Isolation of new aliphatic sulfates and sulfamate as the Daphnia kairomones inducing morphological change of a phytoplankton Scenedesmus gutwinskii

Article abstract of DOI:10.1248/cpb.56.133

New aliphatic sulfates and sulfamates were isolated from Daphnia pulex as the Daphnia kairomones that induced morphological defense of a freshwater phytoplankton Scenedesmus gutwinskii var. heterospina (NIES-802). Their structures were determined by spectroscopic and synthetic studies.

Full text of DOI:10.1248/cpb.56.133

January 2008
Notes
Chem. Pharm. Bull. 56(1) 133—136 (2008)
133
Isolation of New Aliphatic Sulfates and Sulfamate as the Daphnia
Kairomones Inducing Morphological Change of a Phytoplankton
Scenedesmus gutwinskii
a
a
a
a
b
Ko YASUMOTO, Akinori NISHIGAMI, Hiroaki AOI, Chise TSUCHIHASHI, Fumie KASAI,
a
,a
Takenori KUSUMI, and Takashi OOI*
a
Institute of Health Biosciences, The University of Tokushima Graduate School; Tokushima, Tokushima 770–8505, Japan:
b
and National Institute for Environmental Studies; Onogawa, Tsukuba 305–8506, Japan.
Received October 4, 2007; accepted October 26, 2007; published online November 8, 2007
New aliphatic sulfates and sulfamates were isolated from Daphnia pulex as the Daphnia kairomones that in-
duced morphological defense of a freshwater phytoplankton Scenedesmus gutwinskii var. heterospina (NIES-802).
Their structures were determined by spectroscopic and synthetic studies.
Key words Daphnia kairomone; Scenedesmus; aliphatic sulfate; aliphatic sulfamate
1
1
In a series of food chain in aquatic world, phytoplankton is protons at d 1.68—1.23. Interpretation of the H– H COSY
the bottom creature that supports the lives of animals. It has spectrum of 1 led to a gross structure as 7-methyloctyl sul-
been soundly believed that phytoplankton is docile and never fate. To rule out the possibility that isolated substance is less
resists against its fate. This belief has been disputed by Hes- active and it might still be contaminated with a minute
sen and van Donk, who reported that a unicellular green alga, amount of highly active compound, compound 1 was synthe-
Scenedesmus subspicatus, achieved morphological change sized. The Wittig olefination of adipic semialdehyde methyl
into 2-, 4-, and 8-coenobia (colonies) when the water in ester (14) with isopropyltriphenylphosphonium iodide gave
which a crustacean Daphnia magna, a grazer of the alga, had the methyl ester 15. Reduction of the double bond under H2
been cultured (Daphnia water) was added to the cultivation atmosphere over the Pd–C catalyst followed by LiAlH re-
4
1)
medium. They also found that the grazing rate of the coeno-
bium morph was lower than that of the unicellular morph,
owing to the increased size of the former. This metamorpho-
sis was supposed to be a self-defense mechanism acquired by
the green alga and triggered by a kairomone secreted from D.
magna. Their report has aroused the interest of many scien-
2—9)
tists to attempt to identify the kairomone,
Recently, we
reported identification of the Daphnia kairomones that cause
the morphological change in a unicellular green alga
Scenedesmus gutwinskii var. heterospina (NIES-802) at
ꢀ1
3
10,11)
1
0 —10 ng/ml concentrations.
Here we report isolation and structure determination of
aliphatic sulfates and sulfamates 1—13 as the Daphnia
kairomones, which are active at ng—mg/ml concentration. To
the best of our knowledge, compounds 1—3, 5—9 and 11—
1
3 are new compounds. Their activity and structure correla-
tion will be discussed elsewhere.
Frozen Daphnia (10 kg; Aso Tropical Fish Co., Ltd.,
Osaka) was soaked with methanol (20 lꢁ3), and the
methanol solution was evaporated, the residue being treated
with water (9 l). The mixture was successively extracted with
hexane, dichloromethane, and butanol, and the most active
butanol extract was separated by HPLC monitoring the activ-
ity to afford 1 (8.0 mg), 2 (0.5 mg), 3 (0.3 mg), 4 (3.8 mg), 5
Fig. 1. The Structure of Daphnia Kairomones 1—13
The countercations were not identified and expressed as M.
(
(
0.5 mg), 6 (8.0 mg), 7 (0.4 mg), 8 (0.8 mg) 9 (0.6 mg), 10
1.5 mg), 11 (0.1 mg), 12 (0.2 mg), and 13 (1.5 mg).
The molecular formula of 1 was established as C H O S
9
19
4
on the basis of HR-FAB-MS. The presence of a sulfate group
ꢀ
was suggested by a fragment ion peak at m/z 97 (HSO ) in
4
1
the negative ion FAB-MS of 1. The H-NMR spectrum of 1
exhibited two methylene protons bearing a sulfate group at d
4
.03 (2H, t, Jꢂ6.6 Hz; H-1), two doublet methyls at d 0.92
(6H, d, Jꢂ6.6 Hz; H-8, Me-7) and eleven methine/methylene Chart 1. Synthesis of Compound 1
∗
To whom correspondence should be addressed. e-mail: tooi@ph.tokushima-u.ac.jp
© 2008 Pharmaceutical Society of Japan

1
34
Vol. 56, No. 1
duction of the ester, provided 7-methyloctan-1-ol (16). The
alcohol 16 was converted to the sulfate by treatment with C H O NS on the basis of HR-FAB-MS. The H-NMR
The molecular formula of
6
was established as
1
1
0
20
3
pyridine–SO complex in tetrahydrofuran (THF) at room spectrum of 6 exhibited two olefinic protons at d 5.50 (1H,
3
temperature to give the sulfate 1. Natural and synthetic sam- dt, Jꢂ11.0, 7.0 Hz; H-4) and 5.42 (1H, dt, Jꢂ11.0, 7.0 Hz;
ple of compound 1 possessed practically same activity.
H-3), two methylene protons bearing a sulfamate group at d
The molecular formula of 2 was established as C H O S 3.03 (2H, t, Jꢂ7.0 Hz; H-1), a methyl group at d 0.94 (3H, t,
10
19
4
on the basis of HR-FAB-MS. The presence of a sulfate group Jꢂ7.0 Hz; H-10), and twelve methylene protons at d 2.34—
ꢀ
4
1
1
was suggested by a fragment ion peak at m/z 97 (HSO ) in 1.33. The H– H COSY spectrum of the compound was con-
1
the negative ion FAB-MS of 2. The H-NMR spectrum of 2 sistent with the structure of 3-decenyl sulfamate. The geome-
exhibited two olefinic protons at d 5.59 (1H, dtd, Jꢂ15.2, try of the double bonds in 6 was deduced to be 3Z from the
1
3
6.6, 1.2 Hz; H-4) and d 5.47 (1H, dtd, Jꢂ15.2, 6.6, 1.2 Hz;
C-NMR chemical shifts of allylic methylenes, with steri-
H-3), two methylene protons bearing a sulfate group at d cally induced upfield shifts being observed at C-2 (d 28.7)
.01 (2H, t, Jꢂ7.1 Hz; H-1), a methyl group at d 0.94 (3H, t, and C-5 (d 28.3) of 6, NOESY cross-peaks observed be-
Jꢂ7.1 Hz; H-10), and twelve methylene protons at d 2.39— tween H-2/H-5 and H-3/H-4 and the underlined coupling
4
1
1
1
.30. The H– H COSY spectrum of the compound was con- constants of the olefinic protons.
sistent with the structure of 3-decenyl sulfate. The geometry The molecular formula of
of the double bonds in 2 was deduced to be 3E from the un- C H O NS on the basis of HR-FAB-MS. The H-NMR
7
was established as
1
1
2
22
3
derlined coupling constants of the olefinic protons.
The molecular formula of 3 was established as C H O S (4H, m; H-3, 4, 6, 7), two methylene protons bearing a sulfa-
spectrum of 7 exhibited two olefinic protons at d 5.47—5.37
11
21
4
on the basis of HR-FAB-MS. The presence of a sulfate group mate group at d 3.04 (2H, t, Jꢂ7.0 Hz; H-1), a methyl group
ꢀ
was suggested by a fragment ion peak at m/z 97 (HSO ) in at d 0.95 (3H, t, Jꢂ7.0 Hz; H-12), and 12 methylene protons
4
1
1
1
the negative ion FAB-MS of 3. The H-NMR spectrum of 3 at d 2.88—1.33. The H– H COSY spectrum of the com-
exhibited two olefinic protons at d 5.52 (1H, dt, Jꢂ11.5, pound was consistent with the structure of 3,6-dodecadienyl
.1 Hz; H-4) and 5.44 (1H, dt, Jꢂ11.5, 7.1 Hz; H-3), two sulfamate.
methylene protons bearing a sulfate group at d 4.00 (2H, t,
Jꢂ7.2 Hz; H-1), two methyl groups at d 0.92 (6H, d, C H O NS on the basis of HR-TOF-MS. The H-NMR
7
The molecular formula of
8 was established as
1
1
0
22
3
Jꢂ6.6 Hz; H-10, Me-9) and 11 methine/methylene protons at spectrum of 8 exhibited two methylene protons bearing a
d 2.46—1.25. The geometry of the double bonds in 3 was sulfamate group at d 3.01 (2H, t, Jꢂ7.0 Hz; H-1), a methyl
deduced to be 3Z from the underlined coupling constants of group at d 0.94 (3H, t, Jꢂ7.0 Hz; H-9), and twelve methyl-
1
1
the olefinic protons.
ene protons at d 1.58—1.34. The H– H COSY spectrum of
The molecular formula of 4 was established as C H O S the compound was consistent with the structure, decyl sulfa-
10
17
4
on the basis of HR-FAB-MS. The presence of a sulfate group mate.
ꢀ
4
was suggested by a fragment ion peak at m/z 97 (HSO ) in
The molecular formula of 9 was established as C H O NS
9 20 3
1
1
the negative ion FAB-MS of 4. The H-NMR spectrum of 4 on the basis of HR-TOF-MS. The H-NMR spectrum of 9
exhibited four olefinic protons at d 5.45—5.30 (4H, m; H-4, exhibited two methylene protons bearing a sulfamate group
5
4
, 7, 8), two methylene protons bearing a sulfate group at d at d 3.01 (2H, t, Jꢂ7.0 Hz; H-1), a methyl group at d 0.94
.04 (2H, t, Jꢂ6.6 Hz; H-1), a methyl group at d 1.01 (3H, t, (3H, t, Jꢂ7.0 Hz; H-9), and fourteen methylene protons at d
1
1
Jꢂ7.3 Hz; H-10), and eight methylene protons at d 2.85— 1.58—1.34. The H– H COSY spectrum of the compound
1
1
1
.76. The H– H COSY spectrum of the compound was con- was consistent with the structure of nonyl sulfamate.
sistent with the structure of 4,7-decadienyl sulfate. The The molecular formula of 10 was established as
geometry of the two double bonds in 4 was deduced to be C H O NS on the basis of HR-TOF-MS. The H-NMR
1
8
18
3
1
3
4
Z,7Z from the C-NMR chemical shifts of allylic methyl- spectrum of 10 exhibited two methylene protons bearing a
enes, with sterically induced upfield shifts being observed at sulfamate group at d 3.01 (2H, t, Jꢂ7.0 Hz; H-1), a methyl
C-3 (d 24.5), C-6 (d 26.3) and C-9 (d 21.4) of 4, and group at d 0.94 (3H, t, Jꢂ7.0 Hz; H-8), and sixteen methyl-
1
1
NOESY cross-peaks observed between H-3/H-6 and H-6/H- ene protons at d 1.58—1.36. The H– H COSY spectrum of
9. 4 had been reported as an antibacterial and antifungal the compound was consistent with the structure of octyl
compound isolated from the hepatopancreas of the ascidian sulfamate. To the best of our knowledge, compound 10 was
1
2)
Halocynthia roretzi.
first isolated as a natural product which had been reported as
13)
The molecular formula of 5 was established as C H O S a synthetic material.
12
23
4
on the basis of HR-FAB-MS. The presence of a sulfate group
The molecular formula of 11 was established as
ꢀ
1
was suggested by a fragment ion peak at m/z 97 (HSO ) in C H O NS on the basis of HR-FAB-MS. The H-NMR
4
11 24
3
1
the negative ion FAB-MS of 5. The H-NMR spectrum of 5 spectrum of 11 exhibited two methylene protons bearing a
exhibited two olefinic protons at 5.53 (1H, dt, Jꢂ11.0, sulfamate group at d 2.99 (2H, t, Jꢂ7.0 Hz; H-1), two dou-
7
.1 Hz; H-4) and 5.44 (1H, dt, Jꢂ11.0, 7.1 Hz; H-3), two blet methyls at d 0.92 (6H, d, Jꢂ7.0 Hz; H-10, Me-9) and fif-
methylene protons bearing a sulfate group at d 4.00 (2H, t, teen methine/methylene protons at d 1.57—1.19. Interpreta-
1
1
Jꢂ7.2 Hz; H-1), a methyl group at d 0.94 (3H, t, Jꢂ6.6 Hz; tion of the H– H COSY spectrum of 11 led to a gross struc-
H-10), and sixteen methylene protons at d 2.46—1.25. The ture as 9-methyldecyl sulfamate.
1
1
H– H COSY spectrum of the compound was consistent with
The molecular formula of 12 was established as
1
the structure, 3-dodecenyl sulfate. The geometry of the dou- C H O NS on the basis of HR-FAB-MS. The H-NMR
1
0
22
3
ble bonds in 5 was deduced to be 3Z from the underlined spectrum of 12 exhibited two methylene protons bearing a
coupling constants of the olefinic protons. sulfamate group at d 3.01 (2H, t, Jꢂ7.0 Hz; H-1), two dou-

January 2008
135
1
3
blet methyls at d 0.92 (6H, d, Jꢂ7.0 Hz; H-9, Me-8) and thir- d, Jꢂ6.6 Hz). C-NMR (100 MHz, CDCl ) d: 61.3 (C-1), 39.0 (C-6), 32.9
3
(
C-2), 29.7 (C-4), 28.0 (C-7), 27.4 (C-5), 25.8 (C-3), 22.7 (C-8 and Me-7).
The alcohol 16 (24 mg, 0.2 mmol) was converted to the sulfate by treatment
with pyridine–SO complex (104 mg, 0.7 mmol) in tetrahydrofuran (3 ml) at
room temperature for 24 h. The resulting mixture was neutralized with 1 M
teen methine/methylene protons at d 1.58—1.23. Interpreta-
1
1
tion of the H– H COSY spectrum of 12 led to a gross struc-
ture as 8-methylnonyl sulfamate.
3
The molecular formula of 13 was established as sodium hydroxide solution and the aqueous solution was extracted with
1
C H O NS on the basis of HR-FAB-MS. The H-NMR hexane to remove the residual alcohol. The aqueous layer was passed
9
20
3
through an ODS column. Mineral salt was removed by washing the column
spectrum of 13 exhibited two methylene protons bearing a
sulfamate group at d 3.01 (2H, t, Jꢂ7.0 Hz; H-1), two dou-
blet methyls at d 0.92 (6H, d, Jꢂ7.0 Hz; H-8, Me-7) and
eleven methine/methylene protons at d 1.63—1.24. Interpre-
with distilled water and elution with methanol gave 1 (49 mg quant.). The
1
13
H- and C-NMR spectra of this product were identical with those of natu-
ral 1.
3E-Decenyl Sulfate (2) White waxy solid. H-NMR (CD OD) d: 5.59
1
3
1
1
tation of the H– H COSY spectrum of 13 led to a gross (1H, dtd, Jꢂ15.2, 6.6, 1.2 Hz; H-4), 5.47 (1H, dtd, Jꢂ15.2, 6.6, 1.2 Hz; H-
3
(
), 4.01 (2H, t, Jꢂ7.1 Hz; H-1), 2.39 (2H, qd, Jꢂ6.6, 1.2 Hz; H-2), 2.04
2H, q, Jꢂ6.6 Hz; H-5), 1.45—1.30 (8H, envelop, H-6, 7, 8, 9), 0.94 (3H, t,
Jꢂ7.1 Hz; H-10). (Underlined coupling constants support the E-configura-
structure as 7-methyloctyl sulfamate.
Experimental
1
3
High-resolution MS were recorded on a JEOL JMS-SX102A spectrome- tion of the olefin.) C-NMR (CD OD) d: 134.3 (C-4), 126.6 (C-3), 68.9 (C-
ter and Waters LCT Premier. H- and C-NMR spectra were obtained on a 1), 33.7 (C-2 and 5), 32.9 (C-8), 30.5 (C-6), 29.9 (C-7), 23.7 (C-9), 14.4 (C-
Bruker Avance-400 ( H and C at 400 and 100 MHz, respectively). Assign- 10). HR-FAB-MS (ꢀ): m/z 235.0992 (Calcd for C H O S: 235.1004).
9-Methyl-3Z-decenyl Sulfate (3) White waxy solid. H-NMR (CD OD)
d: 5.52 (1H, dt, Jꢂ11.5, 7.1 Hz; H-4), 5.44 (1H, dt, Jꢂ11.5, 7.1 Hz; H-3),
polarimeter. The structures of the compounds were established by COSY, 4.00 (2H, t, Jꢂ7.2 Hz; H-1), 2.46 (2H, q, Jꢂ6.8 Hz; H-2), 2.12 (2H, m ; H-
3
1
13
1
13
1
0
19
4
1
ments of the proton and carbon signals were established by COSY, HSQC
and HMBC spectra. Optical rotations were determined on a JASCO DIP-370
3
HSQC, HMBC, and NOESY spectra. The countercation of natural sulfates
5), 1.57 (1H, nonet, Jꢂ6.6 Hz; H-9), 1.45—1.30 (4H, m, H-6, 7), 1.25 (2H,
m; H-8), 0.92 (6H, d, Jꢂ6.6 Hz; H-10, Me-9). (Underlined coupling con-
stants support the Z-configuration of the olefin.) HR-FAB-MS (ꢀ): m/z
ꢃ
was not identified and expressed as M .
2
Bioassay Each 200 ml of C medium of S. gutwinskii (5.0ꢁ10 cells/ml)
is delivered into the central 30—50 wells of 96-well polystyrene tissue cul- 249.1173 (Calcd for C H O S: 249.1161).
1
1
21
4
12)
1
ture plate (CELLSTAR, Greiner Bio-one Co., Ltd.) containing the test sam-
4Z,7Z-Decadienyl Sulfate (4)
White waxy solid. H-NMR (CD OD)
3
ples (1000—0.01 ng/ml), and the outer wells are filled with distilled water to d: 5.45—5.30 (4H, m; H-4, 5, 7, 8), 4.04 (2H, t, Jꢂ7.0 Hz; H-1), 2.85 (2H,
avoid dehydration of the system. The plate is covered with a plastic lid, and m; H-6), 2.23 (2H, m; H-3), 2.13 (2H, m; H-9), 1.76 (2H, m; H-2), 1.01 (3H,
incubated at 20 °C (12 light/12 dark) for 10 d. A drop of the medium is
placed on a Thoma’s hemacytometer, and the numbers of 1-, 2-, 4-, and 8- consistent with the Z-configurations. C-NMR (CD OD) d: 132.7 (C-4) ,
cell types were counted under a microscope (ꢁ200).
Extraction and Isolation Frozen Daphnia (10 kg; Aso Tropical Fish 24.5 (C-3), 21.4 (C-9), 14.7 (C-10). (Underlined chemical shifts of allylic
Co., Ltd., Osaka) was soaked with methanol (20 lꢁ3), and the methanol so- carbons support the Z-configurations of the olefins.) HR-FAB-MS (ꢀ): m/z
lution was evaporated, the residue being treated with water (9 l). The mixture 233.0859 (Calcd for C H O S: 233.0848). These resonances may be in-
t, Jꢂ7.0 Hz; H-10). NOESY cross-peaks between H-3/H-6 and H-6/H-9 are
1
3
a
3
a
a
a
130.1 (C-8) , 129.8 (C-7) , 128.4 (C-5) , 68.5 (C-1), 30.5 (C-2), 26.3 (C-6),
a
1
0
17
4
was successively extracted with hexane (9 l), dichloromethane (9 l), and bu- terchangeable.
1
tanol (9 l), and the most active butanol extract (18 g) was chromatographed
on a Cosmosil 75C -OPN (25 g), eluting with MeOH–H O in a gradient
3Z-Dodecenyl Sulfate (5) White waxy solid. H-NMR (CD OD) d:
5.53 (1H, dt, Jꢂ11.0, 7.1 Hz; H-4), 5.44 (1H, dt, Jꢂ11.0, 7.1 Hz; H-3), 4.00
3
18
2
manner (1 : 1→10 : 0). The active fractions were further purified by HPLC (2H, t, Jꢂ7.2 Hz; H-1), 2.46 (2H, q, Jꢂ6.8 Hz; H-2), 2.10 (2H, m; H-5),
(
CAPCELLPAK C18 column, 5 mm, 10ꢁ250 mm, MeCN–H O (40 : 60) con-
1.50—1.25 (12H, envelop, H-6, 7, 8, 9, 10, 11), 0.94 (3H, t, Jꢂ6.6 Hz; H-
12). (Underlined coupling constants support the Z-configuration of the
2
taining 250 mM NaClO as mobile phase with the flow rate 1.0 ml/min using
an RI detector) to afford 1 (8.0 mg), 2 (0.5 mg), 3 (0.3 mg), 4 (3.8 mg), 5
4
1
3
olefin.) C-NMR (CD OD) d: 133.5 (C-4), 125.7 (C-3), 68.6 (C-1), 33.1
3
a a a a
(0.5 mg), 6 (8.0 mg), 7 (0.4 mg), 8 (0.8 mg) 9 (0.6 mg), 10 (1.5 mg), 11
(C-10), 30.7 (C-6) , 30.6 (C-7) , 30.4 (C-8) , 30.4 (C-9) , 28.6 (C-3), 28.3
(
0.1 mg), 12 (0.2 mg), and 13 (1.5 mg).
(C-2), 23.7 (C-11), 14.4 (C-12). HR-FAB-MS (ꢀ): m/z 263.1335 (Calcd for
1
a
7
-Methyloctyl Sulfate (1) White waxy solid. H-NMR (CD OD) d: C H O S: 263.1317). These resonances may be interchangeable.
3
12 23
4
1
4
.03 (2H,t, Jꢂ6.6 Hz; H-1), 1.68 (2H, quint, Jꢂ6.6 Hz; H-2), 1.56 (1H,
3Z-Decenyl Sulfamate (6) White waxy solid. H-NMR (CD OD) d:
3
nonet, Jꢂ6.8 Hz; H-7), 1.48—1.30 (6H, m; H-3,4,5), 1.23 (2H, m; H-6),
0
(
5.50 (1H, dt, Jꢂ11.0, 7.0 Hz; H-4), 5.42 (1H, dt, Jꢂ11.0, 7.0 Hz; H-3), 3.03
(2H, t, Jꢂ7.0 Hz; H-1), 2.34 (2H, q, Jꢂ7.0 Hz; H-2), 2.12 (2H, q, Jꢂ7.0 Hz;
H-5), 1.37 (2H, overlapped, H-6), 1.33 (6H, envelop, H-7, 8, 9), 0.94 (3H, t,
1
3
.92 (6H, d, Jꢂ6.6 Hz; H-8, Me-7). C-NMR (CD OD) d: 69.2 (C-1), 40.1
3
a a
C-6), 30.6 (C-2) , 30.5 (C-4) , 29.1 (C-7), 28.4 (C-5), 26.9 (C-3), 23.1 (C-8,
Me-7). HR-FAB-MS (ꢀ): m/z 223.0978 (Calcd for C H O S: 223.1004). Jꢂ7.0 Hz; H-10). NOESY cross-peaks were observed between H-2/H-5 and
9
19
4
a
These resonances may be interchangeable.
Synthesis of Sodium 7-Methyloctyl Sulfate (1) 1.6 M Solution of n-
BuLi in hexane (0.24 ml, 0.7 mmol) was added dropwise to a suspension of
H-3/H-4. (Underlined coupling constants and NOEs support the Z-configu-
1
3
ration of the olefin.) C-NMR (CD OD) d: 132.9 (C-4), 127.5 (C-3), 44.8
3
(C-1), 32.9 (C-8), 30.7 (C-6), 30.0 (C-7), 28.7 (C-2), 28.3 (C-5), 23.7 (C-9),
isopropyltriphenylphosphonium iodide (370 mg, 0.9 mmol) in THF (4 ml). 14.4 (C-10). (Underlined chemical shifts of allylic carbons support the Z-
The mixture was stirred at ꢀ30 °C for 20 min, cooled at ꢀ78 °C and a solu- configurations of the olefins.) HR-FAB-MS (ꢀ): m/z 234.1175 (Calcd for
tion of adipic semialdehyde methyl ester 14 (95 mg, 0.7 mmol; commercially C H O NS: 234.1164).
1
0
20
3
1
available) in THF (2 ml) was added. The mixture was stirred at the same
3,6-Dodecadienyl Sulfamate (7) White waxy solid. H-NMR (CD OD)
3
temperature for 2 h, poured into water, extracted with diethyl ether. The or- d: 5.47—5.37 (4H, m; H-3, 4, 6, 7), 3.04 (2H, t, Jꢂ7.0 Hz; H-1), 2.88 (2H,
ganic layer was dried over Na SO . After concentration, the crude product
was purified by flash chromatography to give the methyl ester 15 (83 mg,
t, Jꢂ7.0 Hz; H-5), 2.37 (2H, q, Jꢂ7.0 Hz; H-2), 2.12 (2H, q, Jꢂ7.0 Hz; H-
8), 1.33 (6H, envelop; H-9, 10, 11), 0.95 (3H, t, Jꢂ7.0 Hz; H-12). HR-FAB-
2
4
1
7
(
(
2
4%). 15; H-NMR (400 MHz, CDCl ) d: 5.09 (2H, m), 3.66 (3H, s), 2.30 MS (ꢀ): m/z 260.1328 (Calcd for C H O NS: 260.1320).
3 12 22 3
1
2H, t, 7.8), 1.98 (2H, q, 7.6), 1.67 (3H, s), 1.62 (2H, m), 1.59 (3H, s), 1.34
Decyl Sulfamate (8) White waxy solid. H-NMR (CD OD) d: 3.01
3
1
3
2H, m). C-NMR (100 MHz, CDCl ) d: 174.1, 131.6, 124.1, 51.5, 34.1, (2H, t, Jꢂ7.0 Hz; H-1), 1.58 (2H, m; H-2), 1.34 (12H, envelop; H-3, 4, 5, 6,
3
1
3
9.4, 27.7, 25.8, 24.7, 17.7. To 75 mg (0.4 mmol) of methyl 7-methyl-6-octe- 7, 8, 9), 0.94 (3H, t, Jꢂ7.0 Hz; H-10). C-NMR (CD OD) d: 45.0 (C-1),
3
a a a a a
nate 15 in MeOH (5 ml) was added Pd–C catalyst (24 mg, 10% w/w). The 33.1 (C-8) 30.9 (C-2) , 30.7 (C-4) , 30.7 (C-5) , 30.6 (C-6) , 30.5 (C-7) ,
a
reaction mixture was stirred at room temperature for 2 h under hydrogen 28.2 (C-3), 23.7 (C-9), 14.5 (C-10). These resonances may be interchange-
atmosphere. The mixture was filtered and the ester (60 mg, 78%) was ob- able. HR-TOF-MS (ꢀ): m/z 236.1321 (Calcd for C H O NS: 236.1320).
1
0
22
3
tained after concentration. To a solution of 60 mg (0.4 mmol) of the ester in
Decyl amine (52 mg, 0.3 mmol) was converted to the sulfate by treatment
THF (3 ml) was added 39 mg (1.0 mmol) of LiAlH . After 1 h, the reaction
with pyridine–SO complex (210 mg, 1.3 mmol) in tetrahydrofuran (3 ml) at
4
3
was quenched by adding 50 ml of H O, 15% NaOH, and then 200 ml of H O.
room temperature for 24 h. The resulting mixture was neutralized with 1 M
The mixture was diluted with EtOAc and filtered through Celite. The filtrate sodium hydroxide solution and the aqueous solution was extracted with
2
2
was concentrated under reduced pressure to furnish 24 mg (0.2 mmol, 48%)
hexane to remove the residual alcohol. The aqueous layer was passed
1
of 7-Methyloctan-1-ol 16. 16; H-NMR (400 MHz, CDCl ) d: 3.65 (2H, t, through an ODS column. Mineral salt was removed by washing the column
3
1
Jꢂ6.6 Hz, H-1), 1.66—1.46 (3H, m), 1.28 (4H, m), 1.16 (2H, m), 0.85 (6H, with distilled water and elution with methanol gave 8 (56 mg, 66%). The H-

1
36
Vol. 56, No. 1
8-Methylnonyl Sulfamate (12) White waxy solid. H-NMR (400 MHz,
13
1
and C-NMR spectra of this product were identical with those of natural 8.
1
Nonyl Sulfamate (9) White waxy solid. H-NMR (CD OD) d: 3.01 CD OD) d: 3.01 (2H, t, Jꢂ7.0 Hz; H-1), 1.58 (3H, m; H-2, 8), 1.34 (8H, en-
3
3
(2H, t, Jꢂ7.0 Hz; H-1), 1.58 (2H, m; H-2), 1.34 (12H, envelop; H-3, 4, 5, 6,
velop; H-3, 4, 5, 6), 1.23 (2H, m; H-7), 0.92 (6H, d, Jꢂ7.0 Hz; H-9, Me-8).
1
3
7
, 8), 0.94 (3H, t, Jꢂ7.0 Hz; H-9). C-NMR (CD OD) d: 45.0 (C-1), 33.1
HR-FAB-MS (ꢀ): m/z 236.1344 (Calcd for C H O NS: 236.1320).
3
10 22
3
a
a
a
a
1
(C-7) 30.9 (C-2) , 30.7 (C-4) , 30.6 (C-5) , 30.4 (C-6) , 28.2 (C-3), 23.7 (C-
7-Methyloctyl Sulfamate (13) White waxy solid. H-NMR (CD OD)
3
a
8
), 14.5 (C-9). These resonances may be interchangeable. HR-TOF-MS d: 3.01 (2H, t, Jꢂ7.0 Hz; H-1), 1.52—1.63 (3H, m; H-2, 7), 1.41—1.30
(ꢀ): m/z 222.1125 (Calcd for C H O NS: 222.1164). (6H, m; H-3, 4, 5), 1.24 (2H, m; H-6), 0.92 (6H, d, Jꢂ7.0 Hz; H-8, Me-7).
9
20
3
1
3
Nonyl amine (57 mg, 0.4 mmol) was converted to the sulfate by treatment
C-NMR (CD OD) d: 45.0 (C-1), 40.2 (C-6), 30.8 (C-2, 4), 29.2 (C-7),
3
with pyridine–SO complex (265 mg, 1.6 mmol) in tetrahydrofuran (3 ml) at 28.5 (C-5), 28.3 (C-3), 23.0 (C-8, Me-7). HR-FAB-MS (ꢀ): m/z 222.1178
3
room temperature for 24 h. The resulting mixture was neutralized with 1 M
sodium hydroxide solution and the aqueous solution was extracted with
hexane to remove the residual alcohol. The aqueous layer was passed References
(Calcd for C H O NS: 222.1164).
9 20 3
through an ODS column. Mineral salt was removed by washing the column
1) Hessen D. O., van Donk E., Hydrobiologia, 127, 129—140 (1993).
2) Lampert W., Rothhaupt K. O., von Elert E., Limnol. Oceanogr., 39,
1543—1550 (1994).
3) von Elert E., Franck A., J. Plankton Res., 21, 789—804 (1999).
4) Wiltshire K. H., Lampert W., Limnol. Oceanogr., 44, 1894—1903
(1999).
1
with distilled water and elution with methanol gave 9 (36 mg, 36%). The H-
13
and C-NMR spectra of this product were identical with those of natural 9.
1
Octyl Sulfamate (10) White waxy solid. H-NMR (CD OD) d: 3.01
3
(
7
2H, t, Jꢂ7.0 Hz; H-1), 1.58 (2H, m; H-2), 1.36 (10H, envelop; H-3, 4, 5, 6,
1
3
), 0.94 (3H, t, Jꢂ7.0 Hz; H-8). C-NMR (CD OD) d: 45.0 (C-1), 33.0 (C-
3
a a
6
) 30.9 (C-2), 30.5 (C-4) , 30.4 (C-5) , 28.3 (C-3), 23.7 (C-7), 14.5 (C-8).
5) Kaler V. L., Bulko O. P., Reshetnikov V. N., Galkovskaya G. A. Russ. J.
Plant Phys., 47, 698—705 (2000).
a
These resonances may be interchangeable. HR-TOF-MS (ꢀ): m/z 208.1050
(
Calcd for C H O NS: 208.1007).
Octyl amine (51 mg, 0.4 mmol) was converted to the sulfate by treatment
6) von Elert E., Verh. Int. Ver. Theor. Angew. Limnol., 27, 2128—2131
(2000).
8
18
3
with pyridine–SO complex (250 mg, 1.6 mmol) in tetrahydrofuran (3 ml) at
7) Lürling M., von Elert E., Limnol. Oceanogr., 46, 1809—1813 (2001).
8) van Holthoon F. L., van Beek T. A., Lürling M., van Donk E., De
Groot A., Hydrobiologia, 491, 241—254 (2003).
3
room temperature for 24 h. The resulting mixture was neutralized with 1 M
sodium hydroxide solution and the aqueous solution was extracted with
hexane to remove the residual alcohol. The aqueous layer was passed
9) Lürling M., Ann. Limnol.-Int. J. Limnol., 39, 85—101 (2003).
through an ODS column. Mineral salt was removed by washing the column 10) Yasumoto K., Nishigami A., Yasumoto M., Kasai F., Okada Y.,
with distilled water and elution with methanol gave 10 (12 mg, 15%). The Kusumi T., Ooi T., Tetrahedron Lett., 46, 4765—4767 (2005).
H- and C-NMR spectra of this product were identical with those of natu- 11) Yasumoto K., Nishigami A., Kasai F., Kusumi T., Ooi T., Chem.
ral 10. Pharm. Bull., 54, 271—274 (2006).
-Methyldecyl Sulfamate (11) White waxy solid. H-NMR (CD OD) 12) Tsukamoto S., Kato H., Hirota H., Fusetani N., J. Nat. Prod., 57,
1
13
1
9
3
d: 2.99 (2H, t, Jꢂ7.0 Hz; H-1), 1.57 (3H, m; H-2, 9), 1.33 (10H, envelop; H-
, 4, 5, 6, 7), 1.19 (2H, m; H-8), 0.92 (6H, d, Jꢂ7.0 Hz; H-10, Me-9). HR-
TOF-MS (ꢀ): m/z 250.1491 (Calcd for C H O NS: 250.1482).
1606—1609 (1994).
13) Nofre C., Pautet F., Bull. Chem. Soc. Fr., 1975(3—4, Pt. 2), 686—688
3
(1975).
1
1
24
3