Synthesis of cinnamoyl glucoside derivatives and their antiproliferation activities against murine melanoma B16-F10 cell line

Article abstract of DOI:10.1016/j.carres.2021.108332

Twelve cinnamoyl glucoside derivatives were prepared by glycosylation of glucosyl trichloroacetimidate and cinnamic acid derivatives, followed by dechloroacetylation with a pyridine/H₂O mixture. Their structures were characterized by ¹H and ¹³C NMR, as well as mass analysis. All the products were tested for their antiproliferation activities against murine melanoma B16-F10 cell line. Compounds 4e-4j were able to inhibit the proliferation of murine melanoma B16-F10 cell line with IC₅₀ values of 17.38 ± 0.07, 9.87 ± 0.09, 9.69 ± 0.12, 29.42 ± 0.04, 32.95 ± 0.08, 25.68 ± 0.09 μM, respectively.

Full text of DOI:10.1016/j.carres.2021.108332

Carbohydrate Research 504 (2021) 108332
Contents lists available at ScienceDirect
Carbohydrate Research
journal homepage: www.elsevier.com/locate/carres
Note
Synthesis of cinnamoyl glucoside derivatives and their antiproliferation
activities against murine melanoma B16-F10 cell line
a
,
*
a
a
a
a
a
Penghua Shu , Mengzhu Yu , Yamin Li , Yuehui Luo , Hao Liu , Huiqing Zhu ,
a
a
b
a,**
Jialong Zhang , Lingxiang Zhang , Xialan Wei , Fugang Xiao
a
Food and Pharmacy College, Xuchang University, Xuchang, China
b
School of Information Engineering, Xuchang University, Xuchang, China
A R T I C L E I N F O
A B S T R A C T
Keywords:
Twelve cinnamoyl glucoside derivatives were prepared by glycosylation of glucosyl trichloroacetimidate and
cinnamic acid derivatives, followed by dechloroacetylation with a pyridine/H O mixture. Their structures were
Cinnamoyl glucoside
Glycosylation
2
characterized by 1H and C NMR, as well as mass analysis. All the products were tested for their anti-
proliferation activities against murine melanoma B16-F10 cell line. Compounds 4e-4j were able to inhibit the
proliferation of murine melanoma B16-F10 cell line with IC50 values of 17.38 ± 0.07, 9.87 ± 0.09, 9.69 ± 0.12,
13
Dechloroacetylation
Antiproliferation
Melanoma
2
9.42 ± 0.04, 32.95 ± 0.08, 25.68 ± 0.09 μM, respectively.
1
. Introduction
Melanoma, a kind of malignant skin tumor, poses a great threat to the
B16-F10 cell line. Among them, the hit compounds 4e-4j showed po-
tential on inhibiting the proliferation of B16-F10 cell line (IC50 values:
17.38 ± 0.07
29.42 ± 0.04
μ
M for 4e, 9.87 ± 0.09 M for 4f, 9.69 ± 0.12
μ
μ
M for 4g,
M for 4j),
quality of human life due to its high malignancy, easy spreading, and
poor prognosis [1]. As one of the highest mortality tumors in the world,
its incidence has been rising at a consistent rate of 3% over the past
thirty years [2]. Despite molecular targeted therapy and immuno-
therapy have achieved some positive results for treatments of metastatic
melanoma, these methods can hardly be widely used in clinical practice
due to their high prices, serious side effects and obvious individual
variations [3]. Therefore, developing new treatment strategies or agents
is still urgent to further improve melanoma prognosis.
μ
M for 4h, 32.95 ± 0.08 μM for 4i, 25.68 ± 0.09
μ
being comparable with the positive control doxorubicin (IC50 value:
2.18 ± 0.04
μ
M). Herein, the synthesis, structure elucidation and anti-
proliferation activity evaluation of the twelve cinnamoyl glucoside de-
rivatives were reported.
2. Results and discussion
2.1. Chemistry
Cinnamoyl glucoside derivatives consist of cinnamic acid derivatives
and glucose. They are widely spread in plants [4–7], fruits [8–11] and
vegetables [12,13] and have antioxidant [14], antiglycative [15],
phytotoxic [16] and inhibition of antigen-stimulated degranulation [9]
activities. Recent study showed that cinnamoyl glucoside derivatives
could inhibit the proliferation of tumor cells; for example, 1-O-p-hy-
droxycinnamoylglucose can significantly inhibit the proliferation of
human HCT116 colorectal cancer cell line in a dose-dependent manner
Reports recommended that the cinnamoyl moiety posed obvious
influence on the bioactivities of cinnamoyl glucosides (Scheme 1). For
example, in DPPH radical scavenging activity assay (DPPH assay), 1-O-
(E)-caffeoyl-β-D-glucopyranose exhibited stronger activity than 1-O-(E)-
cinnamoyl-β-D-glucopyranose, while in superoxide radical scavenging
assay (NBT assay) the latter compound showed more effective than the
former [14]. In the caspase 3/7 activity inhibition tests, cinnamoyl
glucosides had an order of cinnamoyl- > caffeoyl- > p-coumaroyl- [15]
(Scheme 1). Therefore, considering the structural diversity of synthesis,
we set the cinnamoyl moiety as aglycone for 4a-4h, with
electron-donating, electron-withdrawing, and neutral substituents at the
[
17].
In this work, a series of cinnamoyl glucoside derivatives 4a-4l were
synthesized and screened as lead compounds for melanoma treatment
by evaluating their antiproliferation activities against murine melanoma
*
Corresponding authorFood and Pharmacy College, Xuchang University, 88 Bayi Road, Xuchang, China. (P. Shu).
** Corresponding author. (F. Xiao).
E-mail addresses: shupenghua@yeah.net (P. Shu), xfug@163.com (F. Xiao).
https://doi.org/10.1016/j.carres.2021.108332
Received 28 January 2021; Received in revised form 23 March 2021; Accepted 23 April 2021
Available online 29 April 2021
0
008-6215/© 2021 Elsevier Ltd. All rights reserved.

P. Shu et al.
Carbohydrate Research 504 (2021) 108332
ortho-, meta- or para-position of the benzene ring. Additionally, we
designed 4i-4l by small modifications of the structures, for example,
replacing the six-membered aromatic ring with the less bulky
five-membered heterocyclic ring (4i, 4j), or further introducing a methyl
onto the β-position of double bond (4k, 4l).
3. Materials and methods
3.1. General information
All reactions were monitored by thin-layer chromatography over
silica-gel-coated TLC plates (Yantai Chemical Industry Research Insti-
Given the base-sensitive groups in the target compounds 4a-4l,
chloroacetyl was chosen as the key protecting group for glycosyl donor
since it could be smoothly removed under mild conditions and keep the
base-sensitive 1-O-acyl intact [18]. The general synthesis route con-
sisted of two steps as shown in Scheme 2. Initially, glucopyranosyl tri-
chloroacetimidate 1 and various cinnamic acid derivatives 2a-2l were
stirred at room temperature with 10% TMSOTf as catalyst to obtain
protected cinnamoyl glycosides 3a-3l (81–94% yield). Afterwards, 3a-3l
tute). The spots on TLC were visualized by warming 10% H
2 4
SO (10%
H
2
SO in ethanol) sprayed plates on a hot plate. Column chromatog-
4
raphy was performed using silica gel (Qingdao Marine Chemical Inc.,
China). NMR spectra were recorded on a Bruker AM-400 spectrometer
1
13
(400 MHz), and the H and C NMR chemical shifts were referenced to
the solvent or solvent impurity peaks for CDCl
3
at δ
H
7.24 and δ 77.23,
C
2.05 and δ 29.84.
C
for CD OD at δ 3.31 and δ 49.15, for (CD CO at δ
3
H
C
3
)
2
H
were dechloroacetylation with a pyridine/H
2
O mixture, resulting in the
High-resolution electrospray ionization mass spectra (HRESIMS) were
carried out on a Bruker micrOTOF II spectrometer. Compounds 2a-2j
were purchased from Adamas and used without further purification.
Compounds 2k and 2l were synthesized following literature procedures
[19].
corresponding target compounds 4a-4l (90–95% yield). Under mild,
concise reaction conditions, all of the synthetic compounds were ob-
tained in good yields and gave satisfactory analytical data, which
coincided with the structures in Scheme 2. Although the synthesis
approach of cinnamoyl glucoside derivatives 4a and 4b have been re-
ported [18], 4c-4l were synthesized for the first time.
3
.2. Preparation of compounds 3a-3l
2
.2. Anti-proliferation assay
General glycosylation procedure: A solution of the glycosyl donor 1
(
0.12 mmol, 1.2 equiv) and acceptor 2 (0.1 mmol, 1.0 equiv) in dry
For developing novel anti-melanoma agents, the in vitro anti-
CH
2
Cl
2
(2.0 mL) in the presence of 4 Å MS (100 wt%) was stirred for 15
◦
proliferation effects of compounds 4a-4l against murine melanoma
B16-F10 cell line were evaluated. The results were shown in Table 1. A
general tendency was that when the substituent group of benzene ring
changed from electron-donating (4b-4d) to electron-withdrawing (4e-
min at 0 C. After addition of TMSOTf (0.01 mmol, 0.1 equiv), the
mixture was slowly warmed to room temperature and stirred for 5 h.
Molecular sieves were removed by filtration and the solution was pu-
rified by silica gel chromatography to give the desired product 3.
4
h), the anti-proliferation effect became more potent. Further replacing
Compound 3a. 14.8 mg, 0.1 mmol of 2a to yield 3a (56.0 mg, 0.09
the benzene ring (4a) with the five-membered heterocyclic ring (4i, 4j),
the anti-proliferation effect increased obviously. However, introducing a
methyl onto the β-position of double bond (4k) led to a sharp decrease of
its activity against the growth of B16-F10 cell line. Over all, the density
of the double bond and electron density of ring could make big differ-
ences on the anti-proliferation effects of cinnamoyl glucoside de-
1
mmol, 91%) as a white powder. H NMR (CDCl
3
, 400 MHz) δ: 7.73 (1H,
d, J = 16.0 Hz), 7.53–7.49 (2H, m), 7.40–7.38 (3H, m), 6.38 (1H, d, J =
6.0 Hz), 5.89 (1H, d, J = 8.0 Hz, H-1), 5.43 (1H, t, J = 9.2 Hz), 5.29
1
(
1H, dd, J = 9.2, 8.0 Hz), 5.24 (1H, t, J = 9.6 Hz), 4.39 (1H, dd, J = 12.8,
4
3
.4 Hz), 4.28 (1H, dd, J = 12.8, 2.0 Hz), 4.10 (2H, s), 4.01 (2H, d, J =
.2 Hz), 3.99 (2H, s), 3.98 (2H, s), 3.97 (1H, m) [18].
rivatives. When looking at the most potential compounds 4e (17.38
f (9.87 M), 4g (9.69 M), 4h (29.42 M), 4i (32.95 M), 4j (25.68
M), the structures are quite similar but a simple structure-activity
μ
M),
Compound 3b. 17.8 mg, 0.1 mmol of 2b to yield 3b (60.1 mg, 0.09
4
μ
μ
μ
μ
1
mmol, 93%) as a white powder. H NMR (CDCl
3
, 400 MHz) δ: 7.67 (1H,
μ
d, J = 16.0 Hz), 7.45 (2H, d, J = 8.8 Hz), 6.87 (2H, d, J = 8.8 Hz), 6.22
relationship could be got which can guide us towards the preparation
of other more effective analogs in the future. When the electron-
withdrawing effects of substituent groups on benzene ring became
stronger, the antiproliferation activities of cinnamoyl glucosides
increased greatly. In order to see whether these compounds were
selectively toxic to cancer cells, normal human dermal fibroblast (HDF)
cell line was evaluated in parallel. All tested compounds showed no
significant cytotoxicity at a concentration of 40 mM (Table 2), indicating
that they were all nontoxic to the healthy cell line. Those first attempts
to try to understand bioactivity of these compounds using simple in vitro
methods need obviously more in vivo studies to clearly identify their
mode or mechanism of action. The anti-proliferation activities of 4a-4l
were tested for the first time and the results above would shed light on
the design and development of novel anti-melanoma agents.
(
1H, d, J = 16.0 Hz), 5.88 (1H, d, J = 8.4 Hz, H-1), 5.42 (1H, t, J = 9.2
Hz), 5.30 (1H, dd, J = 9.2, 8.4 Hz), 5.24 (1H, t, J = 9.6 Hz), 4.38 (1H, dd,
J = 12.8, 4.0 Hz), 4.28 (1H, dd, J = 12.8, 2.4 Hz), 4.09 (2H, s), 4.00 (2H,
d, J = 3.6 Hz), 3.99 (2H, s), 3.98 (2H, s), 3.97 (1H, m), 3.81 (3H, s,
3
OCH ) [18].
Compound 3c. 16.2 mg, 0.1 mmol of 2c to yield 3c (56.7 mg, 0.09
1
mmol, 90%) as a white powder. H NMR (CDCl
3
, 400 MHz) δ: 7.72 (1H,
d, J = 16.0 Hz), 7.41 (2H, d, J = 8.8 Hz), 7.19 (2H, d, J = 8.8 Hz), 6.33
(
1H, d, J = 16.0 Hz), 5.88 (1H, d, J = 8.0 Hz, H-1), 5.42 (1H, t, J = 9.2
Hz), 5.30 (1H, dd, J = 9.2, 8.4 Hz), 5.24 (1H, t, J = 10.0 Hz), 4.39 (1H,
dd, J = 12.8, 4.4 Hz), 4.28 (1H, dd, J = 12.8, 2.4 Hz), 4.10 (2H, s), 4.02
(
13
2H, d, J = 2.8 Hz), 3.99 (2H, s), 3.97 (1H, m), 3.98 (2H, s), 2.36 (3H, s);
C NMR (CDCl
3
, 100 MHz) δ: 167.2, 167.1, 166.5, 166.3, 164.8, 148.4,
1
7
42.0, 131.2, 130.0, 130.0, 128.7, 128.7, 114.7, 91.5 (C-1), 74.1, 72.3,
1.6, 69.4, 63.0, 40.7, 40.4, 40.4, 40.4, 21.7. HRMS calc. for
+
C
24
H24Cl
4
O
11Na [M+Na] : 652.9941, found: 652.9943.
Compound 3d. 16.2 mg, 0.1 mmol of 2d to yield 3d (59.2 mg, 0.09
1
mmol, 94%) as a white powder. H NMR (CDCl
3
, 400 MHz) δ: 7.71 (1H,
Scheme 1. Cinnamoyl glucosides with different cinnamoyl moieties and their bioactivities.
2

P. Shu et al.
Carbohydrate Research 504 (2021) 108332
Scheme 2. Synthesis of cinnamoyl glucoside derivatives 4a-4l.
d, J = 16.0 Hz), 7.33 (1H, d, J = 1.6 Hz), 7.32 (1H, d, J = 8.0 Hz), 7.27
1H, t, J = 8.0 Hz), 7.21 (1H, d, J = 8.0 Hz), 6.37 (1H, d, J = 16.0 Hz),
.89 (1H, d, J = 8.0 Hz, H-1), 5.42 (1H, t, J = 9.2 Hz, H-1), 5.31 (1H, dd,
d, J = 16.0 Hz), 7.50 (1H, d, J = 8.0 Hz), 7.49 (1H, t, J = 8.0 Hz), 7.06
(1H, d, J = 8.0 Hz), 7.04 (1H, t, J = 8.0 Hz), 6.28 (1H, d, J = 16.0 Hz),
5.88 (1H, d, J = 8.0 Hz, H-1), 5.43 (1H, t, J = 9.6 Hz), 5.29 (1H, t, J =
9.6 Hz), 5.23 (1H, t, J = 9.6 Hz), 4.38 (1H, dd, J = 12.8, 4.0 Hz), 4.28
(
5
J = 9.2, 8.4 Hz), 5.25 (1H, t, J = 9.6 Hz), 4.40 (1H, dd, J = 12.8, 4.0 Hz),
4
3
1
1
6
.30 (1H, dd, J = 12.8, 2.4 Hz), 4.10 (2H, s), 4.02 (2H, d, J = 3.2 Hz),
(1H, dd, J = 12.8, 2.4 Hz), 4.08 (2H, s), 4.01 (2H, s), 3.99 (2H, s), 3.99
.99 (2H, s), 3.99 (1H, m), 3.99 (2H, s), 2.35 (3H, s); 13C NMR (CDCl
,
(1H, m), 3.98 (2H, s); C NMR (CDCl
13
, 100 MHz) δ: 167.1, 167.0, 166.5,
3
3
00 MHz) δ: 167.2, 167.0, 166.5, 166.3, 164.7, 148.5, 139.0, 133.8,
32.2, 129.3, 129.1, 125.9, 115.6, 91.5 (C-1), 74.0, 72.3, 71.6, 69.3,
166.3, 164.5, 146.9, 130.7, 130.6, 130.1, 130.1, 116.5, 116.3, 115.5,
91.5 (C-1), 73.9, 72.2, 71.5, 69.2, 62.9, 40.7, 40.4, 40.4, 40.4. HRMS
+
2.9, 40.7, 40.4, 40.4, 40.4, 21.5. HRMS calc. for C24
H24Cl
4
O
11Na
calc. for C23
H
21
O
11NaCl
4
F [M+Na] : 654.9720, found: 654.9724.
+
[
M+Na] : 652.9941, found: 652.9945.
Compound 3f. 21.6 mg, 0.1 mmol of 2f to yield 3f (58.8 mg, 0.09
1
Compound 3e. 16.6 mg, 0.1 mmol of 2e to yield 3e (53.3 mg, 0.08
mmol, 86%) as a white powder. H NMR (CDCl
3
, 400 MHz) δ: 7.74 (1H,
1
mmol, 84%) as a white powder. H NMR (CDCl
3
, 400 MHz) δ: 7.67 (1H,
d, J = 16.0 Hz), 7.64–7.60 (4H, overlap), 6.45 (1H, d, J = 16.0 Hz), 5.90
3

P. Shu et al.
Carbohydrate Research 504 (2021) 108332
+
Table 1
HRMS calc. for C21
H
20
O
12Cl
4
Na [M+Na] : 626.9607, found: 626.9606.
Anti-proliferation activities of compounds 4a-4l against murine melanoma B16-
Compound 3j. 15.4 mg, 0.1 mmol of 2j to yield 3j (55.4 mg, 0.09
a
1
F10 cell line.
mmol, 89%) as a white powder. H NMR (CDCl
, 400 MHz) δ: 7.82 (1H,
3
Compounds
IC50
(
μ
M)
Compounds
IC50 (μM)
d, J = 16.0 Hz), 7.42 (1H, d, J = 4.8 Hz), 7.29 (1H, d, J = 3.6 Hz), 7.05
(
1H, dd, J = 4.8, 3.6 Hz), 6.15 (1H, d, J = 16.0 Hz), 5.88 (1H, d, J = 8.0
4
4
4
4
4
4
4
a
b
c
d
e
f
195.1 ± 0.15
90.48 ± 0.09
91.92 ± 0.10
112.0 ± 0.08
17.38 ± 0.07
9.87 ± 0.09
9.69 ± 0.12
4h
29.42 ± 0.04
32.95 ± 0.08
25.68 ± 0.09
299.7 ± 0.16
44.13 ± 0.06
2.18 ± 0.04
Hz, H-1), 5.41 (1H, t, J = 9.2 Hz), 5.29 (1H, dd, J = 9.2, 8.0 Hz), 5.23
(1H, t, J = 9.2 Hz), 4.38 (1H, dd, J = 12.4, 4.4 Hz), 4.28 (1H, dd, J =
4i
4j
4k
1
2.4, 2.4 Hz), 4.09 (2H, s), 4.00 (2H, d, J = 3.2 Hz), 3.99 (2H, s), 3.99
4l
13
(
2H, s), 3.98 (1H, m); C NMR (CDCl
3
, 100 MHz) δ: 167.2, 167.0, 166.5,
b
Doxorubicin
1
7
66.3, 164.5, 140.5, 139.0, 132.6, 130.2, 128.5, 114.3, 91.5 (C-1), 74.0,
g
2.2, 71.5, 69.2, 62.9, 40.7, 40.4, 40.4, 40.4. HRMS calc. for
a
b
Results were expressed as means ± SEMs.
+
C
21
H
20
O
11NaSCl
4
[M+Na] : 642.9378, found: 642.9370.
Positive control.
Compound 3k. 16.8 mg, 0.1 mmol of 2k to yield 3k (56.0 mg, 0.09
1
mmol, 88%) as a white powder. H NMR (CDCl
d, J = 4.8 Hz), 7.35 (1H, d, J = 3.6 Hz), 7.04 (1H, dd, J = 4.8, 3.6 Hz),
.18 (1H, s), 5.86 (1H, d, J = 8.4 Hz, H-1), 5.40 (1H, t, J = 9.6 Hz), 5.26
3
, 400 MHz) δ: 7.37 (1H,
Table 2
6
Inhibitory effects of compounds 4a-4l on cell viability of HDF cell line at 40
a
(
1H, dd, J = 9.6, 8.4 Hz), 5.23 (1H, t, J = 9.6 Hz), 4.38 (1H, dd, J = 12.4,
mM.
4
3
.0 Hz), 4.28 (1H, dd, J = 12.4, 2.4 Hz), 4.10 (2H, s), 4.01 (2H, d, J =
Compounds
Viability (%)
Compounds
Viability (%)
13
.6 Hz), 3.99 (2H, s), 3.98 (2H, s), 3.97 (1H, m), 2.59 (3H, s); C NMR
, 100 MHz) δ: 167.2, 167.1, 166.5, 166.3, 164.1, 152.6, 144.8,
4
4
4
4
4
4
4
a
b
c
d
e
f
100.11 ± 0.15
101.95 ± 1.66
100.46 ± 0.37
101.46 ± 1.07
101.18 ± 0.16
102.96 ± 1.12
100.79 ± 0.34
4h
100.36 ± 0.21
100.65 ± 0.32
102.13 ± 0.98
100.10 ± 0.38
99.72 ± 0.38
101.23 ± 0.98
(
CDCl
3
4i
1
28.7, 128.4, 128.1, 111.5, 91.0 (C-1), 74.2, 72.2, 71.5, 69.2, 62.9, 40.7,
4j
[M+Na]+:
4k
40.4, 40.4, 40.4, 18.0. HRMS calc. for C22
22 4
H O11NaSCl
4l
656.9535, found: 656.9537.
b
Doxorubicin
Compound 3l. 16.2 mg, 0.1 mmol of 2l to yield 3l (57.3 mg, 0.09
g
1
mmol, 93%) as a white powder. H NMR (CDCl
(2H, overlap), 7.39–7.35 (3H, overlap), 6.10 (1H, d, J = 1.6 Hz), 5.86
1H, d, J = 8.4 Hz, H-1), 5.40 (1H, t, J = 9.6 Hz), 5.27 (1H, dd, J = 9.6,
.4 Hz), 5.24 (1H, t, J = 9.6 Hz), 4.39 (1H, dd, J = 12.4, 4.0 Hz), 4.30
(1H, dd, J = 12.4, 2.4 Hz), 4.11 (2H, s), 4.01 (2H, d, J = 3.2 Hz), 3.99
(2H, s), 3.98 (2H, s), 3.97 (1H, m), 2.58 (3H, s); 13C NMR (CDCl
, 100
3
, 400 MHz) δ: 7.47–7.45
a
b
Results were expressed as means ± SEMs.
Positive control.
(
8
(
1H, d, J = 8.4 Hz, H-1), 5.44 (1H, t, J = 8.4 Hz), 5.31 (1H, t, J = 8.0 Hz),
.25 (1H, d, J = 9.6 Hz), 4.40 (1H, dd, J = 12.8, 4.0 Hz), 4.30 (1H, dd, J
12.8, 1.2 Hz), 4.09 (2H, s), 4.02 (2H, s), 4.01 (1H, m), 3.99 (2H, s),
.99 (2H, s); 13C NMR (CDCl
, 100 MHz) δ: 167.2, 167.0, 166.5, 166.3,
64.1, 146.2, 137.2, 133.0 (q, J = 32.3 Hz), 128.8, 128.8, 128.8, 126.2
5
3
=
MHz) δ: 167.2, 167.1, 166.5, 166.3, 164.1, 160.9, 141.5, 130.0, 128.9,
128.9, 126.6, 126.6, 114.8, 91.1 (C-1), 74.2, 72.3, 71.6, 69.3, 62.9, 40.8,
40.4, 40.4, 40.4, 18.7. HRMS calc. for C24H24O11NaCl4 [M+Na]+:
650.9970, found: 650.9967.
3
1
3
(
d, J = 4.0 Hz), 126.2 (d, J = 4.0 Hz), 118.5, 91.7 (C-1), 73.9, 72.4, 71.5,
6
9.3, 62.9, 40.7, 40.4, 40.4, 40.4. HRMS calc. for C24
H
22
O11Cl
4
F
3
+
[
M+H] : 682.9868, found: 682.9861.
3.3. Preparation of compounds 4a-4l
Compound 3g. 19.3 mg, 0.1 mmol of 2g to yield 3g (53.6 mg, 0.08
1
mmol, 81%) as a white powder. H NMR (CDCl
3
, 400 MHz) δ: 8.25 (2H,
General dechloroacetylation procedure: Compound 3 (0.08 mmol)
d, J = 8.4 Hz), 7.76 (1H, d, J = 16.0 Hz), 7.68 (2H, d, J = 8.4 Hz), 6.51
was suspended in a 1:1 mixture of pyridine and H O (1.0 mL). After
2
◦
(
1H, d, J = 16.0 Hz), 5.89 (1H, d, J = 8.0 Hz, H-1), 5.43 (1H, t, J = 9.6
stirring at 35 C for 5 h, TLC showed the reaction was complete. The
Hz), 5.32 (1H, dd, J = 9.6, 8.0 Hz), 5.25 (1H, d, J = 9.6 Hz), 4.40 (1H,
dd, J = 12.4, 4.0 Hz), 4.31 (1H, dd, J = 12.8, 2.0 Hz), 4.10 (2H, s), 4.02
solvents were removed under vacuum and the residue was purified by
flash column chromatography to yield 4.
1
3
(
2H, d, J = 2.8 Hz), 3.99 (1H, m), 3.99 (2H, s), 3.99 (2H, s); C NMR
Compound 4a. 49.3 mg, 0.08 mmol of 3a to yield 4a (22.8 mg, 0.07
1
(
CDCl
3
, 100 MHz) δ: 167.1, 167.0, 166.5, 166.3, 163.7, 149.2, 145.1,
mmol, 92%) as a white powder. H NMR (CD OD, 400 MHz) δ: 7.80 (1H,
3
1
6
39.8, 129.3, 129.3, 124.5, 124.5, 120.2, 91.9 (C-1), 73.9, 72.5, 71.5,
d, J = 16.0 Hz), 7.64–7.62 (2H, m), 7.43–7.40 (3H, m), 6.58 (1H, d, J =
16.0 Hz), 5.59 (1H, d, J = 7.6 Hz, H-1), 3.87 (1H, dd, J = 12.4, 2.0 Hz),
3.70 (1H, dd, J = 12.4, 5.2 Hz), 3.52–3.40 (4H, overlap) [18].
Compound 4b. 51.7 mg, 0.08 mmol of 3b to yield 4b (25.6 mg, 0.08
9.3, 62.9, 40.7, 40.4, 40.4, 40.3. HRMS calc. for C23
H
21NO13NaCl
4
+
[
M+Na] : 681.9665, found: 681.9659.
Compound 3h. 18.3 mg, 0.1 mmol of 2h to yield 3h (59.9 mg, 0.09
1
1
mmol, 92%) as a white powder. H NMR ((CD
3
)
2
CO, 400 MHz) δ: 8.12
mmol, 94%) as a white powder. H NMR (CD OD, 400 MHz) δ: 7.75 (1H,
3
(
1H, d, J = 16.0 Hz), 7.93 (1H, d, J = 7.6 Hz), 7.53 (1H, d, J = 8.0 Hz),
d, J = 16.0 Hz), 7.56 (2H, d, J = 8.4 Hz), 6.96 (2H, d, J = 8.4 Hz), 6.42
(1H, d, J = 16.0 Hz), 5.59 (1H, d, J = 7.6 Hz, H-1), 3.86 (1H, dd, J =
12.0, 2.0 Hz), 3.83 (3H, s), 3.70 (1H, dd, J = 12.0, 4.0 Hz), 3.48–3.36
(4H, overlap) [18].
7
.48 (1H, t, J = 8.0 Hz), 7.42 (1H, t, J = 8.0 Hz), 6.60 (1H, d, J = 16.0
Hz), 6.18 (1H, d, J = 8.0 Hz, H-1), 5.71 (1H, t, J = 9.6 Hz), 5.35 (1H, t, J
=
9.2 Hz), 5.33 (1H, d, J = 9.2 Hz), 4.46–4.37 (3H, overlap), 4.32–4.20
(
8H, overlap); 13C NMR ((CD
3
)
2
CO, 100 MHz) δ: 168.4, 168.2, 168.0,
Compound 4c. 50.4 mg, 0.08 mmol of 3c to yield 4c (24.1 mg, 0.07
1
1
9
67.8, 165.4, 143.6, 136.2, 133.8, 133.5, 131.7, 129.9, 129.3, 120.8,
mmol, 93%) as a white powder. H NMR (CD OD, 400 MHz) δ: 7.77 (1H,
3
3.0 (C-1), 74.8, 73.5, 72.9, 71.0, 64.5, 42.2, 42.2, 42.1, 42.0. HRMS
d, J = 16.0 Hz), 7.50 (2H, d, J = 8.0 Hz), 7.23 (2H, d, J = 8.0 Hz), 6.51
(1H, d, J = 16.0 Hz), 5.59 (1H, d, J = 7.6 Hz, H-1), 3.85 (1H, dd, J =
12.0, 1.6 Hz), 3.70 (1H, dd, J = 12.0, 4.4 Hz), 3.47–3.36 (4H, overlap),
+
calc. for C23
H
21
O
11Cl
5
Na [M+Na] : 670.9424, found: 670.9424.
Compound 3i. 13.8 mg, 0.1 mmol of 2i to yield 3i (54.6 mg, 0.09
1
13
mmol, 90%) as a white powder. H NMR (CDCl
m), 7.45 (1H, d, J = 16.0 Hz), 6.68 (1H, m), 6.47 (1H, m), 6.23 (1H, d, J
16.0 Hz), 5.87 (1H, d, J = 8.0 Hz, H-1), 5.41 (1H, t, J = 9.2 Hz), 5.28
1H, t, J = 9.2 Hz), 5.23 (1H, d, J = 9.2 Hz), 4.38 (1H, dd, J = 12.4, 4.0
Hz), 4.28 (1H, dd, J = 12.4, 2.0 Hz), 4.08 (2H, s), 4.00 (2H, d, J = 2.8
Hz), 3.99 (1H, m), 3.97 (2H, s), 3.97 (2H, s); 13C NMR (CDCl
, 100 MHz)
δ: 167.2, 167.0, 166.5, 166.3, 164.7, 150.5, 145.9, 133.9, 116.9, 113.2,
12.8, 91.5 (C-1), 74.0, 72.2, 71.5, 69.3, 62.9, 40.7, 40.4, 40.4, 40.4.
3
, 400 MHz) δ: 7.49 (1H,
2.36 (3H, s); C NMR (CD OD, 100 MHz) δ: 167.5, 147.8, 142.7, 133.0,
3
130.9, 130.9, 129.6, 129.6, 117.2, 96.0 (C-1), 79.0, 78.1, 74.2, 71.2,
+
=
62.5, 21.6. HRMS calc. for C16H20O Na [M+Na] : 347.1107, found:
7
(
347.1109.
Compound 4d. 50.4 mg, 0.08 mmol of 3d to yield 4d (24.6 mg, 0.08
1
3
mmol, 95%) as a white powder. H NMR (CD OD, 400 MHz) δ: 7.77 (1H,
3
d, J = 16.0 Hz), 7.44 (1H, d, J = 1.6 Hz), 7.40 (1H, d, J = 7.6 Hz), 7.30
1
(1H, t, J = 7.6 Hz), 7.24 (1H, d, J = 7.6), 6.55 (1H, d, J = 16.0 Hz), 5.60
4

P. Shu et al.
Carbohydrate Research 504 (2021) 108332
(
1H, d, J = 7.6 Hz, H-1), 3.86 (1H, dd, J = 12.0, 1.6 Hz), 3.70 (1H, dd, J
mmol, 93%) as a white powder. 1H NMR (CD
3
OD, 400 MHz) δ:
1
3
=
12.0, 4.4 Hz), 3.50–3.37 (4H, overlap), 2.37 (3H, s); C NMR
7.54–7.45 (2H, overlap), 7.42–7.35 (3H, overlap), 6.21 (1H, d, J = 1.6
Hz), 5.57 (1H, d, J = 8.0 Hz, H-1), 3.86 (1H, dd, J = 12.4, 2.4 Hz), 3.70
(
CD
3
OD, 100 MHz) δ: 167.3, 148.0, 140.1, 135.7, 132.7, 130.1, 130.1,
1
3
1
26.7, 118.1, 96.1 (C-1), 79.0, 78.1, 74.2, 71.2, 62.5, 21.4. HRMS calc.
(1H, dd, J = 12.4, 4.8 Hz), 3.49–3.37 (4H, overlap), 2.59 (3H, s);
C
+
for C16
H
20
O
7
Na [M+Na] : 347.1107, found: 347.1102.
NMR (CD
3
OD, 100 MHz) δ: 166.7, 159.7, 143.2, 130.7, 129.9, 129.9,
Compound 4e. 50.7 mg, 0.08 mmol of 3e to yield 4e (23.6 mg, 0.07
127.5, 127.5, 117.1, 95.5 (C-1), 78.9, 78.2, 74.1, 71.3, 62.5, 18.5. HRMS
1
+
mmol, 90%) as a white powder. H NMR (CD
d, J = 15.6 Hz), 7.69 (1H, d, J = 8.0 Hz), 7.67 (1H, t, J = 8.0 Hz), 7.17
1H, d, J = 8.0 Hz), 7.15 (1H, t, J = 8.0 Hz), 6.54 (1H, d, J = 16.0 Hz),
.59 (1H, d, J = 7.2 Hz, H-1), 3.85 (1H, dd, J = 12.0, 1.2 Hz), 3.69 (1H,
3
OD, 400 MHz) δ: 7.78 (1H,
calc. for C16
.4. Anti-proliferation assay
Anti-proliferation assay was conducted using the method described
H
20
O
7
Na [M+Na] : 347.1107, found: 347.1113.
(
3
5
1
3
dd, J = 12.0, 4.8 Hz), 3.49–3.38 (4H, overlap); C NMR (CD
3
OD, 100
MHz) δ: 165.6, 144.9, 130.7, 130.7, 130.3, 130.2, 116.8, 115.7, 115.5,
in the literature [20] with slight modifications. B16-F10 murine mela-
noma cell line and normal human dermal fibroblast (HDF) cell line were
obtained from ATCC. B16-F10 cells were cultured in DMEM containing
9
4.6 (C-1), 77.4, 76.6, 72.6, 69.8, 61.0. HRMS calc. for C15
H
17
O
7
FNa
+
[
M+Na] : 351.0856, found: 351.0856.
Compound 4f. 54.7 mg, 0.08 mmol of 3f to yield 4f (27.2 mg, 0.07
1
1
0% FBS, 1 g/L glucose, 1 mmol/L glutamine, 100 U/mL penicillin, and
00 g/mL streptomycin. Murine fibrosarcoma L929 cells were used for
1
mmol, 90%) as a white powder. H NMR (CD
d, J = 16.0 Hz), 7.81 (2H, d, J = 8.0 Hz), 7.71 (2H, d, J = 8.0 Hz), 6.71
1H, d, J = 16.0 Hz), 5.62 (1H, d, J = 7.2 Hz, H-1), 3.86 (1H, dd, J =
2.0, 1.2 Hz), 3.70 (1H, dd, J = 12.0, 4.8 Hz), 3.51–3.37 (4H, overlap);
3
OD, 400 MHz) δ: 7.84 (1H,
μ
cytotoxicity analysis, and were cultured in ATCC medium (McCoy’s 5A
medium with 1.5 mM L-glutamine, 90%; fetal bovine serum, 10%). In-
(
1
1
◦
cubation was carried out at 37 C in a humidified atmosphere of 95% air
3
C NMR (CD
3
OD, 100 MHz) δ: 166.7, 145.7, 139.5, 133.0 (q, J = 32.2
2
and 5% CO .
Hz), 130.0, 130.0, 130.0, 127.1 (d, J = 3.6 Hz), 127.0 (d, J = 3.6 Hz),
Cell viability was assessed by the MTT assay. B16-F10 and HDF cells
1
21.4, 96.2 (C-1), 79.0, 78.1, 74.2, 71.2, 62.5. HRMS calc. for
3
were seeded at a density of 8 × 10 cells/well on 96-well plates. After 24
+
C
16
H
18
O
7
F
3
[M+H] : 379.1005, found: 379.1003.
h incubation, 4a-4l at different concentrations (1.25, 2.5, 5, 10, 20 and
Compound 4g. 52.9 mg, 0.08 mmol of 3g to yield 4g (25.9 mg, 0.07
4
0
μ
M) were dissolved in PBS containing 0.5% DMSO and added to cells
1
mmol, 91%) as a white powder. H NMR (CD
d, J = 8.0 Hz), 7.88 (2H, d, J = 8.0 Hz), 7.86 (1H, d, J = 16.0 Hz), 6.78
1H, d, J = 16.0 Hz), 5.62 (1H, d, J = 8.0 Hz, H-1), 3.86 (1H, dd, J =
3
OD, 400 MHz) δ: 8.28 (2H,
in each well and then incubated for 48 h. Doxorubicin was used as the
positive control. After incubation, 20
μ
L of MTT (5 mg/mL in PBS) was
(
◦
added to each well and incubated for 4 h at 37 C. After the supernatant
1
1
2.0, 1.2 Hz), 3.70 (1H, dd, J = 12.0, 4.0 Hz), 3.49–3.37 (4H, overlap);
was removed, 160 L of DMSO was added to each well to solubilize the
μ
3
C NMR (CD
3
OD, 100 MHz) δ: 166.4, 150.3, 144.8, 141.9, 130.5,
produced fromazan crystals, and measurements were taken at 490 nm
using a Bio-Rad 680 Microplate reader. Results were expressed as a
percentage of the control, and half maximal inhibitory concentration
1
30.5, 125.2, 125.2, 122.8, 96.3 (C-1), 79.1, 78.1, 74.2, 71.3, 62.5.
+
HRMS calc. for C15
H17NO
9
Na [M+Na] : 378.0796, found: 378.0791.
Compound 4h. 52.0 mg, 0.08 mmol of 3h to yield 4h (25.9 mg, 0.08
(
IC50) values were calculated. Cell viability was expressed as a per-
1
mmol, 94%) as a white powder. H NMR (CD
d, J = 16.0 Hz), 7.78 (1H, d, J = 7.6 Hz), 7.46 (1H, d, J = 8.0 Hz), 7.39
1H, t, J = 8.0 Hz), 7.35 (1H, t, J = 8.0 Hz), 6.60 (1H, d, J = 16.0 Hz),
.64 (1H, d, J = 7.6 Hz, H-1), 3.87 (1H, dd, J = 12.0, 1.2 Hz), 3.71 (1H,
3
OD, 400 MHz) δ: 8.15 (1H,
centage of the control culture value, which was considered as 100%
viable.
(
5
Declaration of competing interest
1
3
dd, J = 12.0, 4.4 Hz), 3.51–3.39 (4H, overlap); C NMR (CD
3
OD, 100
MHz) δ: 166.8, 142.9, 136.0, 133.6, 133.0, 131.3, 129.2, 128.7, 121.3,
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
9
6.2 (C-1), 79.0, 78.1, 74.1, 71.2, 62.5. HRMS calc. for C15
H
17
O
7
NaCl
+
[
M+Na] : 367.0561, found: 367.0562.
Compound 4i. 48.5 mg, 0.08 mmol of 3i to yield 4i (22.8 mg, 0.08
1
mmol, 95%) as a white powder. H NMR (CD
m), 7.57 (1H, d, J = 16.0 Hz), 6.81 (1H, m), 6.56 (1H, m), 6.31 (1H, d, J
16.0 Hz), 5.58 (1H, d, J = 8.0 Hz, H-1), 3.85 (1H, dd, J = 12.0, 1.2 Hz),
3
OD, 400 MHz) δ: 7.65 (1H,
Acknowledgements
=
1
3
This work was financially supported by the National Natural Science
Foundation of China (21702178), the Project of Science and Technology
Department of Henan Province (212102311031), National Undergrad-
uate Training Program for Innovation and Entrepreneurship
3
.69 (1H, dd, J = 12.0, 4.8 Hz), 3.48–3.34 (4H, overlap); C NMR
CD
(
3
OD, 100 MHz) δ: 167.3, 152.2, 147.1, 133.9, 117.3, 115.5, 113.8,
9
6.1 (C-1), 79.0, 78.1, 74.1, 71.2, 62.5. HRMS calc. for C13
H
17
O
8
+
[
M+H] : 301.0923, found: 301.0928.
(
202010480003) and Distinguished Young Scholars Fund of Xuchang
Compound 4j. 49.8 mg, 0.08 mmol of 3j to yield 4j (22.8 mg, 0.07
1
University (2019).
mmol, 90%) as a white powder. H NMR (CD
d, J = 16.0 Hz), 7.58 (1H, d, J = 4.8 Hz), 7.41 (1H, d, J = 3.6 Hz), 7.12
1H, dd, J = 4.8, 3.6 Hz), 6.30 (1H, d, J = 16.0 Hz), 5.57 (1H, d, J = 7.6
3
OD, 400 MHz) δ: 7.93 (1H,
Appendix A. Supplementary data
(
Hz, H-1), 3.85 (1H, dd, J = 12.0, 1.2 Hz), 3.69 (1H, dd, J = 12.0, 4.4 Hz),
1
3
NMR spectra of compounds 3a–3l and 4a–4l. This material is
available free of charge via the Internet at XXX.
3
1
6
3
.49–3.37 (4H, overlap); C NMR (CD OD, 100 MHz) δ: 167.1, 140.6,
3
40.3, 133.2, 130.8, 129.6, 116.7, 96.1 (C-1), 79.0, 78.1, 74.1, 71.2,
2.5. HRMS calc. for
39.0508.
C
13
H
16
O
7
NaS [M+Na]+: 339.0514, found:
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6