91, 130.20, 120.11, 119.90, 116.69, 109.52, 62.91, 62.85,
7.21 (d,
J
J
(dd,
J
= 8.1, 1.8 Hz, 1H), 6.39 (d,
J
J =
10.3, 6.2 Hz, 1H), 5.29 (dd,
J
J
J
J
J
J
J
J
J
J
J
J
8.03 (d,
J
= 3.7 Hz, 1H), 7.35 (s, 1H), 7.31 (d,
= 8.4 Hz, 1H), 7.04 (dd, = 8.4, 7.4 Hz, 2H), 6.71 (d,
= 7.3 Hz, 1H), 6.32 (dd,
= 10.2, 6.3 Hz, C19H29N2O5P [M+H]+ :397.1892; found: 397.1908.
= 24.4, 10.2 Hz, 1H), 4.13 – 3.86 (m, 4H), 2.31
4c2: Yield79.27%, 1H NMR (500 MHz, DMSO) δ 11.86 (s, 1H),
(s, 3H), 1.27 – 1.24 (m, 1H), 1.22 (t, = 7.0 Hz, 3H), 1.08 (t, 8.02 (d, = 3.7 Hz, 1H), 7.25 (d, = 9.0 Hz, 1H), 7.13 (dd, = 8.9,
7.0 Hz, 3H).13C NMR (126 MHz, DMSO) δ 161.56, 161.52, 2.6 Hz, 1H), 7.07 (d,
= 2.7 Hz, 1H), 6.85 (d, = 8.3 Hz, 2H), 6.61
147.45, 147.34, 137.53, 137.48, 136.47, 132.10, 131.65, (d, = 8.5 Hz, 2H), 6.06 (dd, = 10.1, 6.6 Hz, 1H), 5.26 (dd,
129.85, 129.34, 127.61, 119.35, 119.32, 117.76, 115.43, 24.6, 10.2 Hz, 1H), 4.12 (dq, = 14.2, 7.1 Hz, 2H), 4.01 – 3.86
113.67, 63.23, 63.18, 62.99, 62.93, 47.98, 46.73, 40.46, 40.38, (m, 2H), 3.76 (s, 3H), 2.09 (s, 3H), 1.23 (t, = 7.0 Hz, 3H), 1.08
40.29, 40.21, 40.12, 40.05, 39.96, 39.79, 39.62, 39.46, 20.76, (t,
= 7.0 Hz, 3H).13C NMR (126 MHz, DMSO) δ 161.22, 161.18,
16.76, 16.72, 16.57, 16.53.ESI-HRMS m/z Calc for C21H25N2O4P 154.78, 145.06, 144.94, 137.19, 137.14, 132.97, 130.46,
[M+Na]+ :423.1450; found: 423.1451.
129.77, 126.28, 120.16, 119.96, 119.94, 116.83, 113.81,
4b5: Yield76.32%, 1H NMR (400 MHz, DMSO) δ 12.05 (s, 1H), 109.33, 63.19, 63.14, 62.95, 62.90, 55.90, 48.33, 47.09, 40.50,
J
= 8.4 Hz, 1H), 40.31, 40.24, 40.15, 39.98, 39.81, 39.65, 39.48, 31.85, 20.17,
7.22 (d,
J
J
J =
16.79, 16.75, 16.64, 16.60, 14.27, 8.41.ESI-HRMS m/z Calc for
7.8 Hz, 2H), 6.56 (t,
1H), 5.30 (dd,
J
J
J
J
J
=
J
J
J
J
J
J
J
J =
J
J
J
8.06 (d,
5.6 Hz, 1H), 7.43 (s, 1H), 7.36 (d,
Hz, 1H), 6.86 (d, = 9.3 Hz, 2H), 5.47 (dd,
4.16 – 3.90 (m, 4H), 2.34 (s, 3H), 1.22 (t,
= 7.0 Hz, 3H).13C NMR (101 MHz, DMSO) δ 161.27, 161.21, 8.04 (d,
153.72, 153.61, 138.23, 138.17, 137.63, 136.63, 132.53, 2.6 Hz, 1H), 7.09 (d,
131.85, 128.47, 127.81, 126.43, 119.14, 119.12, 115.55, (s, 1H), 6.48 (d, = 8.1 Hz, 1H), 6.39 (d,
112.42, 63.42, 63.35, 63.29, 47.73, 46.18, 40.60, 40.39, 40.18, = 10.0, 6.5 Hz, 1H), 5.29 (dd, = 24.5, 10.0 Hz, 1H), 4.11 (dq,
39.97, 39.76, 39.56, 39.35, 20.77, 16.78, 16.73, 16.59, = 14.2, 7.1 Hz, 2H), 4.01 – 3.87 (m, 2H), 3.77 (s, 3H), 2.13 (s,
16.54.ESI-HRMS m/z Calc for C21H24N3O6P [M-H]- :444.1344; 3H), 1.24 (dt, = 7.0 Hz, 3H).13C
= 14.1, 7.0 Hz, 4H), 1.09 (t,
found: 444.1357. NMR (126 MHz, DMSO) δ 161.20, 161.16, 154.80, 147.39,
4b6: Yield84.63%, 1H NMR (400 MHz, DMSO) δ 11.98 (s, 1H), 147.27, 138.35, 137.25, 137.20, 132.99, 130.46, 129.23,
8.03 (d, = 3.5 Hz, 1H), 7.42 – 7.37 (m, 3H), 7.34 (d, = 8.4 Hz, 120.20, 119.97, 118.67, 116.85, 114.44, 110.70, 109.36, 63.21,
1H), 7.24 (d,
J
= 3.4 Hz, 1H), 8.02 (d,
J
= 9.3 Hz, 2H), 7.96 (dd,
J
= 9.3, 40.42, 40.33, 40.26, 40.17, 40.00, 39.83, 39.67, 39.50, 20.45,
J
= 8.4 Hz, 1H), 7.25 (d,
J
= 8.4 16.77, 16.73, 16.59, 16.55.ESI-HRMS m/z Calc for C22H27N2O5P
J
J
= 22.6, 9.4 Hz, 1H), [M+H]+ :431.1736; found: 431.1742.
J
= 7.0 Hz, 3H), 1.11 (t,
4c3: Yield74.36%, 1H NMR (500 MHz, DMSO) δ 11.87 (s, 1H),
= 3.6 Hz, 1H), 7.25 (d, = 8.9 Hz, 1H), 7.14 (dd, = 8.9,
= 2.7 Hz, 1H), 6.92 (t, = 7.8 Hz, 1H), 6.56
= 7.4 Hz, 1H), 6.17 (dd,
J
J
J
J
J
J
J
J
J
J
J
J
J
J
J
J
= 8.4 Hz, 1H), 7.17 (dd,
J
= 9.7, 6.1 Hz, 1H), 6.85 63.16, 62.99, 62.93, 55.91, 48.03, 46.79, 40.48, 40.41, 40.32,
12 | J. Name., 2012, 00, 1-3
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DOI: 10.1039/C7MD00098G
ARTICLE
40.24, 40.15, 39.98, 39.82, 39.65, 39.48, 21.77, 16.77, 16.72, (101 MHz, DMSO) δ 174.78, 161.75, 161.69, 150.38, 143.78,
16.59, 16.55.ESI-HRMS m/z Calc for C22H27N2O5P [M+Na]+ 137.65, 137.58, 135.35, 130.11, 126.35, 113.81, 113.78,
:453.1555; found: 453.1574.
4c4: Yield77%, 1H NMR (500 MHz, DMSO) δ 11.88 (s, 1H), 47.63, 47.47, 31.78, 30.84, 29.56, 29.51, 29.46, 29.31, 29.18,
8.04 (d, = 3.6 Hz, 1H), 7.26 (d, = 8.9 Hz, 1H), 7.14 (dd, = 8.9, 29.06, 27.02, 25.59, 22.58, 20.18, 16.82, 16.77, 16.66, 16.61,
2.7 Hz, 1H), 7.09 (d, = 2.7 Hz, 1H), 7.05 (dd,
= 8.3, 7.5 Hz, 14.42, 14.30.ESI-HRMS m/z Calc for C19H27N2O6P [M+H]+
2H), 6.70 (d, = 8.1 Hz, 2H), 6.57 (t, = 7.3 Hz, 1H), 6.28 (dd,
9.9, 6.6 Hz, 1H), 5.30 (dd, = 24.5, 10.0 Hz, 1H), 4.15 – 4.09 (m,
105.58, 102.20, 95.38, 62.85, 62.78, 62.41, 62.34, 53.17, 51.62,
J
J
J
J
J
J
J
J
=
:411.1585; found: 411.1519.
4d2: Yield66.81%, 1H NMR (500 MHz, DMSO) δ 11.85 (s, 1H),
J
2H), 4.02 – 3.88 (m, 2H), 3.76 (s, 3H), 1.27 – 1.24 (m, 1H), 1.22 7.92 (d,
(t, = 7.0 Hz, 3H), 1.09 (t,
= 7.0 Hz, 3H). 13C NMR (126 MHz, 6.82 (s, 1H), 6.60 (d,
DMSO) δ 161.21, 161.17, 154.81, 147.38, 147.27, 137.31, 10.1, 6.5 Hz, 1H), 5.20 (dd,
137.26, 132.97, 130.30, 129.36, 120.24, 119.96, 119.94, 14.2, 7.1 Hz, 2H), 4.01 – 3.83 (m, 2H), 2.09 (s, 3H), 1.22 (t,
117.78, 116.88, 113.62, 109.36, 63.26, 63.20, 63.04, 62.98, 7.1 Hz, 3H), 1.08 (t,
J
= 3.5 Hz, 1H), 7.08 (s, 1H), 6.85 (d,
= 8.5 Hz, 2H), 6.07 (s, 2H), 6.02 (dd,
= 24.3, 10.1 Hz, 1H), 4.10 (dq,
J = 8.3 Hz, 2H),
J
J
J
J
J
J
=
=
=
J
J
= 7.0 Hz, 3H).13C NMR (126 MHz, DMSO)
55.90, 48.03, 46.79, 40.44, 40.27, 40.11, 39.94, 39.77, 39.60, δ 161.44, 161.40, 150.49, 145.13, 145.01, 143.88, 137.47,
39.44, 16.75, 16.71, 16.58, 16.53.ESI-HRMS m/z Calc for 137.42, 135.47, 129.74, 126.68, 126.18, 113.79, 113.70,
C21H25N2O5P [M+Na]+ :439.1399; found: 439.1403.
113.68, 105.44, 102.22, 95.45, 63.10, 63.05, 62.89, 62.83,
4c5: Yield80.08%, 1H NMR (400 MHz, DMSO) δ 12.03 (s, 1H), 48.14, 46.90, 40.49, 40.42, 40.33, 40.25, 40.16, 39.99, 39.83,
8.07 (d, = 3.4 Hz, 1H), 8.02 (d, = 9.3 Hz, 2H), 7.94 (dd, = 9.1, 39.66, 39.49, 20.45, 16.77, 16.72, 16.59, 16.54.ESI-HRMS m/z
6.1 Hz, 1H), 7.29 (d, = 8.8 Hz, 1H), 7.18 (dt,
= 8.1, 2.6 Hz, Calc for C22H25N2O6P [M+Na]+ :467.1348; found: 467.1381.
2H), 6.85 (d, = 9.2 Hz, 2H), 5.46 (dd, = 22.7, 9.1 Hz, 1H), 4.18
4d3: Yield77.29%, 1H NMR (400 MHz, DMSO) δ 11.94 (s, 1H),
– 3.93 (m, 4H), 3.79 (s, 3H), 1.22 (t, = 7.0 Hz, 3H), 1.11 (t, 7.95 (d, = 3.3 Hz, 1H), 7.10 (s, 1H), 6.89 (dd, = 15.8, 7.0 Hz,
7.0 Hz, 3H).13C NMR (101 MHz, DMSO) δ 160.91, 160.85, 3H), 6.70 (dd,
= 8.9, 4.5 Hz, 2H), 6.30 (dd, = 9.8, 6.6 Hz, 1H),
154.89, 153.69, 153.58, 137.98, 137.93, 137.63, 133.13, 6.08 (s, 2H), 5.19 (dd, = 24.2, 10.0 Hz, 1H), 4.16 – 4.06 (m,
128.91, 126.43, 120.71, 119.76, 119.73, 117.00, 109.47, 63.43, 2H), 3.94 (ddt, = 25.1, 10.0, 7.5 Hz, 2H), 3.06 (d, = 7.2 Hz,
63.38, 63.31, 55.95, 47.82, 46.27, 40.60, 40.39, 40.18, 39.97, 1H), 1.21 (q, = 7.2 Hz, 6H), 1.09 (t,
= 7.0 Hz, 3H).13C NMR
39.77, 39.56, 39.35, 16.78, 16.72, 16.60, 16.55.ESI-HRMS m/z (101 MHz, DMSO) δ 161.46, 161.41, 156.62, 154.31, 150.58,
Calc for C21H24N3O7P [M+Na]+ :484.1250; found: 484.1273.
144.14, 143.98, 143.94, 137.63, 137.57, 135.54, 130.11,
4c6: Yield86.00%, 1H NMR (400 MHz, DMSO) δ 11.90 (s, 1H), 126.31, 115.84, 115.63, 114.53, 114.46, 113.69, 113.66,
8.03 (d, = 3.6 Hz, 1H), 7.26 (d, = 9.0 Hz, 1H), 7.15 (dd, = 8.9, 105.47, 102.28, 95.51, 63.18, 63.12, 62.99, 62.92, 48.40, 46.84,
2.6 Hz, 1H), 7.09 (d, = 2.6 Hz, 1H), 6.90 (t, = 8.9 Hz, 2H), 6.74 45.82, 40.56, 40.35, 40.14, 39.93, 39.73, 39.52, 39.31, 35.59,
– 6.66 (m, 2H), 6.33 (dd, = 9.9, 6.7 Hz, 1H), 5.24 (dd, = 24.4, 31.75, 30.84, 29.45, 29.17, 29.04, 27.01, 25.60, 22.57, 16.78,
10.0 Hz, 1H), 4.15 – 3.84 (m, 4H), 3.77 (s, 3H), 1.23 (t,
J
J
J
J
J
J
J
J
J
=
J
J
J
J
J
J
J
J
J
J
J
J
J
J
J
J
J
= 7.0 16.73, 16.60, 16.54, 14.41, 8.89.ESI-HRMS m/z Calc for
Hz, 3H), 1.08 (t,
J
= 7.0 Hz, 3H).13C NMR (101 MHz, DMSO) δ C22H25N2O6P [M-H]- :443.1372; found: 443.1727.
1
161.21, 161.16, 156.64, 154.79, 154.33, 144.05, 143.90,
137.31, 137.25, 132.99, 130.14, 120.31, 119.90, 119.87, 7.94 (d,
116.88, 115.87, 115.65, 115.43, 115.06, 114.99, 114.53, 2H), 6.82 (s, 1H), 6.70 (d,
114.45, 109.28, 63.23, 63.17, 63.02, 62.95, 55.90, 48.57, 47.02, 6.25 (dd, = 9.9, 6.6 Hz, 1H), 6.07 (s, 2H), 5.23 (dd,
= 14.2, 7.1 Hz, 2H), 4.02 – 3.84 (m, 2H), 1.22
= 7.1 Hz, 3H), 1.08 (t,
= 7.0 Hz, 3H).13C NMR (126 MHz,
DMSO) δ 161.43, 161.38, 150.54, 147.49, 147.38, 143.92,
4d4: Yield77.32%, HNMR (500 MHz, DMSO) δ 11.87 (s, 1H),
J
= 3.4 Hz, 1H), 7.09 (s, 1H), 7.04 (dd,
= 7.8 Hz, 2H), 6.56 (t,
J
= 8.3, 7.5 Hz,
J
J = 7.3 Hz, 1H),
J
J = 24.2, 9.9
40.61, 40.40, 40.19, 39.98, 39.77, 39.56, 39.35, 16.79, 16.74, Hz, 1H), 4.10 (dq,
J
16.60, 16.54.ESI-HRMS m/z Calc for C21H24FN2O5P [M+Na]+ (t,
J
J
:457.1305; found: 457.1323.
4c7: Yield74.08%, 1H NMR (400 MHz, DMSO) δ 11.96 (s, 1H), 137.56, 137.52, 135.52, 129.32, 126.57, 117.65, 113.70,
8.04 (d,
Hz, 1H), 7.15 (ddd,
2H), 5.36 (dd, = 23.8, 9.4 Hz, 1H), 4.15 – 3.89 (m, 4H), 3.78 (s, 39.82, 39.66, 39.49, 16.76, 16.72, 16.59, 16.54.ESI-HRMS m/z
3H), 1.22 (t, = 7.0 Hz, 3H), 1.10 (t,
= 7.0 Hz, 3H).13C NMR Calc for C21H23N2O6P [M+Na]+ :453.1191; found: 453.1169.
(101 MHz, DMSO) δ 161.06, 161.01, 154.84, 150.74, 150.61,
4d5: Yield73.71%, 1H NMR (400 MHz, DMSO) δ 12.02 (s, 1H),
137.55, 137.49, 133.05, 129.59, 126.89, 126.70, 124.20, 8.08 – 7.93 (m, 4H), 7.18 (s, 1H), 6.84 (s, 3H), 6.10 (s, 2H), 5.40
120.49, 119.83, 119.80, 117.57, 117.25, 116.93, 112.96, (dd, = 22.4, 9.2 Hz, 1H), 4.13 – 3.91 (m, 4H), 1.20 (t, = 7.0 Hz,
109.36, 63.32, 63.26, 63.19, 63.12, 55.91, 47.73, 46.18, 40.60, 3H), 1.11 (t,
= 7.0 Hz, 3H). 13C NMR (101 MHz, DMSO) δ
J
= 3.6 Hz, 1H), 7.39 (d,
J
= 8.7 Hz, 2H), 7.27 (d,
J = 8.9 113.68, 113.59, 105.48, 102.25, 95.48, 63.13, 63.08, 62.95,
J
= 13.0, 7.7, 2.6 Hz, 3H), 6.84 (d,
J
= 8.6 Hz, 62.89, 47.83, 46.58, 40.49, 40.42, 40.33, 40.25, 40.16, 39.99,
J
J
J
J
J
J
40.39, 40.18, 39.97, 39.76, 39.56, 39.35, 16.77, 16.72, 16.60, 161.16, 161.10, 153.75, 153.65, 150.89, 144.09, 138.24,
16.54.ESI-HRMS m/z Calc for C21H24F3N2O5P [M+Na]+ 138.18, 137.54, 136.93, 135.78, 126.44, 125.10, 113.55,
:507.1273; found: 507.1252.
113.52, 112.29, 105.67, 102.38, 95.51, 63.34, 63.32, 63.28,
4d1: Yield43.26%, 1H NMR (400 MHz, DMSO) δ 11.80 (s, 63.25, 47.62, 46.06, 40.59, 40.38, 40.17, 39.97, 39.76, 39.55,
1H), 7.92 (d,
2H), 4.44 (d,
J
J
= 3.4 Hz, 1H), 7.17 (s, 1H), 6.83 (s, 1H), 6.10 (s, 39.34, 16.78, 16.72, 16.60, 16.54, -14.99.ESI-HRMS m/z Calc
= 21.4 Hz, 1H), 4.08 (ddd,
= 10.2, 8.7, 5.3 Hz, for C21H22N3O8P [M+Na]+ :498.1042; found: 498.1064.
4d6: Yield85.70%, 1H NMR (400 MHz, DMSO) δ 11.98 (s,1H),
J
2H), 3.97 – 3.82 (m, 2H), 2.50 – 2.34 (m, 2H), 1.40 – 1.22 (m,
10H), 1.08 (t, = 7.0 Hz, 3H), 0.83 (t,
= 7.2 Hz, 3H).13C NMR 7.94 (d,
J
J
J
= 3.2 Hz,1H), 7.38 (d,
J
= 8.6 Hz,2H), 7.15 – 7.06
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 13
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Journal Name
(m,2H), 6.88 – 6.79 (m,3H), 6.08 (s,2H), 5.29 (dd,
Hz,1H), 4.13 – 4.07 (m,2H), 4.02 – 3.86 (m,3H), 1.26 – 1.18 131.19, 131.14, 131.02, 130.94, 127.13, 122.03, 121.45,
(m,6H), 1.09 (t,
= 7.0 Hz,3H).13C NMR (101 MHz, DMSO) δ 121.43, 119.94, 116.98, 110.97, 61.90, 61.85, 55.99, 40.52,
J = 23.5, 9.4 161.58, 155.45, 154.95, 150.38, 150.35, 137.62, 135.01,
J
161.32, 150.72, 144.01, 137.81, 135.65, 126.71, 125.77, 40.45, 40.36, 40.28, 40.19, 40.02, 39.85, 39.69, 39.52, 32.85,
113.63, 112.94, 105.56, 102.32, 95.52, 63.27, 63.20, 63.15, 31.78, 16.70, 16.66.ESI-HRMS m/z Calc for C22H25N2O5P
63.08, 47.54, 45.97, 40.54, 40.33, 40.12, 39.91, 39.70, 39.50, [M+Na]+ : 451.1399; found: 451.1404.
39.29, 29.02, 22.56, 16.76, 16.71, 16.59, 16.53.ESI-HRMS m/z
Calc for C21H22FN2O6P [M+Na]+ :471.1097; found: 471.1118.
5d: Yield70.98%, 1H NMR (400 MHz,
1H), 8.74 (s, 1H), 8.56 (s, 1H), 7.38 (d, J = 8.7 Hz, 2H), 7.32 (dd,
d-DMSO) δ 12.08 (s,
4d7: Yield75.49%, 1H NMR (400 MHz, DMSO) δ 11.94 (s, 1H), J = 8.4, 2.3 Hz, 2H), 7.20 (d, J = 8.0 Hz, 2H), 6.15 (d, J = 8.2 Hz,
7.95 (d,
3H), 6.70 (dd,
6.08 (s, 2H), 5.19 (dd,
2H), 4.03 – 3.83 (m, 2H), 3.06 (d,
Hz, 6H). 13C NMR (101 MHz, DMSO) δ 161.03 (d,
154.84 (s), 150.68 (d, = 12.7 Hz), 137.52 (d,
= 6.0 Hz), 133.05 for C22H23N2O6P [M+H]+ : 443.1372; found: 443.1391.
(s), 129.59 (s), 126.79 (d, = 19.3 Hz), 120.49 (s), 119.82 (d,
3.1 Hz), 117.57 (s), 117.25 (s), 116.93 (s), 112.96 (s), 109.36 (s),
63.22 (dd, = 14.0, 6.9 Hz), 55.91 (s), 47.73 (s), 46.18 (s), 40.60
(s), 40.39 (s), 40.18 (s), 39.97 (s), 39.76 (s), 39.56 (s), 39.35 (s),
J
= 3.3 Hz, 1H), 7.10 (s, 1H), 6.89 (dd,
= 8.9, 4.5 Hz, 2H), 6.30 (dd,
= 9.8, 6.6 Hz, 1H), (t, J = 7.0 Hz, 6H). 13C NMR (101 MHz, DMSO) δ 189.85, 161.90,
= 24.2, 10.0 Hz, 1H), 4.17 – 4.06 (m, 155.37, 153.66, 152.08, 144.53, 142.07, 140.40, 131.17,
= 7.2 Hz, 1H), 1.21 (q, = 7.2 122.90, 121.34, 113.87, 113.23, 107.29, 106.73, 103.01,
= 5.4 Hz), 102.65, 95.37, 61.83, 32.81, 31.44, 16.71.ESI-HRMS m/z Calc
J = 15.8, 7.0 Hz, 2H), 3.97 (dd, J = 8.0, 7.2 Hz, 4H), 3.26 (d, J = 21.5 Hz, 2H), 1.19
J
J
J
J
J
J
J
J
J
J
=
3.2. Biological Assays
The detailed procedures for other experimental methods
are described in the Supplementary data (Part 2). The
materials, instrumentation, and methods for the cytotoxicity
assay, cell cycle analysis, cell apoptosis analysis, western blot
and transfection assays were described in our previous work.26
J
16.66 (dd,
J = 17.6, 5.4 Hz).ESI-HRMS m/z Calc for
C22H22F3N2O6P [M+Na]+ :521.1065; found: 521.1089.
3.1.2 General procedure for compound 5(5a-5d)
2-oxo-quinoline 3-carbaldehyde derivatives
3 (1 mmol),
4. Conclusions
diethyl 4-aminobenzylphosphonate (1.5 mmol) and 5 mL
toluene were mixed in pressure tube and reacted at 120oC for
4h. After the reaction, the mixture was cooled to room
In summary, we have designed and synthesized a set of 2-
oxo quinoline APA derivatives
(4-5) and evaluated the
temperature and filtered to yield compound
5 as pale yellow
cytotoxicity of target compounds on three cancer cell lines
(HepG2, SKOV-3 and NCI-H460). The target compounds
displayed evident anticancer activity with low cytotoxicity
against normal HL-7702 cells. The cell apoptosis-inducing
investigation of representative compound 5b in HpeG2 cells
showed that antitumor activity of this compound may rely on
the apoptosis of cancer cells by regulation of levels of Bax, Bcl-
powder.
1
5a: Yield70.81%, H NMR (400 MHz, DMSO) δ 11.68 (s, 1H),
7.72 (d, J = 2.6 Hz, 1H), 6.86 (s, 1H), 6.69 (t, J = 7.7 Hz, 1H), 6.60
(s, 1H), 6.33 (s, 1H), 6.26 (d, J = 8.0 Hz, 1H), 6.16 (d, J = 7.3 Hz,
1H), 5.98 – 5.90 (m, 1H), 5.85 (s, 2H), 5.01 (dd, J = 24.2, 10.0
Hz, 1H), 3.93 – 3.82 (m, 2H), 3.79 – 3.62 (m, 2H), 1.91 (s, 3H),
0.99 (t, J = 7.0 Hz, 3H), 0.86 (t, J = 7.0 Hz, 3H).13C NMR (101
MHz, DMSO) δ 161.44, 161.38, 150.52, 147.49, 147.35, 143.91,
138.31, 137.56, 137.49, 135.49, 129.20, 126.69, 118.57,
114.40, 113.71, 113.68, 110.68, 105.46, 102.25, 95.46, 63.14,
63.08, 62.94, 62.87, 47.92, 46.36, 46.03, 21.78, 16.78, 16.73,
16.60, 16.54.ESI-HRMS m/z Calc for C21H23N2O4P [M+H]+
:399.1474; found: 339.1476.
2 and cytochrome
c
, intracellular Ca2+ release and ROS
generation, activation of caspase-9 and caspase-3 and
subsequent cleavage of PARP. Cell cycle study revealed that
antitumor activity of compound 5b might be exerted by G2/M
phase arrest that accompanied by the expression of p53, p21
and p27 proteins. The above results confirmed that the
rational design of 2-oxo-quinoline APA derivatives as novel
antitumor agents was feasible.
1
5b: Yield63.13%, H NMR (400 MHz, DMSO) δ 12.10 (s, 1H),
8.79 (s, 1H), 8.59 (s, 1H), 7.67 (s, 1H), 7.42 (dd,
1H), 7.34 (dd, = 8.5, 2.3 Hz, 2H), 7.25 (dd, = 12.0, 8.3 Hz, 3H),
3.97 (dq, = 14.2, 7.1 Hz, 4H), 3.27 (d, = 21.5 Hz, 2H), 2.36 (s,
3H), 1.19 (t,
= 7.0 Hz, 6H).13C NMR (101 MHz, DMSO) δ
J = 8.4, 1.7 Hz,
J
J
J
J
Acknowledgements
J
This study was supported by the National Natural Science
Foundation of China (No. 21501032), Guangxi Natural Science
Foundation of China (Nos. 2016GXNSFAA380300 and
2014GXNSFBA118050) and State Key Laboratory Cultivation
Base for the Chemistry and Molecular Engineering of Medicinal
161.91, 155.52, 150.44, 150.40, 138.36, 137.77, 133.82,
131.91, 131.19, 131.13, 130.97, 130.88, 129.41, 126.80,
121.47, 121.44, 119.29, 115.60, 61.90, 61.84, 40.60, 40.40,
40.19, 39.98, 39.77, 39.56, 39.35, 32.93, 31.59, 20.87, 16.72,
16.66. ESI-HRMS m/z Calc for C22H25N2O4P [M-H]-: 411.1474;
found: 411.1549.
Resources
，Ministry of Science and Technology of China (No.
CMEMR2014-B14).
1
5c: Yield64.65%, H NMR (500 MHz, DMSO) δ 12.06 (s, 1H),
8.80 (s, 1H), 8.64 (s, 1H), 7.46 (d,
8.4, 2.3 Hz, 2H), 7.30 (d, = 8.9 Hz, 1H), 7.24 (dd,
Hz, 3H), 3.96 (dd, = 15.1, 7.1 Hz, 4H), 3.26 (d, = 21.5 Hz, 2H),
1.18 (t,
= 7.0 Hz, 6H).13C NMR (126 MHz, DMSO) δ 190.33,
J
= 2.7 Hz, 1H), 7.34 (dd,
J =
J
J
= 8.9, 2.8
Notes and references
J
J
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14 | J. Name., 2012, 00, 1-3
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Gopalakrishnanb and G. Venkatraman, Bioorg. Med.
View Article Online
DOI: 10.1039/C7MD00098G
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View Article Online
DOI: 10.1039/C7MD00098G
Graphical abstract
Design, synthesis and pharmacological evaluation of new
2ꢀoxoꢀquinoline derivatives containing αꢀaminophosphonates as
potential antitumor agents
YanꢀCheng Yu, WenꢀBin Kuang, RiꢀZhen Huang, YiꢀLin Fang, Ye Zhang, ZhenꢀFeng Chen,
XianꢀLi Ma
O
EtO
P OEt
CH R2
N
H
R1
Improved in vitro
Cytotoxic Activity
N
H
O
CHO
O
R1
Cell Cycle Arrest
O
P
N
OEt
H
Induction of Apotosis
H
C
N
OEt
R1
N
O
H
A series of novel 2ꢀoxoꢀquinoline derivatives containing αꢀaminophosphonates was designed and
synthesized as antitumor agents. Representative compound 5b blocked the HepG2 cell cycle at
G2/M phase by the regulation of p53, p21 and p27 proteins and induced apoptosis through
mitochondrial pathway.
1 / 1