Anti-AGEs and antiparasitic activity of an original prenylated isoflavonoid and flavanones isolated from Derris ferruginea

Article abstract of DOI:10.1016/j.phytol.2013.06.002

A new isoflavonoid, 5-hydroxy-3-(4-hydroxyphenyl)-8-isopropenyl-8,9- dihydro-4H-furo-[2,3-h]-chromen-4-one named derrisisoflavone G (1), four known prenylated flavanones (2-5), four known isoflavonoids (6-9) and two phenolic derivatives (10, 11) have been

Full text of DOI:10.1016/j.phytol.2013.06.002

G Model
PHYTOL 548 1–6
Phytochemistry Letters xxx (2013) xxx–xxx
Contents lists available at SciVerse ScienceDirect
Phytochemistry Letters
journal homepage: www.elsevier.com/locate/phytol
1
2
3
Anti-AGEs and antiparasitic activity of an original prenylated
isoflavonoid and flavanones isolated from Derris ferruginea
a
a
a
a
a
´ ´ ´
Q1 Sylvie Morel , Jean-Jacques Helesbeux , Denis Seraphin , Severine Derbre , Julia Gatto ,
4
5
6
Marie-Christine Aumond a, Yannick Abatuci a, Philippe Grellier b, Mehdi A. Beniddir b,d
,
Patrice Le Pape c, Fabrice Pagniez c, Marc Litaudon d, Anne Landreau a, , Pascal Richomme
a
*
7
8
9
10
a SONAS EA 921, IFR 149, QUASAV UFR des Sciences Pharmaceutiques et d’Inge´nierie de la Sante´, 16 Bd Daviers, 49100 Angers, France
b Muse´um National d’Histoire Naturelle, FRE 3206 CNRS, 61 rue Buffon, F-75231 Paris Cedex 05, France
c Laboratoire de Parasitologie et Mycologie Me´dicale, UPRES EA 1155, Faculte´ de Pharmacie, 1 rue Gaston Veil, 44035 Nantes Cedex, France
d Centre de Recherche de Gif, Institut de Chimie des Substances Naturelles, CNRS, 1, Avenue de la Terrasse, 91198 Gif-sur-Yvette Cedex, France
A R T I C L E I N F O
A B S T R A C T
Article history:
A new isoflavonoid, 5-hydroxy-3-(4-hydroxyphenyl)-8-isopropenyl-8,9-dihydro-4H-furo-[2,3-h]-chro-
men-4-one named derrisisoflavone G (1), four known prenylated flavanones (2–5), four known
isoflavonoids (6–9) and two phenolic derivatives (10, 11) have been isolated from crude extracts of Derris
ferruginea stems and leaves. Compounds 1–11 were identified using spectroscopic methods whereas an
unambiguous structural assignment of 1 was accomplished through hemi-synthesis. Compounds 2–5
exhibited strong in vitro antiparasitic activity against Plasmodium falciparum and Leishmania major but
with poor selectivity, whereas 1–5 significantly inhibited the formation of advanced glycation
endproducts (AGEs).
Received 25 January 2013
Received in revised form 31 May 2013
Accepted 4 June 2013
Available online xxx
Keywords:
Derris ferruginea
Fabaceae
Crown Copyright ß 2013 Phytochemical Society of Europe. Published by Elsevier B.V. All rights reserved.
Flavonoid
Synthesis
Antiparasitic
AGEs (advanced glycation endproducts)
11
12
1. Introduction
from D. ferruginea roots was reported a long time ago (Rao and 29
Seshadri, 1946) and we recently described an efficient method for 30
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
Derris (Fabaceae) is a genus belonging to the tribe Millettieae
(Leguminosae) with about 250 species that are widely distributed
in tropical areas of Africa and Asia (The Angiosperm Phylogeny
Group, 2009). These plants are traditionally used as pesticides and
fish poisons (Zheng and Wing, 2009). Flavonoids, particularly
prenylated flavonoids and isoflavonoids such as rotenoids, are
often found in this genus. These secondary metabolites are
associated with a broad spectrum of biological activities, including
purification of the bioactive cajaflavanone from the stems of this 31
species (Morel et al., 2012). In the course of our investigations on 32
this plant, herein we report the isolation, structural elucidation and 33
hemi-synthesis of a new prenylated isoflavonoid (1) from the 34
CH2Cl2 extract of D. ferruginea stems, together with the identifica- 35
tion of 10 known compounds. Their structures were established by 36
1D and 2D NMR spectroscopy and HRMS analysis. Furthermore, 37
compounds 1–5 exhibited in vitro antiparasitic activity as well as 38
an ability to inhibit the formation of advanced glycation end- 39
a
-glucosidase inhibition (Rao et al., 2007, 2009), insecticidal
(Sreelatha et al., 2009; Yenesew et al., 2009), cytotoxic (Cheen-
pracha et al., 2007; Svasti et al., 2005) and antioxidant activities
(Rao et al., 2007, 2009). Derris ferruginea (Roxburg) Bentham, also
named Robinia ferruginea, is a perennial plant found in sparse
forests and thickets, growing on low elevation mountain slopes
(500–1200 m) and exhibiting densely rust-colored pubescent
branchlets (Rao and Seshadri, 1946). The isolation of rotenone
products (AGEs).
40
2. Results and discussion
41
42
2.1. Phytochemical study of D. ferruginea
Purification of the CH2Cl2 extract from dried stems of D. 43
ferruginea afforded compound 1 whereas 10 known compounds 44
(2–11) were isolated from cyclohexane, CH2Cl2 and EtOAc extracts 45
from stems and cyclohexane extract from dried leaves. They were 46
identified by comparison of their spectroscopic data (UV, IR, NMR, 47
* Corresponding author. Tel.: +33 0241226666; fax: +33 0241226634.
E-mail address: anne.landreau@univ-angers.fr (A. Landreau).
1874-3900/$ – see front matter. Crown Copyright ß 2013 Phytochemical Society of Europe. Published by Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.phytol.2013.06.002
Please cite this article in press as: Morel, S., et al., Anti-AGEs and antiparasitic activity of an original prenylated isoflavonoid and
flavanones isolated from Derris ferruginea. Phytochem. Lett. (2013), http://dx.doi.org/10.1016/j.phytol.2013.06.002

G Model
PHYTOL 548 1–6
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S. Morel et al. / Phytochemistry Letters xxx (2013) xxx–xxx
48
49
50
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[
a
]
D and MS) with those reported in the literature: four flavanones,
5,30,40-trihydroxy-6-( -dimethylallyl)-600,600-dimethylpyrano-
g,g
(200,300:7,8)-flavanone (2) (Shiao et al., 2005), dorsmanine I (3)
(Ngadjui et al., 2000), lonchocarpol A (4) (Innok et al., 2009), 6,8-
diprenyleriodictyol (5) (Harborne et al., 1993) previously isolated
53 Q2 from Derris laxiflora (Young Ho et al., 1995); four isoflavones,
54
55
56
57
58
59
60
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63
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65
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67
68
69
70
71
72
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75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
genistein (6) previously isolated from from Derris scandens
(Mahabusarakam et al., 2004), pratensein (7) (Almeida et al.,
2008), prunetin (8) (Lu et al., 2008) previously isolated from Derris
elliptica (Lu et al., 2008) and D. laxiflora (Lin et al., 1991), 5-
hydroxy-7,30,40-trimethoxyisoflavone (9) (Veitch et al., 2003) and
two phenolic derivatives: coniferyl aldehyde (10) (Miyazawa and
Hisama, 2003) described in patent by Leschot et al. (2007) and 40-
methoxycinnamic acid (11) (Rahman and Moon, 2007).
Compound 1 was obtained as a yellow powder, and its
molecular formula was deduced as C20H16O5 from the analysis
of its NMR and HRMS data (m/z 359.0889 [M+Na]+, calcd for
C20H16O5Na 359.0890). UV data suggested the presence of an
isoflavonoid structure with a characteristic maximum at 266 nm
(Mabry and Markham, 1970). This hypothesis was confirmed in the
1H NMR spectrum of 1 (Table 1), with a typical singlet at dH 7.82
Fig. 1. Structure of derrisisoflavone G (1) with key HMBC (H ! C) correlations.
metabolite BC-1 dehydrate (Tahara et al., 1987). This optical
rotation value suggested a 200R configuration for 1.
90
91
2.2. Hemi-synthesis of compound 1
92
(1H, s, dC 152.0 by HSQC) due to a resonance of H-2. A singlet at dH
13.08 characterized a chelated 5-OH whilst two A2B2 doublets [dH
7.40 (2H, J = 8.5 Hz) and 6.91 (2H, J = 8.5 Hz)] could be associated to
a para-substituted B-ring. The H-6 singlet assignment [dH 6.35
(1H)] was deduced from a HMBC connectivity exhibited between
5-OH and C-6 (dC 94.4) (Fig. 1). The 1H NMR spectrum of 1 also
None of the 2D NMR experiments recorded for 1 allowed us to
unambiguously determine the position of the prenyl appendage.
Therefore the hemisynthesis of this original secondary metabolite
was undertaken to confirm its structure. Our retrosynthetic
analysis highlighted that lupiwighteone (14) was a key interme-
diate. This derivative was prepared via a 4-step C-8 prenylation of
genistein (Al-Maharik and Botting, 2003). Starting from lupiwight-
eone (14), we then achieved a two-step sequence, photooxidation
of the prenyl side chain followed by reduction of the intermediate
hydroperoxide, leading to the secondary allylic alcohol appendage
(16) (Helesbeux et al., 2003). The last step consisted of
intramolecular Mitsunobu condensation of the secondary alcohol
function with the ortho-phenol group yielding the substituted
dihydrofuran ring. Different steps of this hemi-synthesis process
still need to be optimized. However, these reactions allowed us to
synthesize, as a racemic mixture, this original secondary metabo-
lite 1, isolated from D. ferruginea and therefore to confirm its
structure as 5-hydroxy-3-(4-hydroxyphenyl)-7,8-(200-isoprenyldi-
hydrofurano)-4H-chromen-4-one. We eventually propose to name
it derrisisoflavone G.
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97
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101
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103
104
105
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107
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112
exhibited resonances for two ethylenic protons (
and a methyl group [ H 1.78, (s, 3H)], whereas an ABX spin system
H 5.36 (dd, J = 7.3; 9.8 Hz, 1H), 3.43 (dd, J = 9.8; 14.6 Hz, 1H) and
d
H 4.97 and 5.11)
d
[
d
3.08 (dd, J = 7.3; 14.6 Hz, 1H)] was respectively associated with H-
200, H-300a and H-300b. of a dihydrofuran-ring. An isopropenylfuran
moiety derived from a classical prenyl cyclization was thus
identified in 1. The position of this 5-membered ring was then
deduced from the strong HMBC connectivities observed between
H-300a/H-300b and C-8 (
dC 102.8) on one hand and C-8a (dC 163.6) on
the other. This substitution pattern was eventually confirmed by a
NOESY experiment (Table 1). 1 appeared to be optically active with
[
a
]
ꢀ508 (c 0.4, MeOH, 20 8C). Based on a comparison with
D
structurally close compounds such as thonninginisoflavone
(Asomaning et al., 1995), licoagroisoflavone (Li et al., 2001),
lupinisoflavone A, dimethyl-lupinisoflavone A, trimethyl-luteone
2.3. Biological activities
113
Table 1
1H and 13C NMR spectroscopic data (500 MHz, CDCl3) of compound 1.
Crude extracts with four different solvents were tested for
antiparasitic, antioxidant and anti-AGEs activities. The high
antiparasitic potency of the cyclohexane extracts of the leaves
114
115
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123
124
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127
128
129
130
131
132
133
134
Position
d13
dH (J in Hz)
HMBCa
NOESYb
20
C
2
152.0
123.5
180.7
105.9
163.6
94.4
7.82, s
3, 4
10
3
(Plasmodium falciparum: 79.3% inhibition at 10
mg/mL; Leishmania
4
4a
5-OH
6
major: IC50 = 26 ꢁ 6 mg/mL) prompted us to identify the compounds
responsible for this activity. The value obtained for the anti-AGEs
activity (IC50 = 0.28 mg/mL) was also very interesting when com-
pared to the activity of reference compound, aminoguanidine
(IC50 = 1 mg/mL).
Compounds 2–5 showed strong antiparasitic activity against P.
falciparum and L. major according to former results (Batista and
Oliveira, 2009; Kayser et al., 2003). Nevertheless, contrary to
cajaflavanone (Morel et al., 2012), 2–5 exhibited strong cytotoxic-
ity against safe cell strains, as well as tumor cell strains
13.08, s
6.35, s
4a, 6
4a, 8
500, 600
7
166.4
102.8
163.6
123.1
130.4
115.5
155.8
87.9
8
8a
10
20, 60
30, 50
40
200
300
7.40, d (8.5)
6.91, d (8.5)
30, 40, 50
10, 40
2, 30, 50
20, 60
5.36, dd (7.3, 9.8)
3.08, dd (7.3, 14.6)
3.43, dd (9.8, 14.6)
500, 600
200, 8, 400
400, 8a
300
200
30.8
(IC50 < 13.3
mM) (Table 2), which led us to conclude that these
400
500
142.9
112.9
compounds have poor selectivity against the Plasmodium and
Leishmania strains tested. These results are in partial agreement
with those previously reported for lonchocarpol A (4) (Innok et al.,
2009) and 6,8-diprenyleriodictyol (5) (Omisore et al., 2005; Seo
et al., 1997). Alhough exhibiting antiparasitic activity slightly
lower than 2–5, the new isoflavone 1 appeared to be very
4.97, s
5.11, s
1.78, s
200, 600
200, 400
200, 400, 500
5-OH, 600
600
500
600
16.9
a
HMBC correlations (adjusted to 7.5 Hz) are from the specified proton(s) to the
indicated carbons.
b
NOESY correlations are from the specified proton(s) to indicated the proton(s).
Please cite this article in press as: Morel, S., et al., Anti-AGEs and antiparasitic activity of an original prenylated isoflavonoid and
flavanones isolated from Derris ferruginea. Phytochem. Lett. (2013), http://dx.doi.org/10.1016/j.phytol.2013.06.002

G Model
PHYTOL 548 1–6
S. Morel et al. / Phytochemistry Letters xxx (2013) xxx–xxx
3
Table 2
Antiparasitic activity and cytotoxicity of compounds 1–5 compared to cajaflavanone.
Compound
P. falciparuma (IC50
,
mM)
L. majorb (IC50
,
mM)
MRC-5 cells (IC50
,
mM)
KB cells (IC50, mM)
1
>29.8
33.0 ꢁ 1.1
3.3 ꢁ 0.1
5.1 ꢁ 0.5
1.7 ꢁ 0.3
3.7 ꢁ 0.2
40.0 ꢁ 2.0
ND
>29.8
2
11.8 ꢁ 0.2
10.2 ꢁ 0.9
9.9 ꢁ 1.1
5.8 ꢁ 0.8
24.6 ꢁ 0.2
12.8 ꢁ 6.2
6.2 ꢁ 1.4
23.8 ꢁ 0.0
8.0 ꢁ 0.0
>24.6
13.3 ꢁ 7.3
8.3 ꢁ 0.2
8.6 ꢁ 0.2
8.5 ꢁ 0.4
ND
3
4
5
Cajaflavanone
Standards
Chloroquine: 0.097
Pentamidine: 28
Taxotere: 5 ꢂ 10ꢀ10
Taxotere: 2 ꢂ 10ꢀ10
ND: not determined.
a
FcB1/Columbia (n = 3).
b
MHOM/Il/81/BNI (n = 3).
Table 3
Antioxidant and anti-AGEs activities of compounds 1–5 compared to cajaflavanone.
Compound
DPPH (
mM TE/
m
mol)
ORAC (
m
M TE/
m
mol)
AGEs (mM)
1 (derrisisoflavone G)
N.A.
0.36
0.91
0.04
0.99
N.A.
N.A.
N.A.
0.52
0.58
1.25
1.77
1.12
N.A.
5.53
8.68
1.7
2 (5,30,40-trihydroxy-6-(
g,
g
-dimethylallyl)-600,600dimethylpyrano-(200,300:7,8)-flavanone)
1.0
3 (dorsmanine I)
1.9
4 (lonchocarpol A)
1.3
5 (6,8-diprenyleriodictyol)
Cajaflavanone
Naringenina
Genisteina
1.4
0.5
2.0
3.0
Standards
Chlorogenic acid: 1.04
Chlorogenic acid: 4.07
Aminoguanidine: 10
Quercetin: 0.6
N.A.: no activity.
a
Commercial products.
135
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140
141
142
143
144
145
146
147
148
promising because of the absence of toxicity at the tested
concentration on KB cells.
compounds, particularly the atoxic derrisisoflavone G (1), could be 168
useful in AGEs regulation.
169
170
171
None of these compounds exhibited significant antioxidant
potential in DPPH and ORAC bioassays (Table 3). Nevertheless, 1–5
were able to inhibit AGEs formation. Comparison of results
between prenylated and non-prenylated flavonoids (namely
naringenin and genistein) suggests that prenyl side chains have
no influence on the antioxidant potential or anti-AGEs activity.
Flavonoids and isoflavonoids can act either as antioxidant or as a-
dicarbonyl scavengers (Peng et al., 2011). Particularly, as 1
exhibited no significant antioxidant potential in DPPH or ORAC
4. Experimental
4.1. General experimental procedure
HPLC analyses were performed on a Waters 26951 separation 172
module coupled to a Waters1 2996 Photodiode Array Detector 173
using the Empower software package. A Lichrospher1 100 RP18 174
column (150 ꢂ 4.6 mm, 5
mm, Agilent Technologies) using the 175
following gradient: initial mobile phase MeOH/H2O 5/95 reaching 176
100/0 (v/v) in 40 min, with a 1 mL/min flow rate was used for 177
bioassays (Table 3), this compound may scavenge
a
-dicarbonyl
species on C-6 as previously described for catechins (Lo et al., 2006;
Peng et al., 2008) or chalcones (Shao et al., 2008).
fraction analysis.
A Varian OmniSpher C18 column (5 mm, 178
250 ꢂ 4.6 mm) was also used to set up preparative HPLC 179
conditions. Optical rotations were measured by Polartronic D 180
(Eloptron1). IR spectra were recorded on a Bruker Vector22 181
spectrometer. UV data were obtained using a Cary 50 Bio UV 182
149
3. Conclusion
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165
166
167
Herein we reported the identification and hemi-synthesis of the
original derrisisoflavone G (1) together with the isolation of 10
known compounds. To our knowledge, compounds (2–5) are
reported for the first time in the genus Derris. All of these
compounds exhibited significant but poorly selective antiparasitic
activities, whereas they could inhibit AGEs formation. The anti-
AGEs capacity of numerous flavonoids was previously reported
(Matsuda et al., 2003; Wu and Yen, 2005). Comparison of results
between prenylated and non-prenylated flavonoids (namely
naringenin and genistein) suggests that a prenyl side chain have
no influence, neither on antioxidant potential nor on anti-AGEs
activity. It is known that flavonoids and isoflavonoids may act as
either antioxidant or
In the case of 1 which does not exhibit any significant antioxidant
potential in DPPH or ORAC bioassays (Table 3), it is thus assumed
that this compound would scavenge
was previously described for catechin (Lo et al., 2006; Peng et al.,
2008) or chalcone (Shao et al., 2008) derivatives. Therefore, these
Visible Spectrophotometer (Varian).
183
Preparative HPLC was performed on a Varian separation 184
module consisting of an UV/visible ProStar 325 spectrophotome- 185
ter, a PrepStar 218 pump (manual injection, rheodyne valve with a 186
10 mL sample loop) and a ProStar 701 fraction collector. The 187
module was managed with ProStar/Dynamax software. The 188
fractions were injected into an OmniSpher column (C18, 10
mm, 189
250 ꢂ 21.4 mm). Preparative thin-layer-chromatography (PLC) 190
was carried out on silica gel 60F254 (0.25 mm, Merck) and 191
separations were monitored by TLC on 60 F254 (0. 25 mm, Merck) 192
plates.
193
a
-dicarbonyl scavengers (Peng et al., 2011).
CPC was performed using an FCPC 200 (Kromaton) fast 194
centrifugal partition chromatograph. The total volume was 195
275 mL or 1 L. A valve incorporated in the CPC apparatus allowed 196
operation in descending or ascending mode. The system is 197
equipped with a gradient pump, a UV/vis detector, a rheodyne 198
a-dicarbonyl species as this
valve with a 10 mL sample loop and fraction collector.
199
Please cite this article in press as: Morel, S., et al., Anti-AGEs and antiparasitic activity of an original prenylated isoflavonoid and
flavanones isolated from Derris ferruginea. Phytochem. Lett. (2013), http://dx.doi.org/10.1016/j.phytol.2013.06.002

G Model
PHYTOL 548 1–6
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S. Morel et al. / Phytochemistry Letters xxx (2013) xxx–xxx
200
201
202
203
1H NMR, 13C NMR and 2D NMR spectra were recorded in the
appropriate deuterated solvent on a Brucker Avance DRX 500 MHz
spectrometer or Jeol GSX 270 MHz. HR mass spectra were recorded
on LTQ-Orbitrap (ThermoFisher Scientific).
Sephadex gel and preparative HPLC chromatography to give
genistein (6) (3.3 mg). The cyclohexane extract from DfL1 leaves
(30 g) was submitted to FCPC using the R range of the Arizona
heptane/EtOAc/MeOH/H2O (2/1/2/1) system. 10 fractions were
obtained. Purification of fraction 2 over LH-20 Sephadex gel
227
228
229
230
231
232
233
234
235
236
237
204
4.2. Plant material
yielded 40-methoxycinnamic acid (11) (2.0 mg). Fraction
5
was subjected to LH-20 Sephadex gel chromatography followed
by preparative HPLC to give 2 (6.2 mg) and 3 (22.0 mg) or
205
206
207
208
D. ferruginea (ROXBURGH) Bentham was collected at Ha Tinh, in
Vietnam in 1998. It was identified by Dr Nguyen Tien Hiep from the
Hanoi National Herbarium and a voucher specimen was deposited
as no. VN-0452.
over
a Flash silica gel chromatography column to give 4
(11.4 mg). 5 (674.8 mg) was isolated from fraction 7 by LH-20
Sephadex gel .
209
4.3. Extraction and isolation
4.3.1. Compound 1
238
239
240
241
242
243
Pale yellow powder, [
max (log
) 266 (4.65) nm; IR (CHCl3, cmꢀ1): 3245 br (OH), 1649,
1206.
a]
D ꢀ508 (c 0.4, MeOH, 20 8C); UV (MeOH)
210
211
212
213
214
215
216
217
218
219
220
221
222
223
224
225
226
Dried and ground D. ferruginea stems (1200 g) and leaves
(800 g) were successively extracted with cyclohexane, CH2Cl2,
EtOAc and MeOH (8 L, 72 h) in a Soxlhet apparatus to give DfS1-4
extracts (stems) and DfL1-4 extracts (leaves), respectively. The
cyclohexane soluble extract DfS1 (7 g) was separated on MPLC
(silica column) to give 24 fractions. Pratensein (7) (1.2 mg) was
obtained by purification of fraction 11 over LH-20 Sephadex gel
and preparative HPLC. The CH2Cl2 soluble extract DfS2 (3 g) was
also fractionated by MPLC to give 17 fractions. Purification of
fraction 8 over LH-20 Sephadex gel and C18 SPE gave 1 (2.5 mg).
Purification of fraction 9 over LH-20 Sephadex gel generated
coniferyl aldehyde (10). The EtOAc soluble extract DfS3 (20 g)
was fractionated through a FCPC procedure. System: heptane/
EtOAc/MeOH/H2O (10/8/10/6) was used to give 13 fractions.
Prunetin (8) (1.1 mg) and 5-hydroxy-7, 30,40-trimethoxyisofla-
vone (9) (5.4 mg) were obtained from purification of fraction 5
over LH-20 Sephadex gel. Fraction 11 was submitted to LH-20
l
e
1H NMR and 3C NMR: see Table 1. HRMS (EI): m/z 359.0889
[M+Na]+ (calcd for C20H16O5Na 359.0895).
4.4. Hemi-synthesis of 1
244
4.4.1. Preparation of 5,7,40-trihydroxy-8-(3-methylbut-2-en-1-yl)-3-
phenylchromen-4-one (lupiwighteone, 15)
245
246
247
248
249
250
251
252
253
254
Compound 15 (63 mg, 0.19 mmol) was obtained from genistein
6 (0.50 g, 1.85 mmol) as described by (Al-Maharik and Botting,
2003) (Scheme 1).
1H NMR (270 MHz, acetone d6): 1.66 (s, 3H, H-400), 1.81 (s, 3H,
d
H-500), 3.45 (d, 2H, J = 7.0 Hz, H-100), 5.23 (m, 1H, J = 1.5, 7.0 Hz,
H-200), 6.37 (d, 1H, H-6), 6.90 (d, 2H, J = 8.5 Hz, H-30 and H-50), 7.47
(d, 2H, J = 8.5, H-20 and H-60), 8.25 (s, 1H, H-2), 8.60 (7-OH), 8.70
(40-OH), 12.97 (5-OH).
Scheme 1. Hemi-synthesis of compound 1.
Please cite this article in press as: Morel, S., et al., Anti-AGEs and antiparasitic activity of an original prenylated isoflavonoid and
flavanones isolated from Derris ferruginea. Phytochem. Lett. (2013), http://dx.doi.org/10.1016/j.phytol.2013.06.002

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References
312
255
256
257
258
259
260
261
262
263
264
265
266
267
268
269
270
271
272
273
4.4.2. Preparation of 5,7-dihydroxy-3-(4-hydroxyphenyl)-8-
(2-hydroxy-3-methylbut-3-enyl)-4H-chromen-4-one (16)
Al-Maharik, N., Botting, N.P., 2003. Synthesis of lupiwighteone via a para-Claisen– 313
Cope rearrangement. Tetrahedron 59, 4177–4181.
Dried air was bubbled through a 1/1 CH2Cl2/acetone mixture
(15 mL) of compound 15 (63 mg, 0.19 mmol) and tetraphenylpor-
phine (3 mg, 0.005 mmol) as photosensitizer. The reaction mixture
was water-cooled at 15 8C and irradiated with a halogen lamp
(500 W). After total consumption of 15, polymer-supported
triphenylphosphine (16 mg, 1.5 eq.) was added and the solution
was stirred at room temperature for 16 h. The reaction mixture
was then filtered and the resin washed successively with CH2Cl2
(5 mL), a 1/1 mixture of MeOH/CH2Cl2 (5 mL) and CH2Cl2 (5 mL).
After solvent removal under reduced pressure, the crude residue
obtained from the filtrate was submitted to preparative TLC
(MeOH/CH2Cl2 2.5/97.5) to give 16 (42 mg, 42% yield). 1H NMR
314
Almeida, J.G.L., Silveira, E.R., Pessoa, O.D.L., 2008. NMR spectral assignments of a 315
new [C–O–C] isoflavone dimer from Andira surinamensis. Magn. Reson. Chem. 316
46, 103–106.
317
Asomaning, W.A., Amoako, C., Oppong, I.V., Phillips, W.R., Mensah, I.A., Osei-Twum, 318
E.Y., Waibel, R., Achenbach, H., 1995. Pyrano- and dihydrofurano-isoflavones 319
from Milletia thonningii. Phytochemistry 39, 1215–1218.
Batista Jr., R., Oliveira, A.D.J.S., De Oliveira, A.B., 2009. Plant-derived antimalarial 321
agents: new leads and efficient phytomedicines. Part II. Non-alkaloidal natural 322
products. Molecules 14, 3037–3072.
Cheenpracha, S., Karalai, C., Ponglimanont, C., Chantrapromma, K., 2007. Cytotoxic 324
rotenoloids from the stems of Derris trifoliata. Can. J. Chem. 85, 1019–1022.
320
323
325
Derbre, S., Gatto, J., Pelleray, A., Coulon, L., Seraphin, D., Richomme, P., 2009. 326
Automating a 96-well microtiter plate assay for identification of AGEs inhibitors 327
or inducers: application to the screening of a small natural compounds library. 328
Anal. Bioanal. Chem. 398, 1747–1758.
329
(270 MHz, CDCl3): d
1.81 (s, 3H, H-500), 2.92 (dd, 1H, J = 8.0, 15.0 Hz,
Harborne, J.B., Greenham, J., Williams, C.A., Eagles, J., Markham, K.R., 1993. Ten 330
H-100a), 3.11 (dd, 1H, J = 2.5 Hz, 15.0 Hz, H-100b), 4.30 (dd, 1H, J = 2.5,
8.0 Hz, H-200), 6.37 (s, 1H, H-6), 6.90 (d, 2H, J = 8.5 Hz, H-30 and H-
50), 7.47 (d, 2H, J = 8.5, H-20 and H-60), 8.25 (s, 1H, H-2), 8.60 (7-OH),
8.70 (40-OH), 12.97 (5-OH).
isoprenylated and C-methylated flavonoids from the leaves of three Vellozia 331
species. Phytochemistry 34, 219–226.
332
Helesbeux, J.J., Duval, O., Guilet, D., Seraphin, D., Rondeaub, D., Richomme, P., 2003. 333
Regioselectivity in the ene reaction of singlet oxygen with ortho-prenylphenol 334
derivatives. Tetrahedron 59, 5091–5104.
335
Innok, P., Rukachaisirikul, T., Suksamrarn, A., 2009. Flavonoids and pterocarpans 336
from the bark of Erythrina fusca. Chem. Pharm. Bull. 57, 993–996.
Kayser, O., Kiderlen, A.F., Croft, S.L., 2003. Natural products as antiparasitic drugs. 338
Parasitol. Res. 90, 55–62.
Lepape, P., Pagniez, F., Abdala, H., 2003. A new fluorometric method for anti- 340
Leishmania drug screening on axenic amastigotes. Acta Parasitol. 4, 301–305. 341
Li, W., Asada, Y., Koike, K., Hirotani, M., Rui, H., Yoshikawa, T., Nikaido, T., 2001. 342
Flavonoids from Glycyrrhiza pallidiflora hairy root cultures. Phytochemistry 58, 343
337
339
274
275
276
277
278
279
4.4.3. Preparation of 1
65
mL of DIAD (2.5 eq.) and PPh3 (52 mg, 2.5 eq.) were added to
a dry THF solution (3 mL) of 16 (28 mg). The mixture was stirred for
24 h at room temperature. After solvent removal under reduced
pressure, the crude residue was purified by preparative TLC
(MeOH/CH2Cl2 7/93) to generate compound 1 (5 mg, 19%).
595–598.
Lu, H.Y., Liang, J.Y., Yu, P., Qu, W., 2008. Isoflavones and their derivatives from the 345
root of Derris elliptica (Roxb.) Benth. Chin. J. Nat. Med. 6, 265–267.
Mabry, T., Markham, K.R., 1970. The Systematic Identification of Flavonoids, Berlin- Q4347
Heidelberg-New York, p. 354.
344
346
348
280
4.5. Bioassays
Marti, G., Eparvier, V., Moretti, C., Prado, S., Grellier, P., Hue, N., Thoison, O., Delpech, 349
B., Gueritte, F., Litaudon, M., 2010. Antiplasmodial benzophenone derivatives 350
from the root barks of Symphonia globulifera (Clusiaceae). Phtyochemistry 71, 351
281
282
283
284
4.5.1. Antileishmanial activity
Promastigote susceptibility testing was performed with the
Uptiblue1 micromethod described previously (Lepape et al., 2003)
on the L. major strain (MHOM/Il/81/BNI).
964–974.
352
Matsuda, H., Wang, T., Managi, H., Yoshikawa, M., 2003. Structural requirements of 353
flavonoids for inhibition of protein glycation and radical scavenging activities. 354
Bioorg. Med. Chem. 11, 5317–5323.
Miyazawa, M., Hisama, M., 2003. Antimutagenic activity of phenylpropanoids from 356
clove (Syzygium aromaticum). J. Agric. Food Chem. 51, 6413–6422.
355
285
286
287
288
4.5.2. Antiplasmodial activity
Evaluated according to the procedure described previously by
Marti et al. (2010) on P. falciparum strain (FcB1/Columbia).
Chloroquine was used as reference substance.
357
Morel, S., Landreau, A., Nguyen, V.H., Debre, S., Grellier, P., Pape, P.L., Pagniez, F., 358
Litaudon, M., 2012. Preparative isolation, Fast Centrifugal Partition Chromatog- 359
raphy purification and biological activity of cajaflavanone from Derris ferruginea 360
stems. Phytochem. Anal. 23, 152–158.
361
Moret, V., Laras, Y., Cresteil, T., Aubert, G., Ping, D.Q., Di, C., Barthelemy-Requin, M., 362
Beclin, C., Peyrot, V., Allegro, D., Rolland, A., Angelis, F.D., Gatti, E., Pierre, P., 363
Pasquini, L., Petrucci, E., Testa, U., Kraus, J.L., 2009. Discovery of a new family of 364
bis-8-hydroxyquinoline substituted benzylamines with pro-apoptotic activity 365
in cancer cells: synthesis, structure-activity relationship, and action mechanism 366
289
290
291
292
4.5.3. Cytotoxic activities
Cytotoxic activities were evaluated on MRC5 and KB cells in
DMSO at 10 and 1
mg/mL based on the method described by Moret
et al. (2009). Taxotere was used as reference substance.
studies. Eur. J. Med. Chem. 44, 558–567.
367
Ngadjui, B.T., Kouam, S.F., Dongo, E., Kapche, G.W.F., Abegaz, B.M., 2000. Prenylated 368
293
294
295
296
297
4.5.4. AGEs measurement
flavonoids from the aerial parts of Dorstenia mannii. Phytochemistry 55, 915– 369
919.
370
AGEs measurement was evaluated according to the procedure
described previously by Derbre et al. (2009). Aminoganidine was
chosen as reference compound based on previous studies (Reddy
and Beyaz, 2006; Sasaki et al., 2009).
Omisore, N.O.A., Adewunmi, C.O., Iwalewa, E.O., Ngadjui, B.T., Adenowo, T.K., 371
Abegaz, B.M., Ojewole, J.A., Watchueng, J., 2005. Antitrichomonal and antioxi- 372
dant activities of Dorstenia barteri and Dorstenia convexa. Braz. J. Med. Biol. Res. 373
38, 1087–1094.
374
Rahman, A.A., Moon, S.S., 2007. Antimicrobial phenolic derivatives from Den- 375
dranthema zawadskii var. latilobum Kitamura (Asteraceae). Arch. Pharm. Res. 376
298
299
300
301
4.5.5. Scavenging activity of diphenyl-picrylhydrazyl DPPH
radicals and measurement of oxygen radical absorbance
capacity (ORAC)
30, 1374–1379.
377
Rao, N.V.S., Seshadri, T.R., 1946. Chemical examination of plant insecticides. Part I. 378
ChemicalcomponentsofDerris ferruginea. Proc. Indian Nat. Sci. Acad. 24,344–348. 379
Rao, R.R., Tiwari, A.K., Reddy, P.P., Babu, K.S., Ali, A.Z., Madhusudana, K., 2009. New 380
furanoflavonoids, intestinal alpha-glucosidase inhibitory and free-radical 381
(DPPH) scavenging, activity from antihyperglycemic root extract of Derris indica 382
Scavenging activity of diphenyl-picrylhydrazyl DPPH radicals
302 Q3 and measurement of oxygen radical absorbance capacity (ORAC)
303
304
(Lam.). Bioorg. Med. Chem. 17, 5170–5175.
383
were evaluated according to the procedure described previously by
Morel et al. (2012).
Rao, S.A., Srinivas, P.V., Tiwari, A.K., Vanka, U.M., Rao, R.V.S., Dasari, K.R., 2007. 384
Isolation, characterization and chemobiological quantification of alpha-gluco- 385
sidase enzyme inhibitory and free radical scavenging constituents from Derris 386
scandens Benth. J. Chromatogr. B 855, 166–172.
Reddy, V.P., Beyaz, A., 2006. Inhibitors of the Maillard reaction and AGE breakers as 388
therapeutics for multiple diseases. Drug Discov. Today 11, 646–653.
387
305
Acknowledgements
389
´
306
307
308
309
310
311
We are very grateful to G. Aubert from the Museum National
Sasaki, N.A., Garcia-Alvarez, M.C., Wang, Q., Ermolenko, L., Franck, G., Nhiri, N., 390
Martin, M.T., Audic, N., Potier, P., 2009. N-Terminal 2,3-diaminopropionic acid 391
(Dap) peptides as efficient methylglyoxal scavengers to inhibit advanced 392
d’Histoire Naturelle (Paris, France) for the cytotoxicity studies. This
work was supported by a grant from Angers Loire Me´tropole
(France). We also thank David Manley, a professional native
English speaker for correcting the English version of the manu-
script.
glycation endproduct (AGE) formation. Bioorg. Med. Chem. 17, 2310–2320.
393
Seo, E.K., Silva, G.L., Chai, H.B., Chagwedera, T.E., Farnsworth, N.R., Cordell, G.A., 394
Pezzuto, J.M., Kinghorn, A.D., 1997. Cytotoxic prenylated flavanones from 395
Monotes engleri. Phytochemistry 45, 509–515.
396
Please cite this article in press as: Morel, S., et al., Anti-AGEs and antiparasitic activity of an original prenylated isoflavonoid and
flavanones isolated from Derris ferruginea. Phytochem. Lett. (2013), http://dx.doi.org/10.1016/j.phytol.2013.06.002

G Model
PHYTOL 548 1–6
6
S. Morel et al. / Phytochemistry Letters xxx (2013) xxx–xxx
397
398
399
400
401
402
403
404
405
406
407
408
409
410
Shiao, Y.J., Wang, C.N., Wang, W.Y., Lin, Y.L., 2005. Neuroprotective flavonoids from
Flemingia macrophylla. Planta Med. 71, 835–840.
Sreelatha, T., Hymavathi, A., Rao, V.R.S., Devanand, P., Rani, P.U., Rao, J.M., Babu, K.S.,
2009. A new benzil derivative from Derris scandens: structure-insecticidal
activity study. Bioorg. Med. Chem. Lett. 20, 549–553.
Svasti, J., Srisomsap, C., Subhasitanont, P., Keeratichamroen, S., Chokchaichamnan-
kit, D., Ngiwsara, L., Chimnoi, N., Pisutjaroenpong, S., Techasakul, S., Chen, S.T.,
2005. Proteomic profiling of cholangiocarcinoma cell line treated with pomi-
ferin from Derris malaccensis. Proteomics 5, 4504–4509.
Tahara, S., Ingham, J.L., Mizutani, J., 1987. Stereochemical studies on dihydrofurano-
isoflavones. Agric. Biol. Chem. 51, 211–216.
Veitch, N.C., Sutton, P.S.E., Kite, G.C., Ireland, H.E., 2003. Six isoflavones and a
5-deoxyflavonol glycoside from the leaves of Ateleia herbert-smithii. J. Nat. Prod.
66, 2003.
Wu, C.H., Yen, G.C., 2005. Inhibitory effect of naturally occurring flavonoids on
the formation of advanced glycation endproducts. J. Agric. Food Chem. 53,
3167–3173.
Yenesew, A., Twinomuhwezi, H., Kabaru, J.M., Akala, H.M., Kiremire, B.T., Heyden-
reich, M., Peter, M.G., Eyase, F.L., Waters, N.C., Walsh, D.S., 2009. Antiplasmodial
and larvicidal flavonoids from Derris trifoliata. Bull. Chem. Soc. Ethiop. 23,
409–414.
Zheng, X.L., Wing, F.W., 2009. Ethnobotanical study on medicinal plants around Mt.
Yinggeling, Hainan Island, China. J. Ethnopharmacol. 124, 197–210.
411
412
413
414
415
416
417
418
419
420
421
422
The Angiosperm Phylogeny Group, 2009. An update of the Angiosperm Phylogeny
Group classification for the orders and families of flowering plants: APG III. Bot.
J. Linn. Soc. 161, 105–121.
Please cite this article in press as: Morel, S., et al., Anti-AGEs and antiparasitic activity of an original prenylated isoflavonoid and
flavanones isolated from Derris ferruginea. Phytochem. Lett. (2013), http://dx.doi.org/10.1016/j.phytol.2013.06.002