Antiplasmodial benzophenones from the trunk latex of Moronobea coccinea (Clusiaceae)

Article abstract of DOI:10.1016/j.phytochem.2008.10.005

In an effort to find antimalarial drugs, a systematic in vitro evaluation on a chloroquine-resistant strain of Plasmodium falciparum (FcB1) was undertaken on sixty plant extracts collected in French Guiana. The methanol extract obtained from the latex of

Full text of DOI:10.1016/j.phytochem.2008.10.005

Phytochemistry 70 (2009) 75–85
Contents lists available at ScienceDirect
Phytochemistry
journal homepage: www.elsevier.com/locate/phytochem
Antiplasmodial benzophenones from the trunk latex of
Moronobea coccinea (Clusiaceae)
Guillaume Marti a, Véronique Eparvier b, Christian Moretti c, Sophie Susplugas d, Soizic Prado e,
Philippe Grellier d, Pascal Retailleau a, Françoise Guéritte a, Marc Litaudon a,
*
a Institut de Chimie des Substances Naturelles, 1, avenue de la Terrasse, CNRS, 91198 Gif-sur-Yvette, France
b UPS 2561, CNRS, 16, avenue André Aron, 97300 Cayenne, Guyane Française, France
c Unité N° 84 de l’IRD: Biodival, IRD, Technoparc, 5, rue du Carbone, 45072 Orleans Cedex 2, France
d Museum National d’Histoire Naturelle, USM504-EA3335, CP52, 61, rue Buffon, 75231 Paris Cedex 05, France
e Museum National d’Histoire Naturelle, USM502-UMR 5152 CNRS, 63, rue Buffon, 75231 Paris Cedex 05, France
a r t i c l e i n f o
a b s t r a c t
Article history:
In an effort to find antimalarial drugs, a systematic in vitro evaluation on a chloroquine-resistant strain of
Plasmodium falciparum (FcB1) was undertaken on sixty plant extracts collected in French Guiana. The
methanol extract obtained from the latex of Moronobea coccinea exhibited a strong antiplasmodial activ-
Received 1 August 2008
Received in revised form 25 September
2008
ity (95% at 10
cyclic polyprenylated acylphloroglucinols (PPAPs), from which eight showed potent antiplasmodial
activity with IC50 ranged from 3.3 M to 37.2 M.
lg/ml). The phytochemical investigation of this extract led to the isolation of eleven poly-
Available online 4 December 2008
l
l
Keywords:
Moronobea coccinea
Clusiaceae
Ó 2008 Elsevier Ltd. All rights reserved.
Benzophenone
Coccinone
Antiplasmodial
1. Introduction
In an effort to find new naturally antimalarial drugs, we
screened a series of sixty plant extracts prepared from various spe-
cies belonging to the French Guiana biodiversity. The results of this
screening led us to investigate the latex of Moronobea coccinea
Aubl. (Clusiaceae). Indeed, ethyl acetate extracts of trunk bark
and fruits, having a rich latex secretion system, exhibited a strong
antiplasmodial activity (75% and 83% of inhibitory growth at 10 lg/
ml, respectively) whereas the leave and trunk root extracts did not
show any significant activity (4% and 5% of inhibitory growth at
With over 300 millions of new cases each year, resulting in
more than one million deaths annually, malaria remains one of
the most important infectious disease of the developing world.
Fatal cases are generally caused by the most virulent human ma-
laria parasite, Plasmodium falciparum. Symptoms occur during the
asexual parasite development in human red blood cells. Current
clinical treatment involves the use of inexpensive antimalarial
drugs such as chloroquine and their derivatives. However, the
emergence of resistant populations of Plasmodium sp. to this class
of drugs has rendered most of them ineffective (Laufer and Plowe,
2004). Since few years, World Health Organization recommends
the use of a combinatorial therapy to limit resistance phenomenon.
The ‘‘Artemisinin Combination Therapy” is currently the most
effective treatment. Although no clear clinical resistance was yet
reported, there are accumulating evidences for increasing artemis-
inin resistance in vitro of P. faciparum isolates (Gay et al., 1994;
Jambou et al., 2005). There is therefore an urgent need for the dis-
covery of new efficient antimalarial drugs to cure this disease and
to prevent the emergence of resistance.
10
rately by scraping the trunk bark. The methanol extract of the latex
showed 95% of inhibitory growth at 10 g/ml. In this report we de-
lg/ml, respectively). The latex was therefore collected sepa-
l
scribed the isolation, structure elucidation and biological activities
of eleven new polycyclic polyprenylated acylphloroglucinols
(PPAPs) (3–8, 10–14) along with the known isogarcinol, cyclox-
anthochymol and garcinol (1, 2, 9, respectively).
The Clusiaceae family consists of 40 genera and 1200 species
essentially mostly tropical. In French Guyana several species are
well-known in the manufacture of hulls, for the quality of their
wood (Calophyllum brasiliense) and for the healing properties of
their latex (Mahurea, Vismia, Symphonia and Moronobea spp.)
used traditionally for their effectiveness against dermatoses
(Grenand et al., 2004). From a phytochemical point of view, this
family is known to be a rich source of PPAPs with a large spectrum
* Corresponding author. Tel.: +33 1 69 82 30 85; fax: +33 1 69 07 72 47.
E-mail address: marc.litaudon@icsn.cnrs-gif.fr (M. Litaudon).
0031-9422/$ - see front matter Ó 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.phytochem.2008.10.005

76
G. Marti et al. / Phytochemistry 70 (2009) 75–85
of biological activities (Cuesta-Rubio et al., 2005). However, to our
knowledge only guttiferone A has been screened for its antiplasmo-
dial activity (Ngouela et al., 2006). The genus Moronobea consists of
some seven species strictly distributed in Amazonia. Only Morono-
bea pulchra has been investigated and the geranyl benzophenone
marupone was isolated from the trunk wood (Dias et al., 1974).
study on analytical HPLC of these fractions with fraction 6 allowed
us to target those containing the active compounds. Subsequent
preparative HPLC purification resulted in the isolation of com-
pounds 1–14 (Fig. 1).
Compounds 1–14 shared several common spectral characteris-
tics. The UV spectra showed absorption bands at 230 and 280 nm
consistent with aromatic rings and conjugated carbonyl groups.
The IR spectra exhibited bands for hydroxyl (3350 cmꢀ1),
a,b-
2. Results and discussion
unsaturated carbonyl groups (1650, 1668 cmꢀ1), ketone
(1720 cmꢀ1) and aromatic rings (1600 cmꢀ1). The 1H and 13C
NMR spectra recorded in CD3OD appeared quite complex due to
a keto-enol tautomeric balance within the molecules (Fuller
et al., 1999; Gustafson et al., 1992; Williams et al., 2003). In con-
trast, only one set of signals was observed when pyridine-d5 was
used (Table 1). All the structures reported herein follow the IUPAC
convention for numbering bicyclo[3.3.1]nonanes in which C(1)–
C(9) and C(5)–C(9) bonds as bold or hashed wedges according to
One hundred milligram of the latex methanolic extract was fil-
tered on polyamide to remove tannins and 15 mg of the filtered ex-
tract was fractionated on a semipreparative C-18 column to give
nine fractions according to a standardized method (see Section 4)
(Bousserouel et al., 2005). Biological assay on P. falciparum allowed
us to identify the active fraction (fraction 6, tR from 42 to 50 min).
A large amount of the extract (2.5 g) was then submitted to silica
gel chromatography to give 18 fractions (F1–F18). Comparative
Fig. 1. Structures of compounds 1–14.

Table 1
NMR spectroscopic data for compound 1, 3–8 and 10–14 (500 MHz for 1H NMR and 125 MHz for 13C NMR in pyridine-d5.
Position
Compound 1
Compound 3
Compound 4
Compound 5
dC
dH
J (Hz)
dC
dH
J (Hz)
dC
dH
J (Hz)
dC
dH
J (Hz)
1
2
3
4
52.2
171.4
127.2
195.0
69.2
52.2
170.8
129.1
194.9
71.4
48.4
171.3
128.5
195.6
69.4
51.7
171.1
127.0
195.0
69.3
5
6
46.8
46.8
46.8
46.9
7
8
–
47.0
39.8
1.59
2.11
2.43
dt (6.2, 6.1)
dd (14.1, 7.3)
brd (14.1)
42.2
43.1
2.44
1.77
2.43
m
m
m
46.7
42.1
1.62
2.05
2.78
m
47.1
40.0
1.58
2.10
2.48
m
m
m
brd (13.7)
dd (13.7, 5.8)
9
207.9
193.0
130.9
116.6
147.8
153.7
116.5
124.3
26.7
207.3
193.2
131.0
117.1
147.9
153.7
116.5
124.4
25.9
208.6
192.5
130.4
117.1
147.5
153.4
116.1
124.5
26.4
207.9
193.0
130.9
116.7
147.8
153.8
116.6
124.4
26.8
10
11
12
13
14
15
16
17
–
18
19
20
–
8.05
d (2.0)
8.13
brs
8.04
d (2.2)
8.06
d (1.8)
7.28
7.68
2.76
2.95
5.42
d (8.1)
7.31
7.72
2.77
2.97
5.38
brd (7.6)
brd (7.7)
dd (13.6, 4.8)
dd (13.6, 6.4)
brt (6.2)
7.28
7.60
2.81
2.93
5.48
d (8.4)
7.29
7.72
2.79
2.85
5.46
d (8.0)
dd (8.1, 2)
dd (13.7, 5.6)
dd (13.5, 7.6)
brt (6.5)
dd (8.4, 2.2)
dd (13.6, 6.8)
m
dd (8.0, 1.8)
dd (13.3, 7.3)
dd (13.3, 7.3)
brt (5.7)
121.7
134.5
26.6
122.1
134.2
26.2
121.4
134.4
26.5
brt (7.0)
121.7
134.7
26.7
1.57
s
1.55
s
1.76
brs
1.67
s
21
22
23
24
–
25
26
27
28
29
–
30
31
32
–
33
–
34
–
35
–
36
37
38
18.8
27.1
23.1
30.4
1.71
1.05
1.30
1.82
3.21
5.09
s
s
s
18.8
16.6
22.8
28.5
1.73
0.83
1.27
1.82
2.26
5.18
s
s
s
18.7
26.7
22.5
30.2
1.76
1.08
1.28
2.45
3.03
5.15
brs
s
s
m
18.8
27.1
23.1
29.8
1.74
1.08
1.31
2.43
3.19
5.08
s
s
s
ddd (14.2, 9.5, 9.5)
ddd (14.4, 10.7, 9.5)
brt (6.5)
dd (14.1, 8.5)
dd (14.1, 2.5)
brt (6.2)
ma
m
ma
126.4
132.9
26.5
19.0
29.1
123.8
132.9
26.2
18.4
28.5
125.9
133.1
26.1
18.5
29.8
brt (7.0)
126.4
133.4
26.5
19.1
30.4
brt (5.7)
1.74
1.91
1.14
3.27
1.66
s
s
1.62
1.62
1.21
3.28
1.65
s
s
1.71
1.86
1.40
3.08
1.49
s
s
m
1.73
1.86
1.41
3.18
2.30
s
s
dd (13.9, 13.7)
dd (13.9, 3.1)
dt (9.9, 5.0)
dd (13.9, 13.7)
dd (13.6, 3.2)
m
ma
ma
m
dd (14.5, 5.4)
43.8
87.2
21.7
43.8
87.6
21.7
37.1
83.9
48.8
ma
46.0
86.0
22.1
1.23
1.07
s
s
1.14
1.08
s
s
1.04
1.46
ma
ma
1.19
0.99
5.70
5.71
s
29.4
30.4
29.2
30.4
30.6
37.9
26.4
29.4
124.8
144.4
s
1.96
2.42
5.09
brd (14.1)
brd (14.1)
brt (6.5)
1.79
1.96
5.13
m
m
1.03
1.35
1.37
1.89
0.91
0.72
0.97
ma
m
m
m
s
d (15.6)
d (15.6)
122.8
122.9
brt (6.6)
133.7
26.2
18.3
133.7
26.2
18.3
29.0
32.1
27.4
69.9
1.68
1.56
s
s
1.71
1.56
s
s
s
s
31.0b
30.9b
1.43
1.45
s
s
(continued on next page)

Table 1 (continued)
Position
Compound 6
dC
52.3
171.5
127.4
195.1
69.4
Compound 7
Compound 8
Compound 10
dH
J (Hz)
dC
dH
J (Hz)
dC
dH
J (Hz)
dC
dH
J (Hz)
1
2
3
4
52.3
171.2
127.2
195.1
69.5
52.7
171.6
127.5
195.3
69.6
61.5
192.4
120.6
196.2
70.2
5
6
7
8
–
47.4
46.9
40.8
46.9
43.2
40.3
48.1
44.5
45.5
48.2
43.2
45.4
1.54
2.20
2.51
m
2.29
2.28
2.58
ma
2.24
2.47
2.87
m
2.17
1.63
2.40
m
m
m
brd (14.6)
dd (14.6, 7.1)
ma
dd (13.9-5.9)
brd (13.9)
brd (13.3)
9
207.9
193.4
130.9
116.6
147.8
153.8
117.0
124.2
26.6
207.9
193.7
130.9
116.5
147.8
153.9
116.7
125.5
26.7
208.2
193.7
130.9
116.5
147.8
153.8
116.7
121.8
26.4
210.4
195.8
131.1
117.8
147.4
153.3
115.8
125.2
26.2
10
11
12
13
14
15
16
17
–
18
19
20
21
22
23
24
–
25
26
27
28
29
–
30
31
32
–
8.08
d (1.7)
8.08
d (1.8)
8.1
d (1.8)
7.86
brs
7.27
7.70
2.75
2.94
5.42
d (8.2)
7.28
7.73
2.78
2.95
5.43
d (8.0)
7.30
7.71
2.74
2.96
5.45
d (8.0)
7.20
7.61
2.92
3.04
5.55
d (8.1)
brs
m
dd (8.2, 1.7)
dd (13.3, 5.0)
dd (13.3, 7.0)
brt (5.9)
dd (8.0, 1.8)
dd (8.0, 1.8)
dd (13.4, 5.0)
dd (13.4, 7.4)
brt (6.0)
ma
dd (13.5, 7.4)
brt (6.1)
m
121.9
134.4
26.6
18.9
27.1
22.8
30.6
121.8
134.3
26.6
18.8
27.3
23.2
33.7
121.8
134.3
27.0
18.8
26.9
22.9
36.9
122.8
133.3
26.2
18.9
16.7
24.2
29.2
brt (6.0)
1.57
1.71
1.04
1.31
1.80
1.97
5.08
s
s
s
s
m
m
1.57
1.71
1.08
1.43
2.05
2.80
3.80
s
s
s
s
1.58
1.73
1.08
1.36
2.25
2.63
3.69
s
s
s
s
m
m
1.79
1.85
0.88
1.31
1.78
2.15
5.08
s
s
s
s
m
m
dd (13.9, 11.4)
m
brd (10.4)
122.9
133.7
26.2
18.3
29.5
brt (5.9)
78.5
73.4
27.2b
25.0b
29.2
83.1
73.5
26.6b
25.2b
28.9
brd (10.4)
124.5
133.9
26.2
18.4
36.8
brt (6.0)
1.67
1.54
1.14
3.29
1.64
s
s
1.74
1.66
1.14
3.32
1.67
s
1.59
1.73
1.08
3.37
1.64
s
1.59
1.57
2.23
2.33
3.16
s
s
m
m
m
s
s
dd (14.3, 9.3)
dd (14.3, 3.5)
m
ma
ma
dd (14.3, 3.3)
m
dd (14.3, 3.3)
44.0
87.2
21.6
43.9
87.2
21.5
43.9
87.3
21.6
ma
44.9
149.5
113.9
1.22
s
1.22
s
1.13
s
4.82
4.93
1.77
2.28
2.37
5.31
brs
brs
s
m
33
34
–
35
–
29.4
26.9
1.06
2.15
2.43
1.89
1.64
s
29.4
30.4
1.06
1.78
1.98
5.09
s
m
m
29.4
30.3
1.06
1.76
1.94
5.07
s
18.9
33.6
m
m
m
m
ma
m
m
46.6
122.8
brd (6.1)
122.8
brd (6.0)
124.9
brt (6.0)
36
37
38
70.5
30.5
30.2
133.7
26.2
18.3
133.6
26.2
18.3
131.9
26.3
18.6
b
b
1.56
1.56
s
s
1.67
1.54
s
s
1.66
1.52
s
s
1.64
1.68
s
s

Position
Compound 11
Compound 12
Compound 13
Compound 14
1
2
3
4
60.4
193.0
119.5
198.2
68.6
62.2
192.2
119.3
190.3
66.1
62.6
192.2
119.6
196.2
66.2
62.8
191.0
118.0
195.7
66.3
5
6
49.0
49.4
50.2
50.5
7
8
–
47.8
43.1
1.56
2.17
2.48
m
47.9
40.4
1.58
2.22
2.40
m
47.0
42.0
1.51
2.10
2.41
dt (12.5, 5.1)
o
m
47.5
42.2
1.57
2.14
2.43
m
dd (13.9, 6.7)
m
dd (14.0, 7.0)
dd (14.0, 6.9)
dd (14.0, 1.2)
ma
9
206.4
195.7
142.7
116.6
159.3
120.1
129.7
121.4
27.0
211.3
196.7
131.4
117.9
147.3
153.2
116.1
124.4
32.2
210.9
195.9
130.1
118.0
147.4
153.4
116.0
125.9
32.2
210.1
195.0
129.9
117.9
147.4
153.7
115.9
126.0
32.2
10
11
12
13
14
15
16
17
–
18
19
20
21
22
23
24
–
25
26
27
28
29
–
30
31
32
–
7.75
brs
7.95
d (2.0)
7.90
s
7.84
d (2.1)
7.29
7.35
7.53
2.91
2.98
5.60
brd
m
brd
m
7.21
7.70
2.87
2.89
5.81
d (8.4)
dd (8.4, 2.0)
brs
brs
brt (6.0)
7.19
7.59
2.71
2.91
5.67
m
m
7.20
7.53
2.78
2.91
5.69
d (8.2)
dd (8.2, 2.2)
dd (13.8, 5.6)
dd (13.8, 8.0)
brt (7.1)
dd (13.6, 9.1)
dd (13.6, 5.7)
m
m
122.8
133.6
26.7
18.9
27.7
brt (5.6)
122.2
133.8
26.6
18.7
27.8
23.6
30.2
122.5
133.8
26.7
18.6
27.4
23.8
33.3
121.7
134.7
26.7
18.4
27.2
1.84
1.83
1.11
1.36
2.42
2.67
5.05
s
s
s
s
m
m
1.78
1.78
1.15
1.39
2.45
2.78
5.06
ma
1.88
1.79
1.09
1.41
1.59
1.65
5.00
s
s
s
s
m
m
m
1.86
1.81
1.13
1.44
2.43
2.50
5.01
s
s
s
s
ma
s
s
m
m
23.6
33.51
23.7
30.15
dd (15.0, 1.2)
dd (15.0, 9.5)
brt (7.0)
126.4
132.5
26.3
18.5
37.6
brt (5.6)
126.4
132.6
26.4
18.6
29.5
brt (6.0)
125.9
132.7
26.2
18.4
32.2
125.8
132.8
26.3
18.4
30.2
1.62
1.60
2.28
s
s
m
1.62
1.63
2.30
2.40
2.66
s
s
m
m
m
1.61
1.56
2.09
2.56
2.52
s
s
o
m
m
1.63
1.58
3.10
3.14
s
s
d (15.0)
d (15.0)
44.8
149.8
112.7
3.14
m
35.9
137.4
134.0
45.4
147.1
113.5
128.8
125.9
47.4
4.81
4.92
1.78
2.32
2.42
5.31
brs
brs
s
m
5.21
brs
4.81
4.87
1.94
1.59
1.65
1.87
1.94
brs
brs
s
m
m
m
m
1.67
1.74
d (19.3)
d (19.3)
33
34
–
35
–
36
37
–
19.1
27.7
32.3
34.5
18.4
33.3
29.2
37.0
1.62
1.13
1.59
1.78
2.11
0.94
ma
ma
ma
ma
s
1.29
m
m
124.7
brt (5.6)
25.5
36.6
29.3
1.96
2.20
1.86
0.69
m
m
s
131.9
26.3
23.4
31.5
146.7
110.5
21.1
28.3
1.67
1.71
s
s
s
4.72
4.76
1.65
brs
brs
s
s
38
18.7
29.9
0.84
s
22.5
28.9
0.84
s
a
Overlap.
b
Assignments are interchangeable, brt: broad triplet, brs: broad singulet.

80
G. Marti et al. / Phytochemistry 70 (2009) 75–85
Grossman’s recommendations (Ciochina and Grossman, 2006).
While several PPAPs have been isolated in both enantiomeric
forms (Baggett et al., 2005; Gustafson et al., 1992; Iinuma et al.,
1996; Krishnamurthy et al., 1981; Rao and Venkatswamy, 1980;
Roux et al., 2000; Sahu et al., 1989) only few of them have their
absolute configuration confirmed experimentally (Blount and Wil-
liams, 1976; Krishnamurthy et al., 1982). Therefore, it has been
postulated that a positive or negative [a]D value is due to the bicy-
clo[3.3.1]nonane system orientation. PPAPs presented here follow
this rule.
Compound 1 has been isolated as a brown crystal, [
a
]
D
ꢀ168°
(c 1.0, CHCl3). The HREIMS indicated a [M+Na]+ ion peak at
625.3499, which suggested a molecular formula of C38H50O6Na
(calc. 625.3505). Examination of the 1D and 2D NMR spectra asso-
ciated with the value of optical rotation confirmed that 1 was iso-
garcinol (Iinuma et al., 1996; Krishnamurthy et al., 1981).
However, no detailed NMR data were available in the literature.
We reported in Table 1 the complete assignment of 1H and 13C
NMR data of 1. The X-ray diffraction analysis from crystals of the
dibrosylate derivative of 1, (10) allowed us to confirm the absolute
configuration of isogarcinol previously established by Krishnamur-
thy and coworkers (Krishnamurthy et al., 1982; Rogers et al.,
1981). X-ray data are available on the Cambridge Crystallographic
Data Centre (see supplementary material section).
Fig. 2. Key HMBC (arrows) and COSY (bold) correlations for 4.
tion of H-30 and the Me-36 group indicating a cis-junction be-
tween the tetrahydropyrane and the dimethylcyclohexane
rings. From a biogenetic point of view, compound 4 could be
derived from a double intramolecular cyclisation of the
x-isola-
vandulyl side chain at C-1 position of the known xanthochymol,
although this last has not been isolated (Dreyer, 1974). The
trimethylperhydrochromene system has already been described
The spectroscopic data of compound 2 were similar to those of
cycloxanthochymol isolated from Garcinia subelliptica and Garcinia
pyrifera (Iinuma et al., 1996; Roux et al., 2000).
in a type A benzophenone derivative isolated from Clusia
obdeltifolia (Teixeira and Cruz, 2005). Compound 4 was named
coccinone A.
Compound 3, a brown powder, exhibited [
a
]
ꢀ158° (c 1.0,
Compound 5 was isolated as a brown oil. The HREIMS indicated
a molecular formula C38H50O7 deduced from the ion peak at m/z
641.3446 M+Na+ (calc. 641.3454), which suggested the presence
of an additional hydroxy group when compared with 1. Examina-
tion of the 1D and 2D NMR spectral data of compound 5 confirmed
that a benzoylphloglucinol skeleton is associated to a tetrahydro-
pyrane ring. The presence of a 3-hydroxyl-3-methylbutenyl side
chain in position C-30 was suggested by the following evidence.
In the HMBC spectrum, correlations were observed from the qua-
ternary carbon C-36 and the methine C-30 (dC 69.9 and 46.0,
respectively) to the vinylic protons at dH 5.70 and 5.71 (each 1H,
d, J = 15.6 Hz) H-34 and H-35, respectively, and from C-35 at dC
144.4 and C-36 to the methyl protons H3-37 and H3-38 (dH 1.43
and 1.45, respectively). The correlation between H-34 and H-30
(dH 2.30) in the COSY spectrum confirmed the location of the side
chain at position C-30. These data, together with other results of
2D NMR analysis confirmed the structure of compound 5, which
was named coccinone B.
Compound 6 was isolated as a brown oil. The molecular formula
C38H52O7 has been deduced by HREIMS from the ion peak [M+Na]+
m/z 643.3619 (calc. 643.3611) and suggested the presence of two
additional protons when compared with 5. The presence of a satu-
rated side chain was confirmed in the 1H and 13C NMR spectra by
signals for two additional methylene groups instead of two vinylic
carbons. In the HMBC spectrum, correlations were observed from
the quaternary carbon C-36 (dC 70.5) and the methylene C-35 (dC
46.6) to the methyl protons H3-37 and H3-38 (dH 1.56, 6 H, s). In
the COSY spectrum, the correlations between H-35, H-34 and H-
30 confirmed the location on position C-30 of the 3-hydroxy-3-
methylbutyl side chain. This was further confirmed by NOESY
D
CHCl3). The HREIMS indicated an ion peak at m/z 625.3519, which
suggested the same molecular formula as 1 and 2 (C38H50O6Na).
The 1H and 13C NMR spectral data were almost identical to those
of 1. In the 13C NMR, only the signals of carbons C-7, C-8, C-22,
C-23 and C-24 shifted slightly suggesting a b-orientation for the
prenyl side chain located at position 7. According to Ciochina and
Grossman, the orientation of the substituent on C-7 (axial or equa-
torial) can be deduced from the 1H and 13C NMR spectral data (Cio-
china and Grossman, 2006; Hamed et al., 2006; Piccinelli et al.,
2005). The C-22 and C-23 methyl resonate at d 27.1 and 23.1
respectively for isogarcinol (1) having an axial prenyl side chain
on position C-7, whereas the gem-dimethyl group showed chemi-
cals shifts at d 16.6 and d 22.8, respectively in compound 3. The up-
field shift of the C-22ax signal resulted from a c-gauche interaction
between this carbon and the CH2-24 of the prenyl group. In the
NOESY spectrum, nOe interactions between H-8eq, H-7 and Me-
23eq (dH 1.77, 2.44 and 1.27, respectively) confirmed the absolute
configuration (S) for C-7. Compound 3 was thus identified as 7-
epi-isogarcinol.
Compound 4 was isolated as a brown oil. The HREIMS showed
the same ion peak [M+Na]+ m/z 625.3521 as compound 1 indicat-
ing the same formula C38H50O6Na. The 1H and 13C NMR spectra
confirmed the presence of a 1,2,4-trisubstituted benzene ring (Ta-
ble 1) and displayed resonances for nine methyl groups [dH 1.08 (3
H, s, Me-22), 1.28 (3 H, s, Me-23), 0.91 (3 H, s, Me-36), 0.72 (3 H, s,
Me-37), 0.97 (3 H, s, Me-38), 1.76 (6 H, brs, Me-20 and Me-21), 1.71
(3 H, s, Me-27), and 1.86 (3 H, s, Me-28)], and two vinylic protons
[dH 5.48 (1H, brt, J = 7.0 Hz, H1-18), and 5.15 (1H, brt, J = 7.0 Hz, H1-
25)]. The COSY and HMBC spectra revealed the presence of an addi-
tional 1,1-dimethylcyclohexane fused to the tetrahydropyrane ring
on positions C-30 and C-31 instead of a prenyl side chain observed
in 1 (Fig. 2). The relative configuration of the asymmetric carbons
delineated in structure 4 has been determined by nOe experiment.
Resonances for Me-36 with H-29ax (dH 3.08), H-30 (dH 1.49) and H-
32ax (dH 1.46) allowed us to place these groups in the same face of
the trimethylperhydrochromene moiety. On the other hand, reso-
nances of Me-38 with H-35ax (dH 1.89) allowed us to locate these
groups on the opposite face. These correlations suggested a b-posi-
interactions between Me-32 with H -34 (dH 2.43). Compound 6
a
was named coccinone C.
Compound 7, which was isolated as a brown oil, has an [a]
D
value of ꢀ76° (c 0.4, CHCl3). The HREIMS revealed an ion peak
at [M+Na]+ m/z 659.3563 (calc. 659.3560), giving the molecular
formula C38H52O8 suggesting the presence of an additional hy-
droxy group when compared with compound 6. Examination
of the 1D and 2D NMR spectra showed that compound 7 shared
several characteristics with the previous compounds. The

G. Marti et al. / Phytochemistry 70 (2009) 75–85
81
1H NMR spectrum showed only two vinylic protons at dH 5.09
and 5.43 (1H each, brt, J = 6.1 Hz, H1-35 and H1-18, respec-
tively), which suggested the presence of two prenyl side chains.
The location of the first prenyl side chain at C-5 was suggested
by correlations from C-4, C-5, C-6 and C-18 to the methylene
H2-17 (dH 2.78) in the HMBC spectrum. In the COSY spectrum,
correlations between protons at dH 1.14/3.32, 1.67, 1.78/1.98
and 5.09 (H2-29, H-30, H2-34 and H-35, respectively) confirmed
the location of the second prenyl side chain at C-30. The pres-
ence of a 2,3-dihydroxy-3-methylbutyl side chain on position
C-7 was suggested by the following evidence. In the HMBC
spectrum, correlations were observed from the quaternary car-
bon C-26 (dC 73.4) and the methine carbon C-25 (dC 78.5) to
the methyl protons H3-27 and H3-28 (3H, s each, dH 1.74 and
1.66, respectively). In addition, correlations between H-24, H-7
and H-25 in the COSY spectrum allowed to locate the side chain
on position C-7. The chemical shifts of the carbons C-25 and C-
26 observed in the 13C NMR spectrum suggested the presence of
two hydroxy groups. The chemical shifts of the gem-dimethyl
group C-22 and C-23 (dC 27.3 and 23.2, respectively) suggested
an axial position of the side chain. This was confirmed by the
correlations between H-7eq (dH 2.29), H-8eq (dH 2.58) and Me-
22 (dH 1.08) observed in the NOESY spectrum. On the other
hand, despite the presence of correlations between H-25 (dH
the catechol previously observed for compounds 1–9. All other
data are fully similar to those of garcinol (9) indicating that com-
pound 11 was 14-deoxygarcinol.
Compound 12, a yellow oil, exhibited an [
a
]
ꢀ32° (c 0.8,
D
CHCl3). The HREIMS indicated the same molecular formula as gar-
cinol (C38H50O6). The 1H and 13C NMR spectral data (Table 1) were
very close to those of garcinol. The COSY and HMBC spectrum sug-
gested that the
x-isolavandulyle side chain was replaced by a
1,3,3-trimethylcyclohex-1-enyl (Fig. 3). In the HMBC spectrum, a
gem-dimethyl group is deduced from the correlations observed
from the quaternary carbon C-33 (dC 32.3), the methylene carbon
C-34 (dC 34.5) and the methine C-32 (dC 134.0) to the methyl pro-
tons H3-37 and H3-38. Correlations were also observed from the
two methines C-32 and C-30 (dC 35.9) to the methyl protons H3-
36 (dH 2.11) and from the methylene carbon C-35 (dC 25.5) and
the quaternary carbons C-1, C-2 and C-31 (dC 62.2, 192.2 and
137.4, respectively) to the methylene protons H2-29, which con-
firmed the presence of the C10 moiety located at position C-1. In
the NOESY spectrum, the correlation observed between H-30 (dH
2.66) and H-34ax (dH 1.62) suggested a b-orientation for H-30. This
relative configuration was confirmed by correlations between H-
34ax and H3-38 from one side and H3-37 with H-35ax (dH 1.59)
on the other side. These data, together with other results of 2D
NMR analysis confirmed the structure of compound 12, which
was named coccinone F.
3.80) with Me-27, Me-28, H-24 (dH 2.80), H-7eq (dH 2.29) and
a
H-8eq (dH 2.58), we cannot resolve the relative configuration at
C-25. Compound 7 was named coccinone D.
Compound 13 was isolated as a yellow oil. The molecular for-
mula of C38H50O6 has been deduced from the ion peak at
[M+Na]+ m/z = 625.3524 (calc. 625.3505) obtained by HREIMS.
The 1H and 13C NMR spectra suggested the presence of a 1,2,4-tri-
substituted benzene ring (Table 1). In addition, resonances for
eight methyl at dH 1.09 (3 H, s, Me-22), 1.41 (3 H, s, Me-23), 1.94
(3 H, s, Me-33), 1.65 (3 H, s, Me-38), 1.61 (3 H, s, Me-27), 1.56 (3
H, s, Me-28), 1.79 (3 H, s, Me-21) and 1.88 (3H, s, Me-20), two tri-
substituted double bonds at dH 5.67 (1 H, m, H-18) and 5.00 (1 H,
m, H-25), two olefinic exomethylenes at dH 4.81 and 4.87 (2 H,
Compound 8, which was isolated as a brown oil, has an [a]D va-
lue of ꢀ70° (c 0.3, CHCl3). The HREIMS presented the same ion peak
as compound 7 at m/z 659.3560 [M+Na]+ (calc. 659.3560) indicat-
ing the same formula C38H52O8. The 1H and 13C spectra of both
compounds 7 and 8 were very similar except for the ‘‘south region”
implying carbons C-6, C-7, C-8, C-24 and C-25. A close examination
of the 1D and 2D NMR data allowed us to deduce that compound 8
possessed the same skeleton as compound 7 with an axial 2,3-
dihydroxy-3-methylbutyl side chain at position C-7. In contrast
with compound 7, other correlations were observed between the
protons H-25 and Me-27, Me28, H-24b (dH 2.25), H-7eq (dH 2.24)
and Me-23eq (dH 1.36) in the NOESY spectrum. Compound 8 was
named coccinone E.
The preparation of Mosher esters should confirm the absolute
configuration at C-25. Unfortunately the amount of both com-
pounds was not sufficient to obtain the desired products. To our
knowledge, it is the first report of a 2,3-dihydroxy-3-methylbutyl
side chain located at C-7 in the type B PPAPs.
Compound 9 was isolated as yellow crystals. The HREIMS
[M+Na]+ m/z 625.3494 indicated a molecular formula of C38H50O6
(calc. 625.3505). The 1H and 13C NMR (Table 1) data and the value
of [
a
]
ꢀ135° (c 1.0, CHCl3) were identical with those of garcinol
D
isolated from Garcinia xanthochymus (Krishnamurthy et al., 1981;
Rao and Venkatswamy, 1980). A complete set of data obtained
from the X-ray crystallographic analysis is available on Cambridge
Crystallographic Data Centre (see supplementary material section).
Fig. 3. Key HMBC (arrows) and COSY (bold) correlations for 12.
Compound 10, a yellow oil, exhibited [
a]
ꢀ86° (c 0.8, CHCl3).
d
The HREIMS indicated an ion peak at m/z 625.3517, which sug-
gested the same molecular formula as 9 (C38H50O6). The 1H and
13C NMR spectral data were almost identical to those of 9. In the
13C NMR, only the signals of carbons C-7, C-8, C-22, C-23 and C-
24 shifted slightly suggesting a b-orientation for the prenyl side
chain located at position 7. This was confirmed by nOe data (see
discussion for compound 3). Compound 10 was deduced to be
the 7-epi-garcinol.
Compound 11 was purified as a yellow oil, the HREIMS indi-
cated a molecular formula of C38H50O5 [M+Na]+ m/z = 609.3538
(calc. 609,3556). Examination of the 1D and 2D NMR data sug-
gested the presence of a disubstituted benzene ring instead of
Fig. 4. Key HMBC (arrows) and COSY (bold) correlations for 13 and 14.

82
G. Marti et al. / Phytochemistry 70 (2009) 75–85
Fig. 5. X-ray crystal structure for compound 14.
brs, H-32) and dH 4.72 (1 H, m, H-37), 4.76 (1 H, m, H-37), and seven
allylic protons at dH 2.1-2.9 (10 H, m) were observed (Table 1).
These data associated with the correlations observed in the COSY
(Fig. 4). The relative configuration of compound 14 was confirmed
by the X-ray crystallographic analysis (Fig. 5, see supplementary
material section). Compound 14 was named coccinone H. To our
knowledge, coccinones G and H are the first example of PPAPs hav-
ing a C10 side chain attached at C-5.
and the HMBC spectra, suggested the presence of a
and two prenyl side chains (Fig. 4). The location of the prenyl side
chain, -oriented, at position C-7 was deduced from the chemical
x-isolavandulyl
a
shifts of the gem-dimethyl at dC 27.4 and 23.8 (H3-22 and H3-23,
2.1. Biological activity
respectively) observed in the 13C NMR spectra. The location of
the
x
-isolavandulyl side chain at position C-5 was deduced from
The antiplasmodial activity of compounds 1–14 was evaluated
by their ability to inhibit the growth of P. falciparum following
the correlations from the carbons C-4, C-5, C-9 and C-30 to the
methylene protons H2-29 observed in the HMBC spectrum
(Fig. 4). The relative configuration of the methine C-30 remains un-
known. Compound 13 was named coccinone G.
Compound 14 was purified as a yellow crystal. The ion peak in
HREIMS [M+Na]+ m/z 625.3499 indicated the same molecular for-
mula as compound 13, C38H50O6Na (calc. 625.3505). The 1H and
13C NMR spectra showed the same benzoylphloroglucinol skeleton
as the previous one (Table 1). The 2D NMR data were very similar
Table 2
IC50 value of compounds 1–14 tested against FcB1 strain of P. falciparum and MRC5
cells.
Compound
IC50 P.f. FcB1 (
l
M) SD (n = 3) IC50 MRC 5 (lM) (n = 3)
Isogarcinol (1)
Cycloxanthochymol
(2)
3.5 1.1
2.1 0.5
3.5 0.4
3.6 0.6
with those of compound 11, except for the
x-isolavandulyl side
7-epi-isogarcinol (3)
Coccinone A (4)
Coccinone B (5)
Coccinone C (6)
Coccinone D (7)
Coccinone E (8)
Garcinol (9)
7-epi-garcinol (10)
14-deoxygarcinol (11)
Coccinone F (12)
Coccinone G (13)
Coccinone H (14)
Chloroquine
5.1 1.3
4.3 0.5
5.5 0.4
9.0 1.2
7.0 0.9
2.3 0.5
3.3 0.8
11.7 0.6
9.3 1.1
10.9 0.3
9.1 1.3
11.0 0.2
11.3 0.6
19.1 0.2
21.3 2.8
11.5 0.4
10.5 0.6
25 3.8
chain, which is replaced by a 2,4,4-trimethylcyclohex-1-enyl moi-
ety in the case of compound 14. In the HMBC spectrum, an addi-
tional gem-dimethyl group is deduced from the correlations
observed from the quaternary carbon C-33 (dC 29.2) and the meth-
ylenes C-32 and C-34 (dC 47.4 and 37.0, respectively) to the methyl
protons H3-37 and H3-38. In addition, correlations from the car-
bons C-30 (dC 128.8), C-31 (dC 125.9) and C-32 (dC 47.4) to the
methyl protons H3-36, and from the carbons C-32 and C-35 (dC
29.3) to the methylene protons H2-34 confirmed the presence of
a 2,4,4-trimethylcyclohex-1-enyl moiety. Finally, correlations from
the carbons C-4, C-5, C-9, C-30 and C-35 to the methylene protons
H2-29 indicated that the C10 moiety is attached at position C-5
4.9 0.7
12.6 4.8
10.1 4.6
37.2 9.7
17.0 9.4
19.2 5.9
16.6 6.1
0.078 0.006
0.010 0.005
Taxotere
31.5 4.5

G. Marti et al. / Phytochemistry 70 (2009) 75–85
83
the method of Desjardins et al. (1979). The anti-plasmodial activity
4.3. Extraction and isolation procedures
against the chloroquine-resistant strain of P. falciparum FcB1 and
the cytotoxicity on the human MRC-5 cell line are summarized in
Table 2. The results demonstrated that compounds 1–14 exhibited
potent antiplasmodial activity. Compounds 1–8 have shown the
best IC50s between 3.3 and 9.0 lM, indicating that the benzophe-
nones having a tetrahydropyrane ring are the most potent com-
Trunk bark, roots bark, fruits, and leaves (20 g each) were
extracted three times using the automatic extractor DionexÒ
ASE 300 with EtOAc (3 ꢁ 100 ml) followed by MeOH
(3 ꢁ 100 ml) at 40 °C and 100 bar. Each extract were concen-
trated in vacuo at 40 °C to yield 2.69 g (trunk bark, EtOAc),
1.48 g (trunk bark, MeOH), 5.14 g (roots bark, EtOAc) 0.96 g
(trunk bark, MeOH), 2.57 g (fruits, EtOAc), 5.23 g (fruits,
MeOH), 1.71 g (leaves, EtOAc) and 1.13 g (leaves, MeOH). Ali-
quot of 100 mg of each extract and yellow resin were dis-
solved in MeOH and then were filtered on polyamide
pounds. In contrast, compounds 9–14 showed moderate
antiplasmodial activities with IC50s over 10 lM. Moreover, hydro-
xyl position on the benzoyl groups seems to play an important role
for the antiplasmodial activity regarding the IC50 values for garcin-
ol and dehydroxygarcinol. The analysis of the IC50s obtained on
the MRC-5 cell line indicated that compounds having a hydroxyl-
ated side chain (5–8) exhibited a lowest cytotoxicity. Further
mechanistic and structure-activity relationship studies have to be
done in order to better understand the structural features needed
for developing new anti-malarial drugs based on PPAPs.
cartridge before being tested at 10 lg/ml on P. falciparum
FcB1 strain. The resin-filtered extract was then fractionated
on a semi-preparative C-18 column according to a standardized
method previously detailed (Bousserouel et al., 2005). Fraction
6 was shown to possess antiplasmodial activity.
A larger amount of latex (3.51 g) was dissolved in MeOH and fil-
tered on polyamide DC 6 to remove bark residue and tannins. The
filtered extract was dried under vacuum at 40 °C to give a yellow
gummy residue (2.35 g), which was subjected to silica gel chroma-
tography using cyclohexane, mixtures of cyclohexane–ethyl ace-
tate (9:1–1:9) and ethyl acetate as eluents. According to their
TLC profiles, 18 fractions (F1–F18) were obtained. Analytical HPLC
was used to compare these fractions with the active fraction 6 pre-
viously identified and allowed us to identify those containing the
active compounds. Fraction 12 (740 mg) was submitted to a pre-
parative C-18 column using an isocratic mobile phase consisting
of MeCN–H2O 90:10 + 0.1% formic acid, leading to the isolation of
7-epi-isogarcinol 3 (109 mg; w/w 4.6%), isogarcinol 1 (332 mg;
w/w 14.1%), cycloxanthochymol 2 (11 mg; w/w 0.5%), coccinone
A 4 (18 mg; w/w 0.76%), 14-dehydroxygarcinol 11 (4 mg; w/w
0.17%) garcinol 9 (134 mg; w/w 5.7%), 7-epi-garcinol 10 (20 mg;
w/w 0.85%) coccinone F 12 (16 mg; w/w 0.68%), coccinone G 13
(61 mg; w/w 2.6%) and coccinone H 14 (30 mg; w/w 1.4%), with
retention times of 4.7, 5.2, 5.5, 6.0, 9.3, 9.4, 16.5, 13.5 and
12.6 min, respectively. Fraction 13 (100 mg) using an isocratic sys-
tem consisting of MeCN–H2O 70:30 + 0.1% formic acid on the same
column, afforded coccinone B 5 (4 mg; 0.17%), coccinone C 6 (3 mg;
0.12%), coccinone D 7 (3 mg; 0.12%) and coccinone E 8 (4 mg,
0.17%) with retention times of 6.5, 7.2, 10.6 and 11.4 min,
respectively.
3. Conclusions
Phytochemical investigation of M. coccinea led to the isolation
of eleven new PPAPs derivatives. This is the first report on the iso-
lation of PPAPs with a C10 side chain attached at C5. Furthermore,
in vitro antiplasmodial assays reveal that PPAPs having a furan ring
fused to the phloroglucinol moiety exhibited potent biological
activity. Biological and phytochemical investigations of other trop-
ical Clusiaceae species are underway in order to confirm this
hypothesis.
4. Experimental
4.1. General
The NMR spectra were recorded with a Bruker 500 MHz (ad-
vance 500) spectrometer with pyridine-d5 as solvent. ESIMS were
obtained on a Navigator mass Thermoquest. HRESIMS were run
on a MALDI-TOF spectrometer (Voyager-De STR; Perspective Bio-
systems). Kromasil analytic, semi-preparative and preparative C-
18 columns (250 ꢁ 4.5 mm; 250 ꢁ 10 mm and 250 ꢁ 21.2 mm
I.D, 5 l
m ThermoÒ) were used for preparative HPLC separations
using a Waters autopurification system equipped with a binary
pump (Waters 2525), UV–vis diode array detector (190–
a
4.3.1. Isogarcinol 1
600 nm, Waters 2996) and a PL-ELS 1000 ELSD detector Polymer
Laboratory. IR spectra were obtained on a Nicolet FTIR 205 spectro-
photometer. The UV spectra were recorded on a Perkin–Elmer
Lambda 5 spectrophotometer. Specific rotation was obtained in
CHCL3 with a JASCO P-1010 polarimeter. Melting point for isolated
Brown crystal, HR-EIMS [M+Na]+ m/z 625.3499, C38H50O6Na re-
quires 625.3505; [
a
]
D
ꢀ158° (c 1, CHCl3); mp 251 °C; UV kmax
(log ): 317 (3.82), 277 (4.14), 233 (4.07); IR mmax (ns) 3290, 2920,
e
2850, 1730, 1670, 1590, 1520, 1440, 1370, 1290, 1170 cmꢀ1 1H
;
NMR and 13C NMR (Table 1). CCDC (Deposit No: 689070) contains
the supplementary crystallographic data (Krishnamurthy et al.,
1982).
crystals was done on a Büchi B-540. Silica gel 60 (35–70 lm) and
analytical TLC plates (Si gel 60 F 254) were purchased from SDS
(France). Polyamide DC 6 and polyamide cartridge was purchased
from Macherey–Nagel (Chromabond PAÒ, 1 g). All other chemicals
and solvents were of analytical grade and purchased from SDS
(France).
4.3.2. 13-O,14-O-Bis(4-bromobenzoylsulfonyl)-isogarcinol 10
At
a pyridine solution of isogarcinol (32 mg in 350 ll,
0.053 mmol) were added DMAP (3 mg, 0.024 mmol) and 4-bro-
mobenzenesulfonyl chloride (44 mg, 0.17 mmol). After stirring
for 4 h at 0 °C, the solution was extracted with diisopropyl ether
and washed with HCl 1 N. Four products were separated using pre-
parative TLC plates (cyclohexane/EtOAc 7:3): starting materials
(17 mg); 13-O-(4-bromobenzoylsulfonyl)-isogarcinol (5 mg); 14-
O-(4-bromobenzoylsulfonyl)-isogarcinol (9 mg); and the desired
product 13-O,14-O-bis(4-bromobenzoylsulfonyl)-isogarcinol (7
mg). The last was crystallized under MeOH/H20 95:5. mp 138 °C;
CCDC (Deposit No: 688119) contains the supplementary crystallo-
graphic data.
4.2. Plant material
Trunk barks, roots barks, fruits, leaves and latex of M. coccinea
were collected in the dense rain forest of French Guyana by one
of us (G.M). Latex was collected by scoring trunk tissue and then
scraping the latex as it oozed from the wound. This yellow resin
was transferred to Teflon-stoppered glass vials. A herbarium sam-
ple (GM-008) was deposited in the Guyane Herbarium of Cayenne,
and identified by M-F. Prevost (Institut de Recherche pour le
Développement).

84
G. Marti et al. / Phytochemistry 70 (2009) 75–85
4.3.3. Cycloxanthochymol 2
4.3.13. Coccinone F 12
Yellow oil, [
a]
+112° (c 1, CHCl3), HR-EIMS [M+Na]+ m/z
Yellow oil, [
a
]
D
ꢀ32° (c 0.7, CHCl3), HR-EIMS [M+Na]+ m/z
D
625.3512, C38H50O6Na requires 625.3505; UV kmax (log
e): 315
625.3490, C38H50O6Na requires 625.3505; UV kmax (loge): 280
(3.87), 277 (4.18), 229 (4.16); IR mmax 3330, 2920, 1730, 1650,
(4.09), 229 (4.03); IR mmax 3300, 2920, 1720, 1660, 1590, 1440,
1590, 1520, 1440, 1370, 1290, 1190, 1120 cmꢀ1
.
1375, 1290, 1200 cmꢀ1 1H NMR and 13C NMR (Table 1).
;
4.3.4. 7-Epi-isogarcinol 3
4.3.14. Coccinone G 13
Brown powder, [
a]
D ꢀ158° (c 1.0, CHCl3); HR-EIMS [M+Na]+ m/z
Yellow oil, [
a]
ꢀ16° (c 1.0, CHCl3); HR-EIMS (M+Na+) m/z
D
625.3519, C38H50O6Na requires 625.3505; UV kmax (log
e): 319
625.3524, C38H50O6Na requires 625.3505; UV kmax (loge): 279
(3.85), 276 (4.15), 233 (4.12); IR mmax 3320, 2970, 2930, 1730,
(4.18), 233 (4.10); IR mmax 3280, 2920, 2850, 1730, 1640, 1600,
1650, 1590, 1520, 1440, 1370, 1290, 1170 cmꢀ1
NMR (Table 1).
;
1H NMR and 13C
1440, 1380, 1290, 1180 cmꢀ1 1H NMR and 13C NMR (Table 1).
;
4.3.15. Coccinone H 14
4.3.5. Coccinone A 4
Yellow crystal, [
a]
+2° (c 1.0, CHCl3); HR-EIMS (M+Na+) m/z
D
Brown oil, [
a]
D
+28° (c 1.0, CHCl3); HR-EIMS [M+Na]+ m/z
625.3499, C38H50O6Na requires 625.3505; mp 169 °C; UV kmax
625.3521, C38H50O6Na requires 625.3505; UV kmax (log
e
): 276
(loge): 282 (3.96), 232 (3.88); IR mmax 3310, 2940, 2870, 1720,
(3.23), 232 (3.19); IR mmax 3280, 2920, 2850, 1730, 1650, 1550,
1590, 1440, 1380, 1290, 1190 cmꢀ1
1).
;
1H NMR and 13C NMR (Table
1440, 1370, 1290, 1170 cmꢀ1 1H NMR and 13C NMR (Table 1).
;
4.3.6. Coccinone B 5
4.3.16. The crystal data of 14
Brown oil, [
a]
D
ꢀ55° (c 0.3, CHCl3); HR-EIMS [M+Na]+ m/z
C38H50O6,
a = 31.987(5), b = 12.928(2), c = 8.847(2) Å, b = 98.54(2)°, V =
3617.9(11) Å3, Z = 4, Dc = 1.107 g cmꢀ3 (Mo–Ka) = 0.073 mmꢀ1
,
M = 602.78,
monoclinic,
space
group
C2,
641.3446, C38H50O7Na requires calc. 641.3454; UV kmax (loge):
316 (3.71), 278 (4.03), 233 (3.99); IR mmax 3330, 2930, 1730,
,
l
1650, 1590, 1440, 1350, 1290, 1120 cmꢀ1
(Table 1).
;
1H NMR and 13C NMR
F(000) = 1304, prismatic crystal, colorless, size = 0.20 ꢁ 0.15 ꢁ
0.08 mm, 21043 reflections measured [Rint = 0.022], 3376 unique,
wR2 = 0.115 for all data, conventional R = 0.044 [(D/s)max = 0.001]
on F-values of 2587 reflections with I > 2 s(I), GooF = 1.039 for all
data and 407 parameters. Unit cell determination and intensity
data collection (q = 25.1°) were performed on an Enraf–Nonius
KAPPA CCD diffractometer at 293(2) K. Structure solution by direct
methods and refinement by full-matrix least-squares on F2.
Programs: COLLECT, (Nonius, 1997–2000) HKL2000 (Otwinowski
and Minor, 1997), SHELX97 (Sheldrick, 2008). CCDC [Deposit No:
688118] contains the supplementary crystallographic data for this
paper (see supplementary material section).
4.3.7. Coccinone C 6
Brown oil, [
a]
ꢀ60° (c 0.2, CHCl3); HR-EIMS [M+Na]+ m/z
D
643.3619, C38H52O7 (calc. 643.3611); UV kmax (loge): 316 (3.83),
277 (4.15), 233 (4.11); IR mmax 3350, 2930, 1730, 1640, 1590,
1440, 1350, 1290, 1120 cmꢀ1 1H NMR and 13C NMR (Table 1).
;
4.3.8. Coccinone D 7
Brown oil, [
a
]
d
ꢀ76° (c 0.4, CHCl3); HR-EIMS [M+Na]+ m/z
659.3563, C38H52O8 (calc. 659.3560); UV kmax (loge): 313 (3.83),
278 (4.14), 233 (4.08); IR mmax 3410, 2930, 1730, 1590, 1440,
1350, 1290, 1120 cmꢀ1
;
1H NMR and 13C NMR (Table 1).
4.4. Biological activities
4.3.9. Coccinone E 8
Extracts were tested against the chloroquine-resistant FcB1/
Colombia strain of P. falciparum (Frappier et al., 1996). P. falciparum
was maintained continuously in vitro in human erythrocytes accord-
ing to Trager and Jensen (2005). The antiplasmodial activity was
determined according to Desjardins et al. (1979). The extracts were
dissolved in dimethylsulfoxide (DMSO) and tested at a concentration
Brown oil, [
a]
ꢀ70° (c 0.3, CHCl3); HR-EIMS [M+Na]+ m/z
D
659.3560, C38H52O8 (calc. 659.3560); UV kmax (loge): 313 (3.82),
278 (4.14), 233 (4.08); IR mmax 3430, 2930, 1730, 1590, 1440,
1350, 1290, 1120 cmꢀ1 1H NMR and 13C NMR (Table 1).
;
of 10 lg/ml. Compounds showing significant inhibition rates were
4.3.10. Garcinol 9
submitted to serial dilutions with culture medium before being
added to asynchronous parasite cultures (1% parasitemia and 1% final
hematocrite) in 96-well microplates for 24 h at 37 °C. A concentra-
Yellow crystal, [
a
]
D ꢀ135° (c 1.0, CHCl3); HR-EIMS [M+Na]+ m/z
625.3494, C38H50O6Na requires 625.3505; mp 132 °C; UV kmax
(loge): 279 (4.18), 232 (4.04); IR mmax 3300, 2920, 1720, 1640,
tion of 0.5 l
Ci of [3H] hypoxanthine was then added to each well,
1590, 1440, 1370, 1290, 1190 cmꢀ1; 1H NMR and 13C NMR are con-
sistent with published data (Krishnamurthy et al., 1981; Rao and
Venkatswamy, 1980; Sahu et al., 1989). CCDC (Deposit No:
688120) contains the supplementary crystallographic data (see
supplementary material section).
and parasites were maintained for an additional 24 h. The growth
inhibition for each compound concentration was determined by
comparing the radioactivity incorporated in the treated culture with
that in the control culture maintained on the same plate. The concen-
trations causing 50% inhibition of parasite growth (IC50) were calcu-
lated from the drug concentration-response curves.
The human diploid embryonic lung cells MRC-5 were seeded
into 96-well microplates at 2000 cells per well. The cytotoxicity as-
says were performed according to a published procedure (Tempete
et al., 1995). TaxotereÒ was used as a control compound.
4.3.11. 7-epi-garcinol 10
Yellow oil, [
a
]
ꢀ86° (c 0.8, CHCl3); HR-EIMS [M+Na]+ m/z
D
625.3517, C38H50O6Na requires 625.3505; UV kmax (loge): 280
(4.32), 230 (4.17); IR mmax 3300, 2920, 1720, 1640, 1520, 1440,
1375, 1290, 1200 cmꢀ1 1H NMR and 13C NMR (Table 1).
;
4.3.12. 14-deoxygarcinol 11
Supplementary material
Yellow oil, [
a]
D
ꢀ42° (c 0.3, CHCl3); HR-EIMS [M+Na]+ m/z
CCDC 689070 (1), CCDC 688119 (10), CCDC 688120 (9) and CCDC
688118 (14) contain the supplementary crystallographic data for
this paper. These data can be obtained free of charge via http://
609.3538, C38H50O5 (calc. 609.3556); UV kmax (loge): 283 (4.05),
258 (4.03); IR mmax 3290, 2920, 1720, 1640, 1560, 1440, 1370,
1290, 1190 cmꢀ1 1H NMR and 13C NMR (Table 1).
;

G. Marti et al. / Phytochemistry 70 (2009) 75–85
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Acknowledgements
This works was supported by an ICSN–CNRS grant to one of us
(G.M.). We are very grateful to Bernard Delpech and Philippe Nuh-
ant (ICSN) for their helpful advises, Marie-Françoise Prevost (IRD)
for her assistance in the identification and the plant collection,
Marie-Thérèse Martin for her assistance in the structural determi-
nation and Geneviève Aubert (ICSN) who performed the cytotoxic
assays on MRC5 cells.
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