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, 707.2 (2MNa)
, and in the
negative ion mode at m/z 341.4 (M-H)
2
. The molecular weight (342 Da)
and the chemical formula C
23
H
18
O
3
were readily determined by EI HRMS
due to m/z 342.1344. The chemical formulas of 1 and 2 suggested the
presence of 15 double bond equivalents.
In the
1
H NMR spectrum of compound 1, the low-eld signals at d
H
6.42 4 7.55 were pecular to eight aromatic protons. Furthermore, two
olenic protons appeared at d
H
7.08 and 8.28 as two doublets. The congur-
ation of the double bond at C-2 was determined as 2E from the large trans
coupling constants of H-2 (J 16.0 Hz) and H-3 (J 16.1 Hz). This fact
suggested the presence of a trans olen conjugated with a carbonyl. The
aliphatic region of
1
H NMR spectrum could be interpreted as methylene
two nonequivalent geminal protons at d
H
2.81 (dd, J 16.7, 3.4 Hz) and
3.05 (dd, J 16.7, 12.9 Hz), and an oxygen-linked methine proton at d
H
5.49 (dd, J 13.0, 3.0 Hz). In the
13
C NMR and DEPT spectra of 1,
nineteen carbon signals were visible (Table 1) which were assignable to one
carbonyl carbon (d
C
192.00), twelve aromatic carbons (8 CH and 4 C
q
signals), two signals of olenic carbons, one methylene and three oxygen-
bonded (C-2
0
, C-6
0
, C-7a
0
) carbons.
A
1
H-
1
H chemical shift correlation spectroscopy (COSY) experiment
(Fig. 1) demonstrated coupling only from the olenic proton of H-2 at d
H
7.08 to the olenic proton of H-3 (d
H
8.28) and the methylene protons of
H-3
0
at d
H
2.81/3.05 to the methine proton at d
H
5.49 (H-2
0
).
The HMBC experiment (Fig. 1) showed, additionally, a long range
coupling from the methylene protons of H-3
0
to the carbonyl C-1 and from
the olenic protons of H-2 and H-3 to the quaternary C-4
0
and C-1
000
, respect-
ively. Cross peaks were also observed from the aromatic doublets of H-5
0
to
a quaternary (C-4
0
, C-3a
0
) and C-7
0
, and H-7
0
to C-5
0
and C-3a
0
. Similarly, a
coupling was established from the aromatic proton of H-4
0 0 0
to C-2
0 0 0
and
C-6
0 0 0
; from the aromatic protons of H-3
0 0
and H-5
0 0 0
to the quaternary C-1
0 0 0
;
from the aromatic protons of H-3
000
and H-5
0 0
to the quaternary C-1
0 0
and from
H-2
0 0
and H-6
0 0
to C-4
0 0
. The stereochemistry of the furanyl ring of 1 and 2
was elucidated mainly on the basis of NOESY cross-peaks for H-2
0
/H-3
0
b
and H-3
0
a/H-3
0
b. Irradiation of H-2
0
enhanced the signals of H-3
0
a and H-3
0
b, suggesting that H-2
0
was gauche to H-3
0
a (equatorial) and H-3
0
b (axial).
Anti-Inammatory and Antibacterial Agents 347
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Sanionin B (2) was obtained as colorless needles, [a]
D
25
229.18 (c 0.1,
CHCl
3
). The compound 2 showed the same molecular weight (342 Da) and the
chemical formula (C
23
H
18
O
3
) by EI HRMS as 1. The UV-VIS spectrum of 2
showed absorption maxima at 294, 270, and 224 nm by HPLC/diode array
Figure 1. Selected
1
H-
1
H COSY (bold lines) and
1
H-
13
C HMBC (arrows) corre-
lations of sanionin A (1) and sanionin B (2).
Table 1.
1
H and
13
C NMR data of sanionin A (1) and sanionin B (2) in CDCl
3
(500 MHz and 125.76 MHz, TMS as internal standard, chemical shifts in ppm)
Position
1 2
d
H
d
C
d
H
d
C
1 192.00 s 191.22 s
2 7.08 d, J 16.0 Hz 131.09 d 6.59 d, J 12.2 Hz 129.22 d
3 8.28 d, J 16.1 Hz 128.81 d 7.04 d, J 12.3 Hz 131.18 d
2
0
5.49 dd, J 13.0, 3.0 Hz 79.28 d 5.44 dd, J 11.1, 3.6 Hz 79.24 d
3
0
2.81 dd, J 16.7, 3.4 Hz 2.84 dd, J 16.8, 3.1 Hz
3.05 dd, J 16.7, 12.9 Hz 45.85 t 3.06 dd, J 16.8, 12.9 Hz 45.48 t
3a
0
112.55 s 113.00 s
4
0
143.60 s 142.30 s
5
0
6.76 d, J 2.2 Hz 108.76 d 6.39 d, J 2.5 Hz 102.99 d
6
0
164.10 s 161.66 s
7
0
6.42 d, J 2.3 Hz 103.11 d 6.26 d, J 2.5 Hz 112.65 d
7a
0
166.00 s 165.21 s
1
0 0
139.20 s 139.00 s
2
0 0
/6
0 0
7.52 d, J 7.3 Hz 126.50 d 7.08 d, J 7.4 Hz 127.04 d
3
0 0
/5
0 0
7.41 t, J 7.4 Hz 129.29 d 7.12 t, J 7.4 Hz 128.67 d
4
0 0
7.35 t, J 7.4 Hz 129.21 d 7.10 t J 7.4 Hz 129.22 d
1
0 0 0
138.54 s 136.92 s
2
0 0 0
/6
0 0 0
7.55 d, J 7.3 Hz 127.40 d 7.54 d, J 7.3 Hz 127.89 d
3
0 0 0
/5
0 0 0
7.35 t, J 7.4 Hz 129.39 d 7.41 t, J 7.3 Hz 128.90 d
4
0 0 0
7.25 t, J 7.4 Hz 128.64 d 7.35 t, J 7.3 Hz 128.90 d
6
0
-OH 5.42 s 5.44 s
V. Ivanova et al. 348
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analysis. This spectrum demonstrated a decrease in the intensity of the corre-
sponding maximum absorption by comparison with that of 1. The conjugated
carbonyl absorption of 2 was shifted to 294 nm from 318 nm in 1. The major
difference arose from the two olenic protons of the a,b-unsaturated ketone
system, which shifted upeld at d
H
6.59 (H-2) and 7.04 (H-3). The congur-
ation of the double bond (in
1
H NMR spectrum) of compound 2 at C-2 was
determined as 2Z from the cis coupling constants of H-2 (J 12.2 Hz) and
H-3 (J 12.3 Hz), typical for a cis-olen. Sanionin B (2) was identied as
cis isomer of 1. The chemical shifts of all protons and carbon atoms of 1
and 2 are summarized in Table 1.
Sanionins A (1) and B (2) were investigated in vitro for antimicrobial
activity against Gram-positive and Gram-negative bacteria, yeasts and la-
mentous fungi (Table 2). Both substances showed activity against Gram-
positive bacteria, such as Bacillus subtilis ATCC 6633, Staphylococcus
aureus SG 511, Staphylococcus aureus 134/93 (MRSA), Mycobacterium
vaccae 10670, and Enterococcus faecalis 1528 (VRE), but were inactive
against Gram-negative bacteria, yeast, and lamentous fungi such as Escher-
ichia coli SG 458, Pseudomonas aeruginosa SG137, Sporobolomyces salmo-
nicolor 549 and Penicillium notatum JP 36.
The antiproliferative and cytotoxic effects of sanionin A (1) and sanionin
B (2) were determined with L-929 mouse broblast cells, K-562 human
leukemia cells, and HeLa human cervix carcinoma. The GI
50
and CC
50
values are summarized in Table 3.
The inhibition effect of sanionin A and sanionin B on 3a-hydroxysteroid
dehydrogenase was investigated by comparison with the polyenic macrolide
antibiotics, such as pimaricin, hexafungin, fungicidin and the standard
compound indomethacin. The polyene macrolide antibiotics displayed an
Table 2. Diameter of inhibition zones (mm) of sanionin A (1) and sanionin B (2) in
the agar diffusion assay
Test organisms
Diameter of inhibition zones (mm)
1 2
Bacillus subtilis ATCC 6633 15.0 14.5
Staphylococcus aureus SG 511 17.0 13.0
Staphylococcus aureus 134/93 (MRSA) 13.5 13.0
Escherichia coli SG 458 0 0
Pseudomonas aeruginosa SG 137 0 0
Pseudomonas aeruginosa K 799/61 0 0
Enterococcus faecalis 1528 (VRE) 12.5 12.0
Mycobacterium vaccae 10670 19.5 19.0
Sporobolomyces salmonicolor 549 0 0
Penicillium notatum JP 36 0 0
Anti-Inammatory and Antibacterial Agents 349
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inhibitory effect
[10]
comparable to sanionin A (1). The sanionin B inhibited the
enzyme too, but with lower efciency than sanionin A. The IC
50
values are
summarized in Table 4.
On the basis of the obtained data (Table 1, Fig. 1), it was established that
sanionin A (1) [1- (2,3-dihydro-6
0
-hydroxy-2
0
-phenyl-4
0
-benzofuranyl)-3-
phenyl-2 (E)-propen-1-one] and sanionin B (2) [1- (2,3-dihydro-6
0
-hydroxy-
2
0
-phenyl-4
0
-benzofuranyl)-3-phenyl-2 (Z)-propen-1-one] were identical
with pallidisetin A and pallidisetin B, isolated from the moss Polytrichum pal-
lidisetum, which showed cytotoxic activity against two human tumor cell lines
(RPMI-7951 and U-251 MG) (1). The sanionin A (trans-isomer) was with
better biological activity than sanionin B (cis-isomer).
CONCLUSION
Both substances, sanionin A (1) and sanionin B (2), for the rst time, were
isolated from the antarctic moss Sanionia georgico-uncinata
[11,12]
and can
be classied into cinnamoyl bibenzyls. We did not nd data about the
Table 4. Inhibition of 3a-hydroxysteroid dehydro-
genase by sanionin A(1) and sanionin B (2)
Compounds IC
50
(mg/mL)
Sanionin A (1) 70
Sanionin B (2) 130
Pimaricin 76
Hexafungin 76
Fungicidin 80
Indomethacin 24
Table 3. Antiproliferative effect and Cytotoxicity of sanionin A (1) and sanionin B
(2) against cell cultures of L-929, K-562 and HeLa cells
Compounds
Antiproliferative effect
GI
50
(mg/mL)
a
Cytotoxicity
CC
50
(mg/mL)
b
L-929 K-562 HeLa
Sanionin A (1) 32.9 23.1 34.1
Sanionin B (2) 31.1 23.3 32.4
a
Cellular growth inhibition.
b
Cytotoxic efcacy.
V. Ivanova et al. 350
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isolation of secondary metabolites from the moss Sanionia georgico-uncinata
as 3a-hydroxysteroid dehydrogenase inhibitors and with antiproliferative
and cytotoxic effects against L-929, K-562, and HeLa cell lines. These
compounds showed activity against important Gram-positive pathogens like
mycobacteria, multiresistant staphylococci, and vancomycin resistant
enterococci.
ACKNOWLEDGMENTS
Support of this work by the German Research Foundation, Bonn, Germany
(Project 436 BUL 113/107/0-2) is gratefully acknowledged.
REFERENCES
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Anti-Inammatory and Antibacterial Agents 351
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Received September 29, 2006
Accepted November 3, 2006
Manuscript 7502
V. Ivanova et al. 352
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