A cyanoallyl glucoside from Alliaria petiolata, as a feeding deterrent for larvae of Pieris napi oleracea

Article abstract of DOI:10.1021/np000534d

Alliarinoside, a feeding inhibitor against early instar larvae of Pieris napi oleracea, was isolated from the foliage of Alliaria petiolata and characterized as (2Z)-4-(β-D-glucopyranosyloxy)-2-butenenitrile (1) by spectroscopic methods. The structural assignment was confirmed by synthesis of peracetylated alliarinoside (2) and its 2E isomer (3). A sample of synthetic 1 was isolated by preparative HPLC from the hydrolysis of the 2Z acetate. Feeding inhibition assays showed comparable activity for the synthetic and natural glycosides.

Full text of DOI:10.1021/np000534d

4
40
J . Nat. Prod. 2001, 64, 440-443
A Cya n oa llyl Glu cosid e fr om Allia r ia petiola ta , a s a F eed in g Deter r en t for
La r va e of P ier is n a pi oler a cea
,
†
‡
‡
†
‡
Meena Haribal,* Zhicai Yang, Athula B. Attygalle, J . Alan A. Renwick, and J errold Meinwald
Boyce Thompson Institute, Tower Road, Ithaca, New York 14853, and Department of Chemistry and Chemical Biology,
Cornell University, Ithaca, New York 14853
Received November 17, 2000
Alliarinoside, a feeding inhibitor against early instar larvae of Pieris napi oleracea, was isolated from
the foliage of Alliaria petiolata and characterized as (2Z)-4-(â-D-glucopyranosyloxy)-2-butenenitrile (1)
by spectroscopic methods. The structural assignment was confirmed by synthesis of peracetylated
alliarinoside (2) and its 2E isomer (3). A sample of synthetic 1 was isolated by preparative HPLC from
the hydrolysis of the 2Z acetate. Feeding inhibition assays showed comparable activity for the synthetic
and natural glycosides.
Alliaria petiolata Cavara & Grande (Cruciferae) (garlic
mustard), an introduced weed from Europe, is rapidly
replacing many native species in the flora of the north-
eastern United States. From extracts of the leaves of this
plant, we reported previously an apigenin derivative,
upon drying under vacuum. Its IR spectrum (neat) showed
absorptions at 3300 (OHs), 2260 and 2130 (CN), 1680
(unsaturation) 1630, 855, 790, 750, and 720 cm . On the
basis of positive ion accurate-mass spectroscopy, the mo-
lecular formula of alliarinoside was determined to be
-
1
1
isovitexin-6′′-O-â-D-pyranoglucoside, as a feeding deterrent
to fourth instar larvae of Pieris napi oleracea. In addition
to this activity, water and n-butanol extracts of foliage were
found to inhibit feeding of early instars of P. napi oleracea
10 15 6
C H O N.
2
13C NMR and DEPT spectra of 1a showed the presence
of two methylene, six methine, and two quaternary car-
bons. The signals at δ 5.73 (1H, d, J ) 11.2 Hz) and 6.76
(Renwick et al. unpublished). An HPLC separation of these
1
(1H, ddd, J ) 11.2, 6.0 and 6.4 Hz) in the H NMR
extracts, in conjunction with bioassays of the fractions,
revealed the presence of an additional feeding inhibitor,
which we designate as alliarinoside. We now describe the
identification and synthesis of this compound, (2Z)-4-(â-D-
glucopyranosyloxy)-2-butenenitrile (1).
spectrum suggested the presence of an olefinic moiety. The
multiplet signal at δ 6.76 correlated to the proton signals
at δ 4.35 (1H, dd, J ) 6.4 and 14 Hz) and 4.49 (1H, dd,
J ) 6 and 14 Hz) of methylene protons. The signals at δ
4
.20, 2.98, 3.16, 3.13, 3.05, 3.40, and 3.60 together with
signals at δ 4.92, 4.97, and 5.07, which collapsed to produce
broad singlets at δ 4.75, 5.10, and 5.23 on addition of D O,
2
were assigned to a hexose moiety. The coupling constant
of 8 Hz for the anomeric proton at δ 4.20 suggested that
the hexose is an R-substituted sugar. The signal at δ 116.6
in the 13C NMR spectrum due to a quaternary carbon
further complemented the IR spectral evidence for the
presence of a nitrile function. The HMBC spectrum showed
the presence of two isolated spin systems, one of which
could be assigned to a hexose moiety and the other to an
allylic group with a terminal nitrile functional group. Thus,
on the basis of the NMR spectra, we concluded that 1 was
a cyanoallyl glycoside.
The positive-ion ESIMS showed a quasi-molecular ion
+
(
M + H) at m/z 246. Other significant fragment ions were
+
observed at m/z 214 (M + H - 32) [a loss of CH
3
+
OH from
the primary alcohol moiety], 163 ((M + H - 83) [loss of
+
aglycone], and 84 (M + H - 162) [loss of dehydrohexose].
2
Alliarinoside (1) was acetylated with (Ac) O-pyridine to
Resu lts a n d Discu ssion
give a peracetylated derivative (2). The positive-ion ESIMS
+
Alliarinoside (1) was isolated by repeated semiprepara-
tive HPLC of an aqueous extract of young rosette leaves
of A. petiolata. Choice bioassays of this compound with
neonates of P. napi oleracea confirmed the activity of this
compound as a feeding inhibitor (Table 1, Supporting
Information). Compound 1 was highly water-soluble and
formed a hygroscopic, gummy, and transparent material
of the derivative showed a quasi-molecular ion (M + H)
at m/z 414, indicating the presence of four free OH groups
in 1. This was further confirmed by the presence of four
singlets at δ 2.12, 2.06, 2.04, and 2.01 for the COCH
groups in the H NMR spectrum of 2.
3
1
Compound 1 was hydrolyzed with 1 M HCl, and the
products were partitioned between aqueous and organic
solvents. The aqueous layer gave a hexose sugar along with
other compounds. The hexose was identified as glucose by
comparing the GC retention time of its TMS derivative with
that of the derivative of authentic glucose. Subsequent
*
To whom correspondence should be addressed. Tel: 607-254-5414.
Fax: 607-254-2598. E-mail: mmh3@cornell.edu.
†
Boyce Thompson Institute.
Cornell University.
‡
1
0.1021/np000534d CCC: $20.00
© 2001 American Chemical Society and American Society of Pharmacognosy
Published on Web 03/21/2001

A Cyanoallyl Glucoside from Alliaria
J ournal of Natural Products, 2001, Vol. 64, No. 4 441
Sch em e 1
from plants and animals. For example, such glycosides
have been reported from a variety of plant families includ-
7
8
9
ing Celastraceae, Crassulaceae, Sapindaceae, Euphor-
biaceae,10 Gramineae, etc. Recently a related aglycone has
11
1
2
been reported from Cruciferae. Noncyanogenic glycosides
have also been reported from many unpalatable insects,
some of which feed on the above plant families. These
compounds are known to have a range of biological activi-
7
-9
ties to herbivores such as deterrency,
and phytotoxic-
ity;12 thus they may play a role in plant defense. Specialist
herbivores7 and organisms such as powdery mildews
-9
12
have utilized these chemicals for their own defense by
sequestering them.
Exp er im en ta l Section
Gen er a l Exp er im en ta l P r oced u r es. GC-MS analysis
was conducted using a capillary column (DB-5, 30 m × 0.23
mm) installed in a HP5890 GC linked to a Hewlett-Packard
5
970 mass selective detector. UV spectra were recorded on a
Perkin-Elmer Lambda 5 UV/vis spectrophotometer. IR spectra
were recorded on a Perkin-Elmer 297 infrared spectrophotom-
1
13
eter. H NMR spectra were recorded at 399.99 MHz, and
C
NMR at 25.59 MHz in DMSO-d or in acetone-d unless
6
6
otherwise stated, on a INOVA 400 Varian instrument. 2D
NMR spectra were recorded on a 499.99 MHz INOVA 500
Varian instrument using standard software. ESI mass spectra
were recorded on a Micromass (Manchester, UK) Quattro 1
instrument, and the samples were introduced by a direct
infusion method. HPLC was performed on a Varian Vista 5500
NMR spectrometry confirmed this conclusion. As expected,
the organic layer did not yield any appreciable amount of
cyanoallyl alcohol as the aglycone.
Typical coupling constant values for cis protons in 1,2-
disubstituted double bonds are around 10 Hz (the range is
from 6 to 12), whereas those for trans are around 17 Hz
(Limmerick, Ireland) instrument, attached to a HP 1040A
diode array detector (Hewlett-Packard, Palo Alto, CA) moni-
tored at 218 and 254 nm. Melting points were uncorrected.
Optical rotations were measured on a Perkin-Elmer 241MC
polarimeter with a 1 dm cell. High-resolution mass spectra
were recorded on a Micromass Autospec instrument operated
in the positive-ion ESI mode. The positive-ion ESI MS-MS
were recorded on a VG Quattro instrument. Tetra-O-acetyl-
R-glucosyl bromide was purchased from Sigma. TRI-SIL ‘Z’ was
purchased from Pierce Chemical Company (Rockford, IL).
(but range from 12 to 18). The observed value of 11.2 Hz
for olefinic protons did not allow for unambiguous assign-
ment of the configuration. Therefore, we undertook chemi-
cal synthesis of both cis and trans isomers.
Both cis-allyl nitrile 2 and trans-allyl nitrile 4 tetrac-
etates were synthesized as described in Scheme 1. Con-
densation of tetra-O-acetyl glucosyl bromide 5 with allyl
P la n t Ma ter ia l. Seedlings of A. petiolata were collected
from Eastern Heights area in Ithaca (Tompkins County, NY)
in November 1996 and were grown in a greenhouse under
controlled conditions (16:8:L:D photoperiod and RH of 65%).
Extr a ction a n d Isola tion . Young rosette leaves (50 g) of
A. petiolata were extracted with 200 mL of 95% boiling EtOH.
The resulting mixture was homogenized, and the insoluble
plant material was removed by filtration. The extract was
alcohol in the presence of silver triflate provided the
â-glucoside tetracetate 6 in good yield.3,4 Ozonolysis of the
allyl alcohol 6 afforded the aldehyde 7 in excellent yield.
A Horner-Emmons reaction of the aldehyde 7 with diethyl
cyanomethylenephosphonate yielded both cis- and trans-
allyl nitriles 2 and 4 in the ratio of 3:2.5 The 1H NMR
spectrum revealed coupling constants J 2,3 ) 11.2 Hz for 2
and J 2,3 ) 16 Hz for 4, and both MS and NMR data of 2
were congruent with those of the 2,3,4,6-tetra-O-acetyl-
2
concentrated to 50 mL, diluted with an equal amount of H O,
and vortexed. The insoluble material was removed by cen-
trifugation, and the supernatant was evaporated to dryness,
redissolved in water, and diluted to 50 mL. Compound 1 was
collected from the fraction that eluted as a single peak at 11.2
min by HPLC on a reversed-phase column (C18 Bondex 10, 5
µm; 300 × 7.0 mm, Phenomenex, Torrance, CA), using a
water-acetonitrile gradient at a flow rate of 2 mL/min (0-5
min 0% acetonitrile; 25% acetonitrile at 40 min; 100% aceto-
nitrile at 50 min) monitored by a diode-array detector at 218
and 254 nm.
2 3
alliarinoside. Deacetylation with K CO in dry MeOH at
room temperature produced at least two major compounds,
and one of these products showed an HPLC retention time
identical to that of 1. The ESIMS of this product was
indistinguishable from that of the natural 1. This com-
pound (1) was isolated by repeated HPLC and tested for
its activity by a choice bioassay. The activity of the
synthetic 1 was comparable to that of natural 1 (Table 1,
Supporting Information). The 1H NMR spectrum of the
trans peracetylated synthetic compound (4) was very
similar to that of the cis compound, except for the peaks of
trans olefinic protons, which were shifted downfield by
Alliar in oside [(2Z)-4-(â-D-glu copyr an osyloxy)-2-bu ten e-
n itr ile] (1): colorless gummy material; UV (MeOH) λmax 218
nm; IR (as neat and KBR pellet) ν at 3300, 2260, 2130, 1680,
max
1 1
-
1630, 855, 790, 750, and 720 cm ; H NMR (DMSO-d ) δ 6.76
6
1
about 0.15 ppm. In fact, H NMR analysis of 1 also showed
(1H, ddd, J ) 11.2, 6.0, 6.4 Hz, H-3), 5.73 (1H, d, J ) 11.2 Hz,
H-2), 4.49 (1H, dd, J ) 6.0, 14.0 Hz, H-4a), 4.35 (1H, dd, J )
the presence of 2-3% of the trans compound. However, we
were not able to determine whether the E isomer is natural
or an artifact formed during the isolation process.
6
.4, 14.0 Hz, H-4b), 4.20 (1H, d, J ) 8.0 Hz, H-1′), 3.60 (1H, d,
J ) 11.8 Hz, H-6b′), 3.40 (1H, dd, J ) 11.8, 6.0 Hz, H-6a′),
3
.16 (1H, dd, J ) 6.8, 16.4 Hz, H-3′), 3.13 (1H, m, H-4′), 3.05
The presence of cyanogenic glycosides (cyanohydrins) in
13
(
1H, m, H-5′) 2.98 (1H, dd, J ) 8.0, 16.4 Hz, H-2′); C NMR
6
many plant families is well established. In addition, a few
(DMSO-d ) δ 151.8 (C-3), 116.6 (C-1), 103.0 (C-1′), 100.9 (C-
6
noncyanogenic glycosides formed from aglycones with
nongeminal hydroxy and nitrile groups are also known
2), 77.4 (C-4′), 76.8 (C-3′), 73.6 (C-2′), 70.0 (C-5′), 67.3 (C-4a),
67.3 (C-4b), 61.3 (C-6a), 61.3 (C-6b); positive-ion ESIMS m/z

4
42 J ournal of Natural Products, 2001, Vol. 64, No. 4
Haribal et al.
+
+
2
(
46 (M + H) , 214, 168, 145, 84 (M + H - 162) ; HRESIMS
2,3,4,6-Tetr a -O-a cetyla llia r in osid e a n d Its tr a n s-Iso-
m er (2 a n d 4). To a solution of diethyl cynomethylphospho-
nate (100 mg, 0.564 mmol) in THF (15 mL) was added LDA
+
M + Na) , m/z 268.0807; calcd for C10
Acid Hyd r olysis of Allia r in osid e (1). Alliarinoside (5 mg)
was heated with 1 M HCl (2 mL) at 80 °C for 3 h. The reaction
and
15 6
H O NNa 268.0797.
(1.5 M, 0.38 mL) at -78 °C. The reaction mixture was stirred
at -78 °C for 1 h, and then the aldehyde 7 (220 mg, 0.564
mmol) in THF (5 mL) was added at -78 °C. The reaction
mixture was stirred, slowly warmed to 0 °C, and quenched
mixture was extracted with 6 mL (3 × 2 times) of CHCl
3
EtOAc each. The aqueous solution was evaporated to dryness
under reduced pressure and used for preparation of a TMS
derivative. The TMS derivative was prepared by adding 1 mL
of TRI-SIL ‘Z’ to the residue and heating the mixture at 60 °C
for 20 min. The reaction mixture was directly injected into GC.
TMS glucose was confirmed by comparison with an authentic
material prepared in similar a manner. Another batch of
compound (1 mg) was hydrolyzed in the same way, and the
aqueous fraction was directly injected into the mass spectrom-
eter for analysis of hydrolysis products.
with saturated aqueous NH
rated, and the aqueous layer was extracted with ethyl acetate
2 × 20 mL). The combined organic layers were washed with
brine (5 mL), dried (MgSO ), and concentrated in vacuo. The
4
Cl. The organic layer was sepa-
(
4
residue was purified by flash chromatography (EtOAc-hex-
anes, 2:1) to yield the cis-alkenyl nitrile 2 (118 mg, 51%) as
colorless prisms, mp 84-86 °C, [R]
D
-4° (c, 0.4, CHCl
3
):
+
ESIMS 414 (M + H ), 331, 271, 169, 113, 98, 87, 79, 73, 60;
1
HRCIMS m/z 414.1390 (calcd for C18
NMR and 13C NMR identical to that of 2 obtained from natural
. Compound 2 was formed together with the trans-alkenyl
nitrile 4 (80 mg, 34%) as colorless needles, mp 86-88 °C, [R]
24 1
H N O10 414.1400); H
P er a cetyla ted Allia r in osid e (2). Alliarinoside (5 mg) was
heated in Ac O-pyridine (1:1, 3 mL) at 60 °C for 4 h. The
2
1
reaction mixture was worked up in the usual way to obtain 2
D
as a colorless crystalline compound: UV (MeOH) λmax 218 nm;
1
-
23.1° (c, 0.3, CHCl
6.0, 4.0 Hz, H-3), 5.63 (1H, dt, J ) 16.0, 2.4 Hz, H-2), 5.23
1H, dd, J ) 9.6, 9.6 Hz, H-3′), 5.10 (1H, dd, J ) 9.6, 9.6 Hz,
3 3
): H NMR (CDCl ) δ 6.71 (1H, dt, J )
1
H NMR (CDCl ) δ 6.58 (1H, ddd, J ) 11.2, 6.8, 5.6 Hz, H-3),
3
1
5
.48 (1H, dd, J ) 11.2, 1.6 Hz, H-2), 5.22 (1H, dd, J ) 9.6, 9.6
(
Hz, H-3′), 5.10 (1H, dd, J ) 9.6, 9.6 Hz, H-4′), 5.02 (1H, dd,
J ) 9.6, 8.0 Hz, H-2′), 4.59 (1H, ddd, J ) 14.4, 5.6, 1.6 Hz,
H-4a), 4.57 (1H, d, J ) 8.0 Hz, H-1′), 4.53 (1H, dd, J ) 14.4,
H-4′), 5.06 (1H, dd, J ) 9.6, 8.0 Hz, H-2′), 4.56 (1H, d, J ) 8.0
Hz, H-1′), 4.52 (1H, ddd, J ) 16.8, 4.0, 2.4 Hz, H-4a), 4.26 (1H,
dd, J ) 12.4, 4.8 Hz, H-6a′), 4.23 (1H, ddd, J ) 16.8, 4.0, 2.4
Hz, H-4b), 4.16 (1H, dd, J ) 12.4, 2.4 Hz, H-6b′), 3.72 (1H,
ddd, 9.6, 4.8, 2.4 Hz, H-5′), 2.10, 2.07, 2.04, 2.03 (s 3H each
6
.8 Hz, H-4b), 4.26 (1H, dd, J ) 12.4, 4.8 Hz, H-6a′), 4.17 (1H,
dd, J ) 12.4, 2.4 Hz, H-6b′), 3.73 (1H, ddd, J ) 9.6, 4.8, 2.4
1
3
Hz, H-5′), 2.12, 2.06, 2.04, 2.01 (3H, s each, COCH
3
;
C NMR
1
3
COCH
1
7
(
4
3
); C NMR (CDCl
49.0 (C-3), 117.1 (C-1), 100.5 (C-1′), 100.4 (C-2), 72.7 (C-3′),
2.3 (C-2′), 71.2 (C-4′), 68.4 (C-4a), 68.4 (C-4b), 67.5 (C-5′), 61.9
); ESIMS
14 (M + H ), 331, 271, 169, 113, 98, 87, 79, 73, 60; HRCIMS
10 414.1400).
Hyd r olysis of 2 a n d 4. The acetylated compound (2, 5-6
mg) was dissolved in 0.5 mL of dry MeOH, and anhydrous K
CO was added. The mixture was stirred at room temperature
3
) δ 170.8, 170.4, 169.5, 169.4 (CO),
(
(
(
(
4
CDCl ) δ 170.8, 170.4, 170.1, 169.7 (CO), 150.5 (C-3), 116
C-1), 102.1 (C-1′), 100.9 (C-2), 73.5 (C-2′), 72.6 (C-3′), 72.0
C-4′), 69.4 (C-4a), 69.4 (C-4b), 67.9 (C-5′), 62.7 (C-6a′), 62.7
3
C-6a′), 61.9 (C-6b′), 20.9, 20.9, 20.8, 20.8 (COCH
3
3
C-6b′), 20.7, 20.7, 20.6, 20.6 (COCH ); positive-ion ESIMS m/z
+
+
+
+
14 (M + H) , 383 (M + H - 31) , 372 (M + H - 42) , 84
m/z 414.1405 (calcd for C18
24 1
H N O
+
(
M + H - 330) .
-O-Allyl-2,3,4,6-tetr a-O-acetyl-â-D-glu copyr an oside (6).
To a mixture of tetra-O-acetyl-R-glucosyl bromide (5) (206 mg,
.5 mmol), allyl alcohol (44 mg, 0.75 mmol), and 4 Å molecular
sieves (150 mg) in CH Cl (5 mL) was added silver triflate (193
1
2
-
3
0
for 4 h. The reaction was monitored by ESIMS to detect the
formation of 1. The resultant mixture was filtered through
glass wool and separated by repeated HPLC to give 1.
Bioa ssa ys. Choice bioassays for feeding deterrent/inhibitor
activity were conducted with neonate larvae of P. napi olera-
cea. Two 1.5 cm disks of cabbage leaves were used as test
substrate. One disk was treated with the sample to be tested,
and the other was treated with solvent alone. Five to eight
replicates were used for each test. In the case of the natural
compound 0.1 g leaf equivalent of sample dissolved in 20 µL
of MeOH was applied on both sides of each test disk. For the
synthetic compound, although the amount was not verified, a
sample of UV absorbance similar to that for the natural
compound was used (to ensure comparable amount of natural
and synthetic alliarinoside). Two disks were mounted on insect
pins that were affixed to the bottom of waxed paper cups, 10
cm wide and 5 cm deep, lined with moistened filter paper. The
pins were placed in a way that there was a slight overlap of
two disks to enable neonate larvae to move from one disk to
the other. Eggs from P. napi oleracea were collected on a
Parafilm layer so that the hatching larvae had no contact with
the host plants. Either four or five freshly emerged neonates
were transferred to the disks. The cups were covered with an
airtight lid and placed in a larger box lined with moistened
paper napkins to keep the humidity high to prevent the disks
from desiccating. This box was kept in an incubator for 18 h
at 28 °C. Activity was expressed as numerical values based
on the area that was consumed. This was done visually as
follows: 0 ) no feeding; 1 ) <5% of the disk; 2 ) >5% and
<10%; and 3 ) >10%. Ranks were assigned to both control
and treated disks.
2
2
mg, 0.75 mmol) at 0 °C. The reaction mixture was stirred
vigorously at 0 °C for 40 min, and the mixture was filtered
through a pad of silica gel. The filtrate was washed with
saturated NaHCO and brine, dried with MgSO , and concen-
3 4
trated in vacuo. The residue was purified by flash chromatog-
raphy (EtOAc-hexanes, 1:2) to afford the glucoside 6 (106 mg,
5
5%) as colorless prisms, mp 86-88 °C, [R] -22.6° (c, 1.8,
D
1
3 3
CHCl ): H NMR (CDCl ) δ 5.81 (1H, dddd, J ) 17.2, 10.4,
6
.4, 4.8 Hz), 5.24 (1H, ddd, J ) 17.2, 3.2, 1.6 Hz), 5.17 (1H,
ddd, J ) 10.4, 3.2, 1.6 Hz), 5.16 (1H, d, J ) 9.6 Hz), 5.06 (1H,
t, J ) 9.6 Hz), 4.99 (1H, dd, J ) 9.6, 8.0 Hz), 4.53 (1H, d, J )
8
.0 Hz), 4.30 (1H, ddt, J ) 13.2, 4.8, 1.6 Hz), 4.23 (1H, dd,
J ) 12.4, 4.8 Hz), 4.11 (1H, dd, J ) 12.4, 2.4 Hz), 4.06 (1H, tt,
6
2
1
7
.4, 1.6 Hz), 3.66 (1H, ddd, J ) 9.6, 4.8, 2.4 Hz), 2.05 (3H, s),
13
.01 (3H, s), 1.99 (3H, s), 1.97 (3H, s); C NMR (CDCl ) δ
3
70.6, 170.3, 169.4, 169.3, 133.3, 117.6, 99.6, 72.9, 71.8, 71.3,
0.0, 68.4, 61.9, 20.7, 20.7, 20.6, 20.6.
2
-(2′,3′,4′,6′-Tet r a -O-a cet yl-â-D-glu cop yr a n osyl)et h a -
n a l (7). The olefin 6 (24 mg, 0.062 mmol) in CH Cl (2 mL)
2
2
was cooled to -78 °C, and ozone was bubbled into the solution
at -78 °C until the solution appeared blue. The reaction
mixture was purged with nitrogen as it warmed to room
temperature. Dimethyl sulfide (20 µL) was added, and the
reaction mixture was stirred at room temperature overnight.
After washing with water (2 mL) and brine (2 mL), the solution
was dried (MgSO
The residue was purified by chromatography (EtOAc-hexanes,
:1) to afford aldehyde 7 (20 mg, 83%) as colorless oil, [R]
4
), and the solvent was removed in vacuo.
1
-
1
D
1
23.5° (c, 0.7, CHCl
.6, 0.8 Hz), 5.24 (1H, t, J ) 9.6 Hz), 5.10 (1H, dd, J ) 9.6, 8.0
Hz), 5.09 (1H, t, J ) 9.6 Hz), 4.60 (1H, d, J ) 8.0 Hz), 4.28
1H, dd, J ) 17.2, 0.8 Hz), 4.25 (1H, dd, J ) 12.4, 5.2 Hz),
.19 (1H, dd, J ) 17.2, 1.6 Hz), 4.12 (1H, dd, J ) 12.4, 2.4
Hz), 3.72 (1H, ddd, J ) 9.6, 5.2, 2.4 Hz), 2.09 (3H, s), 2.09
3 3
): H NMR (CDCl ) δ 9.68 (1H, dd, J )
Sta tistica l An a lyses. Ranks assigned to disks were com-
pared by Wilcoxon’s nonparametric sign rank test for paired
samples (Siegel and Castallen).13
(
4
Ack n ow led gm en t. This work was supported by NSF
grant no. DEB-9419797 to Dr. Alan Renwick and NIH grant
no. GM53830 to Dr. J errold Meinwald. We thank Dr. Bruce
Ganem for the use of the Perkin-Elmer IR spectrophotometer.
1
3
(
1
6
3H, s), 2.04 (3H, s), 2.02 (3H, s); C NMR (in CDCl
3
) δ 199.8,
70.6, 170.2, 160.4, 160.3, 100.9, 74.1, 72.5, 72.1, 70.9, 68.3,
1.3, 20.7, 20.7, 20.6, 20.6.

A Cyanoallyl Glucoside from Alliaria
J ournal of Natural Products, 2001, Vol. 64, No. 4 443
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