Semisynthesis of apigenin and acacetin-7-O-β-d-glycosides from naringin and their cytotoxic activities

Article abstract of DOI:10.1016/j.carres.2012.05.013

Apigenin-7-O-β-d-glycosides 1-8 and acacetin-7-O-β-d-glycosides 9-16 were semisynthesized from 4′-O-benzyl apigenin 17 and acacetin 18 by glycosidation and deprotection with the corresponding α-acetylglycosyl bromide, respectively. Compounds 17 and 18 were prepared by iodination followed by base-induced elimination, 4′-O-benzylation, or 4′-O-methylation and acid hydrolysis using naringin as starting material which is readily available and cheap. Their cytotoxic potential against five human cancer cell lines (HL-60, SMMC-7721, A-549, MCF-7, and SW480) was evaluated by standard MTT method. The results show that compounds 2, 9, and 19 exhibit moderate cytotoxicity against HL-60, SMMC-7721, A-549, MCF-7, and SW480, while compound 3 exhibits potent cytotoxicity against MCF-7 selectively. Among the synthesized target compounds, 3, 4, 7, 11, 12, 15, and 16 were new compounds, the natural product 8 was the first synthesized and the synthesis of natural products 5, 6, 13, and 14 was efficiently improved by the new synthetic routes.

Full text of DOI:10.1016/j.carres.2012.05.013

Carbohydrate Research 357 (2012) 41–46
Contents lists available at SciVerse ScienceDirect
Carbohydrate Research
journal homepage: www.elsevier.com/locate/carres
Semisynthesis of apigenin and acacetin-7-O-b-D-glycosides from naringin
and their cytotoxic activities
⇑
Jidan Liu, Ling Chen, Shuanglian Cai, Qiuan Wang
College of Chemistry and Chemical Engineering, Hunan University, ChangSha 410082, PR China
a r t i c l e i n f o
a b s t r a c t
-glycosides 9–16 were semisynthesized from 40-
-acet-
Article history:
Apigenin-7-O-b-
D-glycosides 1–8 and acacetin-7-O-b-D
Received 26 March 2012
Received in revised form 14 May 2012
Accepted 15 May 2012
O-benzyl apigenin 17 and acacetin 18 by glycosidation and deprotection with the corresponding
a
ylglycosyl bromide, respectively. Compounds 17 and 18 were prepared by iodination followed by base-
induced elimination, 40-O-benzylation, or 40-O-methylation and acid hydrolysis using naringin as starting
material which is readily available and cheap. Their cytotoxic potential against five human cancer cell
lines (HL-60, SMMC-7721, A-549, MCF-7, and SW480) was evaluated by standard MTT method. The
results show that compounds 2, 9, and 19 exhibit moderate cytotoxicity against HL-60, SMMC-7721,
A-549, MCF-7, and SW480, while compound 3 exhibits potent cytotoxicity against MCF-7 selectively.
Among the synthesized target compounds, 3, 4, 7, 11, 12, 15, and 16 were new compounds, the natural
product 8 was the first synthesized and the synthesis of natural products 5, 6, 13, and 14 was efficiently
improved by the new synthetic routes.
Available online 23 May 2012
Keywords:
Semisynthesis
Apigenin
Acacetin
Glycosidation
Cytotoxic activity
Ó 2012 Elsevier Ltd. All rights reserved.
1. Introduction
available at low cost. Furthermore, all synthesized flavoniod glyco-
sides were evaluated for their cytotoxic potential against myeloid
Flavonoid glycosides constitute an important class of biomole-
cules and are widely distributed in fruits, vegetables, and tradi-
leukemia (HL-60), liver carcinoma (SMMC-7721), lung carcinoma
(A-549), breast carcinoma (MCF-7), and intestinal carcinoma
(SW480) cell lines by standard MTT method.
tional medicinal plants. They exhibit
a
broad spectrum of
-glucoside
biological activities.1,2 For example, apigenin-7-O-b-
D
(cosmosiin) which was isolated from Lonicera gracilipes var. glan-
2. Results and discussion
dulosa can inhibit the growth of various human cancer cells,3
acacetin-7-O-b-D-galactopyranoside, a natural flavonoid isolated
Our present method provides an efficient and easy approach to
from flower heads of Chrysanthemum morifolium, was efficient to
inhibit the replication of HIV in H9 cells.4 In the past decade, the
biological activities of flavonoid glycosides against cancer, anti-vir-
al, and cardiovascular diseases have attracted increasing interest.
To generate useful materials, many attempts of glycosylation of
phenolic hydroxyl groups of flavonoids have been performed since
1938.5
During our continuous medicinal research on flavonoids to sup-
port the investigation of many important physiology processes and
drug discoveries, some attempts have been made in the total syn-
thesis of flavonoids and flavonoid glycosides starting from simple
materials as well as employing many protection strategies.6–8
However, they were achieved with very time-consuming and com-
plicated syntheses approaches. Herein, we reported a short and
access of apigenin-7-O-b-D-glycosides 1–8 and acacetin-7-O-b-D-
glycosides 9–16 in just six steps from naringin, respectively as de-
picted in Scheme 1. This straightforward synthesis is based on the
key intermediates 40-O-benzyl apigenin 17 and acacetin 18 which
are prepared in gram quantities in only three steps, firstly iodin-
ation followed by base-induced elimination to rhoifolin, then 40-
O-benzylation or 40-O-methylation and acid hydrolysis of the neo-
hesperidose residue with naringin as starting material which is
readily available and cheap.
It is well known that sugar moiety could enhance water solubil-
ity and improve the targeting activity of bioactive molecules. For
example, lactose can be recognized by the hepatic asialoglycopro-
tein receptor (ASGP-R) which localizes to liver cells providing an
efficient entry point for lactose-modified molecules.9 The modifica-
tion of flavonoids with lactose may be possible to specifically tar-
get molecules to liver cells, facilitating the application of
bioactive flavonoids to the treatment of hepatitis B, hepatitis C
and liver cancer. On the other hand, the largely hydrophobic char-
acter of flavonoids 17, 18 makes them poorly soluble in aqueous
effective synthesis of apigenin-7-O-b-
D
-glycosides 1–8 and acace-
tin-7-O-b-
D-glycosides 9–16 from naringin which is commercially
⇑
Corresponding author.
E-mail address: WangQA@hnu.edu.cn (Q. Wang).
0008-6215/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.carres.2012.05.013

42
J. Liu et al. / Carbohydrate Research 357 (2012) 41–46
OH
OBn
OH
I2
O
Ο
HO
Ο
O
Ο
neohesperidosyl
(1) BnBr/K2CO3
(2) H+/EtOH
neohesperidosyl
pyridine
OH
O
OH
O
OH
O
17
naringin
rhoifolin
OBn
OH
OH
R1O
Ο
R2O
R1O
Ο
Ο
5% Pd/C, H2
O-acetylglycosyl bromide
aq.NH3.H2O
MeOH rt
0.25 mol/L aq.K2CO3, CHCl3
MeOH/EtOAc
(n-C4H9)4N+Br-)
OH
O
OH
O
OH
O
19-22
1-4
5-8
OCH3
OCH3
OCH3
R2O
aq.NH3.H2O
Ο
HO
Ο
(1) (CH3O)2SO2
R1O
Ο
3% aq.NaOH 10-25 oC
O-acetylglycosyl bromide
rhoifolin
0.25 mol/L aq.K2CO3, CHCl3
( -C H ) N+Br-)
(2) H+/H2O
MeOH rt
OH
O
OH
O
n
4
9 4
OH
O
18
9-12
13-16
OH
OH
HO
OH
O
OH
OAc
OAc
OAc
O
OAc
O
5
R2
=
=
R2
=
=
1
9
7
3
HO
HO
O
O
R1
R1
=
=
13
O
R1
R1
=
=
O
15
AcO
AcO
11
21
AcO
O
OH
19
OH HO
D-lactosyl
OAc
OH
OAc
AcO
OAc
D-glucosyl
OH
OAc
OAc
OAc
OH
OAc
2
OH
OH
O
4
O
10
20
O
R2
R2
AcO
AcO
O
6
8
12
22
O
HO
HO
O
O
AcO
14
16
O
OAc
AcO
OAc
HO
OH
HO
OAc
OH
OH
D-galactosyl
D-maltosyl
Scheme 1. Synthetic route of apigenin and acacetin-7-O-b-D-glycosides from naringin.
media which in some cases limits their therapeutic efficacy and
this has a strong influence on their pharmacokinetic properties.
Then, we turned our attention to the introduction of glucosyl,
galactosyl, lactosyl and maltosyl moieties into bioactive flavonoids,
so 40-O-benzyl apigenin 17 and acacetin 18 were condensed with
All the synthesized apigenin and acacetin-7-O-b-
and their derivatives 1–16 as well as intermediates 19–22 were
tested for cytotoxic activity against five human cancer cell lines
(HL-60, SMMC-7721, A-549, MCF-7, and SW480). Their IC50 (lg/
mL) values were plotted in Table 1. The results showed that com-
pound 2 exhibits potent cytotoxicity against the HL-60, SMMC-
7721, A-549, MCF-7, and SW480 cancer cell lines with IC50 values
of 11.77, 17.60, 16.77, 10.31, and 20.71, respectively. Compound 19
exhibits potent cytotoxicity against the SMMC-7721, A-549, and
D
-glycosides
the corresponding
droxyl groups on C-7 position of flavones, respectively.
a
-acetylglycosyl bromide at more reactive hy-
-acetyl-
a
glycosyl bromide had to be used immediately after preparation
owing to its instability. The glycosidation procedure was modified
by using potassium carbonate in a solvent mixture of CHCl3/H2O
and tetrabutylammonium bromide (TBAB) as a phase transfer cat-
alyst leading to the desired 40-O-benzyl apigenin-7-O-b-
glycosides 1–4 and acacetin-7-O-b-
65–85% yields. Although the yield was moderate, the stereo- and
regioselectivities were perfect with no -glycoside, 5-O-glycoside,
D
-acetyl-
Table 1
D
-acetylglycosides 9–12 in
IC50 values (lg/mL) of apigenin and acacetin-7-O-b-D-glycosides on the human
cancer cell lines
a
Compound
HL-60
SMMC-7721
A-549
MCF-7
SW480
or 5,7-O-biglycoside being detected. Glycosylation was found to be
highly regioselective with glycosylation at the hydroxyl group on
C-7 predomominating over glycosylation at the hydroxyl group
on C-5. This regioselectivity for hydroxyl group on C-7 can be ex-
plained by both steric and electronic effects. The hydroxyl group
on C-7 is more electronically favored toward glycosylation owing
to the fact that the neighboring carbonyl group at C-4 renders
the hydroxyl group on C-5 less reactive toward glycosyl donors.
Moreover the neighboring group participation of 2-acetyl group
in proton coupling (J = 6.4–9.2 Hz) observed for the anomeric pro-
ton of glycose residue (d 5.02–5.78) demonstrated the expected b-
linkage to the apigenin or acacetin moiety.10
1
2
3
4
5
6
7
8
>40
11.77
>40
>40
>40
>40
>40
>40
>40
>40
>40
>40
>40
>40
>40
>40
>40
2.51
>40
17.60
>40
>40
>40
>40
>40
>40
>40
>40
>40
>40
>40
>40
>40
>40
14.99
14.99
>40
16.77
>40
>40
>40
>40
>40
>40
28.07
>40
>40
>40
>40
>40
>40
>40
29.94
13.61
>40
>40
20.71
>40
>40
>40
>40
>40
>40
16.38
>40
>40
>40
>40
>40
>40
>40
>40
18.85
10.31
4.58
>40
>40
>40
4.58
>40
16.63
>40
>40
>40
>40
>40
>40
>40
27.08
18.65
9
10
11
12
13
14
15
16
19
(DPP)a
Removal of the benzyl group, which protects the apigenin 40-
hydroxyl group, by H2 with Pd/C at room temperature furnished
the compounds 19–22. The sugar moiety was deprotected using
aq NH3ꢀH2O in methanol to yield apigenin-7-O-b-
D-glycosides 5–
a
Cisplatin (DDP) was employed as positive control.
8 and acacetin-7-O-b- -glycosides 13–16.
D

J. Liu et al. / Carbohydrate Research 357 (2012) 41–46
43
MCF-7 cancer cell lines with IC50 values of 14.99, 29.94, and 27.08,
respectively. Compound 9 shows cytotoxicity against cancer cell
lines A-549, MCF-7, and SW480 with IC50 values of 28.07, 16.63,
and 16.38, respectively, while compound 3 exhibits significant
selective inhibitory activity against MCF-7 cancer cell line with
IC50 value of 4.58. Biological activities of all the compounds men-
tioned above are similar or even improved compared with the po-
sitive control, cisplatin (DDP).
DMSO-d6) d 12.92 (s, 1H, OH-5), 10.86 (s, 1H, OH-7), 8.03 (d,
J = 8.8 Hz, 2H, H-20, H-60), 7.11 (d, J = 9.2 Hz, 2H, H-30, H-50), 6.87
(s, 1H, H-3), 6.50 (d, J = 2.0 Hz, 1H, H-8), 6.20 (d, J = 2.0 Hz, 1H,
H-6), 3.85 (s, 3H, OCH3); EIMS (m/z): 285 (M+1)+.
3.5. Synthesis of 40-O-benzyl apigenin-7-O-b-
D-acetylglucoside
(19)
A mixture of 17 (0.43 g, 1.2 mmol), TBAB (0.06 g, 0.2 mmol), and
a-acetylglucose bromide (0.98 g, 2.4 mmol) in CHCl3 (25 mL) was
3. Experimental
added to 0.25 mol/L K2CO3 (10 mL, 2.5 mmol), the mixture was
stirred under nitrogen for 12 h at 45 °C, the organic phase was sep-
arated, and the aqueous phase was extracted with CH2Cl2. The or-
ganic extracts were combined, washed with brine, and dried over
anhydrous MgSO4. The solvent was removed under reduced pres-
sure, and the residue was chromatographed on silica gel using
petroleum ether/ethyl acetate (2:1) as eluent to afford the light
yellow solid 0.7 g, yield: 85%. Mp 163–165 °C. 1H NMR (400 MHz,
CDCl3) d 12.83 (s, 1H, OH-5), 7.83 (d, J = 9.2 Hz, 2H, H-20, H-60),
7.45–7.35 (m, 5H, OCH2C6H5), 7.09 (d, J = 9.2 Hz, 2H, H-30, H-50),
6.60 (s, 1H, H-3), 6.57 (d, J = 1.2 Hz, 1H, H-8), 6.44 (d, J = 1.2 Hz,
1H, H-6), 5.32 (d, J = 6.4 Hz, 1H, H-100), 5.30 (s, 2H, OCH2C6H5),
5.19–5.13 (m, 3H, H-200,300,400), 4.31–4.19 (m, 2H, H-600), 3.96–3.92
(m, 1H, H-500), 2.11–2.05 (m, 12H, 4 ꢁ CH3CO); HRMS [M+Na]+
calcd for C36H34O14Na 713.6352, found 713.6363.
3.1. General methods
Melting points were measured on a XRC-I apparatus and were
uncorrected. IR spectra were recorded on a Bruker Tensor-27 spec-
trometer, 1H NMR and 13C NMR spectra were recorded on a Bruker
AM-400 instrument, using tetramethylsilane as an internal stan-
dard, chemical shifts (d) in ppm, and coupling constants (J) in Hz.
Mass spectra were determined with VG Autospec-3000 or ZAB-
HS spectrometer by the FAB or EI method.
3.2. Synthesis of rhoifolin11
A solution of naringin (2.0 g, 3.45 mmol) and iodine (0.89 g,
3.5 mmol) in pyridine (20 mL) was heated to 95 °C for 2 h. The
mixture was cooled to room temperature and poured into cold
water. The resulting precipitate was filtered and washed with sat-
urated sodium thiosulfate and water successively, and then was
dried in vacuum to afford the resulting yellow solid rhoifolin
1.6 g, yield: 80%. Mp 249–252 °C. (lit. 11, 251–253 °C).
3.6. Synthesis of acacetin-7-O-b-D-acetylglucoside (9)
Compound 9 was prepared from 18 and
mide as described for the preparation of 19 from 17 and
a
-acetylglucose bro-
-acetyl-
a
glucose bromide, yellow solid, yield: 83%. Mp 150–152 °C. 1H NMR
(400 MHz, CDCl3) d 12.85 (s, 1H, OH-5), 7.83 (d, J = 8.8 Hz, 2H, H-20,
H-60), 7.03 (d, J = 8.8 Hz, 2H, H-30, H-50), 6.60 (s, 1H, H-3), 6.58 (d,
J = 2.0 Hz, 1H, H-8), 6.44 (d, J = 2.0 Hz, 1H, H-6), 5.33 (d,
J = 7.2 Hz, 1H, H-100), 5.31–5.28 (m, 1H, H-200), 5.20–5.14 (m, 2H,
H-300,400), 4.32–4.19 (m, 2H, H-600), 3.98–3.95 (m, 1H, H-500), 3.90
(s, 3H, OCH3), 2.11–2.05 (m, 12H, 4 ꢁ CH3CO); 13C NMR
(100 MHz, CDCl3) d 20.3, 20.5, 20.6, 56.0, 61.3, 67.6, 71.0, 71.3,
72.1, 94.7, 96.4, 99.6, 102.6, 103.1, 105.2, 116.8, 120.2, 128.4,
156.8, 161.2, 161.3, 161.5, 164.1, 169.2, 169.5, 169.7, 169.8,
181.9; HRMS [M+Na]+ calcd for C30H30O14Na 637.5394, found
637.5381.
3.3. Synthesis of 40-O-benzyl apigenin (17)
To a mixture of rhoifolin (2.31 g, 4.0 mmol) and K2CO3 (0.69 g,
5.0 mmol) in DMF (20 mL) was added benzyl bromide (0.6 mL,
5.0 mmol) and the mixture was stirred under nitrogen for 8 h at
90 °C. The mixture was cooled to room temperature and poured
into cold water (200 mL). The resulting precipitate was filtered
and dissolved in ethanol (50 mL), and then concentrated H2SO4
(3 mL) was added to the mixture slowly. The mixture was heated
under reflux for 2 h, and then cooled to room temperature and fil-
tered, the resulting precipitate was washed with saturated KHCO3
and water and evaporated to dryness successively, and then the
crude solid was chromatographed on silica gel using petroleum
ether/ethyl acetate (4:1) as eluent to afford the yellow solid
0.9 g, yield: 63%. Mp 304–306 °C. 1H NMR (400 MHz, DMSO-d6) d
12.92 (s, 1H, OH-5), 10.93 (s, 1H, OH-7), 8.04 (d, J = 8.8 Hz, 2H, H-
20, H-60), 7.33–7.49 (m, 5H, OCH2C6H5), 7.19 (d, J = 8.8 Hz, 2H, H-
30, H-50), 6.88 (s, 1H, H-3), 6.52 (d, J = 1.6 Hz, 1H, H-8), 6.21 (d,
J = 1.6 Hz, 1H, H-6), 5.23 (s, 2H, OCH2C6H5); EIMS (m/z): 361
(M+1)+.
3.7. Synthesis of 40-O-benzyl apigenin-7-O-b-
D-acetylgalactoside
(20)
Compound 20 was prepared from 17 and
mide as described for the preparation of 19 from 17 and
a
-acetylgalacotse bro-
-acetyl-
a
glucose bromide, yellow solid, yield: 76%. Mp 256–258 °C. 1H NMR
(400 MHz, CDCl3) d 12.76 (s, 1H, OH-5), 7.84 (d, J = 9.2 Hz, 2H, H-20,
H-60), 7.51–7.36 (m, 5H, OCH2C6H5), 7.09 (d, J = 9.2 Hz, 2H, H-30, H-
50), 6.65 (s, 1H, H-3), 6.59 (d, J = 1.2 Hz, 1H, H-8), 6.45 (d, J = 1.2 Hz,
1H, H-6), 5.26 (d, J = 8.0 Hz, 1H, H-100), 5.16 (s, 2H, OCH2C6H5),
4.24–5.14 (m, 6H, H-200,300,400,500,600), 2.20–2.03 (m, 12H,
4 ꢁ CH3CO); HRMS [M+Na]+ calcd for C36H34O14Na 713.6352,
found 713.6338.
3.4. Synthesis of acacetin (18)12
(CH3O)2SO2 (2.3 mL, 24 mmol) was added dropwise to a stirred
solution of rhoifolin (1.38 g, 2.4 mmol) in 3% aq NaOH (50 mL) at
10–25 °C for 30 min, then the reaction mixture was stirred for
6 h at the same temperature, and then concd H2SO4 (5 mL) was
added to the mixture slowly. The mixture was heated under reflux
for 8 h and then cooled to room temperature and filtered, the
resulting precipitate was washed with saturated NaHCO3 and
water and evaporated to dryness successively, and then the crude
solid was chromatographed on silica gel using petroleum ether/
ethyl acetate (3:1) as eluent to afford the yellow solid 0.22 g, yield:
32%. Mp 262–264 °C. (lit. 12, 260–262 °C). 1H NMR (400 MHz,
3.8. Synthesis of acacetin-7-O-b-D-acetylgalactoside (10)
Compound 10 was prepared from 18 and
mide as described for the preparation of 19 from 17 and
a
-acetylgalactose bro-
-acetyl-
a
glucose bromide, yellow solid, yield: 75%. Mp 152–154 °C. 1H NMR
(400 MHz, CDCl3) d 12.82 (s, 1H, OH-5), 7.84 (d, J = 8.8 Hz, 2H, H-20,
H-60), 7.02 (d, J = 8.8 Hz, 2H, H-30, H-50), 6.60 (s, 1H, H-3), 6.59 (d,
J = 2.0 Hz, 1H, H-8), 6.45 (d, J = 2.0 Hz, 1H, H-6), 5.50 (d,
J = 8.0 Hz, 1H, H-100), 5.42–5.49 (m, 1H, H-200), 5.19–5.13 (m, 2H,

44
J. Liu et al. / Carbohydrate Research 357 (2012) 41–46
H-300,400), 4.26–4.20 (m, 1H, H-500), 4.17–4.08 (m, 2H, H-600), 3.90 (s,
3H, OCH3), 2.20–2.03 (m, 12H, 4 ꢁ CH3CO); 13C NMR (100 MHz,
CDCl3) d 20.0, 20.1, 20.3, 56.1, 61.4, 67.5, 71.1, 71.2, 72.5, 94.3,
96.4, 99.7, 102.5, 103.3, 105.4, 116.9, 120.1, 128.5, 156.9, 161.1,
161.2, 161.5, 164.2, 169.5, 169.6, 169.9, 169.4, 182.5; HRMS
[M+Na]+ calcd for C30H30O14Na 637.5394, found 637.5377.
J = 2.0 Hz, 1H, H-6), 5.44 (d, J = 7.6 Hz, 1H, H-100), 5.40–5.33 (m,
2H, H-200,300), 5.16–5.03 (m, 4H, H-400,500,600), 4.52 (d, J = 8.4 Hz,
1H, H-1000), 4.31–4.24 (m, 2H, H-2000, 3000), 4.09–4.06 (m, 2H, H-
4000,5000), 4.00–3.92 (m, 2H, H-6000), 3.90 (s, 3H, OCH3), 2.12–2.02
(m, 21H, 7 ꢁ CH3CO) 2.17–1.98 (m, 21H, 7 ꢁ CH3CO); 13C NMR
(100 MHz, CDCl3) d 20.1, 20.2, 20.5, 21.2, 21.5, 21.7, 22.0, 59.7,
61.5, 64.7, 67.8, 69.6, 72.3, 72.4, 72.7, 73.1, 75.2, 76.5, 96.1,
96.2, 99.4, 102.5, 104.2, 104.4, 115.5, 123.1, 127.2, 157.4,
157.3, 163.1, 164.2, 166.6, 169.1, 169.3, 169.6, 169.8, 169.7,
170.1, 170.4, 182.1; HRMS [M+Na]+ calcd for C42H46O22Na
925.7894, found 925.7883.
3.9. Synthesis of 40-O-benzyl apigenin-7-O-b-
D-acetyllactoside
(21)
Compound 21 was prepared from 17 and
mide as described for the preparation of 19 from 17 and
a
-acetyllactose bro-
-acetyl-
a
glucose bromide, yellow solid, yield: 70%. Mp 158–160 °C. 1H NMR
(400 MHz, CDCl3) d 12.82 (s, 1H, OH-5), 7.83 (d, J = 8.8 Hz, 2H, H-20,
H-60), 7.50–7.38 (m, 5H, OCH2C6H5), 7.09 (d, J = 8.8 Hz, 2H, H-30, H-
50), 6.60 (s, 1H, H-3), 6.55 (d, J = 2.0 Hz, 1H, H-8), 6.42 (d, J = 2.0 Hz,
1H, H-6), 5.38 (d, J = 7.2 Hz, 1H, H-100), 5.31 (t, J = 8.8 Hz, 1H, H-200),
5.20 (t, J = 9.2 Hz, 1H, H-300), 5.18 (s, 2H, OCH2C6H5), 5.17–5.13 (m,
4H, H-400,500,600), 4.50 (d, J = 8.0 Hz, 1H, H-1000), 4.15–4.10 (m, 3H, H-
2000, 3000, 4000), 3.91–3.87 (m, 3H, H-5000, 6000), 2.17–1.98 (m, 21H,
7 ꢁ CH3CO); HRMS [M+Na]+ calcd for C48H50O22Na 1001.8852,
found 1001.8865.
3.13. Synthesis of apigenin-7-O-b-D
-acetylglucoside (1)13
A solution of 19 (0.345 mg, 0.5 mmol) and 5% Pd/C (20 mg) in
dry methanol/ethyl acetate (1:1, V/V, 20 mL) was stirred under
H2 at room temperature overnight. TLC indicated that the starting
material disappeared. After being filtered, the mixture was evapo-
rated under reduced pressure to afford a residual yellow solid
which was chromatographed on silica gel using petroleum ether/
ethyl acetate (1:1) as eluent to afford the yellow solid 256 mg,
yield: 85%. Mp 207–209 °C. (lit. 13, 207–208 °C). 1H NMR
(400 MHz, DMSO-d6) d 13.04 (s, 1H, OH-5), 10.46 (s, 1H, OH-40),
7.97 (d, J = 8.4 Hz, 2H, H-20, H-60), 6.94 (d, J = 8.4 Hz, 2H, H-30, H-
50), 6.90 (s, 1H, H-3), 6.80 (d, J = 2.0 Hz, 1H, H-8), 6.45 (d,
J = 2.0 Hz, 1H, H-6), 5.76 (d, J = 8.0 Hz, 1H, H-100), 5.40 (t,
J = 9.6 Hz, 1H, H-200) 5.10 (t, J = 13.2 Hz, 1H, H-300) 5.02 (t,
J = 10.0 Hz, 1H, H-400), 4.37–4.10 (m, 3H, H-500, 600), 2.03–1.98 (m,
12H, 4 ꢁ CH3CO); 13C NMR (100 MHz, DMSO-d6) d 20.1, 20.4,
20.5, 61.4, 67.7, 70.3, 71.2, 71.8, 95.1, 96.4, 99.2, 103.2, 105.3,
116.2, 120.1, 128.2, 156.5, 161.3, 161.5, 161.7, 164.0, 169.1,
169.4, 169.6, 169.8, 182.1; HRMS [M+Na]+ calcd for C29H28O14Na
624.2996, found 624.2982.
3.10. Synthesis of acacetin-7-O-b-D-acetyllactoside (11)
Compound 11 was prepared from 18 and
mide as described for the preparation of 19 from 17 and
a
-acetyllactose bro-
-acetyl-
a
glucose bromide, yellow solid, yield: 65%. Mp 98–100 °C. 1H NMR
(400 MHz, CDCl3) d 12.85 (s, 1H, OH-5), 7.81 (d, J = 8.8 Hz, 2H, H-
20, H-60), 7.00 (d, J = 8.8 Hz, 2H, H-30, H-50), 6.58 (s, 1H, H-3), 6.54
(d, J = 2.4 Hz, 1H, H-8), 6.40 (d, J = 2.4 Hz, 1H, H-6), 5.38 (d,
J = 7.2 Hz, 1H, H-100), 5.32 (t, J = 8.8 Hz, 1H, H-200), 5.22 (t,
J = 9.2 Hz, 1H, H-300), 5.17–5.11 (m, 4H, H-400,500,600), 4.56 (d,
J = 8.0 Hz, 1H, H-1000), 4.20–4.10 (m, 3H, H-2000, 3000, 4000), 3.96–3.90
(m, 3H, H-5000, 6000), 3.89 (s, 3H, OCH3), 2.18–1.99 (m, 21H,
7 ꢁ CH3CO) 2.17–1.98 (m, 21H, 7 ꢁ CH3CO); 13C NMR (100 MHz,
CDCl3) d 20.1, 20.3, 20.4, 21.3, 21.5, 21.8, 22.0, 58.9, 61.2, 64.6,
67.3, 68.7, 71.1, 72.2, 72.3, 73.2, 75.2, 76.4, 96.2, 96.3, 99.5,
102.8, 104.6, 104.7, 115.9, 123.1, 127.3, 157.8, 159.4, 163.5,
163.8, 165.7, 169.1, 169.2, 169.3, 169.5, 169.7, 170.5, 170.7,
181.3; HRMS [M+Na]+ calcd for C42H46O22Na 925.7894, found
925.7880.
3.14. Synthesis of apigenin-7-O-b-D-acetylgalactoside (2)
Compound 2 was prepared from 20 as described for the prepa-
ration of 1 from 19, yellow solid, yield: 83%. Mp 207–209 °C. 1H
NMR (DMSO-d6, 400 MHz) d 13.05 (s, 1H, OH-5), 10.48 (s, 1H,
OH-40), 7.77 (d, J = 8.0 Hz, 2H, H-20, H-60), 6.98 (d, J = 8.0 Hz, 2H,
H-30, H-50), 6.58 (s, 1H, H-3), 6.56 (d, J = 2.4 Hz, 1H, H-8), 6.45 (d,
J = 2.4 Hz, 1H, H-6), 5.49 (d, J = 7.6 Hz, 1H, H-100), 5.17–5.15 (m,
2H, H-200, 300), 4.31–4.15 (m, 4H, H-400,500, 600), 2.20–2.04 (m, 12H,
4 ꢁ CH3CO); 13C NMR (100 MHz, DMSO-d6) d 20.2, 20.4, 20.6,
61.6, 68.0, 70.4, 71.6, 71.8, 95.3, 96.4, 99.4, 103.2, 105.1, 116.0,
120.9, 128.4, 156.8, 161.3, 161.4, 161.5, 164.2, 169.1, 169.3,
169.5, 169.7, 182.0; HRMS [M+Na]+ calcd for C29H28O14Na
624.2996, found 624.2985.
3.11. Synthesis of 40-O-benzyl apigenin-7-O-b-
D-acetylmaltoside
(22)
Compound 22 was prepared from 17 and
a-acetylmaltose bro-
mide as described for the preparation of 19 from 17 and
a-acetyl-
glucose bromide, yellow solid, yield: 68%. Mp 176–178 °C. 1H NMR
(400 MHz, CDCl3) d 12.84 (s, 1H, OH-5), 7.84 (d, J = 8.8 Hz, 2H, H-20,
H-60), 7.50–7.42 (m, 5H, OCH2C6H5), 7.09 (d, J = 8.8 Hz, 2H, H-30, H-
50), 6.60 (s, 1H, H-3), 6.57 (d, J = 2.4 Hz, 1H, H-8), 6.44 (d, J = 2.4 Hz,
1H, H-6), 5.44 (d, J = 8.4 Hz, 1H, H-100), 5.42 (s, 2H, OCH2C6H5),
5.41–5.33 (m, 2H, H-200,300), 5.26–5.03 (m, 4H, H-400,500,600), 4.52 (d,
J = 9.6 Hz, 1H, H-1000), 4.30–4.24 (m, 2H, H-2000, 3000), 4.11–3.97 (m,
4H, H-4000, 5000 6000), 2.12–2.02 (m, 21H, 7ꢁCH3CO); HRMS [M+Na]+
calcd for C48H50O22Na, 1001.8852, found 1001.8868.
3.15. Synthesis of apigenin-7-O-b-D-acetyllactoside (3)
Compound 3 was prepared from 21 as described for the prepa-
ration of 1 from 19, yellow solid, yield: 78%. Mp 116–118 °C. 1H
NMR (400 MHz, DMSO-d6) d 12.96 (s, 1H, OH-5), 10.26 (s, 1H,
OH-40) 7.87 (d, J = 8.8 Hz, 2H, H-20, H-60), 6.93 (d, J = 8.8 Hz, 2H,
H-30, H-50), 6.76 (s, 1H, H-3), 6.69 (d, J = 1.2 Hz, 1H, H-8), 6.41 (d,
J = 1.2 Hz, 1H, H-6), 5.60 (d, J = 8.0 Hz, 1H, H-100), 5.32 (t,
J = 9.2 Hz, 1H, H-200), 5.27–5.16 (m, 2H, H-300,400), 5.08–4.77 (m,
3H, H-500,600), 4.43 (d, J = 11.2 Hz, 1H, H-1000), 4.23–4.05 (m, 5H, H-
2000, 3000, 4000, 6000), 3.92 (t, J = 9.2 Hz, 1H, H-5000), 2.54–1.92 (m, 21H,
7 ꢁ CH3CO); 13C NMR (100 MHz, DMSO-d6) d 20.0, 20.1, 20.4,
21.2, 21.3, 21.7, 21.8, 59.9, 65.8, 68.3, 69.6, 71.2, 72.3, 72.5, 73.1,
75.0, 76.4, 96.5, 96.8, 99.4, 102.9, 104.5, 104.6, 115.8, 123.0,
127.8, 157.7, 159.3, 163.4, 163.7, 165.4, 169.2, 169.3, 169.6,
169.7, 169.8, 170.1, 170.2, 182.3; HRMS [M+Na]+ calcd for
3.12. Synthesis of acacetin-7-O-b-D-acetylmaltoside (12)
Compound 12 was prepared from 18 and
bromide as described for the preparation of 19 from 17 and
acetylglucose bromide, yellow solid, yield: 71%. Mp 118–120 °C.
a-acetylmaltose
a-
1H NMR (400 MHz, CDCl3)
d 12.82 (s, 1H, OH-5), 7.84 (d,
J = 8.8 Hz, 2H, H-20, H-60), 7.02 (d, J = 8.8 Hz, 2H, H-30, H-50),
6.61 (s, 1H, H-3), 6.57 (d, J = 2.0 Hz, 1H, H-8), 6.44 (d,
C41H44O22Na 911.7629, found 911.7645.

J. Liu et al. / Carbohydrate Research 357 (2012) 41–46
45
3.16. Synthesis of apigenin-7-O-b-
D
-acetylmaltoside (4)
(100 MHz, DMSO-d6) d 60.5, 69.3, 73.2, 76.6, 77.0, 94.3, 99.3,
100.1, 103.2, 105.2, 116.8, 121.4, 128.1, 156.3, 161.4, 162.3,
163.2, 164.5, 180.3; HRMS [M+Na]+ calcd for C21H20O10Na
455.3665, found 455.3677.
Compound 4 was prepared from 22 as described for the prepa-
ration of 1 from 19, yellow solid, yield: 74%. Mp 138–140 °C. 1H
NMR (400 MHz, DMSO-d6) d 12.82 (s, 1H, OH-5), 10.30 (s, 1H,
OH-40) 7.80 (d, J = 7.2 Hz, 2H, H-20, H-60), 6.97 (d, J = 7.2 Hz, 2H,
H-30, H-50), 6.62 (s, 1H, H-3), 6.54 (d, J = 2.0 Hz, 1H, H-8), 6.42 (d,
J = 2.0 Hz, 1H, H-6), 5.45 (d, J = 8.4 Hz, 1H, H-100), 5.32 (t,
J = 8.8 Hz, 1H, H-200), 5.20–5.14 (m, 2H, H-300,400), 5.04–4.75 (m,
3H, H-500,600), 4.50 (d, J = 13.2 Hz, 1H, H-1000), 4.27–4.06 (m, 5H, H-
2000, 3000, 4000, 6000), 3.96 (t, J = 8.8 Hz, 1H, H-5000), 2.12–2.02 (m, 21H,
7ꢁCH3CO); 13C NMR (100 MHz, DMSO-d6) d 20.2, 20.3, 20.5, 21.3,
21.4, 21.6, 21.8, 59.7, 65.4, 68.3, 69.6, 72.2, 72.3, 72.5, 73.3, 75.1,
76.4, 96.0, 96.2, 99.1, 102.3, 104.1, 104.2, 115.8, 123.2, 127.5,
157.7, 158.3, 164.1, 164.2, 165.5, 169.0, 169.4, 169.5, 169.7,
169.7, 170.3, 170.6, 182.0; HRMS [M+Na]+ calcd for C41H44O22Na,
911.7629, found 911.7641.
3.20. Synthesis of acacetin-7-O-b-D
-galactoside (14)16
Compound 14 was prepared from 10 as described for the prep-
aration of 5 from 1, yellow solid, yield: 67%. Mp 252–254 °C. (lit.
16, 251–254 °C); IR (KBr, cmꢂ1) 3437, 2951, 1557, 1524, 1466,
1134, 1041, 896; 1H NMR (400 MHz, DMSO-d6) d 12.93 (s, 1H,
OH-5), 8.07 (d, J = 9.2 Hz, 2H, H-20, H-60), 7.13 (d, J = 9.2 Hz, 2H,
H-30, H-50), 6.98 (s, 1H, H-3), 6.85 (d, J = 2.0 Hz, 1H, H-8), 6.46 (d,
J = 2.0 Hz, 1H, H-6), 5.28 (d, J = 6.4 Hz, 1H, H-100), 5.03 (d,
J = 7.6 Hz, 1H, OH-200), 4.95 (d, J = 5.6 Hz, 1H, OH-300), 4.72 (t,
J = 5.2 Hz, 1H, OH-600), 4.58 (d, J = 4.8 Hz, 1H, OH-400), 3.87 (s, 3H,
OCH3), 3.72–3.68 (m, 2H, H-200, 300), 3.57–3.43 (m, 4H, H-400,500,
600); 13C NMR (100 MHz, DMSO-d6) d 56.2, 60.3, 69.3, 73.1, 76.5,
77.6, 95.2, 98.4, 101.2, 104.3, 105.2, 117.8, 121.2, 128.5, 156.7,
161.8, 162.3, 164.2, 165.6, 181.5; HRMS [M+Na]+ calcd for
3.17. Synthesis of apigenin-7-O-b-D-glucoside (5)
Compound 1 (150 mg, 0.25 mmol) was added to a solution of
30% aq NH3ꢀH2O (5 mL) in CH3OH (20 mL) with stirring. After stir-
ring for 12 h at room temperature, the solvent was removed under
reduced pressure. The residual was chromatographed on silica gel
using ethyl acetate/MeOH (3:1) as eluent to afford the yellow solid
78 mg, yield: 72%. Mp 234–236 °C. (lit. 13, 238–239 °C); IR (KBr,
cmꢂ1) 3440, 2918, 1615, 1563, 1452, 1346, 1273, 1245, 1146,
1103, 1072, 1024, 895; 1H NMR (400 MHz, DMSO-d6) d 12.97 (s,
1H, OH-5), 10.48 (s, 1H, OH-40), 7.96 (d, J = 8.8 Hz, 2H, H-20, H-60),
6.94 (d, J = 8.8 Hz, 2H, H-30, H-50), 6.87 (s, 1H, H-3), 6.83 (d,
J = 1.6 Hz, 1H, H-8), 6.45 (d, J = 1.6 Hz, 1H, H-6), 5.42 (d,
J = 8.0 Hz, 1H, H-100), 5.15 (s, 1H, OH-200), 5.08 (s, 1H, OH-300), 5.06
(s, 1H, OH-400), 4.63 (s, 1H, OH-600), 3.72–3.70 (m, 1H, H-200), 3.49–
3.18 (m, 5H, H-300,400,500, 600); 13C NMR (100 MHz, DMSO-d6) d
60.6, 69.6, 73.1, 76.5, 77.2, 94.8, 99.6, 99.9, 103.1, 105.4, 116.0,
121.1, 128.6, 156.9, 161.1, 161.4, 163.0, 164.3, 180.1; HRMS
[M+Na]+ calcd for C21H20O10Na 455.3665, found 455.3649.
C
22H22O10Na 469.3930, found 469.3947.
3.21. Synthesis of apigenin-7-O-b- -lactoside (7)
D
Compound 7 was prepared from 3 as described for the prepara-
tion of 5 from 1, yellow solid, yield: 62%. Mp 172–174 °C; IR (KBr,
cmꢂ1) 3421, 2976, 1647, 1504, 1476, 1114, 1071, 892; 1H NMR
(400 MHz, DMSO-d6) d 12.97 (s, 1H, OH-5), 10.46 (s, 1H, OH-40),
7.97 (d, J = 8.8 Hz, 2H, H-20, H-60), 6.94 (d, J = 8.8 Hz, 2H, H-30, H-
50), 6.87 (s, 1H, H-3), 6.84 (d, J = 2.0 Hz, 1H, H-8), 6.46 (d,
J = 2.0 Hz, 1H, H-6), 5.57 (d, J = 9.2 Hz, 1H, H-100), 5.17 (d,
J = 8.0 Hz, 1H, OH-200), 5.11 (d, J = 4.4 Hz, 1H, OH-300), 4.83–4.82
(m, 1H, OH-600), 4.80 (d, J = 5.2 Hz, 1H, OH-2000), 4.70–4.66 (m, 2H,
OH-3000,6000), 4.54 (d, J = 3.2 Hz, 1H, OH-4000), 4.25 (d, J = 6.8 Hz, 1H,
H-1000), 3.60–3.43 (m, 12H, sugar-H); 13C NMR (100 MHz, DMSO-
d6) d 65.3, 65.6, 73.5, 75.5, 75.7, 76.1, 77.4, 78.5, 79.8, 81.7, 96.5,
96.8, 104.5, 104.6, 105.8, 106.9, 115.8, 123.0, 127.8, 157.7, 159.3,
163.1, 163.7, 165.4, 182.1; HRMS [M+Na]+ calcd for C27H30O15Na
617.5067, found 617.5082.
3.18. Synthesis of acacetin-7-O-b-D
-glucoside (13)14
Compound 13 was prepared from 9 as described for the prepa-
ration of 5 from 1, yellow solid, yield: 70%. Mp 262–264 °C. (lit. 14,
260–262 °C); IR (KBr, cmꢂ1) 3343, 2961, 1623, 1505, 1444, 1220,
1144, 1081, 886; 1H NMR (400 MHz, DMSO-d6) d 12.93 (s, 1H,
OH-5), 8.07 (d, J = 8.8 Hz, 2H, H-20, H-60), 7.13 (d, J = 8.8 Hz, 2H,
H-30, H-50), 6.96 (s, 1H, H-3), 6.86 (d, J = 1.6 Hz, 1H, H-8), 6.46 (d,
J = 1.6 Hz, 1H, H-6), 5.33 (d, J = 7.4 Hz, 1H, H-100), 5.16 (s, 1H, OH-
200), 5.08 (s, 1H, OH-300), 5.07 (s, 1H, OH-400), 4.64 (s, 1H, OH-600),
3.87 (s, 3H, OCH3), 3.74–3.70 (m, 1H, H-200), 3.49–3.18 (m, 5H, H-
300,400,500, 600); 13C NMR (100 MHz, DMSO-d6) d 56.0, 61.5, 68.7,
72.3, 75.7, 78.3, 95.8, 98.5, 99.7, 102.4, 105.6, 116.1, 122.3, 127.5,
157.6, 162.0, 162.4, 163.1, 164.3, 182.4; HRMS [M+Na]+ calcd for
3.22. Synthesis of acacetin-7-O-b-D-lactoside (15)
Compound 15 was prepared from 11 as described for the prep-
aration of 5 from 1, yellow solid, yield: 63%. Mp 278–280 °C; IR
(KBr, cmꢂ1) 3420, 2917, 1635, 1505, 1457, 1224, 1038, 885; 1H
NMR (400 MHz, DMSO-d6)
d 12.94 (s, 1H, OH-5), 8.08 (d,
J = 8.4 Hz, 2H, H-20, H-60), 7.15 (d, J = 8.4 Hz, 2H, H-30, H-50), 6.98
(s, 1H, H-3), 6.87 (d, J = 1.2 Hz, 1H, H-8), 6.48 (d, J = 1.2 Hz, 1H,
H-6), 5.61 (d, J = 9.2 Hz, 1H, H-100), 5.20 (d, J = 8.0 Hz, 1H, OH-200),
5.14 (d, J = 4.4 Hz, 1H, OH-300), 4.85–4.83 (m, 1H, OH-600), 4.80 (d,
J = 3.2 Hz, 1H, OH-2000), 4.70–4.68 (m, 2H, OH-3000,6000), 4.57 (d,
J = 4.0 Hz, 1H, OH-4000), 4.25 (d, J = 6.8 Hz, 1H, H-1000), 3.87 (s, 3H,
OCH3), 3.80–3.31 (m, 12H, sugar-H); 13C NMR (100 MHz, DMSO-
d6) d 56.4, 65.2, 65.3, 74.2, 75.6, 76.3, 76.5, 78.2, 78.6, 79.4, 81.5,
95.6, 96.4, 103.5, 104.7, 105.5, 106.8, 116.3, 122.3, 125.7, 158.6,
158.9, 162.1, 164.6, 167.6, 181.3; HRMS [M+Na]+ calcd for
C22H22O10Na 469.3930, found 469.3946.
3.19. Synthesis of apigenin-7-O-b-
D
-galactoside (6)15
Compound 6 was prepared from 2 as described for the prepara-
tion of 5 from 1, yellow solid, yield: 65%. Mp 206–208 °C. (lit. 15,
208–210 °C); IR (KBr, cmꢂ1) 3377, 2924, 1627, 1509, 1497, 1152,
1071, 897; 1H NMR (400 MHz, DMSO-d6) d 12.97 (s, 1H, OH-5),
10.40 (s, 1H, OH-40), 7.96 (d, J = 8.8 Hz, 2H, H-20, H-60), 6.94 (d,
J = 8.8 Hz, 2H, H-30, H-50), 6.87 (s, 1H, H-3), 6.82 (d, J = 2.0 Hz, 1H,
H-8), 6.45 (d, J = 2.0 Hz, 1H, H-6), 5.25 (d, J = 8.0 Hz, 1H, H-100),
5.02 (d, J = 7.6 Hz, 1H, OH-200), 4.92 (d, J = 5.6 Hz, 1H, OH-300), 4.70
(t, J = 5.6 Hz, 1H, OH-600), 4.55 (d, J = 4.4 Hz, 1H, OH-400), 3.72–3.68
(m, 2H, H-200,300), 3.60–3.32 (m, 4H, H-400,500,600); 13C NMR
C
28H32O15Na 631.5332, found 631.5348.
3.23. Synthesis of apigenin-7-O-b-
D
-maltoside (8)17
Compound 8 was prepared from 4 as described for the prepara-
tion of 5 from 1, yellow solid, yield: 60%. Mp 252–254 °C. (lit. 17,
251–254 °C); IR (KBr, cmꢂ1) 3523, 2967, 1605, 1555, 1438, 1265,
1026, 816; 1H NMR (400 MHz, DMSO-d6) d 12.97 (s, 1H, OH-5),
10.44 (s, 1H, OH-40), 7.96 (d, J = 8.4 Hz, 2H, H-20, H-60), 6.94 (d,
J = 8.4 Hz, 2H, H-30, H-50), 6.88 (s, 1H, H-3), 6.84 (d, J = 2.0 Hz, 1H,

46
J. Liu et al. / Carbohydrate Research 357 (2012) 41–46
H-8), 6.45 (d, J = 2.0 Hz, 1H, H-6), 5.67 (d, J = 8.4 Hz, 1H, H-100), 5.54
(d, J = 3.2 Hz, 1H, OH-200), 5.48 (d, J = 2.0 Hz, 1H, OH-300), 5.15–4.13
(m, 1H, OH-600), 5.06 (d, J = 7.6 Hz, 1H, OH-2000), 4.97–4.92 (m, 2H,
OH-3000,6000), 4.65 (d, J = 6.0 Hz, 1H, OH-4000), 4.56 (d, J = 6.8 Hz, 1H,
H-1000), 3.65–3.34 (m, 12H, sugar-H); 13C NMR (100 MHz, DMSO-
d6) d 65.4, 65.6, 73.7, 75.2, 75.4, 76.5, 77.6, 78.6, 79.1, 81.2, 96.3,
96.7, 104.1, 104.2, 105.3, 106.6, 115.9, 123.1, 127.8, 157.3, 159.7,
163.2, 163.7, 165.8, 181.6; HRMS [M+Na]+ calcd for C27H30O15Na
617.5067, found 617.5054.
curve was plotted for each compound, and the half maximal inhib-
itory concentration (IC50) for cancer cell lines was recorded.
Acknowledgements
The financial supports from the Personal Training Funds in Na-
tional Basic Science of China (No. J1103312) and the Science &
Technology Plan of Hunan Province (No. 2011FJ3214) are
gratefully.
3.24. Synthesis of acacetin-7-O-b-
D-maltoside (16)
References
1. Janmejaik, K. S.; Sanjay, G. J. Agric. Food Chem. 2007, 55, 9470–9478.
2. (a) Cook, N. C.; Sammam, S. J. Nutr. Biochem. 1996, 7, 66–76; (b) Harbored, J. B.;
Williams, C. A. Photochemistry 2000, 55, 481–504.
3. Masao, K.; Morikco, M. J. Nat. Prod. 1996, 59, 314–315.
4. Hu, C. Q.; Chen, K.; Shi, Q.; Kilkuskie, R. E.; Cheng, Y. C.; Lee, K. H. J. Nat. Prod.
1994, 57, 42–51.
Compound 16 was prepared from 12 as described for the prep-
aration of 5 from 1, yellow solid, yield: 61%. Mp 266–268 °C; IR
(KBr, cmꢂ1) 3456, 2939, 1623, 1508, 1371, 1226, 1057, 839; 1H
NMR (400 MHz, DMSO-d6)
d 12.91 (s, 1H, OH-5), 8.06 (d,
J = 9.2 Hz, 2H, H-20, H-60), 7.12 (d, J = 9.2 Hz, 2H, H-30, H-50), 6.95
(s, 1H, H-3), 6.85 (d, J = 1.2 Hz, 1H, H-8), 6.45 (d, J = 1.2 Hz, 1H,
H-6), 5.66 (d, J = 8.4 Hz, 1H, H-100), 5.54 (d, J = 5.2 Hz, 1H, OH-200),
5.48 (d, J = 6.0 Hz, 1H, OH-300), 5.05–4.03 (m, 1H, OH-600), 5.01 (d,
J = 5.6 Hz, 1H, OH-2000), 4.95–4.92 (m, 2H, OH-3000,6000), 4.55 (d,
J = 6.0 Hz, 1H, OH-4000), 4.56 (d, J = 6.8 Hz, 1H, H-1000), 3.62–3.35
(m, 12H, sugar-H); 13C NMR (100 MHz, DMSO-d6) d 56.3, 64.5,
65.7, 73.4, 75.1, 75.4, 75.5, 78.5, 78.7, 79.2, 82.3, 95.5, 96.8,
103.5, 104.2, 105.6, 106.7, 116.8, 123.1, 128.7, 156.3, 158.8,
163.1, 165.7, 167.5, 182.0; HRMS [M+Na]+ calcd for C28H32O15Na
631.5332, found 631.5317.
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3.25. Assay for cytotoxic activity
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´
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various concentrations (30, 10, 1, 0.1 lg/mL) and 10 lL of the test
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adding 5% MTT in PBS (20
the incubator for 4 h. To each well was added 100
the formazan crystals were dissolved, and the plate was read on
l
L) to each well, the plate was kept in
1219–1222.
lL of 20% SDS,
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393.