Synthesis and antiproliferative activity of helonioside A, 3′,4′,6′-tri-O-feruloylsucrose, lapathoside C and their analogs

Article abstract of DOI:10.1016/j.ejmech.2012.10.034

The first total synthesis of natural phenylpropanoid sucrose esters (PSEs) helonioside A 1, 3′,4′,6′-tri-O-feruloylsucrose 2 and lapathoside C 3 along with 17 unnatural PSE analogs has been successfully accomplished in a short and simple synthetic route.

Full text of DOI:10.1016/j.ejmech.2012.10.034

European Journal of Medicinal Chemistry 58 (2012) 418e430
Contents lists available at SciVerse ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis and antiproliferative activity of helonioside A, 30,40,60-tri-O-
feruloylsucrose, lapathoside C and their analogs
Parthasarathi Panda a, Manjuvani Appalashetti a, Meenubharathi Natarajan b, Chan-Park Mary a,
,
Subbu S. Venkatraman b, Zaher M.A. Judeh a
*
a School of Chemical and Biomedical Engineering, Nanyang Technological University, 62 Nanyang Drive, N1.2-B1-14, Singapore 637459, Singapore
b School of Materials Science and Engineering, Nanyang Technological University, 50 Nanyang Avenue, N4.1-02-06, Singapore 639798, Singapore
a r t i c l e i n f o
a b s t r a c t
The first total synthesis of natural phenylpropanoid sucrose esters (PSEs) helonioside A 1, 30,40,60-tri-O-
feruloylsucrose 2 and lapathoside C 3 along with 17 unnatural PSE analogs has been successfully
accomplished in a short and simple synthetic route. A selected set of 17 synthesized PSEs were evaluated
for the antiproliferative activity against human cervical epithelioid carcinoma (HeLa) cell lines using MTS
assay method. Eleven (11) compounds showed significant antiproliferative activity with their IC50values
Article history:
Received 5 August 2012
Received in revised form
17 October 2012
Accepted 18 October 2012
Available online 27 October 2012
ranging from 0.16 to 6.01
mM. The structureeactivity-relationship studies revealed that the anti-
proliferative activity is influenced by the lipophilicity and number of feruloyl substituents on these
compounds. The preliminary screening indicated that these compounds are potentially very valuable
source for new lead chemotherapeutics.
Keywords:
Helonioside A, 30,40,60-tri-O-feruloylsucrose
Lapathoside C
Ó 2012 Elsevier Masson SAS. All rights reserved.
Phenylpropanoid sucrose esters
Antiproliferative activity
Isopropylidene acetal
Regioselective acylation
Sucrose esters
HeLa cells
1. Introduction
cell lines [4,14]. Yan et al. reported that helonioside A 1 exhibited
cytotoxic effects in a dose-dependent manner against mice lung
O-feruloylsucroses belong to an important class of phenyl-
propanoid sucrose esters (PSEs) and are found in various medicinal
plants with promising biological activities [1]. For example, hel-
onioside A 1 (i.e. 30,60-di-O-feruloylsucrose) (Fig. 1) was isolated
from the extracts of various liliaceous plants of Heloniopsis orientalis
[2], Smilax glabra [3], Smilax china [4], Trillium kamtschaticum [5],
Paris polyphylla var. yunnanensis [6,7], Smilacaceous plants of Smilax
bracteata [8] and Polygonaceous plants of Polygonum perfoliatum [9],
Bistorta manshuriensis [10] and Rumex dentatus [11]. Thewhole plant
P. polyphylla var. yunnanensis (Liliaceae) has been used in traditional
Chinese medicine to treat lung, liver and laryngeal carcinoma [6,12].
The extracts of H. orientalis displayed potent cytotoxicity against
adenocarcinoma cell line (LA 795) [6,7]. Helonioside A 1 also dis-
played potent antioxidant activity against stable DPPH-free radical
at a concentration of 0.02 mM [5]. 30,40,60-tri-O-feruloylsucrose 2
(Fig. 1) was isolated from the ethanol extracts of the rhizomes and
roots of Smilax riparia [15]. Lapathoside C 3 (i.e. 6-mono-O-feruloyl-
30,60-di-O-coumaroylsucrose) (Fig. 1) was isolated from the various
Polygonaceous plant species such as Polygonum lapathifolium [16],
Polygonum sachalinensis [17] and Polygonum cuspidatum [18]. The
Smilax genus of Liliaceous Plants and Polygonum genus of the Pol-
ygonaceous plants and their extracts are widely used in traditional
and folk medicines for the treatment of various disorders and
diseases such as cancer, tumors, jaundice, coughs, carbuncles,
dysentery ... etc. [1].
lung carcinoma cell lines (A549) with IC50 value of 4.6
mg/mL and
colon carcinoma cell lines (Col2) with IC50 of 4.5 g/mL [13]. The S.
china crude extracts were found to have antimutagenic activities
whileits tuber showedsignificant cytotoxicityagainstvarioustumor
m
The entire published research in the area of PSEs focusses on the
extraction, characterization and biological screening of these
compounds [1,19e22]. With the exception of two reports [23,24],
the synthesis of this large class of compounds (about 150 natural
PSEs have been reported to date) has not been reported. In one of
these reports, our group has successfully demonstrated the
synthesis of lapathoside D and its analogs and reported their
* Corresponding author. Tel.: þ65 6790 6738; fax: þ65 6794 7553.
E-mail address: zaher@ntu.edu.sg (Z.M.A. Judeh).
0223-5234/$ e see front matter Ó 2012 Elsevier Masson SAS. All rights reserved.
http://dx.doi.org/10.1016/j.ejmech.2012.10.034

P. Panda et al. / European Journal of Medicinal Chemistry 58 (2012) 418e430
419
Fig. 1. Structure of helonioside A 1, 30,40,60-tri-O-feruloylsucrose 2 and lapathoside C 3.
antiproliferative activities [24]. In continuation to our on-going
2.1.1. Preparation of helonioside A 1, 30,40,60-tri-O-feruloylsucrose 2
and their analogs
At the outset, when di-O-isopropylidene sucrose 4 [24,25] was
reacted with 1.1 equiv of p-acetoxyferuloyl chloride 5 [26] at r.t. for
3 days (Scheme 1), compound 6 was obtained as the major product
in 31% yield along with compounds 8 (11% yield) and 9 (12% yield)
as white solids.
Reaction of di-O-isopropylidene sucrose 4 with 2.2 mol equiv of
p-acetoxyferuloyl chloride 5 at r.t. (Scheme 1) afforded compound 8
as the major product (30% yield) along with compound 6 in 3%
yield. Clearly this result indicated that the use of 2.2 mol instead of
1.1 mol selectively gave the di-acylated product 8 over the mono-
acylated product 6. Attempts to increase the yield by varying the
reaction conditions (time, higher temperature) were not successful.
In order to prepare the tri-acylated compounds, di-O-iso-
propylidene sucrose 4 was reacted with 3.3 mol equiv of p-ace-
toxyferuloyl chloride 5 (Scheme 1). Interestingly, the di-acylated
product 8 was found to be the major reaction product (44% yield)
while the tri-acylated product 9 was obtained in 26% yield,
both as white solids. Likewise, reaction between 4.4 mol equiv of
research efforts for developing practical and viable synthetic
routes and exploring the potential of various PSEs as anticancer
drug lead compounds [24], we became interested in the synthesis
of helonioside A 1, 30,40,60-tri-O-feruloylsucrose 2, lapathoside C 3
and their analogs.
Herein, we report the first total syntheses of helonioside A 1,
30,40,60-tri-O-feruloylsucrose 2, lapathoside C 3 and their unnatural
analogs. We also report on their antiproliferative activities against
human cervical epithelioid carcinoma (HeLa) cell lines and examine
the structural features that contribute to better activities.
2. Results and discussion
2.1. Chemistry
The synthesis of the target compounds, helonioside A 1, 30,40,60-
tri-O-feruloylsucrose 2, lapathoside C 3 and their analogs was
attempted by regio- and chemoselective acylation of the versatile
substrate 2,10:4,6-di-O-isopropylidene sucrose 4 [24,25].
Scheme 1. Regio- and chemoselective acylation of di-O-isopropylidene 4 with p-acetoxyferuloyl chloride 5.

420
P. Panda et al. / European Journal of Medicinal Chemistry 58 (2012) 418e430
p-acetoxyferuloyl chloride 5 and di-O-isopropylidene sucrose 4
(Scheme 1), gave the expected tetra-acylated derivative 10 in 44%
yield along with the tri-acylated product 9 (35% yield). These
results indicated: (1) the reactivity of the OH groups are in the order
of 60-OH > 30-OH > 40-OH > 3-OH; (2) with the correct number of
mole equiv of p-acetoxyferuloyl chloride 5, we can have the choice
to prepare the mono-, di-, tri- and tetra-acylated compounds; and
(3) analogs (or unnatural O-feruloylsucroses) can easily be
prepared by controlling the number and position of p-acetoxyfer-
uloyl chloride 5.
(C-1100 for COCH3) and also carbonyl carbons at
d 168.4e170.5 ppm
(C-1000 for COCH3). Furthermore, the IR peaks corresponding to the
acetyl ester carbonyl group at ca. 1764 cmꢁ1 for compounds 11e13
were missing in the spectra of compounds 1, 2 and 14. The HR-MS
of helonioside A 1 showed m/z 717.1984 [M þ Na]þ (calcd 717.2001
for C32H38O17Na). Furthermore, the structure of helonioside A 1
(Fig. 2) was confirmed by comparison to the data reported for the
isolated natural product [2,6,7] (See Supporting information).
The HR-MS of the 30,40,60-tri-O-feruloylsucrose 2 suggested the
molecular formula C42H46O20 based on the molecular ion peak at
m/z 893.2478 [M þ Na]þ (calcd 893.2475 for C42H46O20Na). The
analysis data for 30,40,60-tri-O-feruloylsucrose 2 (Figs. 3 and 4) did
not completely match with literature data especially in the sucrose
unit [15]. At this point, we presume that the structure of the iso-
lated natural product needs further confirmation (See Supporting
information).
Next, compounds 8e10 were subjected separately to acetal
deprotection using 60% aq. AcOH [24]. The crude products were
purified by recrystallization from EtOAc or column chromatography
(CH2Cl2/EtOAc) to afford compounds 11e13, respectively, as white
solids in good yields ranging from 67% to 89% (Scheme 1). In
comparison to the NMR spectra of compounds 8e10, compounds
11e13 revealed the absence of the characteristic signals for the two
isopropylidene moieties usually observed in the 1H NMR spectra
In the same fashion, deacetylation of compound 9 gave the
conformationally restricted PSE 15 in 71% yield as white solid
(Scheme 1). The 1H and 13C NMR spectra of compound 15 revealed
that the disappearance of the characteristic signals for the acetyl
at
d
1.20e1.53 ppm (4ꢀ s, 12H, 2 (CH3)2C) and in the 13C NMR
spectra at
d
19.0e29.1 ppm (4 ꢀ 2 (CH3)2C) and 99.6e101.8 ppm
(2ꢀ (CH3)2C). The typical value of the chemical shift of the
moieties at
d
2.34 ppm (s, 9H, H-1100 for the COCH3),
d
20.7 ppm (C-
anomeric (H-1) protons for compounds 11e13 shifted downfield at
1100 for COCH3) and at
COCH3).
d
168.6, 168.7 and 168.8 ppm (C-1000 for
d
d
5.50 ppm compared to the same for compounds 8e10 at
6.16 ppm.
The IR spectrum of compound 15 further indicated the
successful deprotection where peaks corresponding to the acetyl
ester carbonyl group of compound 9 at 1766 cmꢁ1 have dis-
appeared. Furthermore, the HR-MS spectrum of compound 15
Successful deacetylation of the phenyl rings of compounds 11e
13 would afford the final compounds helonioside A 1, 30,40,60-tri-O-
feruloylsucrose 2 and the analog 14. Hatfield et al. [26] demon-
strated the successful deacetylation of a similar sugar, the methyl-
displayed m/z 973.3078 [M
C48H54O20Na). This compound was intentionally synthesized to
þ
Na]þ (calcd 973.3101 for
5-O-acetylferuloyl-
a
-L-arabinofuranoside, using piperidine-95%
ethanol in good yield. Gladly, when compounds 11e13 were sub-
jected separately to the Hatfield et al. [26] deacetylation conditions,
helonioside A 1, 30,40,60-tri-O-feruloylsucrose 2 and 3,30,40,60-tetra-
O-feruloylsucrose 14 were successfully obtained as white solids in
68%, 65% and 76% yield, respectively (Scheme 1). The success of
removal of the acetyl protecting group from the feruloyl moiety
selectively is due to the fact that the ferulic ester bond is stronger
than an acetate ester bond, allowing the differentiation between
the two acyl groups [27]. The 1H and 13C NMR spectra of
compounds 1, 2 and 14 indicated the loss of the characteristic
signals for the acetyl moieties on the phenyl ring, represented by
investigate the effect of free phenolic hydroxyl groups and the
presence of additional feruloyl substituents on the antiprofilative
activities.
2.1.2. Preparation of lapathoside C and its analogs
We have previously reported the successful synthesis of 30,60-di-
O-acetoxycinnamoyl sucrose 16 [24] and envisioned it to be an ideal
starting substrate for the planned synthesis of lapathoside C 3
considering that the primary hydroxyl group (6-OH) of compound
16 is more reactive than the rest of the hydroxyl groups [28,29].
Consequently, reaction between compound 16 and p-acetoxyfer-
uloyl chloride 5 (1.1 equiv) gave compound 18 (36% yield) along
with compound 19 (7% yield) both as white solid (Scheme 2).
the characteristic proton signals at
d
2.26e2.32 ppm (s, 3H, H-1100
20.5e20.9 ppm
for the COCH3) and acetyl ester carbon signals at
d
Fig. 2. 1H NMR spectrum of helonioside A 1 (300 MHz, CD3OD).

P. Panda et al. / European Journal of Medicinal Chemistry 58 (2012) 418e430
421
1
Fig. 3. H NMR spectrum of 30,40,60-tri-O-feruloylsucrose 2 (300 MHz, CD3OD).
Compound 18 would be used to prepare lapathoside C 3 at a later
stage. Various analogs can be prepared in a similar fashion. Thus,
reaction between sucrose 16 and p-acetoxycinnamoyl chloride 17
[24,27] (1.3 equiv) afforded compound 20 in just 12% yield (Scheme
2). Likewise, reaction between compound 18 and p-acetox-
ycinnamoyl chloride 17 gave compound 23 as white solid in 27%
yield (Scheme 2).
Unfortunately, the Hatfield deacetylation method [26] did not
work on compounds 18e20 and 23 since the starting materials
were recovered unchanged. Therefore, the deacetylation was per-
formed using pyrrolidine [27] (Scheme 2) which successfully gave
compounds 3, 21, 22 and 24 in 75%, 47%, 71% and 35% yield,
respectively, as white solids. The 1H and 13C NMR spectra of
compounds 3, 21, 22 and 24 indicated the absence of the charac-
teristic signals for the acetyl moieties at 2.23e2.30 ppm (s, 3H, H-
1100 for the COCH3), 20.6e21.1 ppm (C-1100 for COCH3) and
168.8e
d
d
d
169.5 ppm (C-1000 for COCH3). The IR spectra of these compounds
were devoid of peaks corresponding to the acetyl ester carbonyl
groups at ca. 1765e1768 cmꢁ1 observed in the starting compounds
18e20 and 23. The HR-MS of compound 3 showed m/z 833.2283
[M þ Na]þ (calcd 833.2263 for C40H42O18Na). Based on the
spectroscopic data and also by comparison to the reported
literature data for the isolated natural lapathoside
C [16]
(See Supporting information), the structure of compound 3 was
assigned to be lapathoside C 3 (i.e. 6-mono-O-feruloyl-30,60-di-O-
coumaroylsucrose).
Fig. 4. 1H NMR spectrum of lapathoside C 3 (300 MHz, CD3OD).

422
P. Panda et al. / European Journal of Medicinal Chemistry 58 (2012) 418e430
Scheme 2. Preparation of lapathoside C 3 and its analogs.
2.2. Antiproliferative activities
3.22 mM, respectively, compared to their flexible deprotected OH
counterparts 1, 2 and 14 (entries 4 vs 1, 5 vs 2 and 6 vs 9, Table 2). The
The antiproliferative activities of natural PSEs helonioside A 1,
30,40,60-tri-O-feruloylsucrose 2, lapathoside C 3 as well as selected
analogs 6, 8e12, 14, 15, 18e22 and 24 were evaluated against HeLa
cells using our previously reported method [24].
The IC50 values of these compounds along with the standard drug
camptothecin (CPT) are shown in Tables 1 and 2. Helonioside A 1
(Table 2, entry 1) and its analog compound 11 in which the phenolic
groups are acetylated (Table 2, entry 7) did not showanyappreciable
trend of the antiproliferative activity of 30,40,60-tri-O-feruloylsucrose
analogs are: 9 (IC50 ¼ 0.16
m
M) > 12 (IC50 ¼ 1.70
mM) > 15
M). From the above results, it is
(IC50 ¼ 4.20 M) > 2 (IC50 ¼ 22.35
m
m
clear that the completely acetylated di-O-isopropylidene variants
like compound 9 have superior activity while deacetonide (di-O-
isopropylidene-free) 12 or deacetylated product 15 showed lower
activities. Additionally, after cleavage of the acetyl and di-O-iso-
propylidene groups of compound 9 to give compound 2, the activity
of the obtained product was dramatically reduced due to probable
differences in the lipophilicity of these compounds (entries 2 & 5,
Table 2). Increased lipophilicity of molecules was reported to be
responsible for enhanced cytotoxicity [30]. The presence of free
antiproliferative activity upto 100 mM concentration. Hence, acety-
lation of the phenolic groups seems to be ineffective in enhancing
the antiprolifrative activity. Compound 2, a tri-O-feruloyl sucrose,
showed improved antiprolifrative activity with an IC50 value of
22.35
exhibited greater antiprolifrative activity with an IC50 value of
1.62 M (entry 9, Table 2). Therefore, introduction of more feruloyl
groups seems to have positive effect on the activity. Rigid
compounds having di-O-isopropylidene groups with their phenolic
OH protected with an acetyl group 8, 9 and 10 showed significant
antiproliferative activities with their IC50 values of 6.01, 0.16 and
m
M (entry 2, Table 2) whereas tetra-O-feruloyl sucrose 14
feruloyl derivatives is: 14 (IC50 ¼ 1.62
m
M) > 10 (IC50 ¼ 3.22
mM)
(entries 9 & 6, Table 2). It was anticipated that compound 6 with
a mono phenylpropanoid group would show weak activity. Instead,
this compound did not show any antiproliferative activity up to
m
100 mM concentration (entry 3, Table 2). In this case, the positional
effect of the feruloyl group on the sucrose core seems to have a great
effect. We suspect that substituents at the C60 position play no major
role in the antiproliferative activity.
Tetra-phenylpropanoid sucrose esters analogs 18e22 and 24
(entries 3e7, Table 3) showed better antiproliferative activities
compared to the tri-phenylpropanoid sucrose esters analogs 3 and 18
(entries 1 and 2, Table 3). Acetylationof3 to give compound 18 did not
improve the IC50 value (entry 1 vs 2, Table 3). The tetra-coumaroyl PSE
22, tetra-feruloyl PSE 14 and “mixed” PSEs 21 having a combination of
coumaroyl and feruloyl units exhibited almost similar anti-
proliferative activity with their IC50 values of 1.70, 1.62 and 1.67 mM,
respectively (entries 5 & 6, Table 3 and entry 9, Table 2). Replacing one
of thecoumaroyl substituents atC3of24 witha feruloyl groupasin21
improved the IC50 value from 3.12 to 1.67 mM (entry 7 vs 5, Table 3).
Table 1
Regio- and chemoselective acylation of di-O-isopropylidene sucrose 4 with p-ace-
toxyferuloyl chloride 5.
Entry
1
Equiv. of acid
chloride 5
Time
(d)
Product
Yield
(%)
Total yield
(%)
1.1
3
6
7
8
6
8
8
9
9
31
11
12
3
30
44
26
35
44
54
2
3
4
2.2
3.3
4.4
5
2
4
33
70
79
Compound21 (entry5, Table 3) showedsuperioractivitycompared to
its acetylated product 19 (entry 3, Table 3).
10

P. Panda et al. / European Journal of Medicinal Chemistry 58 (2012) 418e430
423
Table 2
In vitro cytotoxicity (IC50 (mM)) of selected feruloyl PSEs against HeLa cells at 48 h of exposure.
Entry
Cpd
IC50
1
2
3
4
5
6
7
8
9
10
15
1
2
6
8
6.01
9
0.16
10
3.22
11
>100
12
1.70
14
1.62
15
4.20
CPT
0.40
a
>100
22.35
>100
a
IC50 (mM): the concentration that induces 50% growth inhibition compared with untreated control cells and were calculated from three independent experiments.
Additionally, compounds 2, 8 and 14 were selected for evaluation
of the cytotoxicityat twodifferent time intervals of drug exposureby
MTS assay (Table 4) in order to examine time-dependent cytotox-
icity. The IC50 values of compounds 2, 8 and 14 were enhanced as the
time of exposure increased from 24 h to 48 h. These compounds
revealed time-dependent antiproliferative activities.
synthesized according to the reported procedures [24]. p-Acetox-
yferuloyl chloride 5 was synthesized according to the reported
literature [26] (see Supporting information).
4.1.1. Acylation of di-O-isopropylidene 4 with p-acetoxyferuloyl
chloride 5
In summary, we can conclude that: (1) di-O-isopropylidene
group proved to be essential for enhancing the cytotoxicity while
acetyl and methoxy groups had much lower effects; (2) Substitu-
ents at the C60 position are not essential for antiproliferative
activity; (3) Lipophilicity of the examined PSEs seems to influence
the cytotoxicity in MTS model [30]. Herein, the position and nature
of phenylpropanoid unit had a little effect on the antiproliferative
activity of PSEs; and (4) The PSEs examined also revealed time-
dependent antiproliferative activities.
4.1.1.1. General procedure. Di-O-isopropylidene 4 was dissolved in
dry pyridine under a nitrogen atmosphere. The solution was then
cooled to 0 ꢂC in an ice bath. p-Acetoxyferuloyl chloride 5 was then
added slowly at the same temperature and the reaction was left to
stir while warming to r.t. Stirring was continued at r.t. until the
reaction was completed (TLC analysis (EtOAc/hexanes 3:1)). The
resulting reaction mixture was poured into vigorously stirred ice-
water (100 mL) and the precipitated white solid was obtained by
vacuum filtration. The precipitates were redissolved in EtOAc
(25 mL) and washed once with 1 N HCl (50 mL). The combined
organic layers were then successively washed with 5% NaHCO3
(50 mL), brine (25 mL) and then dried over anhyd. MgSO4. The
EtOAc solution was evaporated under reduced pressure to give
yellow syrup that was subjected to column chromatography using
a gradient of CH2Cl2/EtOAc as eluent to afford compounds 6e10.
3. Conclusions
We successfully synthesized, for the first time, the natural PSEs
helonioside A 1, 30,40,60-tri-O-feruloylsucrose 2 and lapathoside C
3 along with 17 unnatural PSEs analogs using regio- and che-
moselective acylation of di-O-isopropylidene sucrose 4. The
reactivities of the free OH groups of di-O-isopropylidene sucrose 4
towards acylation using p-acetoxyferuloyl chloride 5 were found
to be in the order of 60-OH > 30-OH > 40-OH > 3-OH. The
synthetic methodology used is wide in scope and applicable to
wide range of PSEs. The MTS results revealed that 11 out of the 17
examined PSEs displayed significant antiproliferative activity
against HeLa cells at 48 h drug exposure with their IC50values
4.1.1.2. 60-mono-O-acetoxyferuloyl-2,10:4,6-di-O-isopropylidene
sucrose 6 and 30-mono-O-acetoxyferuloyl-2,10:4,6-di-O-isopropylidene
sucrose 7. Following the general procedure, the reaction between
di-O-isopropylidene 4 (1.0 g, 2.4 mmol) and p-acetoxyferuloyl
chloride 5 (0.7 g, 2.6 mmol) in dry pyridine (10 mL) for 3 days
afforded compound 6 as a white solid (0.47 g, 31% yield) along with
compounds 7 and 8 in 11% (0.16 g) and 12% (0.25 g) yield, respec-
tively. Analytical data for 6: Rf ¼ 0.12 (EtOAc/CH2Cl2 3:2); mp: 147e
150 ꢂC; FT-IR (KBr) nmax: 3452, 2993, 2941, 1766, 1712, 1636, 1601,
1511, 1417, 1372, 1260, 1199, 1157, 1127, 1069, 1013, 943, 858, 718,
ranging from 0.16 to 6.01
m
M
compared with CPT
(IC50 ¼ 0.40 M). The results also indicated time-dependent
m
antiproliferative activities. Lipophilicity which is influenced by
the presence/absence of di-O-isopropylidene group and the
number of the phenylpropanoid units on the sucrose core directly
influence the antiproliferative activities. Our present results
provide great motivation for the synthesis of other natural and
unnatural PSEs and explore their applications and mechanism of
action against various human cancer cell lines such as colon
cancer, lung carcinoma, breast cancer, skin cancer etc in search for
new lead anticancer drug candidates.
651 cmꢁ1; 1H NMR (300 MHz, CDCl3)
d
(ppm): 1.45, 1.51 (2ꢀ s, 12H,
(CH3)2C), 3.50 (m, 1H, H-10a), 3.60 (m, 1H, H-4), 3.73 (m, 2H, H-2, H-
6a), 3.93 (m, 3H, H-30, H-5, H-6b), 4.05 (m, 1H, H-3), 4.23 (m, 2H, H-
40, H-50), 4.30 (m, 2H, H-10b, H-60a), 4.51 (m, 1H, H-60b), 6.19 (br d,
1H, J ¼ 1.8 Hz, H-1); trans-p-feruloyl units: 2.32 (1s, 3H, H-1100), 3.85
(s, 3H, eOCH3), 6.41 (d, 1H, J ¼ 15.9 Hz, H-800), 7.00e7.19 (m, 3H, H-
200, H-500, H-600), 7.64 (d, 1H, J ¼ 15.9 Hz, H-700); 13C NMR (75.48 MHz,
CDCl3)
d
(ppm): 19.2, 24.2, 25.3, 29.1 (4ꢀ (CH3)2C), 62.3 (C-6), 63.8
(C-5), 65.9 (C-60), 66.5 (C-10), 69.3 (C-3), 73.4 (C-4), 73.9 (C-2), 77.5
(C-40), 79.0 (C-30), 79.7 (C-50), 91.0 (C-1), 100.1, 102.2 (2ꢀ (CH3)2C),
103.7 (C-20); trans-p-feruloyl units: 20.7 (C-1100), 55.9 (eOCH3), 111.3
(C-200), 117.9 (C-800), 121.4 (C-500), 123.2 (C-600), 133.2 (C-100), 141.5 (C-
300), 144.7 (C-700), 151.4 (C-400), 167.0 (C-900), 168.8 (C-1000); LC-MS
(ESI): m/z 663.22 [M þ Na]þ; HR-MS: m/z [M þ Na]þ calcd for
4. Experimental
4.1. Chemistry
2,10:4,6-di-O-isopropylidene sucrose 4, 30,60-di-O-acetox-
ycinnamoyl sucrose 16 and p-acetoxycinnamoyl chloride 17 were
C30H40O15Na: 663.2259, found: 663.2268. Analytical data for 7:
Table 4
IC50 values of selected synthesized PSEs 2, 8 and 14 along with CPT at 24 h and 48 h
of exposure.
Table 3
In vitro cytotoxicity (IC50
(mM)) of selected lapathoside C and its analogs PSEs against
HeLa cells at 48 h of exposure.
No
Compound
IC50 (mM)
24 h
48 h
Entry
Cpd
IC50
1
2
3
4
5
6
7
8
1
2
3
4
2
8
14
CPT
55.07
27.92
7.90
22.35
6.01
1.62
0.40
3
18
>100
19
3.14
20
1.86
21
1.67
22
1.70
24
3.12
CPT
0.40
a
12.61
a
IC50 (mM): the concentration that induces 50% growth inhibition compared with
1.57
untreated control cells and were calculated from three independent experiments.

424
P. Panda et al. / European Journal of Medicinal Chemistry 58 (2012) 418e430
Rf ¼ 0.21 (EtOAc/CH2Cl2 3:2); mp: 130e132 ꢂC; FT-IR (KBr) nmax
:
(m, 3H, H-200, H-500, H-600), 7.66 (d, 1H, J ¼ 15.9 Hz, H-700); R2: 2.34 (s,
3H, H-1100), 3.88 (s, 3H, eOCH3), 6.42 (d,1H, J ¼ 15.9 Hz, H-800), 7.02e
7.13 (m, 2H, H-500, H-600), 7.20e7.22 (m, 1H, H-200), 7.68 (d, 1H,
J ¼ 15.9 Hz, H-700); R3: 2.34 (s, 3H, H-1100), 3.93 (s, 3H, eOCH3), 6.52
(d, 1H, J ¼ 15.9 Hz, H-800), 7.08 (d, 2H, J ¼ 8.7 Hz, H-500, H-600), 7.20e
7.22 (m, 1H, H-200), 7.79 (d, 1H, J ¼ 15.9 Hz, H-700); 13C NMR
2994, 2936,1765,1716,1638,1590,1510, 1467, 1421, 1373, 1333, 1260,
1199, 1156, 1125, 1069, 1033, 1015, 946, 855, 650 cmꢁ1
;
1H NMR
(300 MHz, CDCl3)
d
(ppm): 1.40, 1.44, 1.53 (3ꢀ s, 12H, (CH3)2C), 3.58
(m, 2H, H-10a, H-4), 3.68 (m, 2H, H-6a, H-60a), 3.77 (m,1H, H-2), 3.87
(m, 4H, H-3, H-5, H-6b, H-60b), 4.10 (m, 2H, H-10b, H-50), 4.87 (m,1H,
H-40), 5.03 (d, 1H, J ¼ 7.8 Hz, H-30), 6.22 (d, 1H, J ¼ 3.6 Hz, H-1);
trans-p-feruloyl units: 2.05 (1s, 3H, H-1100), 3.91 (s, 3H, eOCH3), 6.50
(d, 1H, J ¼ 15.9 Hz, H-800), 7.09 (d, 1H, J ¼ 7.8 Hz, H-500), 7.16e7.22 (m,
2H, H-500, H-600), 7.77 (d,1H, J ¼ 15.9 Hz, H-700); 13C NMR (75.48 MHz,
(75.48 MHz, CDCl3)
d
(ppm): 19.1, 24.1, 25.5, 29.0 (4ꢀ (CH3)2C), 62.0
(C-6), 63.9 (C-5), 64.9 (C-60), 66.3 (C-10), 70.2 (C-3), 72.8 (C-4), 73.8
(C-2), 77.3 (C-40), 77.6 (C-30), 80.1 (C-50), 91.4 (C-1), 99.7, 101.8
(2ꢀ (CH3)2C), 104.8 (C-20); trans-p-feruloyl units: R1: 20.7 (C-1100),
55.9 (eOCH3), 111.2 (C-200), 116.5 (C-800), 121.4 (C-500), 123.2 (C-600),
132.9 (C-100), 141.4 (C-300), 144.4 (C-700), 151.3 (C-400), 165.7 (C-900),
168.6 (C-1000); R2: 20.7 (C-1100), 56.0 (eOCH3), 111.3 (C-200), 116.8
(C-800), 121.5 (C-500), 123.3 (C-600), 132.9 (C-100), 141.8 (C-300), 144.8
(C-700), 151.4 (C-400), 165.8 (C-900), 168.7 (C-1000); R3: 20.7 (C-1100),
56.0 (eOCH3), 111.4 (C-200), 117.9 (C-800), 122.1 (C-500), 123.4 (C-600),
133.3 (C-100), 141.9 (C-300), 146.4 (C-700), 151.5 (C-400), 166.3 (C-900),
168.8 (C-1000); LC-MS (ESI): m/z 1099.35 [M þ Na]þ; HR-MS: m/z
[M þ Na]þ calcd for C54H60O23Na: 1099.3418, found: 1099.3401.
CDCl3)
d
(ppm): 19.0, 24.1, 25.3, 29.0 (4ꢀ (CH3)2C), 61.2 (C-6), 61.8
(C-60), 63.9 (C-5), 66.5 (C-10), 70.0 (C-3), 71.4 (C-40), 72.7 (C-2), 73.4
(C-4), 80.0 (C-30), 84.0 (C-50), 91.0 (C-1), 99.9, 102.0 (2ꢀ (CH3)2C),
103.48 (C-20); trans-p-feruloyl units: 20.6 (C-1100), 56.0 (eOCH3),
111.3 (C-200), 116.8 (C-800), 122.0 (C-500), 123.4 (C-600), 132.9 (C-100),
141.9 (C-300), 146.3 (C-700), 151.5 (C-400), 167.2 (C-900), 168.7 (C-1000);
LC-MS (ESI): m/z 663.24 [M þ Na]þ; HR-MS: m/z [M þ Na]þ calcd for
C30H40O15Na: 663.2259, found: 663.2255.
4.1.1.3. 30,60-di-O-acetoxyferuloyl-2,10:4,6-di-O-isopropylidene
sucrose 8. Following the general procedure, the reaction between
di-O-isopropylidene 4 (1.0 g, 2.4 mmol) and p-acetoxyferuloyl
chloride 5 (1.3 g, 5.2 mmol) in dry pyridine (10 mL) for 5 days gave
compound 8 as a white solid (0.6 g, 30% yield) along with
compound 6 in 3% (0.05 g) yield. Analytical data for 8: Rf ¼ 0.61
(EtOAc/CH2Cl2 3:2); mp: 109e110 ꢂC; FT-IR (KBr) nmax: 3486, 2993,
2942, 1766, 1716, 1637, 1601, 1511, 1417, 1372, 1332, 1259, 1198, 1069,
4.1.1.5. 3,30,40,60-tetra-O-acetoxyferuloyl-2,10:4,6-di-O-isopropylidene
sucrose 10. Following the general procedure, the reaction between
di-O-isopropylidene 4 (1.0 g, 2.4 mmol) and p-acetoxyferuloyl
chloride 5 (2.7 g, 10.4 mmol) in dry pyridine (10 mL) for 4 days
afforded compound 10 as a white solid (1.34 g, 44% yield) along
with compound 9 in 35% (0.9 g) yield. Analytical data for 10:
Rf ¼ 0.88 (EtOAc/CH2Cl2 3:2); mp: 133e135 ꢂC; FT-IR (KBr) nmax
:
1032, 1012, 947, 906, 858, 655 cmꢁ1
d
;
1H NMR (300 MHz, CDCl3)
3629, 2993, 2942, 1767, 1721,1637, 1601, 1467, 1419, 1371, 1327, 1259,
1198, 1153, 1123, 1070, 1033, 1010, 944, 903, 856, 832, 796, 727,
(ppm): 1.40, 1.41, 1.53 (3ꢀ s, 12H, (CH3)2C), 3.63 (m, 2H, H-10a, H-
4), 3.74 (m, 1H, H-6a), 3.79 (m, 1H, H-2), 3.88 (m, 2H, H-3, H-5), 3.94
(m, 1H, H-6b), 4.08 (d, 1H, J ¼ 12.3 Hz, H-10b), 4.38 (m, 2H, H-50, H-
60a), 4.46 (m, 1H, H-40), 4.53 (m, 1H, H-60b), 4.92 (d, 1H, J ¼ 6.3 Hz,
H-30), 6.13 (d, 1H, J ¼ 3.3 Hz, H-1); trans-p-feruloyl units: R1: 2.33 (s,
3H, H-1100), 3.87 (s, 3H, eOCH3), 6.43 (d,1H, J ¼ 15.9 Hz, H-800), 7.03e
7.12, 7.20 (2ꢀ m, 3H, H-200, H-500, H-600), 7.66 (d, 1H, J ¼ 15.9 Hz, H-
700), R2: 2.33 (s, 3H, H-1100), 3.92 (s, 3H, eOCH3), 6.48 (d, 1H,
J ¼ 15.9 Hz, H-800), 7.03e7.12, 7.20 (2ꢀ m, 3H, H-200, H-500, H-600), 7.77
649 cmꢁ1; 1H NMR (300 MHz, CDCl3)
d (ppm): 1.20, 1.30, 1.45, 1.49
(4ꢀ s, 12H, (CH3)2C), 3.65 (m, 1H, H-10a), 3.70 (m, 2H, H-4, H-6a),
3.96 (m, 2H, H-2, H-5), 4.07 (m, 1H, H-6b), 4.25 (d, 1H, J ¼ 12.3 Hz,
H-10b), 4.52e4.63 (m, 3H, H-50, H-60a, H-60b), 5.42 (m, 2H, H-3, H-
40), 5.61 (m, 1H, H-30), 6.21 (d, 1H, J ¼ 3.6 Hz, H-1); trans-feruloyl
units: R1: 2.33 (s, 3H, H-1100), 3.86 (s, 3H, eOCH3), 6.38 (d, 1H,
J ¼ 15.9 Hz, H-800), 7.02e7.15 and 7.28e7.33 (2ꢀ m, 3H, H-200, H-500,
H-600), 7.61 (d, 1H, J ¼ 15.9 Hz, H-700), R2: 2.33 (s, 3H, H-1100), 3.88 (s,
3H, eOCH3), 6.42 (d, 1H, J ¼ 15.9 Hz, H-800), 7.02e7.15 and 7.28e7.33
(2ꢀ m, 3H, H-200, H-500, H-600), 7.66 (d, 1H, J ¼ 15.9 Hz, H-700), R3: 2.33
(s, 3H, H-1100), 3.89 (s, 3H, eOCH3), 6.43 (d, 1H, J ¼ 15.9 Hz, H-800),
7.02e7.15 and 7.28e7.33 (2ꢀ m, 3H, H-200, H-500, H-600), 7.69 (d, 1H,
J ¼ 15.9 Hz, H-700), R4: 2.33 (s, 3H, H-1100), 3.92 (s, 3H, eOCH3), 6.57
(d, 1H, J ¼ 15.9 Hz, H-800), 7.02e7.15 and 7.28e7.33 (2ꢀ m, 3H, H-200,
H-500, H-600), 7.93 (d, 1H, J ¼ 15.9 Hz, H-700); 13C NMR (75.48 MHz,
(d,1H, J ¼ 15.9 Hz, H-700); 13C NMR (75.48 MHz, CDCl3)
d (ppm): 19.1,
24.1, 25.4, 29.1 (4ꢀ (CH3)2C), 62.1 (C-6), 63.8 (C-5), 65.7 (C-60), 65.9
(C-10), 70.3 (C-3), 72.9 (C-4), 73.8 (C-2), 76.5 (C-40), 81.3 (C-30), 81.4
(C-50), 90.9 (C-1), 99.9, 101.8 (2ꢀ (CH3)2C), 104.5 (C-20); trans-p-
feruloyl units: R1: 20.7 (C-1100), 55.9 (eOCH3), 111.2 (C-200), 116.5
(C-800), 121.4 (C-500), 123.4 (C-600), 132.8 (C-100), 141.6 (C-300), 144.6
(C-700),151.4 (C-400),166.6 (C-900),168.8 (C-1000); R2: 20.7 (C-1100), 56.1
(eOCH3), 111.4 (C-200), 117.9 (C-800), 122.1 (C-500), 123.4 (C-600), 133.3
(C-100), 142.0 (C-300), 146.6 (C-700), 151.5 (C-400), 167.7 (C-900), 168.þ8
(C-1000); LC-MS (ESI): m/z 881.35 [M þ Na]þ; HR-MS: m/z [M þ Na]
calcd for C42H50O19Na: 881.2839, found: 881.2860.
CDCl3)
d
(ppm): 19.0, 23.9, 25.4, 28.8 (4ꢀ (CH3)2C), 62.1 (C-6), 64.3
(C-5), 65.0 (C-60), 66.2 (C-10), 70.9 (C-3), 71.5 (C-4), 71.9 (C-2), 77.4
(C-40), 77.7 (C-30), 80.3 (C-50), 91.7 (C-1), 99.6, 101.5 (2ꢀ (CH3)2C),
105.0 (C-20); trans-p-feruloyl units: R1: 20.6 (C-1100), 55.9 (eOCH3),
111.2 (C-200), 116.8 (C-800), 121.2 (C-500), 123.0 (C-600), 132.9 (C-100),
141.4 (C-300), 144.1 (C-700), 151.3 (C-400), 165.7 (C-900), 168.7 (C-1000),
R2: 20.6 (C-1100), 55.9 (eOCH3), 111.2 (C-200), 116.9 (C-800), 121.3
(C-500), 123.2 (C-600), 133.3 (C-100), 141.4 (C-300), 144.4 (C-700), 151.4
(C-400), 165.7 (C-900), 168.8 (C-1000), R3: 20.6 (C-1100), 56.0 (eOCH3),
111.3 (C-200), 117.9 (C-800), 121.5 (C-500), 123.2 (C-600), 133.3 (C-100),
141.6 (C-300), 145.7 (C-700), 151.4 (C-400), 166.0 (C-900), 168.8 (C-1000),
R4: 20.6 (C-1100), 56.0 (eOCH3), 112.4 (C-200), 118.2 (C-800), 121.7
(C-500), 123.3 (C-600), 133.4 (C-100), 141.8 (C-300), 146.6 (C-700), 151.4 (C-
400), 166.3 (C-900), 168.8 (C-1000); LC-MS (ESI): m/z 1317.35 [M þ Na]þ;
HR-MS: m/z [M þ Na]þ calcd for C66H70O27Na: 1317.3997, found:
1317.3980.
4.1.1.4. 30,40,60-tri-O-acetoxyferuloyl-2,10:4,6-di-O-isopropylidene
sucrose 9. Following the general procedure, the reaction between
di-O-isopropylidene 4 (1.0 g, 2.4 mmol) and p-acetoxyferuloyl
chloride 5 (2.0 g, 7.8 mmol) in dry pyridine (10 mL) for 2 days
furnished compound 9 as a white solid (0.67 g, 26% yield) along
with compound 8 in 44% (0.89 g) yield. Analytical data for 9:
Rf ¼ 0.73 (EtOAc/CH2Cl2 3:2); mp: 135e138 ꢂC; FT-IR (KBr) nmax
:
3507, 2993, 2942, 1766, 1721, 1638, 16011510, 1467, 1418, 1371, 1332,
1259, 1198, 1155, 1124, 1067, 1032, 1011, 944, 904, 858, 837, 726,
650 cmꢁ1 1H NMR (300 MHz, CDCl3)
; d (ppm): 1.33, 1.41, 1.51, 1.53
(4ꢀ s, 12H, (CH3)2C), 3.62 (m, 2H, H-10a, H-4), 3.70 (m, 1H, H-6a),
3.76 (m, 1H, H-2), 3.84 (m, 2H, H-3, H-5), 4.03 (m, 1H, H-6b), 4.19 (d,
1H, J ¼ 12.0 Hz, H-10b), 4.51 (m, 2H, H-50, H-60a), 4.64 (m,1H, H-60b),
5.38 (d, 1H, J ¼ 5.4 Hz, H-40), 5.60 (br dd,1H, J ¼ 5.1 Hz, 3.6 Hz, H-30),
6.15 (d, 1H, J ¼ 3.3 Hz, H-1); trans-p-feruloyl units: R1: 2.34 (s, 3H,
H-1100), 3.86 (s, 3H, eOCH3), 6.40 (d,1H, J ¼ 15.9 Hz, H-800), 7.02e7.13
4.1.2. Acetal deprotection of the diacetonoides 8e10
4.1.2.1. General procedure. A solution of di-O-isopropylidene fer-
uloyl derivatives 8e10 in 60% aq. AcOH was heated at 80 ꢂC until the
reaction was completed. The reaction was monitored by TLC

P. Panda et al. / European Journal of Medicinal Chemistry 58 (2012) 418e430
425
(EtOAc/CH2Cl2 3:2). The reaction solution was then evaporated to
dryness under reduced pressure by codistillation with toluene
(3 ꢀ 100 mL). The products 11e13 were obtained from recrystalli-
zation in EtOAc and/or by column chromatography using a gradient
of CH2Cl2/EtOAc as eluent.
147.0 (C-700), 152.9 (C-400), 168.0 (C-900), 170.4 (C-1000); LC-MS (ESI):
m/z 1019.23 [M þ Na]þ; HR-MS: m/z [M þ Na]þ calcd for
C48H52O23Na: 1019.2792, found: 1019.2774.
4.1.2.4. 3,30,40,60-tetra-O-acetoxyferuloylsucrose 13. Following the
general procedure, treatment of a solution of compound 10 (0.6 g,
0.4 mmol) with 60% aq. AcOH (32 mL) at 80 ꢂC for 35 min followed
by column chromatographic purification using a gradient of CH2Cl2/
EtOAc as eluent yielded compound 13 as a white solid (0.45 g, 87%
yield). Analytical data for 13: Rf ¼ 0.87 (EtOAc/MeOH 9:1); mp:
127e129 ꢂC; FT-IR (KBr) nmax: 3482, 2963, 2941, 1765, 1717, 1637,
1601, 1510, 1467, 1420, 1371, 1325, 1260, 1154, 1123, 1032, 1008, 945,
4.1.2.2. 30,60-di-O-acetoxyferuloylsucrose 11. Following the general
procedure, a solution of compound 8 (1.1 g, 1.3 mmol) was treated
with 60% aq. AcOH (66 mL) at 80 ꢂC for 20 min. Recrystallization of
the crude product in EtOAc gave compound 11 as a white solid
(0.90 g, 89% yield). Analytical data for 11: Rf ¼ 0.56 (EtOAc/MeOH
9:1); mp: 128e130 ꢂC; FT-IR (KBr) nmax: 3411, 2930, 1762, 1706,
1635,1601,1508,1420,1371,1330,1261,1220,1194, 1159,1125,1060,
1031, 996, 938, 848, 792, 691, 650 cmꢁ1; 1H NMR (300 MHz, CD3OD)
904, 832, 797, 704, 649 cmꢁ1 1H NMR (300 MHz, CDCl3)
; d (ppm):
3.61 (dd, 1H, J ¼ 9.6 Hz, 9.3 Hz, H-4); 3.73 (m, 1H, H-2), 3.84 (m, 3H,
H-10b, H-10a, H-6a), 4.01 (m, 1H, H-6b), 4.12 (m, 1H, H-5), 4.50 (m,
1H, H-50), 4.56 (m, 1H, H-60a), 4.67 (dd, 1H, J ¼ 7.2 Hz, 11.4 Hz, H-
60b), 5.13 (dd, 1H, J ¼ 9.6 Hz, 9.3 Hz, H-3), 5.56 (d, 1H, J ¼ 2.7 Hz, H-
1), 5.59 (d, 1H, J ¼ 5.4 Hz, H-30), 5.71 (app t, 1H, J ¼ 4.8 Hz, H-40);
trans-p-feruloyl units: R1: 2.29 (s, 3H, H-1100), 3.82 (s, 3H, eOCH3),
6.39 (d, 1H, J ¼ 15.9 Hz, H-800), 7.00e7.09 (m, 3H, H-200, H-500, H-600),
7.59 (d, 1H, J ¼ 15.9 Hz, H-700), R2: 2.30 (s, 3H, H-1100), 3.83 (s, 3H, e
OCH3), 6.39 (d, 1H, J ¼ 15.9 Hz, H-800), 7.00e7.09 (m, 3H, H-200, H-500,
H-600), 7.68 (d, 1H, J ¼ 15.9 Hz, H-700), R3: 2.31 (s, 3H, H-1100), 3.83 (s,
3H, eOCH3), 6.42 (d, 1H, J ¼ 15.9 Hz, H-800), 7.00e7.09 (m, 3H, H-200,
H-500, H-600), 7.19 (d, 1H, J ¼ 8.4 Hz, H-200), 7.69 (d, 1H, J ¼ 15.9 Hz, H-
700), R4: 2.32 (s, 3H, H-1100), 3.85 (s, 3H, eOCH3), 6.57 (d, 1H,
J ¼ 15.9 Hz, H-800), 7.00e7.09 (m, 1H, H-600), 7.19 (d, 1H, J ¼ 8.4 Hz, H-
500), 7.24 (br s, 1H, H-200), 7.84 (d, 1H, J ¼ 15.9 Hz, H-700); 13C NMR
d
(ppm): 3.40e3.46 (m, 2H, H-2, H-4), 3.62 (m, 2H, H-10a, H-10b),
3.70 (m, 1H, H-3), 3.80 (m, 1H, H-6a), 3.94 (m, 2H, H-6b, H-5), 4.16e
4.22 (m, 1H, H-50), 4.47 (app t, 1H, J ¼ 7.8 Hz, H-40), 4.53e4.61 (m,
2H, H-60a, H-60b), 5.45 (d, 1H, J ¼ 3.6 Hz, H-1), 5.53 (d, 1H, J ¼ 7.8 Hz,
H-30); trans-p-feruloyl units: R1: 2.27 (s, 3H, H-1100), 3.86 (s, 3H, e
OCH3), 6.58 (d, 1H, J ¼ 16.2 Hz, H-800), 7.07 (d, 1H, J ¼ 8.1 Hz, H-600),
7.22 (dd, 1H, J ¼ 1.5 Hz, 8.4 Hz, H-500), 7.35 (d, 1H, J ¼ 13.8 Hz, H-200),
7.72 (d, 1H, J ¼ 16.2 Hz, H-700), R2: 2.27 (s, 3H, H-1100), 3.86 (s, 3H, e
OCH3), 6.61 (d, 1H, J ¼ 16.2 Hz, H-800), 7.07 (d, 1H, J ¼ 8.1 Hz, H-600),
7.22 (dd, 1H, J ¼ 1.5 Hz, 8.4 Hz, H-500), 7.35 (d, 1H, J ¼ 13.8 Hz, H-200),
7.78 (d, 1H, J ¼ 16.2 Hz, H-700); 13C NMR (75.48 MHz, CD3OD)
d
(ppm): 62.7 (C-6), 65.2 (C-10), 66.5 (C-60), 71.5 (C-4), 73.2 (C-2),
74.5 (C-5), 75.0 (C-40, C-3), 79.5 (C-30), 81.3 (C-50), 93.2 (C-1), 105.1
(C-20); trans-p-feruloyl units: R1: 20.9 (C-1100), 56.6 (eOCH3), 112.7
(C-200), 118.7 (C-800), 122.4 (C-500), 124.3 (C-600), 134.8 (C-100), 143.1 (C-
300), 146.0 (C-700), 153.0 (C-400), 167.6 (C-900), 170.5 (C-1000), R2: 20.9
(C-1100), 56.6 (eOCH3), 113.2 (C-200), 118.9 (C-800), 122.4 (C-500), 124.3
(C-600), 134.8 (C-100), 143.1 (C-300), 146.6 (C-700), 153.1 (C-400), 168.4 (C-
900), 170.5 (C-1000); LC-MS (ESI): m/z 801.25 [M þ Na]þ; HR-MS: m/z
[M þ Na]þ calcd for C36H42O19Na: 801.2213, found: 801.2235.
(75.48 MHz, CDCl3) d
(ppm): 62.5 (C-6), 64.4 (C-10), 64.5 (C-60), 69.6
(C-4), 70.6 (C-2), 73.6 (C-5), 76.5 (C-40), 77.0 (C-3), 78.1 (C-30), 79.8
(C-50), 92.5 (C-1), 105.5 (C-20); trans-p-feruloyl units: R1: 20.7
(C-1100), 55.9 (eOCH3), 111.3 (C-200), 116.4 (C-800), 121.4 (C-500), 123.2
(C-600), 132.8 (C-100), 141.6 (C-300), 145.4 (C-700), 151.3 (C-400), 165.7
(C-900), 168.4 (C-1000), R2: 20.7 (C-1100), 55.9 (eOCH3), 111.4 (C-200),
116.5 (C-800), 121.5 (C-500), 123.2 (C-600), 132.9 (C-100), 141.7 (C-300),
145.6 (C-700), 151.4 (C-400), 165.7 (C-900), 168.7 (C-1000), R3: 20.7
(C-1100), 55.9 (eOCH3), 111.4 (C-200), 117.4 (C-800), 121.6 (C-500), 123.3
(C-600), 133.0 (C-100), 141.8 (C-300), 146.2 (C-700), 151.4 (C-400), 166.7
(C-900), 168.7 (C-1000), R4: 20.7 (C-1100), 56.0 (eOCH3), 112.0 (C-200),
117.4 (C-800), 121.9 (C-500), 123.3 (C-600), 133.0 (C-100), 141.9 (C-300),
147.2 (C-700), 151.5 (C-400), 166.8 (C-900), 168.8 (C-1000); LC-MS (ESI):
m/z 1237.28 [M þ Na]þ; HR-MS: m/z [M þ Na]þ calcd for
4.1.2.3. 30,40,60-tri-O-acetoxyferuloylsucrose
12. Following
the
general procedure, a solution of compound 9 (0.8 g, 0.7 mmol) was
reacted with 60% aq. AcOH (49 mL) at 80 ꢂC for 20 min. Column
chromatographic purification using a gradient of CH2Cl2/EtOAc as
eluent afforded compound 12 as a white solid (0.50 g, 67% yield).
Analytical data for 12: Rf ¼ 0.49 (EtOAc/MeOH 9:1); mp: 128e
130 ꢂC; FT-IR (KBr) nmax: 3425, 2938, 1764, 1713, 1638, 1601, 1510,
1465, 1418, 1371, 1332, 1300, 1260, 1218, 1199, 1155, 1124, 1031, 1012,
C60H62O27Na: 1237.3371, found: 1237.3373.
903, 836, 650 cmꢁ1; 1H NMR (300 MHz, CD3OD)
d (ppm): 3.46e3.52
(m, 2H, H-2, H-4), 3.67 (m, 2H, H-10a, H-3), 3.82 (m, 1H, H-10b), 3.88
(m, 1H, H-6a), 3.99e4.08 (m, 2H, H-6b, H-5), 4.47e4.51 (m, 1H, H-
50), 4.61e4.65 (m, 2H, H-60b, H-60a), 5.49 (d, 1H, J ¼ 3.6 Hz, H-1),
5.82e5.87 (m, 2H, H-30, H-40); trans-p-feruloyl units: R1: 2.26 (s, 3H,
H-1100), 3.74 (s, 3H, eOCH3), 6.47 (d,1H, J ¼ 15.9 Hz, H-800), 6.93e7.10
(m, 2H, H-500, H-600), 7.20e7.25 (m, 1H, H-200), 7.63 (d, 1H, J ¼ 15.9 Hz,
H-700), R2: 2.26 (s, 3H, H-1100), 3.78 (s, 3H, eOCH3), 6.53 (d, 1H,
J ¼ 15.9 Hz, H-800), 6.93e7.10 (m, 2H, H-500, H-600), 7.20e7.25 (m, 1H,
H-200), 7.64 (d, 1H, J ¼ 15.9 Hz, H-700), R3: 2.26 (s, 3H, H-1100), 3.84 (s,
3H, eOCH3), 6.57 (d, 1H, J ¼ 15.9 Hz, H-800), 6.93e7.10, 7.20e7.25
(2ꢀ m, 2H, H-500, H-600), 7.34 (br s, 1H, H-200), 7.76 (d, 1H,
4.1.3. Deacetylation of compounds 9, 11e13
4.1.3.1. General procedure. To a separate suspension of acetoxyfer-
uloyl compounds 9, 11e13 in 95% EtOH, piperidine was added,
whereupon the solution turned yellow. After dissolving the starting
material completely, the reaction was allowed to continue until the
disappearance of the starting material. The reaction was monitored
by TLC analysis (EtOAc/MeOH 9:1). The mixture was quenched with
AcOH and was evaporated to a syrup. It was subjected to silica gel
column chromatography using a gradient of CH2Cl2/EtOAc/MeOH
as eluent. The solvent was evaporated under diminished pressure
to furnish deacetylated products 1, 2, 14 and 15 as a white solid.
J ¼ 15.9 Hz, H-700); 13C NMR (75.48 MHz, CD3OD)
d (ppm): 62.3
(C-6), 64.6 (C-10), 65.8 (C-60), 71.2 (C-4), 73.1 (C-2), 74.6 (C-5), 75.0
(C-3), 77.3 (C-40), 77.4 (C-30), 78.9 (C-50), 93.6 (C-1), 105.8 (C-20);
4.1.3.2. 30,60-di-O-feruloylsucrose (helonioside A, 1). Following the
general procedure, a suspension of compound 11 (0.7 g, 0.9 mmol)
in 95% EtOH (47 mL) was treated with piperidine (335.0 mL, 0.3 g,
trans-p-feruloyl units:
d
¼ R1: 20.5 (C-1100), 56.5 (eOCH3), 112.5
(C-200), 118.1 (C-800), 122.5 (C-500), 124.3 (C-600), 134.4 (C-100), 143.0
(C-300), 146.2 (C-700), 152.9 (C-400),167.2 (C-900), 170.4 (C-1000), R2: 20.5
(C-1100), 56.6 (eOCH3), 112.8 (C-200), 118.3 (C-800), 122.5 (C-500), 124.3
(C-600), 134.5 (C-100), 143.1 (C-300), 147.0 (C-700), 152.9 (C-400), 167.5
(C-900), 170.4 (C-1000), R3: 20.5 (C-1100), 56.6 (eOCH3), 113.2 (C-200),
118.6 (C-800), 122.6 (C-500), 124.3 (C-600), 134.7 (C-100), 143.2 (C-300),
3.4 mmol) for 3 h to afford compound 1 as a white solid (0.40 g, 68%
yield). Analytical data for 1: Rf ¼ 0.49 (EtOAc/MeOH 9:1); mp: 154e
156 ꢂC; FT-IR (KBr) nmax: 3417, 2938, 1720, 1691, 1632, 1519, 1454,
1431,1379,1273,1151,1057,1030, 995, 938, 841, 822, 788, 696 cmꢁ1
;
1H NMR (300 MHz, CD3OD)
d (ppm): 3.39 (m, 1H, H-4), 3.44 (dd, 1H,
J ¼ 3.6 Hz, 6.0 Hz, H-2), 3.61 (m, 2H, H-10b, H-10a), 3.69 (m, 1H, H-3),

426
P. Panda et al. / European Journal of Medicinal Chemistry 58 (2012) 418e430
3.80 (dd, 1H, J ¼ 4.2 Hz, 11.7 Hz, H-6a), 3.88 (m, 1H, H-6b), 3.96 (m,
1H, H-5), 4.16 (m, 1H, H-50), 4.45 (dd, 1H, J ¼ 7.6 Hz, 7.8 Hz, H-40),
4.57 (m, 2H, H-60b, H-60a), 5.45 (d, 1H, J ¼ 3.6 Hz, H-1), 5.50 (d, 1H,
J ¼ 7.8 Hz, H-30), trans-p-feruloyl units: R1: 3.90 (s, 3H, eOCH3), 6.41
(d, 1H, J ¼ 15.9 Hz, H-800), 6.82 (d, 1H, J ¼ 8.1 Hz, H-500), 7.14 (dd, 1H,
J ¼ 1.5 Hz, 8.4 Hz, H-600), 7.24 (d, 1H, J ¼ 1.5 Hz, H-200), 7.66 (d, 1H,
J ¼ 15.9 Hz, H-700), R2: 3.90 (s, 3H, eOCH3), 6.44 (d, 1H, J ¼ 15.9 Hz,
H-800), 6.82 (d, 1H, J ¼ 8.1 Hz, H-500), 7.10 (dd, 1H, J ¼ 1.5 Hz, 8.4 Hz,
H-600), 7.20 (d, 1H, J ¼ 1.5 Hz, H-200), 7.72 (d, 1H, J ¼ 15.9 Hz, H-700);
J ¼ 8.1 Hz, H-500), 6.95e7.08 (m, 2H, H-200, H-600), 7.59 (d, 1H,
J ¼ 15.9 Hz, H-700), R3: 3.87 (s, 3H, eOCH3), 6.42 (d, 1H, J ¼ 15.9 Hz,
H-800), 6.79 (d, 1H, J ¼ 8.1 Hz, H-500), 7.16 (m, 2H, H-200, H-600), 7.60 (d,
1H, J ¼ 15.9 Hz, H-700), R4: 3.88 (s, 3H, eOCH3), 6.46 (d, 1H,
J ¼ 15.9 Hz, H-800), 6.81 (d,1H, J ¼ 8.1 Hz, H-500), 6.95e7.08 (m,1H, H-
600), 7.28 (br d, 1H, J ¼ 1.8 Hz, H-200), 7.74 (d, 1H, J ¼ 15.9 Hz, H-700);
13C NMR (75.48 MHz, CD3OD)
d
(ppm): 62.0 (C-6), 64.5 (C-10), 65.7
(C-60), 69.3 (C-4), 71.7 (C-2), 74.8 (C-5), 77.0, 77.1 (C-3, C-30, C-40),
78.8 (C-50), 93.7 (C-1), 105.8 (C-20); trans-p-feruloyl units: R1: 56.4
(eOCH3), 111.6 (C-200), 114.4 (C-800), 116.0 (C-500), 124.0 (C-600), 127.5
(C-100), 146.9 (C-700), 149.3 (C-300), 150.6 (C-400), 168.1 (C-900), R2: 56.5
(eOCH3), 111.8 (C-200), 114.8 (C-800), 116.5 (C-500), 124.3 (C-600), 127.6
(C-100), 147.5 (C-700), 149.4 (C-300), 150.7 (C-400), 168.2 (C-900), R3: 56.5
(eOCH3), 111.8 (C-200), 115.0 (C-800), 116.5 (C-500), 124.4 (C-600), 127.8
(C-100), 147.5 (C-700), 149.4 (C-300), 150.7 (C-400), 168.8 (C-900), R4: 56.6
(eOCH3), 112.7 (C-200), 115.0 (C-800), 116.6 (C-500), 124.5 (C-600), 127.9
(C-100), 148.3 (C-700), 149.4 (C-300), 150.9 (C-400), 169.3 (C-900); LC-MS
(ESI): m/z 1069.25 [M þ Na]þ; HR-MS: m/z [M þ Na]þ calcd for
13C NMR (75.48 MHz, CD3OD)
d
(ppm): 62.7 (C-6), 65.2 (C-10), 66.2
(C-60), 71.4 (C-4), 73.2 (C-2), 74.4 (C-5), 75.0 (C-40, C-3), 79.2 (C-30),
81.3 (C-50), 93.1 (C-1), 105.1 (C-20); trans-p-feruloyl units: R1: 56.5
(eOCH3), 111.7 (C-200), 114.9 (C-800), 116.5 (C-500), 124.3 (C-600), 127.7
(C-100), 147.3 (C-700), 149.4 (C-300), 150.7 (C-400), 168.3 (C-900), R2: 56.5
(eOCH3), 112.1 (C-200), 115.1 (C-800), 116.5 (C-500), 124.3 (C-600), 127.7
(C-100), 147.8 (C-700), 149.4 (C-300), 150.7 (C-400), 169.1 (C-900); LC-MS
(ESI): m/z 717.21 [M þ Na]þ; HR-MS: m/z [M þ Na]þ calcd for
C
32H38O17Na: 717.2001, found: 717.1984. Spectral data of helonio-
side A 1 was the same as reported for the isolated natural product
C52H54O23Na: 1069.2948, found: 1069.2926.
[2,6].
4.1.3.5. 30,40,60-tri-O-feruloyl-2,10:4,6-di-O-isopropylidene sucrose 15.
Following the general procedure, a suspension of compound 9
(0.3 g, 0.3 mmol) in 95% EtOH (23 mL) was treated with piperidine
4.1.3.3. 30,40,60-tri-O-feruloyl sucrose 2. Following the general
procedure, a suspension of compound 12 (0.4 g, 0.4 mmol) in 95%
EtOH (28 mL) was treated with piperidine (232.0
mL, 0.2 g,
(176.0 mL, 0.2 g, 1.8 mmol) for 4 h to give compound 15 as a white
2.3 mmol) for 4 h to afford compound 2 [15] as a white solid (0.22 g,
65% yield). Analytical data for 2: Rf ¼ 0.46 (EtOAc/MeOH 9:1); mp:
99e101 ꢂC; FT-IR (KBr) nmax: 3355, 2969, 1715, 1699, 1653, 1595,
solid (0.20 g, 71% yield). Analytical data for 15: Rf ¼ 0.27 (EtOAc/
hexanes 3:1); mp: 145e147 ꢂC; FT-IR (KBr) nmax: 3067, 2991, 2939,
1715, 1632, 1593, 1516, 1465, 1431, 1383, 1271, 1211, 1154, 1065, 1031,
1517, 1457, 1430, 1272, 1157, 1034, 992, 845, 815 cmꢁ1
;
1H NMR
942, 856, 816, 724, 655, 603 cmꢁ1 1H NMR (300 MHz, CDCl3)
;
(300 MHz, CD3OD)
d
(ppm): 3.52 (m, 2H, H-2, H-4), 3.73 (m, 2H, H-
d
(ppm): 1.31, 1.37, 1.49 (3ꢀ s, 12H, (CH3)2C); 3.62 (m, 3H, H-10a, H-4,
10b, H-10a), 3.80 (m, 1H, H-3), 3.90 (m, 1H, H-6a), 4.05 (m, 2H, H-6b,
H-5), 4.47 (m, 1H, H-50), 4.62 (m, 2H, H-60b, H-60a), 5.51 (d, 1H,
J ¼ 3.3 Hz, H-1), 5.81 (m, 2H, H-30, H-40), trans-p-feruloyl units: R1:
3.72 (s, 3H, eOCH3), 6.29 (m, 1H, H-800), 6.74 (m, 1H, H-500), 6.94 (m,
2H, H-200, H-600), 7.54 (d, 1H, J ¼ 15.9 Hz, H-700), R2: 3.77 (s, 3H, e
OCH3), 6.29 (m, 1H, H-800), 6.74 (m, 1H, H-500), 7.08 (br d, 1H,
J ¼ 8.4 Hz, H-600), 7.17 (br s, 1H, H-200), 7.54 (d, 1H, J ¼ 15.9 Hz, H-700),
R3: 3.83 (s, 3H, eOCH3), 6.41 (d, 1H, J ¼ 15.9 Hz, H-800), 6.74 (m, 1H,
H-500), 6.94 (m, 2H, H-200, H-600), 7.70 (d, 1H, J ¼ 15.9 Hz, H-700); 13C
H-6a); 3.75 (m, 1H, H-2); 3.87 (m, 2H, H-3, H-5), 4.02 (m, 1H, H-6b),
4.14 (m, 1H, H-10b), 4.49 (m, 2H, H-50, H-60a), 4.57 (m, 1H, H-60b),
5.35 (br d, 1H, J ¼ 5.1 Hz, H-40), 5.59 (br s, 1H, H-30), 6.16 (m, 1H, H-
1); trans-p-feruloyl units: R1: 3.84 (s, 3H, eOCH3), 6.24 (d, 1H,
J ¼ 15.9 Hz, H-800), 6.83e7.08 (m, 3H, H-200, H-500, H-600), 7.59 (d, 1H,
J ¼ 15.9 Hz, H-700), R2: 3.87 (s, 3H, eOCH3), 6.26 (d, 1H, J ¼ 15.9 Hz,
H-800), 6.83e7.08 (m, 3H, H-200, H-500, H-600), 7.59 (d, 1H,
J ¼ 15.9 Hz, H-700), R3: 3.92 (s, 3H, eOCH3), 6.37 (d, 1H, J ¼ 15.9 Hz,
H-800), 6.83e7.08 (m, 3H, H-200, H-500, H-600), 7.71 (d, 1H, J ¼ 15.9 Hz,
NMR (75.48 MHz, CD3OD)
d
(ppm): 62.4 (C-6), 64.8 (C-10), 65.7 (C-
H-700); 13C NMR (75.48 MHz, CDCl3)
d (ppm): 19.1, 24.1, 25.5, 28.9
60), 1.2 (C-4), 73.2 (C-2), 74.6 (C-5), 75.0 (C-3), 77.1 (C-30, C-40), 78.9
(C-50), 93.6 (C-1), 105.8 (C-20); trans-p-feruloyl units: R1: 56.4 (e
OCH3), 111.6 (C-200), 114.4 (C-800), 116.5 (C-500), 124.3 (C-600), 127.4
(C-100), 148.2 (C-700), 149.2 (C-300), 150.6 (C-400), 168.2 (C-900), R2: 56.4
(eOCH3), 111.8 (C-200), 114.6 (C-800), 116.5 (C-500), 124.4 (C-600), 127.5
(C-100), 147.4 (C-700), 149.2 (C-300), 150.7 (C-400), 168.7 (C-900), R3: 56.5
(eOCH3), 112.1 (C-200), 114.9 (C-800), 116.4 (C-500), 124.4 (C-600), 127.6
(C-100), 147.4 (C-700), 149.2 (C-300), 150.7 (C-400), 167.9 (C-900); LC-MS
(ESI): m/z 893.23 [M þ Na]þ; HR-MS: m/z [M þ Na]þ calcd for
(4ꢀ (CH3)2C), 62.0 (C-6), 63.9 (C-5), 64.9 (C-60), 66.3 (C-10), 70.2
(C-3), 72.8 (C-4), 73.8 (C-2), 77.1 (C-40), 77.3 (C-30), 79.9 (C-50), 91.4
(C-1), 99.8, 101.8 (2ꢀ (CH3)2C), 104.7 (C-20); trans-p-feruloyl units:
R1: 55.8 (eOCH3), 109.4 (C-200), 113.6 (C-800), 114.8 (C-500), 123.2
(C-600), 126.5 (C-100), 145.2 (C-700), 146.8 (C-300), 148.0 (C-400), 166.2
(C-900), R2: 55.9 (eOCH3), 109.4 (C-200), 113.9 (C-800), 114.9 (C-500),
123.4 (C-600), 126.6 (C-100), 146.5 (C-700), 146.8 (C-300), 148.4 (C-400),
166.4 (C-900), R3: 56.0 (eOCH3), 109.5 (C-200), 113.9 (C-800), 114.9
(C-500), 124.1 (C-600), 126.9 (C-100), 146.9 (C-300), 147.2 (C-700), 148.5
(C-400), 166.8 (C-900); LC-MS (ESI): m/z 973.27 [M þ Na]þ; HR-MS:
m/z [M þ Na]þ calcd for C48H54O20Na: 973.3101, found: 973.3078.
C
42H46O20Na: 893.2475, found: 893.2478.
4.1.3.4. 3,30,40,60-tetra-O-feruloyl sucrose 14. Following the general
procedure, a suspension of compound 13 (0.2 g, 0.2 mmol) in 95%
4.1.4. Synthesis of lapathoside C and its analogous
EtOH (16 mL) was reacted with piperidine (150.0
m
L, 0.1 g,
4.1.4.1. Preparation of 6-mono-O-acetoxyferuloyl-30,60-di-O-acetox-
ycinnamoylsucrose 18 and 3,6-di-O-acetoxyferuloyl-30,60-di-O-ace-
toxycinnamoylsucrose 19. 30,60-di-O-acetoxycinnamoyl sucrose 16
(1.1 g, 1.5 mmol) was dissolved in dry CH2Cl2 (21 mL) to which 4 Å
molecular sieves powder and dry pyridine (1.1 g, 1.2 mL, 14.4 mmol)
were added. The solution was then cooled to 0 ꢂC in an ice bath. p-
Acetoxyferuloyl chloride 5 (0.4 g, 1.6 mmol) was then added slowly
at the same temperature and the reaction was left to stir while
warming to r.t. Stirring was continued until the reaction was
completed as indicated by TLC analysis (EtOAc/hexanes 3:1). After
24 h, the resulting mixture was poured into vigorously stirred ice-
water (100 mL) and the white solid precipitated was obtained by
filtration. The precipitate was redissolved in EtOAc (25 mL) and
1.5 mmol) for 4 h to afford compound 14 as a white solid (0.15 g, 76%
yield). Analytical data for 14: Rf ¼ 0.73 (EtOAc/MeOH 9:1); mp:
123e125 ꢂC; FT-IR (KBr) nmax: 3423, 2964, 2940, 1708, 1631, 1594,
1515, 1452, 1431, 1372, 1271, 1158, 1032, 1003, 846, 819, 703,
603 cmꢁ1; 1H NMR (300 MHz, CD3OD)
d (ppm): 3.65e3.95 (m, 5H,
H-10b, H-10a, H-2, H-4, H-6a), 4.03e4.07 (m, 1H, H-6b), 4.18e4.23
(m, 1H, H-5), 4.46e4.50 (m, 1H, H-50), 4.58e4.64 (m, 2H, H-60b,
H-60a), 5.45 (app t, 1H, J ¼ 9.6 Hz, H-3), 5.58 (d, 1H, J ¼ 3.3 Hz, H-1),
5.86 (m, 2H, H-30, H-40); trans-p-feruloyl units: R1: 3.76 (s, 3H, e
OCH3), 6.29 (d, 1H, J ¼ 15.9 Hz, H-800), 6.71 (d, 1H, J ¼ 8.1 Hz, H-500),
6.95e7.08 (m, 2H, H-200, H-600), 7.57 (d, 1H, J ¼ 15.9 Hz, H-700), R2:
3.81 (s, 3H, eOCH3), 6.34 (d, 1H, J ¼ 15.9 Hz, H-800), 6.76 (d, 1H,

P. Panda et al. / European Journal of Medicinal Chemistry 58 (2012) 418e430
427
washed with 1 N HCl (2 ꢀ 50 mL). The aqueous layer was extracted
with EtOAc (25 mL). The combined organic layers were then
successively washed with 5% NaHCO3 (2 ꢀ 50 mL), brine (25 mL)
and then dried over anhyd. MgSO4. The EtOAc solution was
concentrated to residue that was subjected to column chromatog-
raphy using a gradient of CH2Cl2/EtOAc as eluent and further,
purified by PTLC afforded compound 18 as a white solid (0.50 g, 36%
yield) along with compound 19 in 7% (0.12 g) yield. Analytical data
for 18: Rf ¼ 0.06 (EtOAc/hexanes 3:1); mp: 108e110 ꢂC; FT-IR (KBr)
nmax: 3457, 3423, 2925, 2852, 1767, 1710, 1636, 1602, 1508, 1457,
1419, 1371, 1323, 1282, 1261, 1206, 1165, 1056, 1015, 946, 912, 836,
4.1.4.2. Preparation of6-mono-O-feruloyl-30,60-di-O-coumaroylsucrose
3 and 3,6-di-O-feruloyl-30,60-di-O-coumaroylsucrose 21
4.1.4.2.1. General procedure. Pyrrolidine were added to
a suspension of compounds 18 and 19 in 95% EtOH. Consequently, it
caused the solution to turn yellow. The starting material typically
dissolved within 15 min and the reaction was allowed to continue
until the disappearance of the starting material as indicated by TLC
analysis (EtOAc). This mixture was directly added to a column of
strongly acidic ion-exchange resin [Amberlite IRA-120 (Hþ) washed
and packed in 95% EtOH. The appropriate fractions were concen-
trated under diminished pressure to a residue that was subjected to
column chromatography using a gradient of CH2Cl2/EtOAc/MeOH
afforded compounds 3 and 21.
794, 754, 649, 595 cmꢁ1; 1H NMR (300 MHz, CDCl3)
d (ppm): 3.32
(m, 1H, H-4), 3.54 (m, 2H, H-2, H-10a), 3.68 (m, 2H, H-10b, H-3), 4.11
(m, 1H, H-5), 4.24 (m, 1H, H-50), 4.46 (m, 1H, H-40), 4.55 (m, 4H, H-
6b, H-6a, H-60b, H-60a), 5.30 (d, 1H, J ¼ 5.7 Hz, H-30), 5.42 (br s, 1H,
H-1); trans-p-feruloyl units: R1: 2.28 (s, 3H, H-1100), 3.77 (s, 3H, e
OCH3), 6.41 (d, 1H, J ¼ 15.9 Hz, H-800), 6.94 (d, 1H, J ¼ 8.1 Hz, H-500),
7.00e7.09 (m, 2H, H-200, H-600), 7.65 (d, 1H, J ¼ 15.9 Hz, H-700); trans-
p-coumaroyl units: R2: 2.23 (s, 3H, H-1100), 6.39 (d, 1H, J ¼ 15.9 Hz,
H-800), 7.00e7.09 (m, 2H, H-300, H-500), 7.40 (d, 2H, H-200, H-600), 7.56
(d, 1H, J ¼ 15.9 Hz, H-700), R3: 2.26 (s, 3H, H-1100), 6.29 (d, 1H,
J ¼ 15.9 Hz, H-800), 7.00e7.09 (m, 2H, H-300, H-500), 7.47e7.53 (m, 2H,
H-200, H-600), 7.47e7.53 (m, 1H, H-700); 13C NMR (75.48 MHz, CDCl3)
4.1.4.2.2. 6-mono-O-feruloyl-30,60-di-O-coumaroylsucrose (lapa-
thoside C, 3). Following the general procedure, a suspension of
compound 18 (0.2 g, 0.2 mmol) in 95% EtOH (10 mL) was treated
with pyrrolidine (155.0 mL, 0.1 g, 1.9 mmol) for 90 min to furnish
lapathoside C 3 as a white solid (0.13 g, 75% yield). Analytical data
for 3: Rf ¼ 0.55 (EtOAc/MeOH 9:1); mp: 125e127 ꢂC; FT-IR (KBr)
nmax: 3447, 3421, 2956, 2926, 2362, 2340, 1700, 1636, 1559, 1540,
1517, 1457, 1445, 1374, 1328, 1266, 1170, 1059, 997, 946, 831,
668 cmꢁ1; 1H NMR (300 MHz, CD3OD)
d (ppm): 3.32 (m, 1H, H-4),
3.35 (m, 1H, H-5), 3.48 (m, 1H, H-2), 3.60 (m, 2H, H-10b, H-10a), 3.65
(m, 1H, H-3), 4.18 (m, 1H, H-50), 4.29 (m, 1H, H-6a), 4.55 (m, 2H, H-
60b, H-60a), 4.65 (m, 1H, H-40), 4.71 (m, 1H, H-6b), 5.50 (m, 1H, H-1),
5.54 (m, 1H, H-30); trans-p-feruloyl units: R1: 3.84 (s, 3H, eOCH3),
6.48 (d, 1H, J ¼ 15.9 Hz, H-800), 6.75e6.82 (m, 1H, H-500), 7.02 (d, 1H,
J ¼ 7.5 Hz, H-600), 7.21 (br s, 1H, H-200), 7.62 (d, 1H, J ¼ 15.9 Hz, H-700);
trans-p-coumaroyl units: R2: 6.43 (d, 1H, J ¼ 15.9 Hz, H-800), 6.75e
6.82 (m, 2H, H-300, H-500), 7.52 (d, 2H, J ¼ 8.4 Hz, H-200, H-600), 7.73
(d, 1H, J ¼ 15.9 Hz, H-700), R3: 6.24 (d, 1H, J ¼ 15.9 Hz, H-800), 6.75e
6.82 (m, 2H, H-300, H-500), 7.34 (d, 2H, J ¼ 8.4 Hz, H-200, H-600), 7.62 (d,
d
(ppm): 64.3 (C-10), 64.5 (C-6), 64.8 (C-60), 70.3 (C-4), 71.0 (C-5),
71.7 (C-2), 73.9 (C-3), 74.7 (C-40), 79.8 (C-30), 80.9 (C-50), 91.6 (C-1),
104.5 (C-20); trans-p-feruloyl units: R1: 20.6 (C-1100), 55.9 (eOCH3),
111.5 (C-200), 116.7 (C-800), 121.6 (C-500), 123.2 (C-600), 133.1 (C-100),
141.5 (C-300), 146.1 (C-700), 151.3 (C-400), 167.4 (C-900), 169.5 (C-1000);
trans-p-coumaroyl units: R2: 21.1 (C-1100), 117.4 (C-800), 122.1 (C-300,
C-500), 129.4 (C-200, C-600), 131.6 (C-100), 144.5 (C-700), 152.2 (C-400),
166.9 (C-900), 168.9 (C-1000), R3: 21.1 (C-1100), 117.5 (C-800), 122.2 (C-300,
C-500), 129.8 (C-200, C-600), 131.8 (C-100), 145.1 (C-700), 152.4 (C-40þ0),
167.2 (C-900), 169.2 (C-1000); LC-MS (ESI): m/z 959.14 [M þ Na] ;
HR-MS: m/z [M þ Na]þ calcd for C46H48O21Na: 959.2580, found:
959.2549. Analytical data for 19: Rf ¼ 0.11 (EtOAc/hexanes 3:1); mp:
114e116 ꢂC; FT-IR (KBr) nmax: 3483, 2925, 2855, 2363, 2340, 1766,
1713, 1637, 1602, 1508, 1467, 1419, 1371, 1321, 1260, 1203, 1164, 1123,
1H, J ¼ 15.9 Hz, H-700); 13C NMR (75.48 MHz, CD3OD)
d (ppm): 65.4
(C-10), 65.8 (C-6, C-60), 72.1 (C-4), 72.3 (C-5), 73.1 (C-2), 74.8 (C-3),
75.0 (C-40), 79.0 (C-30), 81.1 (C-50), 92.5 (C-1), 104.8 (C-20); trans-p-
feruloyl units: R1: 56.4 (eOCH3), 111.5 (C-200), 115.3 (C-800), 116.3 (C-
500), 124.5 (C-600), 127.7 (C-100), 147.2 (C-700), 149.3 (C-300), 150.6 (C-
400), 169.3 (C-900); trans-p-coumaroyl units: R2: 114.6 (C-800), 116.8
(C-300, C-500), 127.1 (C-100), 131.5 (C-200, C-600), 147.6 (C-700), 161.4 (C-
400), 168.4 (C-900), R3: 114.8 (C-800), 116.8 (C-300, C-500), 127.1 (C-100),
131.2 (C-200, C-600), 146.8 (C-700), 161.3 (C-400), 168.9 (C-900); LC-MS
(ESI): m/z 833.13 [M þ Na]þ; HR-MS: m/z [M þ Na]þ calcd for
1061, 1032, 1011, 909, 834, 652 cmꢁ1
d
;
1H NMR (300 MHz, CDCl3):
(ppm): 3.54 (m, 1H, H-4), 3.74 (m, 3H, H-2, H-10a, H-10b), 4.24e
4.28 (m, 2H, H-5, H-50), 4.46e4.60 (m, 5H, H-40, H-6b, H-6a, H-
60b, H-60a), 5.25 (m, 1H, H-3), 5.29 (d, 1H, J ¼ 7.2 Hz, H-30), 5.57 (br s,
1H, H-1), trans-p-coumaroyl units: R1: 2.28 (s, 3H, H-1100), 6.40 (d,
1H, J ¼ 16.2 Hz, H-800), 6.98e7.16 (m, 2H, H-300, H-500), 7.47 (d, 2H,
H-200, H-600), 7.53e7.64 (m, 1H, H-700), R2: 2.28 (s, 3H, H-1100), 6.46 (d,
1H, J ¼ 16.2 Hz, H-800), 6.98e7.16 (m, 2H, H-300, H-500), 7.53e7.64
(m, 3H, H-200, H-600, H-700); trans-p-feruloyl units: R3: 2.30 (s, 3H,
H-1100), 3.81 (s, 3H, eOCH3), 6.52 (d, 1H, J ¼ 16.2 Hz, H-800), 6.98e
7.16 (m, 3H, H-200, H-500, H-600), 7.53e7.64 (m, 1H, H-700), R4: 2.30
(s, 3H, H-1100), 3.81 (s, 3H, eOCH3), 6.51 (d, 1H, J ¼ 16.2 Hz, H-800),
6.98e7.16 (m, 3H, H-200, H-500, H-600), 7.79 (d, 1H, J ¼ 16.2 Hz, H-700);
C40H42O18Na: 833.2263, found: 833.2283. Spectral data of lapa-
thoside C 3 was the same as reported for the isolated natural
product [16].
4.1.4.2.3. 3,6-di-O-feruloyl-30,60-di-O-coumaroylsucrose
21.
Following the general procedure, a suspension of compound 19
(0.1 g, 0.1 mmol) in 95% EtOH (5 mL) was reacted with pyrrolidine
(130.0 mL, 0.1 g, 1.6 mmol) for 3 h to afford compound 21 as a white
solid (0.04 g, 47% yield). Analytical data for 21: Rf ¼ 0.74 (EtOAc/
MeOH 9:1); mp: 135e138 ꢂC; FT-IR (KBr) nmax: 3363, 2924, 2852,
2363, 2340, 1698, 1635, 1604, 1517, 1457, 1455, 1337, 1277, 1169,
13C NMR (75.48 MHz, CDCl3)
d
(ppm): 63.7 (C-6), 64.5 (C-60), 64.6
(C-10), 69.1 (C-4), 70.6 (C-2), 71.4 (C-5), 74.4 (C-40), 76.6 (C-3), 80.5
(C-30), 80.6 (C-50), 91.9 (C-1), 104.5 (C-20); trans-p-coumaroyl units:
R1: 21.1 (C-1100), 116.4 (C-800), 122.1 (C-300, C-500), 129.4 (C-200, C-600),
131.6 (C-100); 144.7 (C-700), 152.2 (C-400), 166.9 (C-900), 168.8 (C-1000),
R2: 21.1 (C-1100), 116.4 (C-800), 122.1 (C-300, C-500), 129.9 (C-200, C-600),
131.9 (C-100), 145.3 (C-700), 152.5 (C-400), 167.3 (C-900), 168.8 (C-1000);
trans-p-feruloyl units: R3: 20.7 (C-1100), 55.9 (eOCH3), 111.4 (C-200),
117.4 (C-800), 121.5 (C-500), 123.2 (C-600), 133.1 (C-100), 141.6 (C-300),
145.6 (C-700), 151.4 (C-400), 167.8 (C-900), 169.1 (C-1000), R4: 20.7
(C-1100), 55.9 (eOCH3), 111.4 (C-200), 117.4 (C-800), 121.5 (C-500),
123.2 (C-600), 133.1 (C-100), 141.6 (C-300), 146.5 (C-700), 151.4 (C-40þ0),
168.2 (C-900), 169.1 (C-1000); LC-MS (ESI): m/z 1177.11 [M þ Na] ;
HR-MS: m/z [M þ Na]þ calcd for C58H58O25Na: 1177.3159, found:
1177.3159.
1128, 1087, 1031, 987, 932, 831, 666 cmꢁ1 1H NMR (300 MHz,
;
CD3OD) d
(ppm): 3.56 (m, 1H, H-4), 3.65e3.74 (m, 3H, H-10a, H-10b,
H-2), 4.15e4.21 (m, 1H, H-50), 4.24e4.33 (m, 1H, H-60b), 4.41 (dd,
1H, J ¼ 8.7 Hz, 9.0 Hz, H-5), 4.52e4.59 (m, 2H, H-6b, H-6a), 4.67e
4.74 (m, 2H, H-40, H-60a), 5.34 (dd, 1H, J ¼ 9.9 Hz, 9.0 Hz, H-3),
5.57e5.62 (m, 2H, H-1, H-30); trans-p-coumaroyl units: R1: 6.26 (d,
1H, J ¼ 16.2 Hz, H-800), 6.74e6.82 (m, 2H, H-300, H-500), 7.36 (d, 2H,
J ¼ 8.7 Hz, H-200, H-600), 7.54e7.65 (m, 1H, H-700); R2: 6.49 (d, 1H,
J ¼ 16.2 Hz, H-800), 6.74e6.82 (m, 2H, H-300, H-500), 7.54e7.65 (m, 2H,
H-200, H-600), 7.75 (d, 1H, J ¼ 16.2 Hz, H-700), trans-p-feruloyl units:
R3: 3.84 (s, 3H, eOCH3), 6.42 (d, 1H, J ¼ 16.2 Hz, H-800), 6.74e6.82
(m, 1H, H-500), 7.01 (dd, 1H, J ¼ 1.5 Hz, 7.8 Hz, H-600), 7.18 (dd,
1H, J ¼ 9.9 Hz, 1.2 Hz, H-200), 7.54e7.65 (m, 1H, H-700), R4: 3.88 (s,

428
P. Panda et al. / European Journal of Medicinal Chemistry 58 (2012) 418e430
3H, eOCH3), 6.48 (d, 1H, J ¼ 16.2 Hz, H-800), 6.74e6.82 (d, 1H,
J ¼ 7.8 Hz, H-500), 7.07 (dd, 1H, J ¼ 1.5 Hz, 7.8 Hz, H-600), 7.18 (dd, 1H,
J ¼ 9.9 Hz, 1.2 Hz, H-200), 7.54e7.65 (m, 1H, H-700); 13C NMR
LC-MS (ESI): m/z 1117.15 [M þ Na]þ; HR-MS: m/z [M þ Na]þ calcd
for C56H54O23Na: 1117.2948, found: 1117.2918.
4.1.4.3.2. Preparation of 3,6,30,60-tetra-O-coumaroylsucrose 22.
Compound 20 (0.05 g, 0.05 mmol) was suspended in 95% EtOH
(75.48 MHz, CD3OD)
d
(ppm): 65.3 (C-10), 65.6 (C-6), 65.7 (C-60),
70.5 (C-4), 71.6 (C-2), 72.5 (C-5), 74.7 (C-40), 77.0 (C-3), 79.0 (C-30),
81.2 (C-50), 92.7 (C-1), 105.4 (C-20), trans-p-coumaroyl units: R1:
114.7 (C-800), 116.9 (C-300, C-500), 127.1 (C-100), 131.3 (C-200, C-600), 146.9
(C-700), 161.5 (C-400), 168.7 (C-900), R2: 114.9 (C-800), 116.9 (C-300, C-500),
127.3 (C-100), 131.7 (C-200, C-600), 147.0 (C-700), 161.5 (C-400), 169.0
(C-900); trans-p-feruloyl units: R3: 56.5 (eOCH3), 111.8 (C-200), 115.3
(C-800), 116.4 (C-500), 124.1 (C-600), 127.7 (C-100), 147.4 (C-700), 149.4
(C-300), 150.7 (C-400), 169.3 (C-900), R4: 56.5 (eOCH3), 111.8 (C-200),
115.8 (C-800), 116.6 (C-500), 124.6 (C-600), 127.9 (C-100), 147.8 (C-700),
149.5 (C-300), 150.8 (C-400), 169.3 (C-900); LC-MS (ESI): m/z 1009.12
[M þ Na]þ; HR-MS: m/z [M þ Na]þ calcd for C50H50O21Na:
1009.2737, found: 1009.2740.
(4.0 mL) and pyrrolidine (70.0 mL, 0.1 g, 0.9 mmol) was added
(which caused the solution to turn yellow). The starting material
typically dissolved within 15 min and the reaction was allowed to
continue for 15 min. Reaction was monitored by TLC analysis
(EtOAc). The mixture was directly added to a column of strongly
acidic ion-exchange resin [Amberlite IRA-120 (Hþ) washed and
packed in 95% EtOH]. The appropriate fractions were concentrated
under diminished pressure to a residue that was subjected to
column chromatography using a gradient of CH2Cl2/EtOAc/MeOH
to furnish compound 22 as a white solid (0.03 g, 71% yield).
Analytical data for 22: Rf ¼ 0.76 (EtOAc/MeOH 9:1); mp: 149e
151 ꢂC; FT-IR (KBr) nmax: 3415, 2957, 2922, 2850, 2363, 2340,
1700, 1635, 1604, 1559, 1516, 1441, 1369, 1327, 1266, 1169, 1060,
4.1.4.3. Synthesis of 3,6,30,60-tetra-O-coumaroylsucrose 22
1004, 864, 831 cmꢁ1; 1H NMR (300 MHz, CD3OD)
d (ppm): 3.59 (m,
4.1.4.3.1. Preparation of 3,6,30,60-tetra-O-acetoxycinnamoylsucrose
20. 30,60-di-O-acetoxycinnamoyl sucrose 16 (1.1 g, 1.5 mmol) was
dissolved in dry CH2Cl2 (21 mL) to which 4 Å molecular sieves
powder and dry pyridine (1.2 g, 1.2 mL, 15.3 mmol) were added.
The solution was cooled to 0 ꢂC in an ice bath and p-acetox-
ycinnamoyl chloride (17, 0.5 g, 2.0 mmol) was added slowly at the
same temperature and the reaction mixture was left to stir while
warming to r.t. Stirring was continued for 24 h. After this time, the
starting material was completely disappeared as indicated by TLC
(EtOAc/hexanes 3:1). The resulting mixture was poured into
vigorously stirred ice-water (100 mL) and a white solid precipi-
tated was obtained after decantation and filtration. The precipitate
was redissolved in EtOAc (25 mL) and washed with 1 N HCl
(2 ꢀ 50 mL). The aqueous layer was extracted with EtOAc (25 mL).
The combined organic layers were then successively washed with
5% NaHCO3 (2 ꢀ 50 mL) and brine (25 mL) and then dried over
anhyd. MgSO4. The EtOAc solution was concentrated to residue
that was subjected to column chromatography using a gradient of
CH2Cl2/EtOAc as eluent followed by PTLC afforded compound 20
as a white solid (0.20 g, 12% yield). Analytical data for 20: Rf ¼ 0.43
(EtOAc/hexanes 3:1); mp: 113e115 ꢂC; FT-IR (KBr) nmax: 3475,
2934, 2362, 2337, 1768, 1718, 1704, 1636, 1602, 1559, 1540, 1507,
1457, 1419, 1371, 1322, 1283, 1207, 1169, 1058, 1009, 946, 911, 836,
1H, H-4), 3.68 (m, 2H, H-10a, H-10b), 3.73 (m,1H, H-2), 4.14e4.19 (m,
1H, H-50), 4.30 (m, 1H, H-6b), 4.38 (m, 1H, H-5), 4.50e4.57 (m, 2H,
H-6a, H-60b), 4.62e4.70 (m, 2H, H-40, H-60a), 5.33 (app t, 1H,
J ¼ 9.6 Hz, H-3), 5.57 (d, 1H, J ¼ 7.8 Hz, H-30), 5.62 (d, 1H, J ¼ 3.3 Hz,
H-1), trans-p-coumaroyl units: R1: 6.29 (d, 1H, J ¼ 15.9 Hz, H-800),
6.73e6.81 (m, 2H, H-300, H-500), 7.38e7.46 (m, 2H, H-200, H-600), 7.54e
7.66 (m, 1H, H-700), R2: 6.38 (d, 1H, J ¼ 15.9 Hz, H-800), 6.73e6.81 (m,
2H, H-300, H-500), 7.38e7.46 (m, 2H, H-200, H-600), 7.54e7.66 (m,1H, H-
700), R3: 6.40 (d, 1H, J ¼ 15.9 Hz, H-800), 6.73e6.81 (m, 2H, H-300, H-
500), 7.38e7.46 (m, 2H, H-200, H-600), 7.54e7.66 (m, 1H, H-700), R4: 6.48
(d, 1H, J ¼ 15.9 Hz, H-800), 6.73e6.81 (m, 2H, H-300, H-500), 7.54e7.66
(m, 2H, H-200, H-600), 7.76 (d, 1H, J ¼ 15.9 Hz, H-700); 13C NMR
(75.48 MHz, CD3OD)
d
(ppm): 65.7 (C-60), 65.8 (C-6), 65.9 (C-10),
70.8 (C-4), 72.0 (C-2), 72.8 (C-5), 74.9 (C-40), 77.3 (C-3), 79.4 (C-30),
81.6 (C-50), 93.2 (C-1), 105.6 (C-20), trans-p-coumaroyl units: R1:
114.8 (C-800), 117.4 (C-300, C-500), 127.2 (C-100), 131.6 (C-200, C-600), 147.2
(C-700), 162.3 (C-400), 169.2 (C-900), R2: 115.0 (C-800), 117.4 (C-300, C-500),
127.3 (C-100), 131.7 (C-200, C-600), 147.4 (C-700), 162.4 (C-400), 169.4
(C-900), R3: 115.1 (C-800), 117.5 (C-300, C-500), 127.4 (C-100), 131.9 (C-200,
C-600), 147.4 (C-700), 162.5 (C-400), 169.8 (C-900), R4: 115.6 (C-800), 117.6
(C-300, C-500), 127.4 (C-100), 132.1 (C-200, C-600), 148.4 (C-700), 162.6
(C-400), 169.9 (C-900); LC-MS (ESI): m/z 949.14 [M þ Na]þ; HR-MS:
m/z [M þ Na]þ calcd for C48H46O19Na: 949.2526, found: 949.2494.
792, 649 cmꢁ1; 1H NMR (300 MHz, CDCl3)
d (ppm): 3.60 (m, 1H, H-
4), 3.68 (m, 1H, H-10a), 3.78 (m, 2H, H-2, H-10b), 4.26 (m, 3H, H-5,
H-50, H-6a), 4.53 (m, 4H, H-40, H-6b, H-60b, H-60a), 5.27 (dd, 1H,
J ¼ 9.0 Hz, 9.3 Hz, H-3), 5.34 (d, 1H, J ¼ 6.9 Hz, H-30), 5.56 (br s, 1H,
H-1); trans-p-coumaroyl units: R1: 2.27 (s, 3H, H-1100), 6.38 (d, 1H,
J ¼ 15.9 Hz, H-800), 7.05 (d, 2H, H-300, H-500), 7.44e7.49 (m, 2H, H-200,
H-600), 7.61 (m, 1H, H-700), R2: 2.27 (s, 3H, H-1100), 6.40 (d, 1H,
J ¼ 15.9 Hz, H-800), 7.05 (d, 2H, H-300, H-500), 7.44e7.49 (m, 2H, H-200,
H-600), 7.62 (m, 1H, H-700), R3: 2.27 (s, 3H, H-1100), 6.44 (d, 1H,
J ¼ 15.9 Hz, H-800), 7.05 (d, 2H, H-300, H-500), 7.44e7.49 (m, 2H, H-200,
H-600), 7.64 (m, 1H, H-700), R4: 2.27 (s, 3H, H-1100), 6.51 (d, 1H,
J ¼ 15.9 Hz, H-800), 7.05 (d, 2H, H-300, H-500), 7.57 (m, 2H, H-200, H-
600), 7.76 (d, 1H, J ¼ 15.9 Hz, H-700); 13C NMR (75.48 MHz, CDCl3)
4.1.4.4. Synthesis of 6-mono-O-feruloyl-3,30,60-tri-O-coumaroylsucrose
24
4.1.4.4.1. Preparation of 6-mono-O-acetoxyferuloyl-3,30,60-tri-O-
acetoxycinnamoylsucrose 23. Compound 18 (0.4 g, 0.4 mmol) was
dissolved in dry CH2Cl2 (7 mL) to which 4 Å molecular sieves
powder followed by dry pyridine (0.3 g, 0.3 mL, 3.8 mmol) was
added. The solution was then cooled to 0 ꢂC in an ice bath and then
p-acetoxycinnamoyl chloride 17 (0.08 g, 0.35 mmol) was added
slowly at the same temperature and the reaction mixture was left to
stir while warming to r.t. Stirring was continued until the disap-
pearance of the starting material as indicated by TLC (EtOAc/
hexanes 3:1). After 24 h, the resulting mixture was poured into
vigorously stirred ice-water (100 mL) and a white solid precipitated
was obtained after decantation and filtration. The precipitate was
redissolved in EtOAc (25 mL) and washed with 1 N HCl (2 ꢀ 50 mL).
The aqueous layer was extracted with EtOAc (25 mL). The combined
organic layers were then successively washed with 5% NaHCO3
(2 ꢀ 50 mL) and brine (25 mL) and then dried over anhyd. MgSO4.
The solvent was concentrated to residue that was subjected to
column chromatography using a gradient of CH2Cl2/EtOAc as eluent
followed by PTLC furnished compound 23 as a white solid (0.12 g,
27% yield). Analytical data for 23: Rf ¼ 0.43 (EtOAc/hexanes 3:1);
mp: 113e115 ꢂC; FT-IR (KBr) nmax: 3428, 2923, 2362, 2340, 1765,
d
(ppm): 63.8 (C-6), 64.5 (C-60), 64.7 (C-10), 69.0 (C-4), 70.6 (C-2),
71.4 (C-5), 74.5 (C-40), 76.5 (C-3), 80.2 (C-30), 80.6 (C-50), 92.0 (C-1),
104.5 (C-20); trans-p-coumaroyl units: R1: 21.1 (C-1100), 116.6
(C-800), 122.1 (C-300, C-500), 129.4 (C-200, C-600), 131.7 (C-100), 144.6
(C-700), 152.2 (C-400), 166.9 (C-900), 169.2 (C-1000); R2: 21.1 (C-1100),
117.3 (C-800), 122.1 (C-300, C-500), 129.4 (C-200, C-600), 131.8 (C-100),
144.8 (C-700), 152.2 (C-400), 167.4 (C-900), 169.2 (C-1000); R3: 21.1
(C-1100), 117.5 (C-800), 122.1 (C-300, C-500), 129.5 (C-200, C-600), 131.8
(C-100), 145.0 (C-700), 152.2 (C-400), 167.6 (C-900), 169.2 (C-1000); R4:
21.1 (C-1100), 117.5 (C-800), 122.1 (C-300, C-500), 129.9 (C-200, C-600),
131.9 (C-100), 146.3 (C-700), 152.4 (C-400), 168.1 (C-900), 169.2 (C-1000);

P. Panda et al. / European Journal of Medicinal Chemistry 58 (2012) 418e430
429
1718,1636,1602,1559,1540,1507,1457,1419,1374,1320,1281,1260,
1205, 1165, 1054, 1009, 987, 913, 839, 792, 649 cmꢁ1 1H NMR
(C-200), 115.9 (C-800), 116.0 (C-500), 124.2 (C-600), 127.5 (C-100), 146.6 (C-
700), 148.7 (C-300), 150.1 (C-400), 167.8 (C-900); LC-MS (ESI): m/z 979.13
[M þ Na]þ; HR-MS: m/z [M þ Na]þ calcd for C49H48O20Na: 979.2631,
found: 979.2612.
;
(300 MHz, CDCl3)
d (ppm): 3.61 (m, 1H, H-4), 3.78 (m, 3H, H-2, H-
10a, H-10b), 4.27 (m, 2H, H-5, H-50), 4.55 (m, 5H, H-40, H-6b, H-6a, H-
60b, H-60a), 5.25 (m, 1H, H-3), 5.30 (m, 1H, H-30), 5.57 (br s, 1H, H-1);
trans-p-coumaroyl units:
d
¼ R1: 2.28 (s, 3H, H-1100), 6.40 (d, 1H,
4.2. Antiproliferative activity of selected PSEs against HeLa cells
J ¼ 15.9 Hz, H-800), 6.93e7.17 (m, 2H, H-300, H-500), 7.47 (d, 2H, H-200,
H-600), 7.57e7.67 (m, 1H, H-700), R2: 2.29 (s, 3H, H-1100), 6.41 (d, 1H,
J ¼ 15.9 Hz, H-800), 6.93e7.17 (m, 2H, H-300, H-500), 7.47 (d, 2H, H-200,
H-600), 7.57e7.67 (m, 1H, H-700), R3: 2.29 (s, 3H, H-1100), 6.46 (d, 1H,
J ¼ 15.9 Hz, H-800), 6.93e7.17 (m, 2H, H-300, H-500), 7.57e7.67 (m, 2H,
H-200, H-600), 7.57e7.67 (m, 1H, H-700); trans-p-feruloyl units: R4:
2.30 (s, 3H, H-1100), 3.81 (s, 3H, eOCH3), 6.52 (d, 1H, J ¼ 15.9 Hz, H-
800), 6.93e7.17 (m, 3H, H-200, H-500, H-600), 7.77 (d, 1H, J ¼ 15.9 Hz, H-
Antiproliferative activities of the selected PSEs against HeLa cell
lines were evaluated using MTS assay method [24] at 24 and 48 h of
drug exposure (For the details see the Supporting information).
Acknowledgements
This research was supported by a grant from the National
Medical Research Council (NMRC), Singapore (EDG10nv043).
700); 13C NMR (75.48 MHz, CDCl3):
d
¼ R1: 63.7 (C-6), 64.5 (C-60),
64.7 (C-10), 69.0 (C-4), 70.6 (C-2), 71.4 (C-5), 74.4 (C-40), 76.6 (C-3),
80.5 (C-30), 80.6 (C-50), 91.9 (C-1), 104.5 (C-20); trans-p-coumaroyl
units: R1: 21.1 (C-1100),117.4 (C-800), 122.1 (C-300, C-500),129.4 (C-200, C-
600), 131.6 (C-100), 144.7 (C-700), 152.2 (C-400), 166.9 (C-900), 168.8
(C-1000), R2: 21.1 (C-1100), 117.4 (C-800), 122.1 (C-300, C-500), 129.4 (C-200,
C-600), 131.8 (C-100), 145.2 (C-700), 152.3 (C-400), 167.3 (C-900), 168.8
(C-1000), R3: 21.1 (C-1100), 117.4 (C-800), 122.1 (C-300, C-500), 129.9 (C-200,
C-600), 131.9 (C-100), 145.3 (C-700), 152.5 (C-400), 167.8 (C-900), 169.1
(C-1000); trans-p-feruloyl units: R4: 20.7 (C-1100), 55.9 (eOCH3), 111.4
(C-200), 116.5 (C-800), 121.6 (C-500), 123.2 (C-600), 133.1 (C-100), 141.6
(C-300), 146.5 (C-700), 151.4 (C-400), 168.2 (C-900), 169.1 (C-1000); LC-MS
(ESI): m/z 1147.17 [M þ Na]þ; HR-MS: m/z [M þ Na]þ calcd for
Appendix A. Supplementary data
Supplementary data associated with this article can be found in
the online version, at http://dx.doi.org/10.1016/j.ejmech.2012.10.
034. These data include MOL files and InChiKeys of the most
important compounds described in this article.
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C
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4.1.4.4.2. Preparation of 6-mono-O-feruloyl-3,30,60-tri-O-coumar-
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3428, 2923, 2340, 2362, 2337, 1765, 1718, 1734, 1699, 1653, 1636,
1602, 1559, 1540, 1507, 1457, 1419, 1374, 1320, 1281, 1260, 1205,
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d (ppm):
3.69 (m, 3H, H-4, H-10a, H-10b), 3.80 (m, 1H, H-2), 4.27 (m, 1H, H-50),
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5.62 (d, 1H, J ¼ 3.0 Hz, H-1); trans-p-coumaroyl units: R1: 6.36 (d,
1H, J ¼ 15.9 Hz, H-800), 6.83e6.91 (m, 2H, H-300, H-500), 7.50e7.67 (m,
3H, H-200, H-600, H-700), R2: 6.37 (d, 1H, J ¼ 15.9 Hz, H-800), 6.83e6.91
(m, 2H, H-300, H-500), 7.50e7.67 (m, 3H, H-200, H-600, H-700), R3: 6.49 (d,
1H, J ¼ 15.9 Hz, H-800), 6.83e6.91 (m, 2H, H-300, H-500), 7.50e7.67
(m, 3H, H-200, H-600, H-700); trans-p-feruloyl units: R4: 3.89 (s,
3H, eOCH3), 6.56 (d, 1H, J ¼ 15.9 Hz, H-800), 6.83e6.91 (d, 1H,
J ¼ 8.4 Hz, H-500), 7.13 (d, 1H, J ¼ 6.9 Hz, H-600), 7.36 (br s, 1H, H-200),
7.77 (d, 1H, J ¼ 15.9 Hz, H-700); 13C NMR (75.48 MHz, (CD3)2CO)
d
(ppm): 64.9 (C-6), 65.3 (C-60), 65.6 (C-10), 70.0 (C-4), 71.4 (C-2),
72.2 (C-5), 74.7 (C-40), 76.8 (C-3), 79.3 (C-30), 81.0 (C-50), 92.4 (C-1),
104.8 (C-20); trans-p-coumaroyl units: R1: 115.1 (C-800), 116.7 (C-300,
C-500), 126.9 (C-100), 130.9 (C-200, C-600), 145.5 (C-700), 160.6 (C-400),
167.3 (C-900), R2: 115.2 (C-800), 116.7 (C-300, C-500), 127.0 (C-100), 131.1
(C-200, C-600), 145.9 (C-700), 160.7 (C-400), 167.5 (C-900), R3: 115.8 (C-800),
116.8 (C-300, C-500), 127.0 (C-100), 131.4 (C-200, C-600), 146.1 (C-700), 160.8
(C-400), 167.7 (C-900); trans-p-feruloyl units: R4: 56.3 (eOCH3), 111.3

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