Polar steroidal compounds from the Far-Eastern starfish Lethasterias fusca

Article abstract of DOI:10.1007/s11172-008-0030-y

Nine steroidal compounds including three new steroidal glycosides, viz., sodium (24S)-3,24-di-O-(β-D-xylopyranosyl)-5α-cholestane-3β, 6β,8,15α,24-pentol 15-sulfate (fuscaside A), (24S)-3,24-di-O-(β- D-xylopyranosyl)-5α-cholestane-3β,6β,8,15α,24-pentol (fuscaside B), and (22E,24R)-24-O-(β-D-xylopyranosyl)-5α-cholest-22- ene-3β,6α,8,15β,24-pentol (desulfated minutoside A); three previously known glycosides, viz., distolasterosides D₁ and D ₂ and pycno-podioside A; two previously known polyhydroxysteroids, viz., 5α-cholestane-3β,6α,8,15β,16β,26-hexaol and 5α-cholestan-3β,4β,6α,78,15β,16β,26-octol; and the known sodium 24,25-dihydro-marthasterone 3-sulfate were isolated from the Far-Eastern starfish Lethasterias fusca. The structures of these compounds were elucidated by NMR spectroscopy and mass spectrometry.

Full text of DOI:10.1007/s11172-008-0030-y

Russian Chemical Bulletin, International Edition, Vol. 57, No. 1, pp. 204—208, January, 2008
204
Polar steroidal compounds from the FarꢀEastern starfish
Lethasterias fusca

N. V. Ivanchina, T. V. Malyarenko, A. A. Kicha, A. I. Kalinovskii, and P. S. Dmitrenok
Pacific Institute of Bioorganic Chemistry, FarꢀEastern Branch of the Russian Academy of Sciences,
159 prosp. 100 let Vladivostoku, 690022 Vladivostok, Russian Federation.
Fax: +7 (423 2) 31 4050. Eꢀmail: ivanchina@piboc.dvo.ru
Nine steroidal compounds including three new steroidal glycosides, viz., sodium (24S)ꢀ3,24ꢀdiꢀ
Oꢀ(βꢀDꢀxylopyranosyl)ꢀ5αꢀcholestaneꢀ3β,6β,8,15α,24ꢀpentol 15ꢀsulfate (fuscaside A),
(24S)ꢀ3,24ꢀdiꢀOꢀ(βꢀDꢀxylopyranosyl)ꢀ5αꢀcholestaneꢀ3β,6β,8,15α,24ꢀpentol (fuscaside B), and
(22E,24R)ꢀ24ꢀOꢀ(βꢀDꢀxylopyranosyl)ꢀ5αꢀcholestꢀ22ꢀeneꢀ3β,6α,8,15β,24ꢀpentol (desulfated
minutoside A); three previously known glycosides, viz., distolasterosides D1 and D2 and pycnoꢀ
podioside A; two previously known polyhydroxysteroids, viz., 5αꢀcholestaneꢀ3β,6α,8,15β,16β,26ꢀ
hexaol and 5αꢀcholestanꢀ3β,4β,6α,7α,8,15β,16β,26ꢀoctol; and the known sodium 24,25ꢀdihydroꢀ
marthasterone 3ꢀsulfate were isolated from the FarꢀEastern starfish Lethasterias fusca. The strucꢀ
tures of these compounds were elucidated by NMR spectroscopy and mass spectrometry.
Key words: starfish, Lethasterias fusca, steroidal glycosides, polyhydroxylated steroids, NMR
spectra.
Unlike other echinoderms, starfishes contain a diverꢀ
result, nine steroidal compounds (1—9) were isolated, in
particular, three new steroidal glycosides, fuscasides A (1)
sity of highly oxidized steroidal compounds. As a rule,
these include polyhydroxysteroids, polyhydroxysteroid
monoꢀ and biosides, and toxic steroidal oligoglycosides
called asterosaponins.1,2 The oxidized steroids are
encountered in both free and sulfated forms. In continuꢀ
ation of our studies of physiologically active steroidal
metabolites from FarꢀEastern starfishes,3 we studied the
steroid composition of the starfish Lethasterias fusca
(order Forcipulatida, family Asteriidae) collected at the
experimental Marine station of the Pacific Institute of
Bioorganic Chemistry (Possiet Bay, Japanese Sea). As a
3: ∆22, R = H; 4: R = A; 5: ∆22, R = A; 6: R = H
R = OSO3Na (1), OH (2)
R = R´ = H (7), OH (8)
Published in Russian in Izvestiya Akademii Nauk. Seriya Khimicheskaya, No. 1, pp. 196—200, January, 2008.
1066ꢀ5285/08/5701ꢀ0204 © 2008 Springer Science+Business Media, Inc.

Russ.Chem.Bull., Int.Ed., Vol. 57, No. 1, January, 2008
205
Steroidal compounds from Lethasterias fusca
and B (2) and desulfated minutoside A (3), three known
glycosides, viz., distolasterosides D1 (4) and D2 (5) and
pycnopodioside A (6), two known polyhydroxysteroids,
viz., 5αꢀcholestanꢀ3β,6α,8,15β,16β,26ꢀhexol (7) and 5αꢀ
cholestanꢀ3β,4β,6α,7α,8,15β,16β,26ꢀoctol (8), and
known native aglycon of the asterosaponin series, viz.,
sodium 24,25ꢀdihydromarthasterone 3ꢀsulfate (9).
ing no hydrogen. The chemical shifts of the anomeric
protons at δ 4.22 and 4.36 (J = 7.6 Hz), anomeric carbon
atoms at δ 103.2 and 105.1, carbon atoms bound to oxyꢀ
gen at δ 66.7, 66.8, 71.3 (×2), 74.2, 75.0, 75.3, 76.9,
77.9 (×2), 78.0, 80.0, and 86.2 also attested to the presꢀ
ence of two pentoside fragments and a pentasubstituted
steroidal aglycon in the molecule. Using 2D NMR experiꢀ
ments, the signals of all protons and carbon atoms were
assigned (Table 1). Analysis of NMR data of glycoside 1
demonstrated that the proton and carbon chemical shifts
and the corresponding spinꢀspin coupling constants of
the protons of rings A and B in the steroidal core and one
monosaccharide residue almost coincided with the corꢀ
responding signals of asteriidoside L from the Antarctic
starfish of the family Asteriidae4 containing 3β,6βꢀhyꢀ
droxy groups and the βꢀDꢀxylopyranose residue attached
to O(3) of the aglycon. The proton chemical shifts
and spinꢀspin coupling constants and the carbon chemiꢀ
cal shifts of rings C and D for the steroidal core, side
chain, and the second monosaccharide residue were in
good agreement with the data for coscinasteroside B from
the starfish Coscinasterias tenuispina.5 Thus it was demonꢀ
strated that compound 1 also has 8,15α,24ꢀhydroxy
groups, C(15) bears a sulfate group, and the O(24) atom
of the aglycon bears a βꢀDꢀxylopyranose residue. In the
HMBC spectrum, the attachment sites of monosacchaꢀ
ride residues were confirmed by correlations of the
anomeric protons HC(1´) and HC(1″) with C(3) and
C(24), respectively. We ascribed the Sꢀconfiguration to
the asymmetric center C(24) by analogy with coscinꢀ
asteroside B (see Ref. 5) and other 24ꢀglycosylated steꢀ
roids found in starfishes.1 Thus, new fuscaside A (1) was
identified as sodium (24S)ꢀ3,24ꢀdiꢀOꢀ(βꢀDꢀxylopyranoꢀ
syl)ꢀ5αꢀcholestanꢀ3β,6β,8,15α,24ꢀpentol 15ꢀsulfate.
The highꢀresolution (+)ꢀMALDIꢀTOF mass spectrum
of steroidal glycoside 2 contained a pseudomolecular ion
peak with m/z 739.4297 [M + Na]+, which corresponded
to the molecular formula C37H64O13Na. A comparison of
Results and Discussion
Pure steroidal compounds 1—9 were isolated from a
water—ethanolic extract of the starfish L. fusca by
repeated column chromatography on Polychrom 1,
Sephadex LHꢀ20, silica gel, and florisil and HPLC on
Zorbax ODS, Diaspherꢀ110ꢀC18, Kromasil 100AꢀC18,
and Cosmosil 5C18ꢀARꢀ2. The structures of new comꢀ
pounds were determined in 1H—1H COSY, HSQC,
HMBC, and DEPT NMR experiments and data from
mass spectrometry. Known compounds were identified
by comparison of the NMR spectra, mass spectra, and
physical parameters with published data.
The highꢀresolution (+)ꢀMALDIꢀTOF mass spectrum
of compound 1 contained a pseudomolecular ion peak
with m/z 841.3692 [M + Na]+, which corresponded to
the molecular formula C37H63O16Na2S, and the
(–)ꢀMALDIꢀTOF mass spectrum of the same compound
exhibited a pseudomolecular ion peak with m/z 795 [M –
cation]–. The (+)ꢀLSI mass spectrum contained, apart
1
the H NMR data (see Table 1) of compound 2 with the
data obtained for fuscaside A (1) showed that glycoside
2 differed from fuscaside A by only the lack of the sulfate
group at C(15). Indeed, the Hꢀ15 signal is observed in
a higher field (δ 4.26) than that for 1 (δ 4.89). In addition,
the chemical shifts and the spinꢀspin coupling constants
of the protons of the steroidal core and one monosacchaꢀ
ride residue were in good agreement with the correspondꢀ
ing data for asteriidoside L from the starfish of the family
Asteriidae,4 which also confirmed the presence of
the 3β,6β,8,15αꢀtetrahydroxysteroidal core and the
βꢀDꢀxylopyranose residue attached to the O(3) atom of
the aglycon. Mild solvolysis of fuscaside A (1) gave
from the pseudomolecular ion peak with m/z 841 [MNa
+
Na]+, also peaks with m/z 721, 691, and 571 correspondꢀ
ing to elimination of the sulfate group [MNa + Na –
NaHSO4]+, a pentose residue [MNa + Na – C5H10O5]+,
and simultaneous loss of the sulfate group and the pentose
residue [MNa + Na – NaHSO4 – C5H10O5]+. The
(–)ꢀLSI mass spectrum of compound 1 contained a
pseudomolecular ion peak with m/z 795 [M – cation]–, a
peak with m/z 645 corresponding to the loss of the penꢀ
tose residue [M – cation – C5H10O5]–, and a peak for the
sulfate group with m/z 97 [HSO4]–. These data indicated
that 1 is a sulfated bioside. The 13C NMR and DEPT
spectra of compound 1 contained signals for 37 carbon
atoms including signals for five methyl, eleven methylene,
and eighteen methine groups and three carbon atoms bearꢀ
1
a desulfated derivative the H NMR spectrum of which
was identical to the spectrum of glycoside 2. In its
13C NMR spectrum, the C(15) signal resonates in a higher
field (δ 70.1) than in the spectrum of glycoside 1 (δ 78.0).

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Ivanchina et al.
Table 1. 1H and 13C NMR spectra of compound 1 and 1H NMR spectrum of compound 2 (CD3OD, δ, J/Hz)a
Atom
1b
2
c
δ
HMBC
δ
δ
H
H
C
1
2
1.71 (m)
0.98 (m)
1.81 (m)
41.4 CH2
30.1 CH2
1.61 (m)
3
4
3.70 (m)
1.82 (m)
80.0 CH
33.0 CH2
3.70 (m)
1.75 (m)
5
6
7
1.19 (m)
3.86 (m)
2.54 (dd, 3.1, 15.0)
1.55 (dd, 6.0, 15.0)
48.7 CН
74.2 CH
44.7 CH2
3.86 (m)
2.38 (dd, J = 3.0, J = 14.7)
1.60 (m)
C(5),C(6), C(8), C(9)
8
9
10
11
76.9 C
56.9 CH
36.8 C
0.95 (m)
1.84 (m)
1.53 (m)
1.96 (m)
1.25 (m)
19.9 CH2
12
42.7 CH2
13
14
15
16
44.7 C
64.6 CH
78.0 CH
38.6 CH2
1.37 (d, 9.5)
4.89 (dt, 2.8, 9.5)
2.20 (dd.d., 3.0, 8.5, 15.0)
1.90 (dt, 15.0, 9.5)
1.31 (m)
C(13), C(15), C(18)
C(13)
4.26 (dt, J = 3.6, J = 10.0)
17
18
19
20
21
22
56.0 CH
15.3 CH3
15.8 CH3
36.1 CH
18.8 CH3
32.4 CH2
0.97 (s)
1.16 (s)
1.39 (m)
0.89 (d, 6.5)
1.58 (m)
C(12), C(13), C(14), C(17)
C(1), C(5), C(9), C(10)
0.95 (s)
1.16 (s)
C(17), C(20), C(22)
0.90 (d, J = 6.6)
1.01 (m)
23
1.59 (m)
28.8 CH2
1.37 (m)
24
25
26
27
1´
2´
3´
4´
5´
3.34 (m)
1.84 (m)
86.2 CH
31.9 CH
18.3 CH3
18.5 CH3
103.2 CH
75.0 CH
77.9 CH
71.3 CH
66.8 CH2
0.91 (d, 6.5)
0.92 (d, 6.5)
4.36 (d, 7.6)
3.13 (dd, 7.7, 9.4)
3.30d
C(24), C(25), C(27)
C(24), C(25), C(26)
C(3)
0.93 (d, J = 6.9)
0.92 (d, J = 6.9)
4.35 (d, J = 7.6)
3.13 (dd, J = 7.7, J = 9.4)
3.32d
C(1´), C(3´), C(4´)
3.47 (m)
3.47 (m)
3.82 (dd, 5.3, 11.4)
3.18 (dd, 9.4, 11.3)
4.22 (d, 7.6)
3.19 (dd, 7.4, 8.9)
3.27 (t, 8.8)
3.52 (m)
3.87 (dd, 5.3, 11.3)
3.13 (dd, 9.4, 11.3)
C(1´), C(3´), C(4´)
3.82 (dd, 5.3, J = 11.4)
3.18 (dd, 9.4, J = 11.4)
4.22 (d, J = 7.6)
3.18 (dd, J = 7.5, J = 8.9)
3.32d
3.52 (m)
3.87 (dd, 5.3, J = 11.4)
3.13 (dd, 9.4, J = 11.4)
1″
2″
3″
4″
5″
C(24)
C(1″), C(3″), C(4″)
C(2″), C(4″)
105.1 CH
75.3 CH
77.9 CH
71.3 CH
66.7 CH2
C(1″), C(3″), C(4″)
a The 1H and 13C NMR spectra were recorded at 500 and 125.8 MHz, respectively.
b The signals were assigned by 1H—1H COSY and HSQC 2D NMR techniques.
c The signal multiplicity was determined using the DEPT spectrum.
d The signals overlap with the solvent signals.
Conversely, the signals for the C(14) and C(16) atoms
carbon occur in a lower field (d 66.5 and 41.6) than in the
case of compound 1 (d 64.6 and 38.6, respectively). These
data were also indicative of the fact that fuscaside B (2) is

Russ.Chem.Bull., Int.Ed., Vol. 57, No. 1, January, 2008
207
Steroidal compounds from Lethasterias fusca
a native desulfated derivative of fuscaside A (1) having the
structure of (24S)ꢀ3,24ꢀdiꢀOꢀ(βꢀDꢀxylopyranosyl)ꢀ5aꢀ
cholestaneꢀ3b,6b,8,15a,24ꢀpentol.
internal standard (δH 3.30, δC 49.0). Optical rotation was measured
on a Perkin—Elmer 343 polarimeter.
MALDIꢀTOF mass spectra were run on a Bruker Biflex III
mass spectrometer (Germany, N2 laser, 337 nm). The sample was
dissolved in MeOH (1 mg mL–1) and a 1 µL aliquot was analyzed
using 2,5ꢀdihydroxybenzoic acid as the matrix. The LSI mass spectra
were recorded on an Intectra AMDꢀ604S mass spectrometer (Gerꢀ
many), Cs+ ion energy 10 keV, accelerating voltage 8 kV. Glycerol
(Sigma) was used as the matrix. HPLC was carried out on an
Agilent 1100 Series chromatograph (refractometer as the detector)
with Zorbax ODS (5 µ, 250×9.4 mm), Diaspherꢀ110ꢀC18 (Sꢀ5 µ,
250×4 mm), Kromasil 100AꢀC18 (5 µ, 250×4.6 mm), and Cosmosil
5C18ꢀARꢀ2 columns (5 µ, 250×4.6 mm).
Column chromatography was perfomed using Polychrom 1
(Teflon powder, Biolar, Latvia), KSK silica gel (50—160 µm,
Sorbpolimer, Krasnodar, Russia), and Florisil (200—300 mesh,
Aldrich). Thin layer chromatography was carried out on Sorbfil
plates (4.5×6.0 сm, Sorbpolimer, Krasnodar, Russia) with a foilꢀ
supported STKhꢀ1A silica gel layer (5—17 µm) in a butanol : ethanol
: water system (4 : 1 : 2). The substances were visualized by spraying
the chromatograms with concentrated H2SO4 followed by heating
at 110 °C for 10 min.
The highꢀresolution (+)ꢀMALDIꢀTOF mass spectrum
of compound 3 contained a pseudomolecular ion peak
with m/z 605.3614 [M + Na]+ corresponding to the moꢀ
lecular formula C32H54O9Na. The chemical shifts and the
spinꢀspin coupling constants for the protons and carbon
1
atoms of the steroidal core observed in the H and 13C
NMR и DEPT spectra of glycoside 3 were in good agreeꢀ
ment with those known for pycnopodioside A from the
starfish Pycnopodia helianthoides.6 One could suggest that
steroid 3 contains a 3β,6α,8,15βꢀtetrahydroxysteroidal
core. The NMR chemical shifts of the side chain and the
monosaccharide residue in 3 were similar to those for
minutoside A isolated from the starfish Anasterias minuta,7
which might be indicative of the presence of (24R)ꢀ24ꢀOꢀ
(βꢀDꢀxylopyranosyl)ꢀcholestꢀ22ꢀene side chain in glycoꢀ
side 3. The spinꢀspin coupling constant J22,23 = 15.4 Hz
corresponded to the transꢀconfiguration of the 22(23)ꢀ
double bond. The chemical shifts of all protons and carꢀ
bon atoms in 3 were determined by NMR spectroscopy.
Data of 2D NMR experiments confirmed the structure of
glycoside 3 as (22E,24R)ꢀ24ꢀOꢀ(βꢀDꢀxylopyranosyl)ꢀ5αꢀ
cholestꢀ22ꢀeneꢀ3β,6α,8,15β,24ꢀpentol. Glycoside 3 is
desulfated minutoside A being found for the first time as a
natural compound. Simultaneously, we isolated this comꢀ
pound in a repeated study of the FarꢀEastern starfish
Distolasterias nipon.8
The NMR and mass spectra of compounds 4 and 5
were found to be almost identical to those reported for
distolasterosides D1 and D2, respectively, which were
found previously in the starfish Distolasterias nipon.9,10
Similarly, glycoside 6 was identified as pycnopodioside A
detected previously in the starfish Pycnopodia
helianthoides.6 In addition, we isolated two known polyꢀ
hydroxysteroids, viz., 5αꢀcholestanꢀ3β,6α,8,15β,16β,26ꢀ
hexol (7) found previously in the starfish Halityle
regularis11 and 5αꢀcholestanꢀ3β,4β,6α,7α,8,15β,16β,26ꢀ
octol (8) first found in the starfish Asterina pectinifera,12
and sodium 24,25ꢀdihydromarthasterone 3ꢀsulfate (9)
known as native sulfated aglycon of marthasteroside C
from the starfish Marthasterias glacialis13 and isolated first
in a pure state from the starfish Aphelasterias japonica.14
Glycosides 1 and 2 refer to a rather scarce structural
group of natural glycosylated steroidal polyols containing
monosaccharide residues attached to both O(3) and O(24)
atoms of the aglycon.
The starfish samples were collected in August, 2002, in the
Possiet Bay of the Japanese Sea at the Marine Experimental Station
of the Pacific Institute of Bioorganic Chemistry, FarꢀEastern Branch
of the Russian Academy of Sciences, and identified by C. Sh. Dautov
(Institute of Marine Biology, FarꢀEastern Branch of the Russian
Academy of Sciences, Vladivostok, Russia).
Isolation of compounds 1—9. Crushed starfishes (1.14 kg) were
extracted twice with 70% ethanol (3 L kg–1) with heating on a water
bath and the precipitate was separated by filtration through cotton.
Nonpolar compounds (sterols and lipids) were removed by extracting
the filtrate twice with hexane (1 mL per 3 mL of the filtrate). The
water—ethanol layer was concentrated in vacuo, the dry residue was
divided into two parts, and each part was dissolved in water
(500 mL) and passed separately through a Polychrom 1 column
(7.5×18 cm). The column was washed with water until the eluate
contained no chloride ions and then with ethanol, and the ethanolic
eluate was concentrated. The resulting overall fraction of steroidal
compounds (8.2 g) was successively chromatographed on a Sephadex
LHꢀ20 column (3.5×30 cm) in an ethanol : water system (2 : 1) and
a silica gel column (6×19 cm) in a chloroform : ethanol system
(stepwise gradient, 8 : 1 → 1 : 1). The fractions thus formed were
subjected to HPLC on a Zorbax ODS column using 65% ethanol as
the eluent and rechromatographed on Diaspherꢀ110ꢀC18, Kromasil
100AꢀC18, and Cosmosil 5C18ꢀARꢀ2. This gave 13 mg of compound
1 (Rf = 0.59), 0.5 mg of compound 2 (Rf = 0.69), 6 mg of compound
3 (Rf = 0.62), 1 mg of compound 4 (Rf = 0.76), 4 mg of compound 5
(Rf = 0.71), 2.5 mg of compound 6 (Rf = 0.86), 6 mg of compound 7
(Rf = 0.91), 3.5 mg of compound 8 (Rf = 0.83), and 0.5 mg of
compound 9 (Rf = 0.80).
Fuscaside A (1). Amorphous compound, [α]D +0.4 (c 0.3,
MeOH). 1H and 13C NMR spectra are presented in Table 1.
HR (+)ꢀMALDIꢀTOF MS: found m/z 841.3692 [MNa + Na]+.
C37H63O16Na2S. Calculated: M = 841.3632. MS (–)ꢀMALDIꢀ
Experimental
TOF, m/z: 795 [M – cation]–. MS (+)ꢀLSI, m/z: 841 [MNa
+
Na]+, 721 [MNa + Na – NaHSO4]+, 691 [MNa + Na – C5H10O5]+
and 571 [MNa + Na – NaHSO4 – C5H10O5]+. MS (–)ꢀLSI,
m/z: 795 [M – cation]–, 645 [M – cation – C5H10O5]– and 97
[HSO4]–.
1H and 13C NMR spectra were recorded on Bruker DPX 300
(300.13 and 75.5 MHz) and Bruker DRX 500 (500.13 and
125.8 MHz, respectively) spectrometers using CD3OD as the

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Ivanchina et al.
Fuscaside B (2). Amorphous compound, [α]D –9.3 (c 0.05,
MeOH). The 1H NMR spectra are presented in Table 1.
HR (+)ꢀMALDIꢀTOF MS: found m/z 739.4297 [MNa + Na]+.
C37H64O13Na. Calculated: M = 739.4247.
Desulfation of compound 1. Compound 1 (3 mg) was heated for
4 h with 1 mL of a dioxane—pyridine mixture (1 : 1) at 100 °C. The
reaction mixture was concentrated in vacuo and the dry residue was
chromatographed on a column with silica gel (1×4 cm) in
a chloroform—ethanol system (3 : 2) to give 1.5 mg of desulfated
derivative. The 1H NMR spectrum was identical to the spectrum of
compound 2 (see Table 1). 13C NMR (CD3OD), δ: 41.4 (C(1));
30.1 (C(2)); 80.0 (C(3)); 33.0 (C(4)); 48.7 (C(5)); 74.2 (C(6)); 45.4
(C(7)); 77.1 (C(8)); 57.1 (C(9)); 36.8 (C(10)); 19.8 (C(11)); 42.8
(C(12)); 45.5 (C(13)); 66.5 (C(14)); 70.1 (C(15)); 41.6 (C(16));
56.0 (C(17)); 15.3 (C(18)); 15.8 (C(19)); 36.2 (C(20)); 19.0 (C(21));
32.8 (C(22)); 28.6 (C(23)); 86.1 (C(24)); 31.9 (C(25)); 18.3 (C(26));
18.4 (C(27)); 103.2 (C(1´)); 75.0 (C(2´)); 77.9 (C(3´)); 71.3 (C(4´));
66.9 (C(5´)); 104.9 (C(1″)); 75.3 (C(2″)); 78.1 (C(3″)); 71.3 (C(4″));
66.5 (C(5″)).
This study was supported by the Russian Academy of Sciences
(program of the Presidium of the RAS “Molecular and Cell
Biology”, grants of the FarꢀEastern Branch of the RAS Nos. 06ꢀIIꢀ
CMꢀ05ꢀ001 and 06ꢀIIIꢀBꢀ05ꢀ128), the Russian Foundation for
Basic Research (Project No. 05ꢀ04ꢀ48246), and the Council for
Grants at President of the Russian Federation (State Support
Program for Leading Scientific Schools, grant NShꢀ6491.2006.4).
Desulfated minutoside A (3). Amorphous compound, [α]D +3.3
(c 0.2, MeOH). 1H NMR (CD3OD), δ: 0.84 (m, 1 H, H(9)); 0.86
(d, 3 H, Me(26)C, J = 6.6 Hz); 0.93 (d, 3 H, Me(27)C, J = 6.6 Hz);
0.97 (m, 1 H, H(1)); 0.98 (s, 3 H, Me(19)C); 1.01 (d, 3 H, Me(21)C,
J = 6.6 Hz); 1.02 (d, 1 H, H(14), J = 5.5 Hz); 1.04 (m, 1 H, H(17));
1.05 (m, 1 H, H(5)); 1.19 (both m, 1 H each, Heq(4), H(12)); 1.27
(m, 1 H, Hax(7)); 1.29 (s, 3 H, Me(18)C); 1.39 (m, 1 H, H(16));
1.48 (m, 1 H, Heq(2)); 1.51 (m, 1 H, H(11)); 1.71 (m, 1 H, H´(1));
1.73 (m, 1 H, Hax(2)); 1.81 (m, 1 H, H´(11)); 1.82 (m, 1 H, H(25));
1.97 (m, 1 H, H´(12)); 2.18 (m, 1 H, Hax(4)); 2.23 (m, 1 H, H(20));
2.24 (m, 1 H, H´(16)); 2.36 (dd, 1 H, Heq(7), J = 12.2 Hz, J =
4.0 Hz); 3.13 (dd, 1 H, H(5´), J = 11.5 Hz, J = 10.4 Hz); 3.16 (dd,
1 H, H(2´), J = 7.4 Hz, J = 8.5 Hz); 3.26 (t, 1 H, H(3´), J = 8.7 Hz);
3.44 (m, 1 H, H(4´)); 3.48 (m, 1 H, H(3)); 3.69 (both m, 1 H each,
H(6), H(24)); 3.79 (dd, 1 H, H(5″), J = 11.5 Hz, J = 5.3 Hz); 4.21
(d, 1 H, H(1´), J = 7.4 Hz); 4.39 (br.t, 1 H, H(15), J = 5.6 Hz); 5.35
(dd, 1 H, H(23), J = 15.4 Hz, J = 7.7 Hz); 5.39 (dd, 1 H, H(22), J =
15.4 Hz, J = 7.2 Hz). 13C NMR (CD3OD), δ: 39.4 (C(1)); 31.4
(C(2)); 72.2 (C(3)); 32.3 (C(4)); 53.8 (C(5)); 67.6 (C(6)); 49.4
(C(7)); 77.5 (C(8)); 57.4 (C(9)); 38.0 (C(10)); 19.7 (C(11)); 43.2
(C(12)); 44.2 (C(13)); 62.6 (C(14)); 71.1 (C(15)); 43.4 (C(16));
57.6 (C(17)); 16.7 (C(18)); 14.1 (C(19)); 40.8 (C(20)); 20.9 (C(21));
141.2 (C(22)); 128.1 (C(23)); 89.2 (C(24)); 33.8 (C(25)); 18.3
(C(26)); 19.3 (C(27)); 104.3 (C(1´)); 75.4 (C(2´)); 78.1 (C(3´));
71.2 (C(4´)); 67.0 (C(5´)). HMBC (CD3OD): H(18)/C(12), C(13),
C(14), C(17); H(21)/C(17), C(20), C(22); H(26)/C(24), C(25),
C(27); H(27)/C(24), C(25), C(26); H(1´)/C(24); H(2´)/C(1´),
C(3´); H(3´)/C(2´), C(4´); H(5´)/C(3´), C(4´). HR (+)ꢀMALDIꢀ
TOF MS: found m/z 605.3614 [M + Na]+. C32H54O9Na.
Calculated: M = 605.3660.
References
1. L. Minale, R. Riccio, and F. Zollo, in Progress in the Chemistry
of Organic Natural Products, Eds W. Herz, G. W. Kirby, R. E.
Moore, W. Steglich, and Ch. Tamm, SpringerꢀVerlag, Wien—
New York, 1993, 62, 234 pp.
2. V. A. Stonik, Usp. Khim., 2001, 70, 763 [Russ. Chem. Revs,
2001, 70, 673 (Engl. Transl.)].
3. N. V. Ivanchina, A. A. Kicha, A. I. Kalinovsky, P. S. Dmitreꢀ
nok, E. L. Chaikina, V. A. Stonik, M. Gavagnin, and
G. Cimino, J. Nat. Prod., 2006, 69, 224.
Distolasteroside D1 (4). Amorphous compound, [α]D –5.3
(c 0.1, MeOH), cf. Ref. 9: [α]D –17.9 (c 0.24, MeOH). The 1H and
13C NMR spectra were identical to the spectra described preꢀ
4. S. De Marino, M. Iorizzi, E. Palagiano, F. Zollo, and
C. Roussakis, J. Nat. Prod., 1998, 61, 1319.
5. R. Riccio, M. Iorizzi, and L. Minale, Bull. Soc. Chim. Belg.,
1986, 95, 869.
+
viously.9,10 (+)ꢀMALDIꢀTOF MS, m/z: 739 [M + Na] .
Distolasteroside D2 (4).. Amorphous compound, [α]D –0.7
(c 0.2, MeOH), cf. Ref. 9: [α]D –10.8 (c 0.27, MeOH). The 1H and
13C NMR spectra were identical to the spectra described preꢀ
6. I. Bruno, L. Minale, and R. Riccio, J. Nat. Prod., 1989, 52,
1022.
7. H. Chludil and M. Maier, J. Nat. Prod., 2005, 68, 1279.
8. A. A. Kicha, I. I. Kapustina, N. V. Ivanchina, A. I. Kalinovskii,
P. S. Dmitrenok, V. A. Stonik, N. V. Pal´yanova, T. M.
Pankova, and M. V. Starostina, Bioorgan. Khim., 2008, 34, 129
[Russ. J. Bioorg. Chem., 2008, 34 (Engl. Transl.)].
9. I. I. Kapustina, A. I. Kalinovskii, S. G. Polonik, and V. A.
Stonik, Khim. Prirod. Soedin., 1987, 2, 250 [Chem. Nat.
Comp., 1987 (Engl. Transl.)].
+
viously.9,10 (+)ꢀMALDIꢀTOF MS, m/z: 737 [M + Na] .
Pycnopodioside A (6). Amorphous compound, [α]D –1.1
(c 0.25, MeOH), cf. Ref. 6: [α]D –2.0 (MeOH). The 1H and 13
C
NMR spectra were identical to the spectra described previously.6
+
(+)ꢀMALDIꢀTOF MS, m/z: 607 [M + Na] .
5αꢀCholestanꢀ3β,6α,8,15β,16β,26ꢀhexol (7).
Amorphous compound, [α]D +15.7 (c 0.1, MeOH), cf. Ref. 11:
[α]D 0 (MeOH). The 1H and 13C NMR spectra were identical to the
spectra described previously.11 (+)ꢀMALDIꢀTOF MS, m/z: 491
10. M. Iorizzi, L. Minale, R. Riccio, and T. Yasumoto, J. Nat.
Prod., 1993, 56, 1786.
11. M. Iorizzi, L. Minale, R. Riccio, M. Debray, and J. L. Menou,
+
–
[M + Na] . MS (–)ꢀMALDIꢀTOF, m/z: 467 [M – H] .
5αꢀCholestaneꢀ3β,4β,6α,7α,8,15β,16β,26ꢀoctol (8).
Amorphous compound, [α]D +2.3 (c 0.05, MeOH), cf. Ref. 12:
[α]D +26.4 (c 1.04, MeOH). The 1H and 13C NMR spectra were
identical to the spectra described previously.12 (+)ꢀMALDIꢀTOF
J. Nat. Prod., 1986, 49, 67.
12. R. Higuchi, Y. Noguchi, T. Komori, and T. Sasaki, Liebigs
Ann. Chem., 1988, 1185.
13. A. Dini, F. A. Mellon, L. Minale, C. Pizza, R. Riccio, R. Self,
and F. Zollo, Comp. Biochem. Physiol. B, 1983, 76, 839.
14. A. A. Kicha, N. V. Ivanchina, A. I. Kalinovskii,
P. S. Dmitrenok, and V. A. Stonik, Izv. Akad. Nauk. Ser. Khim.,
2001, 695 [Russ. Chem. Bull., Int. Ed., 2001, 50, 724].
+
MS, m/z: 523 [M + Na] .
24,25ꢀDihydromarthasteron 3ꢀsulfate, sodium salt (9). Amorꢀ
phous compound, [α]D +5.6 (c 0.06, MeOH), cf. Ref. 14: [α]D +4.0
(c 0.2, MeOH). The 1H and 13C NMR spectra were identical to the
spectra described previously.13,14 (–)ꢀMALDIꢀTOF MS,
–
m/z: 495 [M – H] .
Received July 12, 2007