M. Y. Putra et al. / Bioorg. Med. Chem. Lett. 22 (2012) 2723–2725
2725
Politecnica delle Marche and was partially financed by 7th Frame-
work Programme of the European Community (REGPOT-2008-1-
NatPharma contract). Mass and NMR spectra were recorded at
‘
Centro di Servizio Interdipartimentale di Analisi Strumentale’,
Università di Napoli ‘Federico II’.
Supplementary data
of the most important compounds described in this article.
References and notes
1.
Coval, S. J.; Patton, R. W.; Petrin, J. M.; James, L.; Rothofsky, M. L.; Lin, S. L.; Patel,
M.; Reed, J. K.; Mc Phail, A. T.; Bishop, W. R. Bioorg. Med. Chem. Lett. 1996, 6, 909.
2. Lindel, T.; Jensen, P. R.; Fenical, W.; Long, B. H.; Casazza, A. M.; Carboni, J.;
Fairchild, C. R. J. Am. Chem. Soc. 1997, 119, 8744.
Figure 3. Effect of different concentrations of sinularioside (2) and cerebroside 1 on
ꢁ
the production of NO2 by J774 macrophages stimulated with LPS (1
lg/mL). Each
bar represents the mean of three separate experiments run in triplicate.
3. El Sayed, K. A.; Hamann, M. T.; Waddling, C. A.; Jensen, C.; Lee, S. K.; Danstan, C.
A.; Pezzuto, J. M. J. Org. Chem. 1998, 63, 7449.
4
5
6
7
8
.
.
.
.
.
Fattorusso, E.; Romano, A.; Taglialatela-Scafati, O.; Bavestrello, G.; Bonelli, P.;
Calcinai, B. Tetrahedron Lett. 2006, 47, 2197.
Fattorusso, E.; Romano, A.; Taglialatela-Scafati, O.; Achmad, M. J.; Bavestrello,
G.; Cerrano, C. Tetrahedron 2008, 64, 3141.
Fattorusso, E.; Romano, A.; Taglialatela-Scafati, O.; Irace, C.; Maffettone, C.;
Bavestrello, G.; Cerrano, C. Tetrahedron 2009, 65, 2898.
Fattorusso, E.; Luciano, P.; Putra, M. Y.; Taglialatela-Scafati, O.; Ianaro, A.;
Panza, E.; Bavestrello, G.; Cerrano, C. Tetrahedron 2011, 67, 7983.
Putra, M. Y.; Bavestrello, G.; Cerrano, C.; Renga, B.; D’Amore, C.; Fiorucci, S.;
Colonies of Sinularia sp. (580 g wet weight) were collected in January 2010 in
the Bunaken Marine Park of Manado along the coasts of the small island of
Siladen (North Sulawesi, Indonesia) at a depth of 2–5 m. A small voucher
sample is deposited at the Dipartimento per lo Studio del Territorio e delle sue
Risorse, Università di Genova. The colonies have been repeatedly extracted
NO by iNOS has been implicated in the pathogenesis of the inflam-
1
9
matory response.
The production of NO2 (a stable metabolite of NO) is a recog-
nised parameter to measure macrophages activation and iNOS
ꢁ
induction. Unstimulated J774 cells (murine monocyte/macro-
ꢁ
phages) generated undetectable (<5 nmol/mL) amounts of NO2
,
while the stimulation of the cells with LPS (1
l
g/mL) for 24 h pro-
9
.
ꢁ
duced a dose-dependent release of NO2 (15.5 ± 0.3 nmol/mL).
ꢁ
NO2 levels in culture media from J774 macrophages were mea-
sured 24 h after LPS with the Griess reaction as previously de-
2
0
scribed.
When the cells were incubated with different
3
with MeOH and CHCl at room temperature and the obtained material (6.8 g)
concentrations (10, 30, and 100
cant dose-dependent (p <0.001) inhibition of LPS-induced NO2
l
M) of sinularioside (2) a signifi-
has been chromatographed by MPLC over silica gel eluting with a gradient
system of increasing polarity from n-hexane to EtOAc to MeOH. Fractions
eluted with EtOAc were further purified by normal phase HPLC (n-hexane/
EtOAc 5:95 as eluent) to afford sinularioside 2 (3.5 mg); fractions eluted with
EtOAc/MeOH 9:1 were further purified by RP18 HPLC (MeOH/H O 95:5 as
2
eluent) to afford cerebroside 1 (1.7 mg).
ꢁ
production was observed, with 58% inhibition at 30
lM (Fig. 3).
Cerebroside 1 was consistently less active, showing only a slight
ꢁ
inhibition of NO2 production (16% inhibition at 30
l
M).
1
1
0. Cheng, S.-Y.; Wen, Z.-H.; Chiou, S.-F.; Tsai, C.-W.; Wang, S.-K.; Hsu, C.-H.; Dai,
C.-F.; Chiang, M. Y.; Wang, W.-H.; Duh, C.-Y. J. Nat. Prod. 2009, 72, 465.
1. Costantino, V.; Fattorusso, E.; Mangoni, A.; Teta, R.; Panza, E.; Ianaro, A. Bioorg.
Med. Chem. 2010, 18, 5310.
The anti-inflammatory potential of marine cerebrosides, related
to the inhibitory effect of NO release, has been first evidenced a
decade ago for a fucosylated triglycosylceramide from the sponge
2
1
12. Striegler, S.; Haslinger, E. Monatsh. Chem. 1996, 127, 755.
Aplysinella rhax. Interestingly, all the following investigations,
25
13. Sinularioside (2). Amorphous white solid; ½
a +35.9 (c 0.3, CHCl
ꢀ
3
); (+)ESI-MS
1
7
D
either starting from the isolation of new natural analogues or
from the preparation of synthetic derivatives,22 agreed in indicat-
ing the key role played by the presence of fucose residues within
the glycosidic moiety. Fucosylated glycolipids appear to be active
even when the ceramide portion is replaced by a greatly simplified
+
(
positive ions) m/z 627 [M+Na] . HR-ESIMS: m/z 627.3362 (calcd. for
H52NaO12 627.3356); H NMR (CDCl , 500 MHz): d 5.27 (1H, dd, J = 10.8,
30 3
.3 Hz, H-3 ), 5.18 (1H, dd, J = 10.8, 3.5 Hz, H-2 ), 5.15 (1H, d, J = 3.5 Hz, H-1 ),
.02 (1H, d, J = 3.5 Hz, H-1 ), 4.88 (1H, dd, J = 10.0, 3.5 Hz, H-2 ), 4.15 (1H, dd,
J = 10.0, 3.3 Hz, H-3 ), 4.07 (1H, overlapped, H-4 ), 4.05 (1H, overlapped H-5 a),
4.04 (1H, overlapped, H-4 ), 3.78 (1H, bd, J = 13.0 Hz, H-5 a), 3.72 (1H, br d,
J = 12.0 Hz, H-5 b), 3.66 (1H, dt, J = 9.8, 6.8 Hz, H-1a), 3.55 (1H, dd, J = 13.0,
.5 Hz, H-5 b), 3.38 (1H, m, H-1b), 2.13 (3H, s, 3 -OAc), 2.06 (3H, s, 2 -OAc),
1
C
3
5
0
0
00
0
0
0
0
0
00
00
0
0
2
2
00
aglycone, thus suggesting the limited importance of the non-
polar portion of the molecule. In this context, the very low activity
of cerebroside 1 is not surprising and gives support to the previ-
ously obtained results.
0
00
00
1
2
0
.03 (3H, s, 2 -OAc), 1.60 (2H, m, H
2
-2), 1.33-1.25 (22H, m, H
(3H, t, J = 9.0 Hz, H -14); C NMR (CDCl , 125 MHz): d 171.4 (3 -OCOCH ),
2
-3 to H
2
-13), 0.88
1
3
00
3
3
3
00
0
00
00
0
170.9 (2 -OCOCH
3
), 170.1 (2 -OCOCH
3
), 96.8 (C-1 ), 96.2 (C-1 ), 74.2 (C-2 ),
0
0
00
0
00
7
5
1
4.1 (C-4 ), 73.8 (C-2 ), 71.1 (C-4 ), 68.6 (C-3 ), 68.3 (C-1), 68.2 (C-3 ), 62.1 (C-
On the other hand, the inhibitory activity exhibited by sinula-
rioside (2), a non-fucose containing glycolipid, clearly indicates
for the first time that glycolipids possessing a poly-acetylated su-
gar residues could have an anti-inflammatory potential worthy of
further investigations. These could also provide interesting insights
into the still poorly understood structure–activity relationships for
anti-inflammatory glycolipids.
0
00
), 59.4 (C-5 ), 31.5 (C-12), 30.3 (C-2), 29.5 (C-4 to C-11), 25.7 (C-3), 22.3 (C-
0
0
0
0
3), 22.1 (2 -OCOCH ), 21.1 (22 -OCOCH ), 20.8 (3 -OCOCH ), 14.2 (C-14).
3
3
3
14. Wojnar, J. M.; Northcote, P. T. J. Nat. Prod. 2011, 74, 69.
15. Fattorusso, E.; Mangoni, A. Fortschr. Chem. Org. Naturst. 1997, 72, 215.
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1
2
2
2
9. Ianaro, A.; Ialenti, A.; Sautebin, L.; Di Rosa, M. Naunyn Schmiedebergs Arch.
Pharmacol. 1998, 358, 489.
0. Ianaro, A.; Ialenti, A.; Maffia, P.; Sautebin, L.; Rombolà, L.; Carnuccio, R.; Iuvone,
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Acknowledgments
This research was partially conduced during the Master Course
‘
Tropical Marine Biodiversity and Natural Products’ of Università