Design, synthesis, and biological evaluation of novel tetrahydroisoquinoline derivatives as potential antitumor candidate

Article abstract of DOI:10.1111/cbdd.12873

A novel class of tetrahydroisoquinoline derivatives was designed and synthesized as antitumor agents and evaluated for their in vitro and in vivo biological activities. The antiproliferative activities of all the target compounds on HUVEC, MCF-7, and HT-2

Full text of DOI:10.1111/cbdd.12873

Received Date : 12-Jun-2016
Revised Date : 04-Aug-2016
Accepted Date : 29-Aug-2016
Article type
: Research Article
Design, Synthesis and Biological Evaluation of Novel
Tetrahydroisoquinoline Derivatives as Potential Antitumor Candidate
Panhu Zhu, Wenfeng Ye, Jiaming Li*, Yanchun Zhang†, Weijun Huang, Mohan Cheng,
Yujun Wang, Yang Zhang, Huicai Liu, Jian Zuo
(Department of Medicinal Chemistry, Anhui University of Chinese Medicine, 103 Meishan Road, Hefei 230031,
PR China)
Abstract
A novel class of tetrahydroisoquinoline derivatives were designed and synthesized as
antitumor agents and evaluated for their in vitro and in vivo biological activities. The
antiproliferative activities of all the target compounds on HUVEC, MCF-7 and HT-29 were
evaluated. Compared with Colchicine (1.04×10-2 μM), 17d and 17e exhibited outstanding
*Corresponding author: lijiaming2004@sina.com
†Corresponding author: yczhang2017@163.com
This article has been accepted for publication and undergone full peer review but has not
been through the copyediting, typesetting, pagination and proofreading process, which may
lead to differences between this version and the Version of Record. Please cite this article as
doi: 10.1111/cbdd.12873
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activity on MCF-7 with IC50 values 0.26×10-2 μM and 0.89×10-3 μΜ in cell cytotoxicity
assay. The tubulin polymerization assay demonstrated that 17d and 17e exhibited better
inhibition rate. In the MCF-7-xenograft mouse model which were treated with 17d and 17e
by intraperitoneal injection, the tumor weight was decreased at same rate with Tamoxifen,
and relative tumor proliferation rates were 59.48 % and 41.33 %, while Tamoxifen was 45.08
% with a daily dose of 20 mg/kg, which were demonstrated potent in vivo efficacy.
Keywords: tetrahydroisoquinoline derivatives, MCF-7, selectivity, antitumor
1. Introduction
Tubulin, a globular protein, is consists of five distinct families, α, β, δ, ε and ζ, among
which α- tubulin and β- tubulin build the microtubule [1, 2]. Microtubules are long, hollow
cylinders of tubulin dimmers, playing critical roles in cell division, formation and
maintenance of cell shape, motility, cell signaling, secretion, and intracellular transport [3-5]. It
is why molecules acting on the cellular microtubule dynamics form one of the largest groups
of effective chemotherapeutics used against cancer.
Podophyllotoxin was a non-alkaloid toxic lignin extracted from the roots and rhizomes
of Podophyllum species. Podophyllotoxin had been extensively researched as a cytotoxic
agent, exhibiting potent anticancer activity toward numerous cancer cell lines by inhibit both
tubulin polymerization and DNA topoisomerase-II. However, the systemic toxicity of
Podophyllotoxin limited its clinical application as a medicinal drug [6]. According to the
biological properties of Podophyllotoxin, lots of derivatives were semi-synthesized [7, 8], such
as Etoposide and Teniposide (semis synthetic derivatives of Podophyllotoxin), and used in
cancer chemotherapy against various cancer [9]. According to the structure of microtubule
assembly inhibitor Podophyllotoxin 3 and Topo II inhibitor Etoposide [10], Akira’s team
developed compounds 1 and 2（Fig 1）with the de-hydroxyl C ring, exhibited cytotoxicity due
to inhibition of microtubule assembly [11]. However, compound 1 did not inhibit Topo II
activity. It was reported that the aza-podophyllotoxin, based on the deoxidized derivatives of
Podophyllotoxin (Compound 3, Fig 1) exhibited tubulin polymerization inhibition ability[12,
13]
.
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Fig 1.
The structure of Podophyllotoxin and its derivatives
Isoquinoline alkaloid was a class of natural products that possess a broad spectrum of
pharmacological actions, particularly anticancer activity [14]. Tetrahydroisoquinoline, one of
the most important heterocyclic scaffolds, can be found in various pharmaceutically
important antitumor antibiotics. Tetrahydroisoquinoline derivatives (Fig 2, compound 4, 5)
which were developed by Ramanivas et al. had shown significant activity against human
prostate cancer cell line (DU-145) with IC50 value 0.72 and 1.23 μM respectively [15]
Comparing with CA-4, compounds 6, 7, 8 developed by Leese et al. manifest better
.
anti-proliferation activity against human burkitt lymphoma cells [16]
.
Fig 2. The structure of tetrahydroisoquinoline derivatives
In recent years, it was found that the phenyl acrylic acid series ferulic acid [17], hydroxy
cinnamic acid [18] and cinnamon acid [19] which have preferable antitumor effect, inducing
apoptosis, sensitization effect of chemotherapy, radiation protection, etc. may correlated with
benzene acrylic.
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Fig 3. Design of the tetrahydroisoquinoline derivatives
Based on the above structural requirements that the A and E rings of the
Podophyllotoxin and tetrahydroisoquinoline were important to the activity, we postulated that
the hydroxyl on C ring and D ring was not the essential group for antineoplastic activity of
Podophyllotoxin. Therefore, we design and synthesis deoxypodophyllotoxin derivatives with
benzene acrylic group, which can also be referred as opened D rings, to promote efficacy and
selectivity to tumor.
2. Chemistry
The target compounds were reached as follows. Compounds 12(a-h) were prepared by
substituted benzoic acid and aromatic amine, in the presence of SOCl2 [20]. Intermediates
13(a-h) were gained by reacting 12(a-h) with substituted benzaldehyde in the presence of
POCl3 (Bischler-Napieralsk [21] reaction), which were then reduced by NaBH4 to achieve
14(a-h). Compounds 16(a-l) were obtained from different substituted cinnamic acid via
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SOCl2-catalyzed amidation reaction [22] followed by hydrolization to achieve compounds
17(a-k). Target compounds 19(c-e) were prepared by etherification with the commercially
available amine in the presence of KI [23]
.
Scheme 1. Synthesis of Compounds 10(a-h)
Reagents and conditions: (a) SOCl2, toluene 77 °C, 4h.
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Reagents and conditions: (a) NaOH (aq), CH2Cl2, 0 °C, 2.5h; (b) POCl3, Toluene 115 °C, 7h; (c) NaBH4, 0 °C, CH3OH, 4h; (d) Et3N,
CH2Cl2, 0 °C, 2.5h; (e) NaOH (aq), CH3OH, r.t, 24h.
In the preparation of cinnamic acid chloride, because homemade acetylated cinnamic
acid was cloudy in a solvent would result the reaction yield significantly reduced, DMF was
used as co-solvent solution.
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Scheme 2. Synthesis of Compounds 17(a-k)
During Bischler-Napieralski [21] reaction, the POCl3 should be added below 95
°C, otherwise, the reaction process would produce a large quantity
of thick black gum, leading to the failure of the reaction.
Scheme 3. Synthesis of Compounds 19(c-e)
Reagents and conditions: (a) K2CO3, KI, isopropanol, r.t, 3h
Scheme 4. Synthesis of Compounds 21(c-f)
Reagents and conditions: (a) Et3N, CH2Cl2, 0 °C, 2.5 h.
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3. Biological evaluation
3.1 In vitro antiproliferative activities
To test the anticancer activities of the synthesized compounds, firstly we evaluated
antiproliferative activities of compounds against HUVEC (Umbilical Vein Endothelial Cells),
HT-29 (Colonic Carcinoma Cells) and MCF-7 (Breast Cancer Cells). Colchicine was taken as
the positive control. The results were summarized in (Table 1). From the results of the SRB
assay, it was found that compound 19c showed better potency against HUVEC and HT-29,
compounds 17d and 17e showed the extraordinary activity against MCF-7. The results
expressed as IC50 values, are reported in Table 1.
Table 1. In vitro inhibitory activity of target compounds against human cancer cells ( x s , n=3)
Compound
IC50/μM
MCF-7
HUVEC
7.00 3.50
2.86 0.36
334.00 13.04
5.86 2.74
11.14 2.89
2.45 0.74
10.74 7.00
11.97 3.83
4.59 0.62
12.03 4.39
81.08 7.34
1.71 0.39
HT-29
17a
17b
17c
17d
17e
17f
17g
17h
17i
6.00 1.02
5.53 0.49
2.98 0.10
5.22 0.15
4.53 0.63
2.54 0.19
6.43 0.99
2.09 0.15
3.37 0.44
9.83 2.84
12.05 7.08
10.59 5.63
7.95 0.96
1.77 0.72
135.97 7.32
0.26×10-2 0.40×10-3
0.89×10-3 0.28×10-3
10.63 0.57
13.58 0.91
23.50 5.00
22.44 3.52
17j
145.16 25.16
74.41 27.78
2.44 0.44
17k
19d
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19e
19c
22d
22e
22f
22c
1.78 0.62
0.78 0.19
3.91 1.29
2.47 0.59
1.54 0.79
0.35 0.08
39.01 6.12
12.38 2.64
47.7 12.49
5.90 0.96
149.20 35.52
16.78 13.17
19.52 8.04
23.72 19.09
49.31 3.80
28.83 4.01
124.41 6.32
53.43 12.49
3.22×10-2 1.30×10-
1.88 0.43
1.04×10-2 0.28×10-2
Colchicine
2
The anti-proliferative activities of compounds against all the tested cell lines were determined using the SRB assay, colchicine was taken as the
positive control. The results were expressed as the IC50
.
3.2 Inhibition of tubulin polymerization
To investigate the mechanism of antiproliferative activity of compound 17d and 17e, the
inhibition of tubulin polymerization were evaluated. CA-4 was taken as a positive control. As
shown in (Table 2), compounds 17d and 17e which exhibit similar inhibitory effect to CA-4,
proved to be better inhibitors of tubulin polymerization, in a dose-dependent manner.
Table 2. Tubulin polymerization inhibitory effect of compounds
Group
α-tubulin
0.68
Negative control
Solvent control
17d (50 nM)
17d (500 nM)
17d (1000 nM)
17e (50 nM)
17e (500 nM)
0.63
0.52
0.21
0.13
0.33
0.15
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17e (1000 nM)
CA-4 (50 nM)
0.11
0.53
0.34
0.22
CA-4 (500 nM)
CA-4 (1000 nM)
Data values of 17d, 17e and CA-4 were determined from the tubulin polymerization assay, CA-4 was taken as a positive control.
3.3 Antitumor activity in vivo
17d and 17e were selected for further in vivo antitumor activity studies on human breast
cancer MCF-7 xenograft model, while tamoxifen was chosen as reference. As showed in Fig
4, remarkable antitumor effects were observed in mice treated with compounds 17d and 17e
which were administered intraperitoneally (i.p.) for 30 consecutive days. Furthermore,
compound 17e showed better in vivo antitumor potency than tamoxifen.
The relative tumor proliferation rates of 17d and 17e were 59.48 % and 41.33 %
separately at the dose of 20 mg/kg while 45.08 % of tamoxifen at the same dose. In addition,
no significant body weight loss or any other obvious signs of toxicity were observed for all of
the 17d and 17e treated mice during the whole research which means that just like tamoxifen,
the compounds 17d and 17e had weak influence on normal cell.
Table 3. In vivo Anti-tumor activity on female BALB/c nude mice/MCF-7 xenograft mode ( x s , n= 6)
Control
Tamoxifen
0.098 0.024
0.154 0.030
1.592 0.157
91.41%
17d
17e
1
2
Tumor volume
Tumor volume
0.098 0.026
0.171 0.057
1.742 0.234
0.099 0.046
0.161 0.087
1.627 0.233
93.38%
0.098 0.041
0.141 0.048
1.480 0.150*
84.95%
RTV
T/C
Tumor volume
3
0.631 0.276
6.382 1.415
0.445 0.116
4.608 1.115*
72.21%
0.494 0.164
0.355 0.074*
5.547 2.343 3.909 0.860**
RTV
T/C
86.91%
61.25%
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Tumor volume
RTV
4
5
1.564 0.452
0.822 0.189**
8.760 2.877**
53.62%
0.972 0.315*
10.904 4.294
66.74%
0.684 0.186**
7.758 3.395**
47.49%
16.337 4.647
T/C
Tumor volume
3.748 1.197
1.672 0.260**
18.025 5.342**
45.08%
1.992 0.302**
23.785 11.251
59.48%
1.457 0.167**
16.526 4.809**
41.33%
RTV
T/C
39.988 14.875
P 0.05
P 0.01; RTV: relative tumor volume, T/C: relative tumor proliferation rates
Fig 4. Anti-tumor activity of 17d and 17e assessed using female BALB/c nude mice/MCF-7 xenograft model. Mice were assessed for tumor growth.
17d, 17e and Tamoxifen were administrated at a dose of 20 mg/kg once daily for 30 days. Tamoxifen as a positive control. (Measure once every six
days, Significance levels p <0.05 and p <0.05 as compared with the respective control)
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Fig 5. Anti-tumor activity of 17d and 17e assessed using female BALB/c nude mice/MCF-7 xenograft model. Mice were assessed for body weight.
17d, 17e and Tamoxifen were administrated at a dose of 20 mg/kg once daily for 30 days. Tamoxifen as a positive control. (Measure once every six
days, Significance levels p >0.05 and p >0.05 as compared with the respective control)
4. Discussion
(1) According to the biological evaluation of Podophyllotoxin deritives, we had proved
that the D ring of Podophyllotoxin moiety could be replaced with phenyl acrylic acid, which
promote activity and target selectivity. (2) During the antiproliferative activities in vitro, it
was found that when the substituent on the E ring was the electron-withdraw group, 4-NO2,
the antiproliferative activities of compounds were lower, or even missing, mean that the
introduction of electron-withdraw group on the E ring decreases the antiproliferative activity.
(3) On the E ring, the electron-donating group may have improved the inhibition of tubulin.
When the substitutions on the E ring are electro-donating group such as 3, 4-(OCH3)2 and
3-OCH3, the inhibitory activity is higher, and selectively inhibit MCF-7 cells. (4) In addition,
R4 and R5 also play an important role on the F ring, for example, the pharmacological
activity of ferulic acid series was better than that of the series of cinnamon acid and hydroxy
cinnamic acid. Thus, the substituent on the benzene ring is an important factor in determining
the activity of compounds. (5) After western blot, flow cytometry analysis was also
performed, while the data was not enclosed. It is proved that the anti-mitotic drugs arrest cell
cycle at G2/M phase in cancer cells due to destruction of the microtubular cytoskeleton, so
we can conclude that the cytotoxicity of 17d and 17e might not be because of tubulin
inhibition. So the mechanism of their cytotoxicity in very low concentration cannot be tubulin
inhibition. Above all, the tumor selectivity to MCF-7 cell and mechanism of 17d and 17e
need further confirmation and research.
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5. Conclusions
In summary, we had developed a series of tetrahydroisoquinoline derivatives and
explored their antiproliferative activity against a panel of three cancer cell lines. Most of the
compounds displayed inhibition activity, especially 17d and 17e，which exhibit selectivity
to MCF-7 cell. In addition, compounds 17d and 17e demonstrated potent in vivo efficacy and
had very little influence on body weight. These results demonstrate that 17d and 17e are
promising lead compounds and could be taken for further exploration.
6. Experimental section
6.1 General information. All reagents and anhydrous solvents were obtained from
commercial sources and used without further purification unless noted otherwise. Melting
points were taken on a WRS-1B melting point apparatus, uncorrected and reported in degrees
Centigrade. 1HNMR spectra were recorded in CDCl3 on a Bruker DRX-400 (400 MHz) using
TMS as internal standard. 13C NMR were recorded in CDCl3 on a Bruker 100 MHz
spectrometer, a Bruker 150 MHz spectrometer, using TMS as internal standard.
6.2 Abbreviations used. CDCl3, Deuterochloroform; DMF, N, N-dimethylformamide;
ESI-MS, Electron spray ionization mass spectrum; h, Hour; IR, Infrared absorption spectrum;
min, Minute; mp, Melting point; NMR, Nuclear magnetic resonance; SRB, Sulforhodamine
B; TLC, Thin layer chromatography; DMSO, dimethylsulfoxide; δ(ppm), chemical shift.
Synthesis of N-(2-(benzo[d][1,3]dioxol-5-yl)ethyl)-4-methoxybenzamide (12d) A mixture
of compound 9d (7.6 g, 50 mmol), SOCl2 10 mL，toluene 50 mL were added to a round
bottom flask. The reaction mixture was heated to 77 °C for 4 h with stirring, then evaporated
with toluene under reduced pressure to remove excess thionyl chloride to give compound
10d.
In a three-neck flask, compound 11 (7.4 g, 45 mmol) was added, NaOH (2.0 g, 50 mmol)
and CH2Cl2 50 mL, Compound 10d dissolved in CH2Cl2 (10 mL) solution was added drop
wise under ice-cooling. After the addition was complete, the reaction was continued for 2.5
h. The reaction mixture was washed with 1% dilute hydrochloric acid (150 mL × 3), 1 mol/L
NaOH aqueous solution (150 mL × 3), saturated sodium chloride solution (100 mL × 3) after
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washing separately, died with sodium sulphate and evaporated in vacuo. The residues were
recrystallized with anhydrous ethanol to afford white solid (10.5 g, yield 78.1 %). mp
120.2-122.1 °C. 1H NMR (400 MHz, CDCl3) δ 7.66 (d, J = 8.8 Hz, 2H, ArH), 6.89 (d, J = 8.8
Hz, 2H, ArH), 6.75 (d, J = 8.0 Hz, 1H, ArH), 6.72 (s, 1H, ArH), 6.67 (d, J = 8.0 Hz, 1H,
ArH), 6.13 (s, 1H, NH), 5.03 (s, 2H, OCH2O), 3.83 (s, 3H, OCH3), 3.67-3.62 (m, 2H,
NHCH2CH2), 2.83 (t, J = 6.8 Hz, 2H, NHCH2CH2); 13C NMR (100 MHz, CDCl3) δ 166.9,
162.1, 147.9, 146.2, 132.7, 128.6, 126.9, 121.6, 113.7, 109.0, 108.3, 100.9, 55.3, 41.2, 35.5;
IR (KBr, cm-1) υ: 3348.8, 2971.8, 2880.9, 1640.0, 1547.4, 1501.4, 1253.9, 928.1, 847.7,
815.7; ESI-Mass for C17H17NO4: m/z (M+H)+ 300.04.
5-(4-methoxyphenyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline (14d).
A solution of the prepared Compound 12d (6.0 g, 20 mmol), POCl3 (12.3 mL, 40 mmol)
in toluene 40 mL was stirred at 115 °C for 7 h. The reaction mixture was evaporated in
vacuo. The residues were partitioned between CH2Cl2 and water. The organic phase was
dried over sodium sulphate and evaporated in vacuo to give the 13d (4.6 g, yield 81.7 %).
A mixture of 13d (4.6 g, 16 mmol) was dissolved in CH3OH (20 mL), and NaBH4 (1.1
g, 29 mmol) was added portion-wise under ice-cooling, and stirred 4h at 0 °C . The reaction
mixture was evaporated. The residues were partitioned between CH2Cl2 and water. The
organic phase was dried over sodium sulphate and evaporated in vacuo. The residue
was recrystallized from 40 mL of methanol to give a white solid (3.5 g, yield 76.0 %). mp
98.5-100.5 °C. 1H NMR (400 MHz, CDCl3) δ 7.17 (d, J = 8.4 Hz, 2H, ArH), 6.64 (d, J = 8.4
Hz, 2H, ArH), 6.58 (s,1H, ArH), 6.21 (s, 1H, ArH), 5.83 (s, 2H, OCH2O), 4.95 (s, 1H,
CHNH), 3.78 (s, 3H, OCH3), 3.23-3.1 (m, 1H, NHCH2CH2), 3.04-2.97 (m, 1H,
NHCH2CH2), 2.95-2.88 (m, 1H, NHCH2CH2), 2.73-2.67 (m, 1H, NHCH2CH2), 2.58 (br, 1H,
NH); 13C NMR (100 MHz, CDCl3) δ 159.0, 146.0, 145.6, 136.6, 131.2, 129.9, 128.4, 113.8,
108.4, 107.9, 100.5, 61.3, 55.2, 42.0, 29.6; IR (KBr, cm-1) υ: 3238.5, 2929.6, 2874.5, 1610.1,
1509.9, 1479.0, 1374.8, 123.1, 1033.0, 933.0, 813.3; ESI-Mass for C17H17NO3: m/z (M+H) +
283.97.
(E)-2-methoxy-4-(3-(5-(4-methoxyphenyl)-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5
H)-yl)-3-oxoprop-1-en-1-yl)phenyl acetate (16d). A mixture of 14d (1.0 g, 3.4 mmol), Et3N
(3.5 mL, 25.3mmol) in CH2Cl2 (30 mL), compound 15a (1.30g, 5.1mmol) dissolved
in CH2Cl2 (10 mL) solution was added drop wise under ice-cooling. After the addition was
complete, the mixture was stirred for 2.5 h and monitored by TLC. The reaction mixture
was washed with 1 % dilute hydrochloric acid (150 mL × 3), 1 mol/L NaOH aqueous
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solution (150 mL × 3), saturated sodium chloride solution (100 mL × 3) after washing
separately, died with sodium sulphate and evaporated in vacuo. The residues were
recrystallized with anhydrous ethanol to afford yellow solid (1.1g, yield 60.1 %). mp
95.8-97.5 °C. 1H NMR (400 MHz, CDCl3) δ 7.69 (d, J = 15.2 Hz, 1H, CH=CH-Ar), 7.21 (d,
J = 8.0 Hz, 1H, ArH), 7.13 (d, J = 7.6 Hz, 2H, ArH), 7.06 (s, 1H, ArH), 7.02 (d, J = 8.4 Hz,
1H, ArH), 6.90 (s, 1H, ArH), 6.85-6.80 (m, 3H, CH=CH-Ar, ArH), 6.54 (s, 1H, ArH), 5.93
(d, J = 8.4 Hz, 1H, OCH2O), 3.95-3.90 (m, 1H, NCH2CH2), 3.86 (s, 3H, OCH3), 3.77 (s, 3H,
OCH3), 3.54 -3.47 (m, 1H, NCH2CH2), 2.99-2.87 (m, 1H, NCH2CH2), 2.78-2.65 (m, 1H,
NCH2 CH2), 2.32 (s, 3H, CH3); 13C NMR (100 MHz, CDCl3). δ: 168.8, 165.0, 158.8, 151.2,
146.7, 146.2, 142.5, 140.7, 134.5, 134.3, 129.9, 128.57, 128.50, 127.4, 123.1, 120.3, 117.6,
113.9, 113.5, 111.5, 108.5, 108.1, 100.9, 55.9, 55.2, 55.0, 39.6, 29.4, 20.6; IR (KBr, cm-1) υ:
3003.3, 2932.5, 2362.7, 1764.6, 1646.8, 1604.2, 1434.4, 1242.2, 1035.3, 941.2, 826.7;
ESI-Mass for C29H27NO7: m/z (M+H) + 501.98.
(E)-3-(4-hydroxy-3-methoxyphenyl)-1-(5-(4-methoxyphenyl)-7,8-dihydro-[1,3]dioxolo[4,
5-g]isoquinolin-6(5H)-yl)prop-2-en-1-one (17d). To a mixture of 16d (1.1 g, 2.2 mmol) in
methanol 30 mL, 10 mL NaOH (9.4 × 10-2 g, 24 mmol) in aqueous methanol was added. The
reaction mixture was stirred at room temperature for 24 h, then evaporated. The residue was
diluted with 3% HCl (liq.) was slowly added drop wise to pH = 4-5 to give a
yellow solid, filtration and drying. The crude product was purified on silica using (Ethyl
acetate/PE = 2:1) to provide the title compound as an off-white solid (0.58 g, yield 58 %). mp
106.5-108.5 °C. 1H NMR (400 MHz, CDCl3) δ 7.68 (d, J = 16.0Hz, 1H, CH = CH-Ar),
7.22-7.09 (m, 3H, ArH), 6.98 (br, 1H, ArH), 6.91 (d, J = 8.0Hz, 2H, ArH), 6.81 (d, J =
8.0Hz, 2H, ArH), 6.73(d, J = 16.0, 1H, CH = CH-Ar), 6.64 (s, 1H, ArH), 6.55 (s, 1H, CHN),
5.93 (d, J = 7.2Hz, 2H, OCH2O), 3.98-3.94 (m, 1H, NCH2CH2), 3.92 (s, 3H, OCH3), 3.77 (s,
3H, OCH3), 3.53-3.46 (m, 1H, NCH2CH2), 2.98-2.94 (m, 1H, NCH2CH2), 2.78-2.64 (m, 1H,
NCH2CH2); 13C-NMR (100 MHz, CDCl3) δ 165.5, 158.9, 147.3, 146.7, 146.3, 143.4, 134.7,
129.9, 128.7, 128.6, 127.9, 127.6, 121.9，114.8, 113.9, 113.6, 110.0, 108.6, 108.2, 101.0,
56.0, 55.3, 55.0, 39.6, 29.4; IR (KBr, cm-1) υ: 3423.0, 2932.1, 1640.2, 1510.9, 1433.9,
1239.4, 924.2, 817.2; ESI-Mass for C27H25NO6: m/z (M-H) + 458.25.
(E)-1-(5-(4- methylphenyl)-7,8-dihydro-[1, 3] dioxole and [4,5-g] isoquinoline-6(5H)-yl)-
3- (4-hydroxy-3-methoxyphenyl) prop-2-en-1-one (17a). Compound 17a was prepared
from 16a according to procedure used for 17d synthesis. White solid, yield 60 %, mp
110.7-112.9 °C. 1HNMR (400 MHz, CDCl3) δ 7.71 (d, J = 15.2 Hz, 1H, CH=CH-Ar), 7.18
(d, J = 7.2 Hz, 1H, ArH), 7.11-7.092 (m, 4H, ArH), 6.98(s, 1H, ArH), 6.92 (d, J = 8.4 Hz,
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2H, ArH), 6.77 (d, J = 15.2 Hz, 1H, CH=CH-Ar), 6.64 (s, 1H, ArH), 6.56 (s, 1H, CHN), 5.94
(d, J = 7.8 Hz, 2H, OCH2O), 4.03-3.99 (m, 1H, NCH2CH2), 3.92 (s, 3H, OCH3), 3.54-3.49
(m, 1H, NCH2CH2), 2.95-2.91 (m, 1H, NCH2CH2), 2.78-2.69 (m, 1H, NCH2CH2), 2.31 (s,
3H, CH3); 13C NMR (100 MHz, CDCl3) δ 166.9, 147.4, 146.7, 143.4, 139.5, 137.1, 128.3,
128.9, 128.6, 127.8, 127.6, 127.3, 121.9, 114.8, 114.7, 110.0, 108.6, 108.1, 101.0, 56.0, 39.8,
29.4, 21.1; IR (KBr, cm-1) υ: 3173.1, 2590.9, 1640.7, 1575.1, 1511.1, 1454.3, 1279.2, 1034.4,
921.5, 813.8; ESI-Mass for C27H25NO5: m/z (M+H)+ 444.10.
(E)-1-(5-(4-chlorophenyl)-7,8-dihydro-[1,3]dioxole and[4,5-g]isoquinoline-6(5H )-yl)-3-(
4-hydroxy-3-methoxyphenyl)prop-2-en-1-one (17b). Compound 17b was prepared from
16b according to procedure used for 17d synthesis White solid, yield 57 %, mp 108.7-110.9
°C. 1H NMR (400 MHz, CDCl3) δ 7.72 (d, J = 15.2 Hz, 1H, CH=CH-Ar), 7.26-7.24 (m, 4H,
ArH), 7.11 (d, J = 8.0 Hz, 1H, ArH), 6.99(s, 1H, ArH), 6.92 (d, J = 8.4 Hz, 2H, ArH), 6.76
(d, J = 15.2 Hz, 1H, CH=CH-Ar), 6.65 (s, 1H, ArH), 6.54 (s, 1H, CHN), 5.94(d, J = 7.9Hz,
2H, OCH2O), 3.98-3.95 (m, 1H, NCH2CH2), 3.92 (s, 3H, OCH3), 3.52-3.45 (m, 1H,
NCH2CH2), 2.95-2.91 (m, 1H, NCH2CH2), 2.77-2.73 (m, 1H, NCH2CH2); 13C NMR (100
MHz, CDCl3) δ 165.7, 147.4, 146.9, 146.7, 146.3, 143.7, 140.8, 133.2, 129.9, 128.7, 128.3,
127.9, 127.7, 121.9, 114.7, 114.3, 110.0, 108.4, 108.3, 101.0, 55.9, 55.0, 39.9, 29.2; IR (KBr,
cm-1) υ: 3182.2, 2933.6, 1640.8, 1590.3, 1511.2, 1430.9, 1275.1, 1235.9, 1035.9, 922.3,
844.9, 812.3; ESI-Mass for C26H22Cl NO5: m/z (M+H) + 464.06.
(E)-1-(5-(4-nitrophenyl)-7,8-dihydro-[1, 3]dioxole and[4,5-g]isoquinoline-6(5H)-yl)-3-(4-
hydroxy-3-methoxyphenyl) prop-2-en-1-one (17c). Compound 17c was prepared from 16c
according to procedure used for 17d synthesis. Yellow solid, yield 68 %. mp 104.3-105.7 °C.
1H NMR (400 MHz, CDCl3) δ 8.15(d, J = 7.2 Hz, 2H, ArH), 7.74 (d, J = 16.0 Hz, 1H,
CH=CH-Ar), 7.48 (d, J = 7.2 Hz, 2H, ArH), 7.13 (d, J = 8.0 Hz, 1H, ArH), 7.00 (s, 1H,
ArH), 6.96 (s, 1H, ArH), 6.93 (d, J = 7.2 Hz, 1H, ArH), 6.78 (d, J = 16.0 Hz, 1H,
CH=CH-Ar), 6.69 (s, 1H, ArH), 6.56 (s, 1H, CHN), 5.97 (d, J = 5.2 Hz, 2H, OCH2O),
4.03-3.99 (m, 1H, NCH2CH2), 3.93 (s, 3H, OCH3), 3.56-3.51(m, 1H, NCH2CH2), 3.01-2.97
(m, 1H, NCH2CH2), 2.78-2.72 (m, 1H, NCH2CH2); 13C NMR (100 MHz, CDCl3) δ 166.1,
149.6, 147.7, 147.3, 147.2, 146.8, 146.6, 144.3, 129.3, 127.9, 127.6, 127.2, 123.5, 122.1,
114.9, 114.0, 110.1, 108.5, 108.3, 101.2, 56.0, 55.3, 40.5, 29.1; IR (KBr, cm-1) υ: 3440.1,
2931.4, 1641.4, 1596.5, 1515.0, 1430.2, 1346.9, 1274.5, 1238.4, 1036.1, 924.6, 815.5;
ESI-Mass for C26H22N2O7: m/z (M-H) + 473.21.
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(E)-1-(5-(3,4-dimethoxyphenyl)-7,8-dihydro-[1, 3]dioxole and[4,5-g]isoquinolinemorpho
line-6(5H)-yl)-3-(4-hydroxy-3-methoxyphenyl)prop-2-en-1-one (17e). Compound 17e was
prepared from 16e according to procedure used for 17d synthesis. White solid, yield 59 %,
mp 120.7-123.9 °C. 1H NMR (400 MHz, CDCl3) δ 7.70 (d, J = 15.2 Hz, 1H, CH=CH-Ar),
7.11 (d, J = 8.0 Hz, 1H, ArH), 6.99 (s, 2H, ArH), 6.91 (d, J = 8.0 Hz, 2H, ArH), 6.77-6.73
(m, 2H, ArH, CH=CH-Ar), 6.66 (d, J = 8.0 Hz, 2H, ArH), 6.57 (s,1H, CHN), 5.95 (d, J = 6.4
Hz, 2H, OCH2O), 3.99-3.94 (m, 1H, NCH2CH2), 3.92 (s, 3H, OCH3), 3.84-3.76 (m, 6H,
OCH3×2), 3.54-3.47 (m, 1H,NCH2CH2), 2.98-2.91 (m, 1H, NCH2CH2), 2.78-2.74 (m, 1H,
NCH2CH2); 13C NMR (100 MHz, CDCl3) δ 165.6, 148.9, 148.4, 147.5, 146.8, 146.2, 143.4,
135.2, 135.1, 128.5, 127.8, 127.6, 121.8, 121.0, 114.9, 114.7, 112.3, 110.5, 110.1, 108.6,
108.2, 100.9, 56.0, 55.9, 55.3, 39.7, 29.4; IR (KBr, cm-1) υ: 3425.2, 2932.3, 1639.8, 1590.6,
1511.8, 1443.7, 1265.7, 1236.3, 1032.9, 927.8, 817.9; ESI-Mass for C28H27NO7: m/z (M-H) +
488.18.
(E)-3-(4-hydroxy-3-methoxyphenyl)-1-(5-(3-hydroxy-4-methoxyphenyl)-7,8-dihydro-[1,3
]dioxolo[4,5-g]isoquinolin-6(5H)-yl)prop-2-en-1-one(17f). Compound 17f was prepared
from 16f according to procedure used for 17d synthesis. White solid, yield 45 %，mp
132.4-134.9°C. 1H NMR (400 MHz, CDCl3) δ 7.67 (d, J = 15.2 Hz, 1H, CH=CH-Ar), 7.09
(d, J = 8.0 Hz, 1H, ArH), 6.97 (s, 1H, ArH), 6.90 (d, J = 8.4 Hz, 1H, ArH), 6.86-6.82 (m, 2H,
ArH), 6.79 (s, 1H, ArH), 6.76-6.69 (m,2H, ArH,CH=CH-Ar), 6.62 (s, 1H, ArH), 6.54-6.46
(m,1H, ArH), 5.93 (br, 2H, OCH2O), 4.14-4.09 (m, 1H, NCH2CH2), 3.91 (s, 3H, OCH3),
3.85 (s, 3H, OCH3), 3.55-3.51 (m, 1H, NCH2CH2), 2.95-2.86 (m, 1H, NCH2CH2), 2.76-2.63
(m, 1H, NCH2CH2); 13C NMR (100 MHz, CDCl3) δ 147.7, 147.3, 146.6, 146.2, 145.8, 145.2,
143.4, 135.7, 128.5, 127.8, 127.6, 127.5, 121.6, 120.6, 114.9, 114.7, 110.3, 109.9, 108.6,
108.1, 100.9, 55.9, 55.0, 39.7, 29.3; IR (KBr, cm-1) υ: 3400.0, 2936.1, 1639.4, 1587.3,
1510.9, 1436.2, 1272.1, 1124.5, 1033.5, 924.4, 815.8; ESI-Mass for C27H25NO7: m/z (M+H)
+ 476.18.
(E)-1-(1-(4-methoxyphenyl)-6,7-dimethoxy-3,4-dihydro-isoquinoline-2(1H)-yl)-3- (4-hyd
roxy-3-methoxyphenyl)prop-2-en-1-one (17g). Compound 17g was prepared from 16g
according to procedure used for 17d synthesis. White solid, yield 51 %, mp 120.7-122.9 °C.
1H NMR (400 MHz, CDCl3). δ 7.71 (d, J = 15.2 Hz, 1H, CH=CH-Ar), 7.22(d, J = 8.4 Hz,
1H, ArH), 7.12 (d, J = 8.0 Hz, 2H, ArH), 6.99 (s, 2H, ArH ), 6.92(d, J = 8.0 Hz, 1H, ArH),
6.82 (d, J = 8.0 Hz, 2H, ArH), 6.78(d, J = 15.6 Hz, 1H, CH=CH-Ar), 6.66 (s, 1H, ArH), 6.56
(s, 1H, CHN), 4.01-3.97 (m, 1H, NCH2CH2), 3.93 (s, 3H, OCH3), 3.89 (s, 3H, OCH3), 3.78
(s, 6H, OCH3×2), 3.50-3.44 (m, 1H, NCH2CH2), 2.99-2.95 (m, 1H, NCH2CH2), 2.80-2.76
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(m, 1H, NCH2CH2); 13C NMR (100 MHz, CDCl3) δ 165.5, 158.8, 148.0, 147.9, 147.6, 147.2,
143.4, 134.8, 130.0, 128.8, 127.5, 127.4, 126.4, 121.9, 115.2, 114.5, 113.5, 111.3, 111.1,
110.3, 56.0, 55.9, 55.2, 54.6, 39.9, 29.0; IR (KBr, cm-1) υ: 3399.0, 3000.3, 2935.9, 2835.7,
1640.3, 1603.7, 1512.8, 1443.4, 1252.6, 1173.5, 1115.8, 1029.6, 976.7, 818.5; ESI-Mass for
C28H29NO6: m/z (M+-H) 474.00.
(E)-1-(5-(4-methoxyphenyl)-7,8-dihydro-[1, 3]dioxole and[4,5-g]isoquinolin-6(5H)-yl)-3-
(4-hydroxyphenyl)prop-2-en-1-one (17j). Compound 17j was prepared from 16j according
to procedure used for 17d synthesis. White solid, yield 50 %, mp 120.6-122.7 °C. 1H NMR
(400 MHz, CDCl3) δ 7.69 (d, J = 14.8 Hz, 1H, CH=CH-Ar), 7.39 (d, J = 7.2 Hz, 2H, ArH),
7.20 (d, J = 8.4 Hz, 1H, ArH), 7.12 (d, J = 8.0 Hz, 1H, ArH), 6.90 (s, 1H, ArH), 6.84-6.79
(m, 4H, ArH), 6.75 (d, J = 14.8 Hz, 1H, CH=CH-Ar), 6.64 (s, 1H, ArH), 6.54 (s, 1H, CHN),
5.93 (d, J = 8.4 Hz, 2H, OCH2O), 3.98-3.93 (m, 1H, NCH2CH2), 3.77 (s, 3H, OCH3),
3.53-3.46 (m, 1H, NCH2CH2), 2.96-2.89 (m, 1H, NCH2CH2), 2.78-2.73 (m, 1H, NCH2CH2);
13C NMR (150 MHz, CDCl3) δ 166.0, 158.8, 158.3, 146.6, 146.2, 143.8, 134.3, 129.8, 129.7,
129.6, 128.5, 128.4, 127.4, 115.9, 113.9, 113.5, 108.5, 108.1, 100.9, 55.27, 55.22, 39.7, 29.3;
IR (KBr, cm-1)υ: 3158.7, 2952.8, 1639.4, 1240.1, 1169.3, 977.9, 825.3; ESI-Mass for
C26H23NO5: m/z (M+H) +430.17.
(E)-1-(5-(3,4-dimethoxyphenyl)-7,8-dihydro-[1, 3]dioxole and[4,5-g]isoquinolinemorpho
line-6(5H)-yl)-3-(4-hydroxyphenyl)prop-2-en-1-one (17k). Compound 17k was prepared
from 16k according to procedure used for 17d synthesis. White solid, yield 52 %, mp
125.6-127.7 °C. 1H NMR (400 MHz, CDCl3) δ 7.68(d, J = 15.2 Hz, 1H, CH=CH-Ar), 7.38
(d, J = 8.0 Hz, 2H, ArH), 6.99 (s, 1H, ArH), 6.90 (s, 1H, ArH), 6.85 (d, J = 8.0 Hz, 2H,
ArH), 6.74 (m, 2H, ArH,CH=CH-Ar), 6.64 (br, 2H, ArH), 6.55 (s, 1H, CHN), 5.93 (d, J =
10.8 Hz, 2H, OCH2O), 3.98-3.94 (m, 1H, NCH2CH2),3.83 (s, 3H, OCH3), 3.81 (s, 3H,
OCH3), 3.54-3.49 (m, 1H, NCH2CH2), 2.99-2.92 (m, 1H, NCH2CH2), 2.79-2.75 (m,1H,
NCH2CH2); 13C NMR (100 MHz, CDCl3) δ 165.2, 148.8, 148.3, 146.6, 146.1, 143.1, 135.2,
135.0, 129.6, 128.8, 127.7, 121.0, 117.2, 112.0, 110.2, 108.7, 108.5, 108.1, 107.9, 100.9,
55.9, 55.8, 55.2, 39.6, 29.4; IR (KBr, cm-1) υ: 3159.4, 2935.2, 1639.3, 1444.5, 1236.1,
1032.6, 925.5, 824.5; ESI-Mass for C26H23NO5: m/z (M+H) +460.10.
(E)-1-(5-(3,4,5-trimethoxyphenyl)-7,8-dihydro-[1, 3]dioxole and[4,5-g]isoquinoline-6(5H
)-yl)-3-(4-hydroxyphenyl)prop-2-en-1-one (17h). Compound 17h was prepared from 16h
according to procedure used for 17d synthesis. White solid, yield 44 %, mp 128.6-130.1 °C.
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1H NMR (400 MHz, CDCl3) δ 7.67(d, J =15.6 Hz, 1H, CH=CH-Ar), 7.36 (d, J = 8.0 Hz, 2H,
ArH), 6.87-6.85 (m, 3H, ArH), 6.76 (d, J = 15.2 Hz, 1H, CH=CH-Ar), 6.67-6.65 (m,1H,
ArH), 6.56 (s, 1H, ArH), 6.51-6.41 (m, 2H, ArH, CHN), 5.94 (d, J = 8.0 Hz, 2H, OCH2O),
4.01-3.98 (m, 1H, NCH2CH2), 3.81 (s, 3H, OCH3), 3.75 (s, 6H, OCH3×2), 3.57-3.49 (m, 1H,
NCH2CH2), 3.00-2.92 (m, 1H, NCH2CH2), 2.81-2.73 (m, 1H, NCH2CH2); 13C NMR (150
MHz, CDCl3) δ 166.1, 158.5, 152.9, 146.6, 146.2, 143.8, 134.3, 129.8, 129.6, 128.5, 127.4,
115.9, 113.9, 113.5, 108.5, 108.1, 100.9, 55.7, 55.2, 55.22, 39.7, 29.3; IR (KBr, cm-1) υ:
3159.7, 2937.0, 1639.3, 1125.9, 1038.0, 924.0, 826.5; ESI-Mass for C28H27NO7: m/z
(M+H)+490.00.
(E)-1-(5-(4-nitrophenyl)-7,8-dihydro-[1,3]dioxole and[4,5-g]isoquinoline-6(5H)-yl)-3-(4-
hydroxyphenyl)prop-2-en-1-one (17i). Compound 17i was prepared from 16i according to
procedure used for 17d synthesis. Yellow solid, yield 62 %, mp 107.7-109.8 °C. 1H NMR
(400 MHz, CDCl3) δ: 8.13 (d, J = 8.4 Hz, 2H, ArH), 7.72(d, J = 15.6 Hz, 1H,
CH=CH-Ar), 7.46 (d, J = 8.0 Hz, 2H, ArH), 7.40 (d, J = 7.6 Hz, 2H, ArH ), 6.94 (s,1H,
ArH), 6.84 (d, J = 7.6 Hz, 2H, ArH), 6.76 (d, J = 15.2 Hz, 1H, CH=CH-Ar), 6.68 (s, 1H,
ArH), 6.55 (s, 1H, CHN), 5.97 (d, J = 6.4 Hz, 2H, OCH2O), 4.00-3.91 (m, 1H, NCH2CH2),
3.55-3.50 (m, 1H, NCH2CH2), 2.99-2.93 (m, 1H, NCH2CH2), 2.78-2.74 (m, 1H, NCH2CH2);
13C NMR (100 MHz, CDCl3) δ 166.6, 158.3, 149.3, 147.3, 147.1, 146.6, 144.5, 129.7, 129.2,
127.8, 127.0, 123.5, 116.0, 113.3, 109.4, 108.5, 108.3, 101.2, 55.6, 40.7, 29.0; IR (KBr, cm-1)
υ: 3134.2, 1719.6, 1639.2, 1605.7, 1514.3, 1346.6, 1238.4, 1168.1, 923.6, 825.9; ESI-Mass
for C25H20N2O6: m/z (M+H) + 445.14.
Synthesis of Compound 19d. To a mixture of 18 (0.20 g, 1.3 mmol), K2CO3 (0.5 g, 3.6
mmlol), KI (0.1 g, 0.6 mmlol) in isopropanol 10 mL were stirred at room temperature for 3 h,
which were add to the mixture of compounds 17d (0.46 g, 1 mmol), K2CO3 (0.5 g, 3.6
mmol), and isopropanol 10 mL in a single neck flask. The mixture were reacted at 85 °C for
4 h then evaporated in vacuo to give the crude product. The crude product was purified on
silica using MeOH/CH2Cl2 6 % to provide the title compound. Recrystallized with EtOH/PE
= 10:1 to give a yellow solid (0.26 g, yield 45 %)
(E)-3-(4-(3-(dimethylamino)propoxy)-3-methoxy-phenyl)-1-(5-(4-methoxyphenyl) -7,8-di
hydro-[1, 3]dioxole[4,5-g]isoquinoline-6(5H)-yl)prop-2-en-1-one (19d).
Yellow solid, yield 45 %, mp 150.4-153.9 °C, 1H NMR (400 MHz, CDCl3) δ 7.68(d, J = 15.2
Hz, 1H, CH=CH-Ar), 7.21 (d, J =8.0 Hz, 2H, ArH), 7.09 (d, J = 7.6 Hz, 1H, ArH), 7.02 (s,
1H, ArH), 6.91-6.87 (m, 2H, ArH), 6.81 (d, J = 8.4 Hz, 2H, ArH), 6.75(d, J = 15.2 Hz, 1H,
CH=CH-Ar), 6.63 (s, 1H, ArH), 6.55 (s, 1H, CHN), 5.92 (d, J = 7.6 Hz, 2H, OCH2O), 4.10
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(t, J = 6.8 Hz, 2H, OCH2CH2), 3.98-3.94 (m, 1H, NCH2CH2), 3.89 (s, 3H, OCH3), 3.77 (s,
3H, OCH3), 3.53-3.47 (m, 1H, NCH2CH2), 2.99-2.91 (m, 1H, NCH2CH2), 2.77-2.73 (m, 1H,
NCH2CH2), 2.47 (t, J = 6.8 Hz, 2H, CH2CH2CH2N), 2.25 (s, 6H, CH3×2), 2.06-1.99 (m, 2H,
CH2CH2CH2N); 13C NMR (100MHz, CDCl3) δ 165.3, 158.7, 150.0, 149.3, 146.6, 146.2,
143.2, 134.6, 130.4, 129.8, 128.7, 128.6, 128.2, 127.5, 121.7, 114.9, 113.5, 112.5, 110.5,
108.5, 108.1, 100.9, 67.2, 56.2, 56.0, 55.2, 54.9, 45.4, 39.5, 29.4, 27.3; IR (KBr, cm-1) υ:
3077.6, 2944.3, 2823.0, 1641.3, 1510.8, 1456.4, 1259.7, 1170.0, 1032.3, 922.0, 801.2;
ESI-Mass for C32H36N2O6: m/z (M+H) + 545.37.
(E)-3-(4-(3-(dimethylamino)propoxy)-3-methoxy-phenyl)-1-(5-(3,4-dimethoxyphenyl)-7,
8-dihydro-[1, 3]dioxole and[4,5-g]isoquinoline-6(5H)-yl)prop-2-en-1-one (19e).
Compound 19e was prepared from 17e according to procedure used for 19d synthesis.
Yellow solid, yield 42 %，mp 148.7-149.8 °C. 1H NMR (400 MHz, CDCl3) δ 7.67 (d, J =
15.6Hz, 1H, CH=CH-Ar), 7.11-7.09 (m, 1H, ArH), 7.03 (s, 1H, ArH), 6.99 (s, 1H, ArH),
6.88 (d, J = 8.4, 1H, ArH), 6.80 (s, 1H, ArH), 6.76-6.73 (m, 2H, ArH, CH=CH-Ar), 6.65 (d,
J = 8.8Hz, 2H, ArH), 6.56 (s, 1H, CHN), 5.93 (d, J = 10.4Hz, 2H, OCH2O), 4.11 (t, J =
6.5Hz, 2H, OCH2CH2), 4.00-3.97 (m, 1H, NCH2CH2), 3.89 (s, 3H, OCH3), 3.83-3.82 (m,
6H, OCH3×2), 3.79-3.76 (m, 1H, NCH2CH2), 3.53-3.45 (m, 1H, NCH2CH2), 2.79-2.75 (m,
1H, NCH2CH2), 2.69 (t, J = 7.2Hz, 2H, CH2CH2CH2N), 2.40 (s, 6H,CH3×2), 2.15-2.08 (m,
2H, CH2CH2CH2); 13C NMR (100 MHz, CDCl3) δ 165.4, 149.3, 146.6, 143.0, 136.2, 131.1,
128.5, 128.4, 127.6, 127.5, 122.0, 121.6, 120.9, 115.2, 112.9, 112.1, 110.5, 108.1,100.9,
100.6, 68.8, 66.9, 56.1, 56.0, 55.9, 55.8, 51.4, 44.8, 43.8, 29.4; IR (KBr, cm-1) υ: 3443.6,
2938.0, 1641.7, 1594.0, 1511.1, 1479.6, 1333.9, 1233.3, 1138.3, 1031.9, 932.9, 865.3;
ESI-Mass for C33H38N2O7: m/z (M+H)+ 575.24.
(E)-3-(4-(3-(dimethylamino)propoxy)-3-methoxy-phenyl)-1-(5-(4-nitrophenyl)-7, 1,8-hyd
rogen-[1, 3]dioxole[4,5-g]isoquinoline-6(5H)-yl)prop-2-en-1-one (19c). Compound 19c
was prepared from 17c according to procedure used for 19d synthesis. Yellow solid, yield 50
%，mp 141.7-143.8 °C. 1H NMR (400 MHz, CDCl3) δ 8.13 (d, J = 8.0Hz, 2H, ArH), 7.72
(d, J = 15.2Hz, 1H, CH=CH-Ar), 7.47 (d, J = 8.0Hz, 2H, ArH), 7.12(d, J = 8.0Hz, 1H, ArH
), 7.02 (s, 1H, ArH), 6.95 (s, 1H, ArH), 6.89 (d, J = 8.0Hz, 1H, ArH), 6.78 (d, J = 16.0Hz,
1H, CH= CH-Ar), 6.68 (s, 1H, ArH), 6.55 (s, 1H, CHN), 5.97 (d, J=5.6Hz, 2H, OCH2O),
4.15 (t, J = 6.0Hz, 2H, OCH2CH2), 4.01-3.98(m, 1H, NCH2CH2),3.89 (s, 3H, OCH3),
3.55-3.49 (m, 1H, NCH2CH2), 2.97 (t, J = 6.8Hz, 3H, CH2CH2CH2N,NCH2CH2), 2.79-2.75
(m, 1H, NCH2CH2), 2.60 (s, 6H, CH3×2), 2.30-2.24 (m, 2H, CH2CH2CH2); 13C NMR (100
MHz, CDCl3) δ 149.6, 149.4, 146.6, 143.9, 131.9, 130.0, 129.3, 128.5, 127.8, 127.0, 123.7,
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123.5, 122.7, 121.7, 118.3, 114.6, 113.1, 110.7, 108.5, 108.3, 103.5, 101.2, 66.6, 56.1, 56.0,
55.2, 44.2, 29.1, 25.7; IR (KBr, cm-1) υ: 3131.2, 3068.4, 2930.6, 2362.9, 1648.3, 1597.5,
1513.4, 1426.8, 1345.5, 1246.3, 1138.3, 1034.5, 812.6; ESI-Mass for C31H33N3O7: m/z
(M-H)+ 558.26.
(E)-1-(5-(4-methoxyphenyl)-7,8-dihydro-[1, 3]dioxole and[4,5-g]isoquinolin-6(5H)-yl)-3-
phenyl-2-en-1-one (21d). Compound 21d was prepared from 14d according to procedure
used for 16d synthesis. White solid, yield 70 %, mp 105.7-107.8 °C. 1H NMR (400 MHz,
CDCl3) δ 7.74 (d, J = 15.4 Hz, 1H, CH=CH-Ar), 7.53-7.51 (m, 2H, ArH), 7.39-7.35 (m, 3H,
ArH), 7.21 (d, J = 6.8 Hz, 2H, ArH ), 7.14 (s, 1H, ArH), 6.90 (t, J = 6.4 Hz, 1H, ArH),
6.83-6.80 (m, 2H, CH=CH-Ar, ArH), 6.63 (s, 1H, ArH), 6.54 (s, 1H, CHN), 5.91 (d, J = 6.8
Hz, 2H, OCH2O), 4.00-3.93 (m, 1H, NCH2CH2), 3.76 (s, 3H, OCH3), 3.53-3.45 (m, 1H,
NCH2CH2), 2.97-2.88 (m, 1H, NCH2CH2), 2.77-2.72 (m, 1H, NCH2CH2); 13C NMR
(100MHz, CDCl3) δ 158.9, 146.7, 146.3, 143.1, 135.3, 134.6, 129.9, 129.6, 128.8, 128.6,
128.5, 127.7, 127.5, 117.4, 113.6, 108.5, 108.1, 100.9, 55.2, 55.0, 39.6, 29.4; IR (KBr, cm-1)
υ: 3015.2, 2897.8, 2836.4, 1645.0, 1595.2, 1481.6, 1236.9, 1173.3, 1033.9, 918.6, 818.5;
ESI-Mass for C26H23NO4: m/z (M+H) + 414.12.
(E)-1-(5-(3,4-dimethoxyphenyl)-7,8-dihydro-[1,3]dioxole and[4,5-g]isoquinolinemorphol
ine-6(5H)-yl)-3-phenyl-2-en-1-one (21e). Compound 21e was prepared from 14e according
to procedure used for 16d synthesis. White solid, yield 71 %, mp 107.6-109.8°C. 1HNMR
(400 MHz, CDCl3). δ: 7.74(d, J =15.2 Hz, 1H, CH=CH-Ar), 7.54-7.52(m, 2H, ArH),
7.37-7.35(m, 3H, ArH), 7.00(s, 1H, ArH), 6.93-6.90(m, 2H, ArH, CH=CH-Ar), 6.74(d, J
=8.0 Hz, 1H, ArH), 6.67(s, 1H, ArH), 6.64(s, 1H, ArH),6.56(s, 1H, CHN), 5.93(d, J =10.0
Hz, 2H, OCH2O), 3.99-3.95(m, 1H, NCH2CH2), 3.84(s, 6H, OCH3×2), 3.54-3.48(m, 1H,
NCH2CH2), 2.99-2.91(m, 1H, NCH2CH2), 2.79-2.71(m, 1H, NCH2 CH2); 13C-NMR (150
MHz, CDCl3) δ:165.2, 148.8, 148.3, 146.6, 146.1, 143.1, 135.2, 135.0, 129.6, 128.8, 127.7,
127.5, 121.0, 117.2, 112.0, 110.2, 108.7, 108.5, 108.1, 107.9, 100.9, 55.9, 55.8, 55.2, 39.6,
29.4; IR (KBr, cm-1) υ: 3014.4, 2955.5, 2925.7, 1643.3, 1483.2, 1454.7, 1230.5, 1191.3,
973.2, 921.1, 846.6; ESI-Mass for C27H25NO5: m/z (M+H) +444.06.
(E)-1-(5-(3,4,5-trimethoxyphenyl)-7,8-dihydro-[1, 3]dioxole and[4,5-g]isoquinoline-6(5H
)- yl)-3-phenyl-2-en-1-one (21f). Compound 21f was prepared from 14h according to
procedure used for 16d synthesis. White solid, yield 68 %, mp 109.6-110.8 °C. 1H NMR (400
MHz, CDCl3) δ 7.74 (d, J = 16.0 Hz, 1H, CH=CH-Ar), 7.54-7.51 (m, 2H, ArH), 7.41-7.36
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(m, 3H, ArH), 6.95-6.87 (m, 2H, ArH,CH=CH-Ar), 6.64 (s,1H, ArH), 6.57 (s, 1H, ArH),
6.52 (s, 1H, ArH), 6.42 (s, 1H, CHN), 5.93(d, J = 14.8 Hz, 1H, OCH2O), 4.01-3.97 (m, 1H,
NCH2CH2), 3.82 (s, 3H, OCH3), 3.77 (s, 6H, OCH3×2), 3.57-3.51 (m, 1H, NCH2CH2),
2.98-2.91 (m, 1H, NCH2CH2), 2.80-2.75 (m, 1H, NCH2CH2); 13C NMR (100 MHz, CDCl3) δ
158.9, 146.7, 146.3, 143.1, 135.3, 134.6, 129.9, 129.6, 128.8, 128.6, 128.5, 127.7, 127.5,
117.4, 113.6, 108.5, 108.1, 100.9, 55.2, 55.0, 39.6, 29.4; IR (KBr, cm-1) υ: 2934.9, 2834.6,
1645.8, 1501.8, 1423.5, 1235.9, 1125.7, 923.4, 848.8; ESI-Mass for C28H27NO6: m/z (M+H)
+474.30.
(E)-1-(5-(4-nitrophenyl)-7,8-dihydro-[1, 3]dioxole and[4,5-g]isoquinoline-6
(5H)-yl)-3-phenyl-2-en-1-one (21c). Compound 21c was prepared from 14c according to
procedure used for 16d synthesis. Yellow solid, yield 75 %, mp 113.6-114.9 °C. 1H NMR
(400 MHz, CDCl3) δ 8.14(d, J = 8.4 Hz, 2H, ArH), 7.78 (d, J =15.2 Hz, 1H, CH=CH-Ar),
7.53 (d, J = 4.4 Hz,2H, ArH), 7.47 (d, J = 8.4 Hz, 2H, ArH), 7.39-7.37 (m, 3H, ArH), 6.95 (s,
1H, ArH), 6.91 (d, J = 15.2 Hz, 1H, CH=CH-Ar), 6.69 (s, 1H, ArH), 6.55 (s, 1H, CHN), 5.97
(d, J = 6.4 Hz, 2H, OCH2O) , 4.01-3.97 (m, 1H, NCH2CH2), 3.55-3.48 (m, 1H, NCH2CH2),
2.99-2.93 (m, 1H, NCH2CH2), 2.78-2.74 (m, 1H, NCH2CH2); 13C NMR (150 MHz, CDCl3) δ
165.6, 149.4, 147.2, 147.0, 146.5, 144.0, 134.9, 129.9, 129.3, 128.8, 127.87, 127.80, 126.9,
123.5, 116.4, 55.2, 40.4, 29.0; IR (KBr, cm-1) υ: 2890.1, 1644.3, 1492.1, 1442.4, 981.5,
856.4; ESI-Mass for C25H20N2O5: m/z (M+H) +429.11.
6.3 In vitro anti-tumor activity
6.3.1 Anti-tumor activity by SRB assay
The anti-proliferation of compounds against HUVEC (human umbilical vein
endothelial cells), MCF-7 (human breast cancer cells) and HT-29 (human colon cancer cells)
were evaluated by SRB (sulforhodamine B) assay. Cells were seeded in 96-well culture plates
(1×104 cells per well) and cultured for 12 h. Incubated (5 % CO2, 95 % humidity, 37 °C) the
cells with following treatment with the test compounds at increasing concentrations in the
presence of 10 % FBS for 72 h. Then TCA was added and incubated at 4 °C for 1 h to fix the
cells. After washing the distilled water, 100 μL of 0.057 % SRB solution was added to each
well and incubated at 37 °C for 30 min and then quickly rinse the plates four times with 1 %
（vol/vol）acetic acid to remove unbound dye. Use a blow dryer to dry the plates. 100 μL(10
mM)Tris-Base solution were added to each well and shake mechanically for 30 min, and then
absorbance values at a wavelength of 515 nm were determined with the Spectramax
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Microtiter Plate Luminometer (Molecular Devices, USA). The percentage of cell-inhibition
was calculated using the formula below.
Inhibition rate (%) = (OD control -OD administration) / OD control × 100%，And
calculate the IC50.
6.3.2 Western blot
MCF-7 cells cultured overnight in a 12-well plate were treated with 17d, 17e and CA-4
for 24 h. Subsequently, cells were lysed using Lysis Buffer (Kengen Biotech, China), and
protein concentration was measured with coomassie brilliant blue G250 protein assay kit
(Kengen Biotech, China). The lysate was subjected to a 15 % SDS-PAGE, and transferred to
a WHATMAN membrane using iBlot™system (Invitrogen, Carlsbad, CA). Membranes were
blocked with 3 % BSA in 1× Tris-buffered saline (TBS) at room temperature for 1 h and then
incubated with primary antibodies at 4 °C overnight, followed by incubation with anti-rabbit
IRDye 800CW secondary antibodies for 1 h at room temperature. All the blots were reported
with total β-actin antibody as control. The signal of target proteins was detected by BIO-RAD
Gel Doc XR and Bio-Rad 164-5051.
6.4 Antitumor activity in vivo
Four weeks old, female BALB/c nude mice (19–22 g) were purchased from the animal
breeding laboratories of Academy of Military Medical Sciences. Thirty female BALB/c nude
mice were randomly divided into four groups, 6 mice per group: a vehicle control group, a
tamoxifen (20 mg/kg) treatment group, and 17d, 17e (20 mg/kg) treatment group. Human
breast cancer xenografts were established by intraperitoneal injection of MCF-7 cells in right
axillary subcutaneous of nude mice. When tumor volume reached 100 mm3, treatment started
with 17d and 17e injected intraperitoneally once daily at doses of 20 mg/kg or with
tamoxifen injected intraperitoneally at doses of 20 mg/kg every day for 30 days. Vehicle
control group were intraperitoneal injection of saline once daily. Tumor burden was
measured every 6 days with a caliper.
(calculated volume (mm3) = 1/2*length*width*width). After completing the treatment
schedule, tumor-bearing mice were euthanized.
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Statistical Analysis
Biological results are reported as means standard error. Statistical analysis was
performed by using one-way analysis of variance. A P value of less than 0.05 was considered
to be statistically significant.
Author Contributions
All authors have given approval to the final version of the manuscript.
Conflict of interest
The authors declare no competing financial interest.
Acknowledgements
This study was supported by grants from Anhui Education Department Science key project
(KJ2016A414), National Undergraduate Training Programs for Innovation and Entrepreneurship
(2015108), the National innovative drug incubation base project (2012ZX09401066).
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