Enzymatic asymmetric hydroxylation of unnatural substrates with soybean lipoxygenase

Article abstract of DOI:10.1016/S0957-4166(01)00381-0

Surrogate substrates mimicking the natural substrate (linoleic acid) bearing a spacing prosthetic group with a non-ionic hydroxyl terminus undergo asymmetric hydroxylation with soybean lipoxygenase. The prosthetic modifier supplies the missing structural features needed for enzymatic recognition and controls the regiochemical outcome of the reaction by its high hydrophobic content. The effect of pH on the regiochemistry clearly shows that all the substrates can arrange themselves at the active site of soybean lipoxygenase in only one orientation leading to formation of hydroperoxides by oxygenation at the ω-6 carbon.

Full text of DOI:10.1016/S0957-4166(01)00381-0

TETRAHEDRON:
ASYMMETRY
Pergamon
Tetrahedron: Asymmetry 12 (2001) 2129–2135
Enzymatic asymmetric hydroxylation of unnatural substrates
with soybean lipoxygenase
J. S. Yadav,* S. Nanda and A. Bhaskar Rao
Organic Division, Indian Institute of Chemical Technology, Hyderabad 500007, India
Received 13 July 2001; accepted 29 August 2001
Abstract—Surrogate substrates mimicking the natural substrate (linoleic acid) bearing a spacing prosthetic group with a non-ionic
hydroxyl terminus undergo asymmetric hydroxylation with soybean lipoxygenase. The prosthetic modifier supplies the missing
structural features needed for enzymatic recognition and controls the regiochemical outcome of the reaction by its high
hydrophobic content. The effect of pH on the regiochemistry clearly shows that all the substrates can arrange themselves at the
active site of soybean lipoxygenase in only one orientation leading to formation of hydroperoxides by oxygenation at the v-6
carbon. © 2001 Published by Elsevier Science Ltd.
1. Introduction
are a single v-6-(Z,Z)-1,4-pentadienyl moiety and an
appropriately spaced proximal carboxyl group as char-
acterized by the natural substrate linoleic acid.3 It had
long been considered that the carboxyl group in the
fatty acid was important for enzymatic recognition and
binding.4 However, Harris et al.5 reported that glycerol
esters of linoleic acid bearing an ionic phosphatidyl-
choline group are also substrates for the enzyme.
Despite its high degree of stereo- and regioselectivity,
soybean lipoxygenase has attracted little attention from
a synthetic point of view.6 As part of our continuing
effort7 to explore the synthetic utility of SBLOX, we
describe herein an enzymatic method for the asymmet-
ric hydroxylation of unnatural synthetic substrates 1a–
The lipoxygenases are a group of non-heme iron-con-
taining dioxygenases which catalyze the addition of
molecular oxygen to polyunsaturated fatty acids in a
stereospecific and regiospecific way1 (Scheme 1). They
are involved in the biosynthesis of inflammatory media-
tors in cell differentiation and atherogenesis.2 Previous
studies of modified substrates with soybean lipoxyge-
nase suggest that the essential structural requirements
1c bearing
a spacing prosthetic modifier, which
contains a non-ionic hydroxyl terminus as shown below
(Scheme 2). The primary purpose of the prosthetic
modifier is to supply the missing structural features
needed for enzymatic recognition and to influence the
regiochemistry of the reaction by its high hydrophobic
content.
2. Results and discussion
2.1. Synthesis
The substrates 1a–1c were synthesised from a common
precursor 3-butyn-1-ol, which was alkyated with LiNH2
and bromopentane to give 3-nonyn-1-ol. Hydrogena-
tion over Lindlar catalyst gives (Z)-3-nonen-1-ol in
90% yield, which upon treatment with iodine,
triphenylphosphine and imidazole8 gives the corre-
Scheme 1. Substrate recognition by SBLO.
* Corresponding author. Fax: +91-40-7170512; e-mail: yadav@
iict.ap.nic.in
0957-4166/01/$ - see front matter © 2001 Published by Elsevier Science Ltd.
PII: S0957-4166(01)00381-0

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J. S. Yada6 et al. / Tetrahedron: Asymmetry 12 (2001) 2129–2135
Scheme 2. Asymmetric hydroxylations of unnatural substrates 1a–1c with soyben lipoxygenase. Reagents and conditions: (a)
SBLO, O2, 0.2 M sodium borate buffer (pH 9), 0°C, 1 h; (b) TPP, ether; (c) KOH, MeOH, rt, 12 h.
sponding iodo compound in 85% yield. The iodo com-
pound when reacted with triphenylphosphine in reflux-
ing acetonitrile yields the phosphonium salt (Scheme 3).
The dienoic alcohols with (Z,Z) geometry were pre-
pared by condensation of the ylide derived from the
phosphonium iodide 8 with the appropriate aldehydic
partner 9 and subsequent removal of the tetra-
hydropyranyl (THP) protecting groups with p-toluene-
sulphonic acid in methanol (Scheme 4). All the dienoic
alcohol products have >95% (Z,Z) geometry, as deter-
mined by GC analysis.
tiomeric purity through lipoxygenase-catalyzed trans-
formation of unnatural substrates. These chiral diols
6a–6c can be efficiently employed for the total synthesis
of some unsaturated hydroxy fatty acids (Scheme 6).
Currently, we are actively engaged in the total synthesis
of some hydroxy fatty acids and their analogs, which
have tremendous biological importance.
In all of the above cases oxygenation occurs at the
appropriate olefinic site yielding the product 2, whereas
unwanted formation of 3 by attack of oxygen at the
other olefinic site is minimal. This clearly indicates that
substrates 1a–1c preferentially arrange themselves at
the active site of SBLO-1 in an orientation leading to
Coupling of the dienols with mono p-methoxybenzyl
protected acids derived from the appropriate diols
under DCC/DMP conditions afforded the coupled
esters 11 in 80% yield. Removal of the 4-methoxybenzyl
group with DDQ9 furnished the lipoxygenase substrates
1a–1c in 80% yield (Scheme 5). All the products were
1
characterised by H and 13C NMR spectroscopy.
For the enzymatic oxidation reactions, solutions of
substrates 1a–1c in 0.2 M sodium borate buffer (pH 9)
at 0°C were treated with lipoxygenase type-1 (activity=
127,000 units/mg of protein). The reaction mixture was
then stirred under a stream of oxygen for 1 h. The
reaction was stopped by addition of ethanol and water,
extracted with ether and evaporated to give the crude
hydroperoxide, which was then converted to alcohol by
treatment with triphenylphosphine. No isomerization
was observed in the reduction of the hydroperoxides to
the corresponding alcohols 5. The prosthetic group
then was removed by hydrolysis with KOH/MeOH to
give the chiral diols, which have comparable specific
rotation values to those previously reported.12
Scheme 3. Reagents and conditions: (a) LiNH2; THF/HMPA
(6:1), C5H11Br; (b) H2, Lindlar catalyst; (c) I2, TPP/imidazole;
acetonitrile:ether (1:3); (d) TPP, acetonitrile, (reflux, 48 h).
2.2. Regiochemistry
Scheme 4. Reagents and conditions: (a) DHP/TsOH 1; (b)
IBX/DMSO, rt 6 h; (c) 8, n-BuLi, THF–HMPA (10:1), 0°C,
2 h, MeOH, TsOH, 80%.
The present study shows that synthetically useful chiral
diols can be obtained in high chemical yield and enan-

J. S. Yada6 et al. / Tetrahedron: Asymmetry 12 (2001) 2129–2135
2131
Scheme 5. Reagents and conditions: (a) 4-Methoxybenzylbromide, NaH, n-Bu4N+I−; (b) CrO3, H2SO4 (8N); (c) 10a–10c DCC,
DMAP; (d) DDQ, DCM–H2O (19:1), rt.
formation of 2 by oxygenation at the v-6 carbon,
whereas formation of 3 by attack of oxygen at the v-10
carbon is possible only if the substrate arranges itself
head to tail (i.e. in opposite orientation) at the active
site of SBLO-1.
orientation of the substrate is solely determined by the
size of the binding pocket, whereby the enzyme prefer-
entially binds the group that has proper pocket fit.
2.3. Effect of pH
The effect of pH on the regiochemistry (Table 2) also
proves this hypothesis. It was reported previously3,10
that the difference in the hydrophobic content between
the proximal and the distal unit significantly influences
the regioselectivity of the oxygenation reaction. The
current study also shows the relevance of hydrophobic
interactions to the regioselectivity of oxygenation reac-
tions with lipoxygenase-1. From Table 1 it is clear that
all of the three substrates show excellent regiospecific-
ity, since the hydrophobicity of the C5H11 (log p=+2.8)
is much higher than that of the hydroxylated terminus12
(log p=−3.7). The reactions of compounds 1a–1c dis-
played similar regioselectivity to that of the natural
substrate linoleic acid because the proximal units are of
The natural substrate, linoleic acid, undergoes SBLO-
catalyzed oxygenation at pH 9.0, to afford a product
ratio of (13S)-HPOD and (9S)-HPOD of 49:1. As the
pH was decreased below 9, the proportion of (9S)-
HPOD increased linearly until at pH 6.0 it constituted
about 25% of the product mixture.11 When substrates
1a–1c are subjected to oxygenation with SBLO at dif-
ferent pH values (6–9), it was observed that the regiose-
lectivity was unaffected by the change in pH (Table 2),
suggesting that substrates 1a–1c can arrange themselves
at the active site of SBLO in only one orientation, when
compared to linoleic acid, which can arrange itself in
either (normal or reverse) orientation (Scheme 7)
depending on the pH of the medium.
,
similar lengths (10.6 and 10.9 A). It indicates that the
Scheme 6. Proposed synthetic plans for unsaturated hydroxy fatty acids from 6a–6c.
Table 1. SBLO-catalyzed oxidation of modified substrates 1a–1c
Regiospecificitya (4:5)
Yield of 6 (%)
[h]2D5 of 6b (°), config.
(S:R)c
1a
1b
1c
49:1
49:1
97:3
40
43
45
+52, (S)
+28, (S)
+39, (S)
49:1
97:3
49:1
a Values were determined by normal-phase HPLC at 235 nm.
b All rotations were measured as solutions in CHCl3, c=1 at 25°C.
c (S:R) ratios of 6 were determined by chiral HPLC (Diacel, Chiral OD column, hexane:isopropanol, 9:1.

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J. S. Yada6 et al. / Tetrahedron: Asymmetry 12 (2001) 2129–2135
Table 2. Effect of pH on the regioisomeric ratio 2a:3a
(m, 2H), 2.1 (t, J=7 Hz, 2H), 2.4 (t, J=7 Hz, 2H), 3.6
(t, J=7 Hz, 2H). EIMS (m/z): 140 (M+).
pH Buffer
Total hydroperoxide Regiospecificity (2a:3a)
(%)
4.3. (3Z)-3-Nonen-1-ol
6.0
6.5
7.0
7.5
8.0
8.5
9.0
Acetate
Acetate
Phosphate
Phosphate
Borate
42
51
50
52
65
70
72
9:1
9:1
3-Nonyl-1-ol (1 g, 7.14 mmol) in absolute ethanol (10
mL) was dissolved in a small round bottom flask.
Lindlar catalyst (50 mg, Pd on CaCO3) and few drops
of quinoline were added and hydrogen was supplied
from balloons. The reaction was monitored by TLC
(visualization was carried out by dipping the plates in
Eckerts reagent and heating it at 120°C), where the
olefinic product moves a little faster than the starting
material. After completion of the reaction the mixture
was filtered to remove the catalyst. Ethanol was
removed on a rotary evaporator. Dichloromethane was
added to the residue and it was washed with 5% aq.
HCl, brine, dried (Na2SO4) and evaporated. The result-
ing product was pure enough for the next step. IR (TF):
92:8
19:1
97:3
97:3
49:1
Borate
Borate
3. Conclusion
Our present study demonstrates a synthetically useful
enzymatic method for asymmetric hydroxylation of
unnatural alkenes. It also reveals two other important
points, firstly the prosthetic group having a non-ionic
hydroxy terminus can recognise the enzyme and sec-
ondly the regiochemical outcome appears to be strongly
influenced by the high hydrophobic content of the
proximal unit. These results lead us directly onto the
design of new unnatural substrates and the study of
their activity with lipoxygenase is currently under
investigation.
1
3390, 1630, 980 cm−1; H NMR: 0.9 (t, J=7 Hz, 3H),
1.35 (brs, 6H), 2.1 (q, J=7 Hz, 2H), 2.35 (q, J=7 Hz,
2H), 3.6 (t, J=7 Hz, 2H), 5.25–5.4 (m, 2H); EIMS
(m/z): 142 (M+).
4.4. (3Z)-3-Nonenyliodide
(3Z)-Nonen-1-ol (2 g, 14 mmol) was dissolved in aceto-
nitrile: ether (3:1, 75 mL). Imidazole (1.44 g, 21 mmol)
was added at rt, followed by addition of I2 (5.36 g, 21
4. Experimental
4.1. General
Unless otherwise stated, materials were obtained from
commercial suppliers and used without further purifica-
tion. THF was distilled from sodium benzophenone
ketyl. HMPA was distilled from BaO and stored over 3
,
A molecular sieves. Dichloromethane (DCM) was dis-
tilled from calciumhydride. Soybean lipoxygenase
(type-I) was obtained from Sigma co. 1H and 13C NMR
were obtained on a Varian VXR 200 (200 and 50
MHz). Chemical shifts are reported as ppm downfield
of tetramethylsilane. Infrared spectra were recorded on
a Perkin–Elmer 1420 spectrometer. Optical rotations
were measured on a Jasco Dip 360 digital polarimeter.
The abbreviation TF denotes thin film.
4.2. 3-Nonyl-1-ol
To a stirred suspension of lithium amide (prepared
from 2.8 g, 0.4 g atom of Li) in liquid ammonia (300
mL) was added 3-butyn-1-ol (14 g, 200 mmol) over 30
min. The mixture was stirred for an additional 1 h. A
solution of 1-bromopentane (30.2 g, 200 mmol) in
THF/HMPA (60 mL, 6:1) was added and stirred for a
further 6 h. The reaction was quenched with solid
NH4Cl (20 g) and excess ammonia was then allowed to
evaporate. The residue was dissolved in water and
extracted with ether. The combined organic extract was
washed with water, brine, and dried (Na2SO4). The
crude product was purified by column chromatography
to afford 3-nonyl-1-ol (20 g, 70%). IR (TF): 3400, 1090
Scheme 7. Orientation of natural substrate and synthetic
substrates in the active site of SBLOX.
1
cm−1; H NMR: 0.9 (t, J=7 Hz, 3H), 1.3 (m, 4H), 1.5

J. S. Yada6 et al. / Tetrahedron: Asymmetry 12 (2001) 2129–2135
2133
mmol) and triphenylphosphine (5.55 g, 21 mmol). The
reaction mixture was stirred for 45 min at rt. The
mixture was filtered and the filtrate was concentrated to
give a semi-solid residue, which was purified through
column chromatography to give the pure iodide (3 g,
85%). IR (TF): 2825–3000 cm−1 (CꢀCH, alkane CH);
1H NMR: 0.9 (t, J=7 Hz, 3H), 1.35 (brs, 6H), 2.1 (q,
J=7 Hz, 2H), 2.65 (q, J=7 Hz, 2H), 3.1 (t, J=7 Hz,
2H), 5.26–5.4 (m, 2H); EIMS (m/z): 252 (M+).
1.54 g) was added to the solution portionwise at 0°C.
The reaction mixture was stirred at 0°C for 1 h under a
nitrogen atmosphere. Tetrabutylammonium iodide (cat-
alytic) was added followed by the addition of 4-
methoxybenzylbromide (9.2 g, 45.6 mmol) and the
mixture was stirred for a further 2 h at rt. Water was
added to the reaction mixture and extracted with
EtOAc. The organic extract was washed with brine and
dried (Na2SO4). Purification by means of column chro-
matography gave the product (6.8 g, 80%). IR (TF):
1
4.5. (3Z)-3-Nonenyl-1-triphenylphosphoniumiodide 8
3400, 3000 cm−1; H NMR: 1.7 (m, 6H), 3.4 (t, J=6.5
Hz, 2H), 3.5 (t, J=6.5 Hz, 2H), 3.8 (s, 3H), 4.4 (s, 2H),
6.85 (d, J=6 Hz, 2H), 7.2 (d, J=6 Hz, 2H); EIMS
(m/z): 224 (M+).
(3Z)-3-Nonenyliodide (3 g, 12 mmol) was dissolved in
dry acetonitrile (50 mL). Triphenylphosphine (3.11 g,
12 mmol) was added and the reaction mixture was
heated at 60°C for 48 h under a nitrogen atmosphere.
Acetonitrile was evaporated and dry ether (30 mL) was
added to the semi-solid residue and shaken vigorously.
Evaporation of the ether gave the phosphonium salt (6
g) as a white solid.
4.9. 5-(4-Methoxybenzyloxy)pentanoic acid
5-(4-Methoxybenzyloxy)-1-pentanol (5 g, 22 mmol) was
dissolved in distilled acetone (50 mL) at 0°C. Freshly
prepared Jones reagent was added dropwise at the same
temperature until the orange brown color persisted. The
reaction mixture was allowed to warm to rt and stirred
for a further 30 min, water was added and the mixture
was extracted with EtOAc. Purification by chromatog-
raphy gave the acid as a white solid (4 g, 75%). IR
4.6. 3-Tetrahydropyranylpropanal 9a
3-Tetrahydropyranyl-1-propanol (1.5 g, 9.4 mmol) was
dissolved in DMSO (10 mL). 2-Iodoxybenzoic acid
(IBX, 5.25 g, 18.75 mmol) was added and the reaction
mixture stirred for 4 h at rt. Water was added to the
solution and 2-iodobenzoic acid was filtered off. The
filtrate was extracted several times with ether, dried
(Na2SO4) and concentrated. Purification of the residue
by column chromatography gave 3-tetrahydro-
pyranylpropanal (1.2 g, 80%). IR (TF): 2860, 1705
1
(KBr): 3000–2800, 1700 cm−1; H NMR: 1.7 (m, 4H),
2.35 (t, J=6.5 Hz, 2H), 3.5 (t, J=6.5 Hz, 2H), 3.8 (s,
3H), 4.4 (s, 2H), 6.85 (d, J=6 Hz, 2H), 7.2 (d, J=6 Hz,
2H). 13C NMR: 21.5, 28.9, 33.6, 55.2, 69.4, 72.5, 114,
129, 130, 159.2, 180. EIMS (m/z): 238 (M+).
4.10. (3Z,6Z)-3,6-Dodecadienyl (4-methoxybenzyloxy)-
pentanoate 11a
1
cm−1; H NMR: 1.45–1.88 (m, 6H), 2.5 (t, J=6.8 Hz,
2H), 3.5 (m, 2H), 3.82 (m, 2H), 4.63 (t, J=3.9 Hz, 1H),
9.6 (1H); EIMS (m/z): 158 (M+).
5-(4-Methoxybenzyloxy)pentanoic acid (400 mg, 1.7
mmol) was dissolved in dichloromethane (4 mL). DCC
(352 mg, 1.7 mmol) was added, followed by the addi-
tion of DMAP (10 mg) at 0°C. The reaction mixture
was stirred for 5 min and a solution of (3Z,6Z)-3,6-
dodecadiene-1-ol (367 mg, 2 mmol) in dichloromethane
(2 mL) was added dropwise to the mixture. The reac-
tion was stirred for a further 3 h at rt. Dicyclohexylurea
was removed by filtration and the filtrate was washed
with water, brine and dried (Na2SO4). After chromato-
graphic separation 13a was obtained (486 mg, 60%). IR
(TF): 3030, 1740 cm−1; PMR: 0.85 (t, J=7 Hz, 3H),
1.35 (brs, 6H), 1.7 (m, 4H), 2.1 (q, J=7 Hz, 2H), 4.4 (s,
2H), 5.25–5.6 (m, 4H), 6.85 (d, J=6 Hz, 2H), 7.2 (d,
J=6 Hz, 2H). 13CNMR: 14, 21.5, 25.7, 27.3, 29.4, 30.9,
31.6, 60.2, 69.4, 72.5, 114, 125.5, 127.7, 129, 130, 130.8,
131.5, 159.2, 179. FABMS: 402 (M+).
4.7. (3Z,6Z)-3,6-Dodecadiene-1-ol 10a
A solution of 8 (1.5 g, 3 mmol) in THF: HMPA (6:1, 30
mL) was treated with a solution of n-BuLi (1.2 mL, 2.5
M in hexane) at 0°C, the resulting orange colored
solution was stirred at the same temperature for 1 h.
3-Tetrahydropyranyl propanal 9a (0.46 g, 3 mmol) in
THF (5 mL) was added and the reaction mixture stirred
for a further 2 h at the same temperature. The mixture
was quenched with saturated NH4Cl solution and
extracted with ether. The organic extract was washed
with water, brine and dried (Na2SO4). The crude
product was dissolved in methanol (15 mL) and cata-
lytic PTSA was added. The mixture was stirred for 1 h
at rt. Evaporation of methanol and purification by
column chromatography afforded the (Z,Z) dienol in
1
60% yield. IR (TF): 3380, 1090 cm−1; H NMR: 0.9 (t,
4.11. (3Z,6Z)-3,6-Dodecadienyl 5-hydroxypentanoate
1a
J=7 Hz, 3H), 1.25 (m, 6H), 2.1 (q, J=7 Hz, 2H), 2.2
(brs, 1H, -OH), 2.33 (q, J=7 Hz, 2H), 2.8 (t, J=7 Hz,
2H), 3.62 (t, J=7 Hz, 2H), 5.25–5.6 (m, 4H); 13C
NMR: 14.09, 22.62, 25.8, 27.3, 29.3, 30.9, 31.57, 62.3,
125.4, 127.5, 130.6, 131.5; EIMS (m/z): 182 (M+).
Compound 11a (220 mg, 0.54 mmol) was dissolved in
dichloromethane:water (19:1, 6 mL), DDQ (186 mg, 0.8
mmol) was added and the resulting solution stirred for
1 h at rt. The reaction mixture was filtered and the
filtrate was washed with a 5% NaHCO3 solution, brine
and dried (Na2SO4). Purification by chromatography
4.8. 5-(4-Methoxybenzyloxy)-1-pentanol
1
1,5-Pentanediol (4 g, 38 mmol) was dissolved in dry
THF (80 mL) and NaH (60% dispersion in mineral oil,
gave 1a (125 mg, 80%). IR (TF): 3380, 1745 cm−1; H
NMR: 0.9 (t, J=7 Hz, 3H), 1.3 (brs, 6H), 1.5–1.8 (m,

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J. S. Yada6 et al. / Tetrahedron: Asymmetry 12 (2001) 2129–2135
4H), 2.1 (q, J=7 Hz, 2H), 2.4 (m, 4H), 2.8 (t, J=7 Hz,
2H), 3.6 (t, J=7 Hz, 2H), 4.1 (t, J=7 Hz, 2H), 5.2–5.6
(m, 4H). 13C NMR: 13.9, 21, 22.4, 25.6, 26.8, 27.1, 29.2,
31.4, 32, 33.8, 62.1, 63.7, 124.6, 127, 130.5, 131, 172.
EIMS (m/z): 282 (M+).
3H), 1.25–1.45 (m, 8H), 2.1 (q, J=7 Hz, 4H), 2.2 (brs,
1H, -OH), 2.8 (t, J=7 Hz, 2H), 3.65 (t, J=7 Hz, 2H),
5.3–5.6 (m, 4H). 13C NMR: 14, 22.6, 24, 25.9, 27.2, 29.2,
31, 31.7, 63, 125.6, 128, 130.5, 131. EIMS (m/z): 196
(M+).
4.12. 7-Hydroxy-(3Z,5E,7S)-3,5-dodecadienyl 5-hydroxy-
pentanoate 4a
4.16. (5Z,8Z)-5,8-Tetradecadiene-1-ol 10c
1
IR (TF): 3385, 1095 cm−1; H NMR: 0.9 (t, J=7 Hz,
To a homogeneous solution of 1a (100 mg, 0.35 mmol)
in sodium borate buffer (0.2 M, pH 9, 20 mL) at 0°C
was added Lipoxygenase type-1 (50 mg), while O2 was
bubbled through the solution at such a rate to produce
the minimum of foaming. After 1 h the reaction mixture
was acidified with citric acid to pH 5.0, and then
extracted several times with either, dried (NaSO4) and
evaporated to afford the crude hydroperoxide.
3H), 1.25 (m, 6H), 1.4–1.6 (m, 4H), 2.15 (q, J=7 Hz,
4H), 2.25 (brs, 1H, -OH), 2.75 (t, J=7 Hz, 2H), 3.68 (t,
J=7 Hz, 2H), 5.28–5.6 (m, 4H). 13C NMR: 13.8, 22.7,
24.5, 25, 26.1, 27.2, 29, 31, 31.8, 62.5, 125.4, 128, 130.2,
131. EIMS (m/z): 210 (M+).
4.17. (4Z,7Z)-4,7-Tridecadienyl (4-methoxybenzyloxy)-
butanoate 11b
1
The crude hydroperoxide (105 mg) was dissolved in
diethyl ether (2 mL) and the solution cooled in an ice
bath. Triphenylphosphine (TPP, 2 mmol) was added
and the mixture was stirred overnight in an ice bath.
Ether was evaporated to give the crude diol. Chromato-
graphic purification afforded the diol (44 mg, 40%). IR
IR (TF): 1735 cm−1; H NMR: 0.85 (t, J=7 Hz, 3H),
1.35 (brs, 6H), 1.6–1.75 (m, 4H), 2.12 (q, J=7 Hz, 4H),
2.4 (t, J=7 Hz, 2H), 2.75 (t, J=7 Hz, 2H), 3.5 (t, J=7
Hz, 2H), 3.8 (s, 3H), 4.1 (t, J=7 Hz, 2H), 4.5 (s, 2H),
5.2–5.45 (m, 4H), 6.9 (d, J=6 Hz, 2H), 7.2 (d, J=6 Hz,
2H). 13C NMR: 14, 21.3, 22.6, 25.5, 26.7, 27.2, 29.2,
31.5, 32, 34, 62.3m, 64.3, 69.5, 73, 115, 125.8, 128, 129.2,
131, 131.4, 159.2, 179. FABMS: 402 (M+).
1
(TF): 3400, 1740 cm−1; H NMR: 0.9 (t, J=7 Hz, 3H),
1.3 (brs, 6H), 1.5–1.8 (m, 4H), 2.3 (t, J=7 Hz, 2H), 2.4
(t, J=7 Hz, 2H), 2.5 (brs, 2H, -H), 3.6 (t, J=7 Hz, 2H),
4.05 (t, J=7 Hz, 2H), 4.1 (q, J=7 Hz, 1H), 5.46 (dd,
J=7, 10.4 Hz, 1H), 5.73 (dd, J=7, 15 Hz, 1H), 6.15 (dd,
J=10, 10.4 Hz, 1H), 6.52 (dd, J=10, 15 Hz, 1H). 13C
NMR: 14, 20.9, 22.5, 26, 26.9, 27.1, 29.4, 31.6, 34, 62.1,
64, 72, 124, 127.4, 128.5, 130.2, 173. EIMS (m/z): 298
(M+), [h]2D5=+11.0 (c=0.5, CHCl3).
4.18. (5Z,8Z)-5,8-Tetradecadienyl (4-methoxybenzyl-
oxy)propanoate 11c
IR (TF): 1740 cm−1; 1H NMR: 0.9 (t, J=7 Hz, 3H), 1.35
(m, 6H), 1.65 (m, 4H), 2.1 (q, J=7 Hz, 4H), 2.5 (t, J=7
Hz, 2H), 2.78 (t, J=7 Hz, 2H), 3.6 (t, J=7 Hz, 2H), 3.8
(s, 3H), 4.1 (t, J=7 Hz, 2H), 4.5 (s, 2H), 5.25–5.5 (m,
4H), 6.9 (d, J=6 Hz, 2H), 7.2 (d, J=6 Hz, 2H). 13C
NMR: 13.8, 21.4, 22.5, 25.7, 26.4, 27.4, 29.6, 31.5, 32,
33.8, 61.9, 64.7, 69.2, 70.8, 114.6, 125.65, 127.2, 128.5,
129.8, 131.1, 131.6, 160, 180.1. FABMS: 402 (M+).
4.13. (3Z,5E,7S)-3,5-Dodecadiene-1,7-diol 6a
Compound 4a (44 mg, 0.147 mmol) was dissolved in
methanol (2 mL) and the solution treated with KOH (1
g). The mixture was stirred for 12 h at rt and then
diluted with water and extracted several times with
EtOAc. Evaporation and purification by column chro-
matography gave the diol as a yellow oil (30 mg, 90%).
4.19. (4Z,7Z)-4,7-Tridecadienyl 4-hydroxybutanoate 1b
IR (TF): 3385, 1740 cm−1; H NMR: 0.9 (t, J=7 Hz,
1
1
IR (TF): 3400, 980 cm−1; H NMR: 0.85 (t, J=7 Hz,
3H), 1.3 (brs, 6H), 1.6–1.8 (m, 4H), 2.1 (m, 4H), 2.4 (t,
J=7 Hz, 2H), 2.78 (t, J=7 Hz, 2H), 3.65 (t, J=7 Hz,
2H), 4.09 (t, J=7 Hz, 2H), 5.2–5.45 (m, 4H). 13C NMR:
14, 21.1, 22.5, 25.6, 26.4, 27.1, 29.2, 31.4, 32, 33.8, 62.1,
64, 125, 127, 130.5, 131.2, 172. EIMS (m/z): 282 (M+).
3H), 1.26 (m, 6H), 1.44 (m, 2H), 2.2 (brs, 2H, -OH), 2.4
(q, J=7.0 Hz, 2H), 3.69 (t, J=7 Hz, 2H), 4.1 (q J=7
Hz, 1H), 5.46 (dd, J=7, 10.4 Hz, 1H), 5.73 (dd, J=7,
15 Hz, 1H), 6.15 (dd, J=10, 10.4 Hz, 1H), 6.52 (dd,
J=10, 15 Hz, 1H). 13C NMR: 13.9, 22.5, 26, 27, 29.5,
32, 62.5, 65, 124.3, 127.5, 128.6, 130.3. HRMS calcd for
C12H22O2 198.1619 obsd. (m/z) 198.1592 (M+).
4.20. (5Z,8Z)-5,8-Tetradecadienyl 3-hydroxypropanoate
1c
1
4.14. (3R,4E,6Z)-4,6-Dodecadiene-1,3-diol 7a
IR (TF): 3400, 1740 cm−1; H NMR: 0.9 (t, J=7 Hz,
3H), 1.2–1.8 (m, 10H), 2.0 (m, 4H), 2.5 (t, J=7 Hz, 2H),
2.75 (t, J=7 Hz, 2H), 3.8 (t, J=7 Hz, 2H), 4.15 (t, J=7
Hz, 2H), 5.2–5.5 (m, 4H). 13C NMR: 13.9, 21.2, 22.6,
25.5, 26, 27.1, 29.2, 31.4, 32, 34, 34, 62.2, 65.1, 125.1,
127, 131, 131.6, 173. EIMS (m/z): 282 (M+).
1
IR (TF): 3400, 1740 cm−1; H NMR: 0.85 (t, J=7 Hz,
3H), 1.25 (brs, 6H), 1.8 (m, 2H), 2.2 (q, J=7 Hz, 2H),
2.5 (brs, 2H, -OH), 3.8 (m, 2H), 4.4 (q, J=7 Hz, 1H),
5.48 (dd, J=7, 10.4 Hz, 1H), 5.65 (dd, J=7, 15 Hz, 1H),
5.98 (dd, J=10, 10.4 Hz, 1H), 6.52 (dd, J=10, 15 Hz,
1H).
4.21. 8-Hydroxy-(4Z,6E,8S)-4,6-tridecadienyl 4-hydroxy-
butanoate 4b
4.15. (4Z,7Z)-4,7-Tridecadiene-1-ol 10b
1
IR (TF): 3390, 1742 cm−1; H NMR: 0.9 (t, J=7 Hz,
1
IR (TF): 3400, 1090 cm−1; H NMR: 0.9 (t, J=7 Hz,
3H), 1.35 (brs, 6H), 1.6–1.8 (m, 4H), 2.35 (t, J=7 Hz,

J. S. Yada6 et al. / Tetrahedron: Asymmetry 12 (2001) 2129–2135
2135
2H), 2.4 (t, J=7 Hz, 2H), 3.65 (t, J=7 Hz, 2H), 4.1 (t,
References
J=7 Hz, 2H), 4.15 (q, J=7 Hz, 1H), 5.48 (dd, J=7,
10.4 Hz, 1H), 5.75 (dd, J=7, 15 Hz, 1H), 6.15 (dd,
J=10, 10.4 Hz, 1H), 6.54 (dd, J=10, 15 Hz, 1H). 13C
NMR: 14.1, 21, 22.6, 25.9, 27, 27.2, 29.1, 31.5, 33.8,
62.1, 64.8, 72, 124.1, 127.6, 129, 130.2, 173. EIMS
(m/z): 298 (M+), [h]D25=+6.2 (c=1.1, CHCl3).
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4.22. 9-Hydroxy-(5Z,7E,9S)-5,7-tetradecadienyl
3-hydroxypropanoate 4c
1
IR (TF): 3300, 1740 cm−1; H NMR: 0.9 (t, J=7 Hz,
3H), 1.2–1.8 (m, 12H), 2.2 (t, J=7 Hz, 2H), 2.5 (t, J=7
Hz, 2H), 3.8 (t, J=7 Hz, 2H), 4.15 (t, J=7 Hz, 2H),
4.18 (q, J=7 Hz, 1H), 5.48 (dd, J=7, 10.4 Hz, 1H),
5.75 (dd, J=7, 15 Hz, 1H), 6.15 (dd, J=10, 10.4 Hz,
1H), 6.55 (dd, J=10, 15 Hz, 1H). 13C NMR: 14, 21.2,
22.3, 26, 26.8, 27.1, 29, 31.1, 33.7, 62.5, 65, 71, 124.2,
127.8, 128.5, 131, 173. EIMS (m/z): 298 (M+), [h]D25=
+9.4 (c=1.0, CHCl3).
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4.23. (4Z,6E,8S)-4,6-Tridecadiene-1,8-diol 6b
1
IR (TF): 3400, 1000 cm−1; H NMR: 0.88 (t, J=7 Hz,
3H), 1.32–1.75 (m, 10H), 2.2 (q, J=7 Hz, 2H), 3.65 (t,
J=7 Hz, 2H). 4.15 (q, J=7 Hz, 1H), 5.50 (dd, J=7,
10.4 Hz, 1H), 5.73 (dd, J=7, 15 Hz, 1H), 6.18 (dd,
J=10, 10.4 Hz, 1H), 6.52 (dd, J=10, 15 Hz, 1H). 13C
NMR: 14, 22.6, 24, 26.3, 29, 31.2, 63, 65.6, 124.5, 128,
128.6, 130.6. HRMS calcd for C13H24O2 212.1777 obsd.
(m/z) 212.1762 (M+).
4.24. (5Z,7E,9S)-5,7-Tetradecadiene-1,9-diol 6c
1
IR (TF): 3395, 990 cm−1; H NMR: 0.89 (t, J=7 Hz,
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3H), 1.3–1.6 (m, 12H), 2.4 (q, J=7 Hz, 2H), 3.7 (t,
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Hz, 1H), 5.75 (dd, J=7, 15 Hz, 1H), 6.15 (dd, J=10,
10.4 Hz, 1H), 6.55 (dd, J=10, 15 Hz, 1H). 13C NMR:
13.9, 22.5, 24.3, 25, 26.2, 27.3, 29.2, 31, 62.3, 65.2,
124.3, 128.2, 128.5, 131. HRMS calcd for C14H26O2
226.1934 obsd. (m/z) 226.1927 (M+).
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