Hydroxy-methoxybenzoic methyl esters: Synthesis and antifeedant activity on the pine weevil, Hylobius abietis

Article abstract of DOI:10.1515/znb-2004-0714

The pine weevil Hylobius abietis (L.) (Coleoptera: Curculionidae) feeds on the bark of coniferous seedlings and is the economically most important forestry restocking pest in large parts of Europe. Substances with an antifeedant effect may offer an environmentally friendly alternative to insecticides for the protection of planted seedlings. Bioassays were performed on commercial and synthetic methyl hydroxy-methoxybenzoates in order to determine their possible antifeedant activity. Two methyl hydroxy-methoxybenzoates were synthesized by esterification and mono-O-methylation of two dihydroxybenzoic acids. A regioselective protection-deprotection strategy was used in the synthetic pathway of the other non-commercial esters, esterification and selective pivaloylation of the less-hindered hydroxyl group of other commercial dihydroxybenzoic acids gave diester intermediates, which then were O-methylated before methanolysis of the pivaloyl group which yielded the desired non-commercial methyl hydroxy-methoxybenzoates. The five synthesized methyl hydroxy-methoxybenzoic esters were complemented with commercial samples of the five other isomers of methyl hydroxy-methoxybenzoate and spectrometric data were collected for the complete set of isomers. All ten isomers were tested for their antifeedant effect on the pine weevil. The effect varied considerably among the hydroxy-methoxybenzoic esters. Methyl 2-hydroxy-3-methoxybenzoate showed the highest effect and may thus be a candidate for practical use in pine weevil pest management.

Full text of DOI:10.1515/znb-2004-0714

Hydroxy-Methoxybenzoic Methyl Esters: Synthesis and Antifeedant
Activity on the Pine Weevil, Hylobius abietis
Sacha Legranda, Go¨ran Nordlanderb, Henrik Nordenhemb, Anna-Karin Borg-Karlsonc,
and C. Rikard Uneliusa
a Department of Chemistry and Biomedical Sciences, University of Kalmar,
SE-391 82 Kalmar, Sweden
b Department of Entomology, Swedish University of Agricultural Sciences,
P.O. Box 7044, SE-750 07 Uppsala, Sweden
c Department of Chemistry, Organic Chemistry, Royal Institute of Technology,
SE-100 44 Stockholm, Sweden
Reprint requests to Associate Prof. C. Rikard Unelius. Fax: +46 480 44 62 62.
E-mail: rikard.unelius@hik.se
Z. Naturforsch. 59b, 829 – 835 (2004); received December 15, 2003
The pine weevil Hylobius abietis (L.) (Coleoptera: Curculionidae) feeds on the bark of coniferous
seedlings and is the economically most important forestry restocking pest in large parts of Europe.
Substances with an antifeedant effect may offer an environmentally friendly alternative to insecti-
cides for the protection of planted seedlings. Bioassays were performed on commercial and synthetic
methyl hydroxy-methoxybenzoates in order to determine their possible antifeedant activity.
Two methyl hydroxy-methoxybenzoates were synthesized by esterification and mono-O-methyl-
ation of two dihydroxybenzoic acids. A regioselective protection-deprotection strategy was used in
the synthetic pathway of the other non-commercial esters, esterification and selective pivaloylation of
the less-hindered hydroxyl group of other commercial dihydroxybenzoic acids gave diester interme-
diates, which then were O-methylated before methanolysis of the pivaloyl group which yielded the
desired non-commercial methyl hydroxy-methoxybenzoates.
The five synthesized methyl hydroxy-methoxybenzoic esters were complemented with commercial
samples of the five other isomers of methyl hydroxy-methoxybenzoate and spectrometric data were
collected for the complete set of isomers. All ten isomers were tested for their antifeedant effect on
the pine weevil. The effect varied considerably among the hydroxy-methoxybenzoic esters. Methyl
2-hydroxy-3-methoxybenzoate showed the highest effect and may thus be a candidate for practical
use in pine weevil pest management.
Key words: Methyl Hydroxy-methoxybenzoates, Antifeedant Activity, Hylobius abietis
Introduction
pounds related to benzoic acid. In this study, we
investigated the potential of hydroxy-methoxy acid
methyl esters as antifeedants useful for the protec-
tion of planted seedlings against pine weevil damage.
There are 10 possible isomers of methyl hydroxy–
methoxybenzoate (Scheme 1). The esters 1 – 5 had to
be synthesized while the esters 6 – 10 were commer-
cially available.
The esters 1, 2, 3 and 4 are intermediates in the
total synthesis of compounds with important biolog-
ical effects and their synthesis have been reported
previously [4]. The synthesis of methyl 5-hydroxy-2-
methoxybenzoate (5) was published in 1983 by Har-
wood [5].
Adult pine weevils, Hylobius abietis (L.), frequently
kill planted conifer seedlings by their feeding on the
stem bark. Unprotected seedlings commonly suffer
over 80% mortality in regions with managed conif-
erous forests [1]. To protect the seedlings it is com-
mon practice in many European countries to routinely
treat transplants with an insecticide. Because of envi-
ronmental hazards and health risks for forest workers
the usage of insecticides is seriously questioned today.
Possibly, antifeedant substances applied to transplants
could offer an alternative to insecticides [2].
Recently we have shown that various benzoate
derivatives have strong antifeedant effect on the pine
The methyl benzoic esters 1 and 2 were synthe-
weevil [3]. This encouraged further studies of com- sized based on the method described by Chakraborty
c
0932–0776 / 04 / 0700–0829 $ 06.00 ꢀ 2004 Verlag der Zeitschrift fu¨r Naturforschung, Tu¨bingen · http://znaturforsch.com
Brought to you by | Purdue University Libraries
Authenticated
Download Date | 10/19/15 8:48 AM

830
S. Legrand et al. · Hydroxy-Methoxybenzoic Methyl Esters
Scheme 1. All isomers of methyl hydroxy-methoxybenzoate (1 – 10).
et al. [4b]. The corresponding acids of 1 and 2 were
esterified and mono-O-methylated (Scheme 2). The
regioselective protection presented by Dornhagen and
Scharf in their synthesis of the dichloroisoeverninic
acid [6] was used as a basis for our synthesis of the
methyl benzoates 3, 4 and 5 (Scheme 3). Our synthesis
started by esterification of the benzoic acids 15a, 15b
and 15c. Acylation of the synthesized esters 16a, 16b
and 16c occurred only at the less-hindered hydroxyl
group (OH group meta or para to the ester group). O-
Scheme 2. Reaction conditions: (i) DCC, ◦MeOH, DMAP,
methylation of the ortho-OH group, followed by de-
protection of the diesters gave the desired methyl ben-
zoic esters 3, 4 and 5.
CH2Cl2, RT; (ii) MeI, K2CO3, DMF, 35 C; (iii) MeOH,
H2SO4, reflux; (iv) MeI, MeOH, K2CO3, RT.
were more effective in this case since the carboxylic
acid was converted to a compound with a better
leaving group. It was noted that the treatment of the
meta disubstituted benzoic acid 13 with an excess of
MeOH and a catalytic amount of H2SO4 afforded the
ester 14 in good yield, due to the absence of resonance
effects with the COOH group. The products 12 and
14 were then mono-O-methylated by use of methyl
iodide in the presence of a weak base.
Results and Discussion
Methyl 2-hydroxy-6-methoxybenzoate (1) and
methyl 3-hydroxy-5-methoxybenzoate (2) were syn-
thesized from the commercially available benzoic
acids 11 and 13 (Scheme 2). It was found that the
esterification of the carboxylic acid 11 with MeOH
and H2SO4 as reactants gave the ester 12 in very
low yield. The low reactivity of the COOH group
A regioselective protection [6] was the key step
in 11 is presumably due to the resonance effect of in the syntheses of the methyl benzoates 3, 4 and 5
two hydroxyl groups ortho to COOH. The yield of (Scheme 3). After esterification of the commercially
this esterification reaction was improved when the available benzoic acids 15a, 15b or 15c, it was found
compound 11 was treated with dicyclocarbodiimide that the esters 16a, 16b or 16c when reacted with
(DCC) and dimethylaminopyridine (DMAP) in a trimethylacetyl chloride, selectively yielded the inter-
MeOH/CH2Cl2 mixture. The esterification conditions mediates 17a, 17b or 17c.
Brought to you by | Purdue University Libraries
Authenticated
Download Date | 10/19/15 8:48 AM

S. Legrand et al. · Hydroxy-Methoxybenzoic Methyl Esters
831
Table 1. Effect of the ten ben-
zoates on bark feeding by the
pine weevil Hylobius abietis,
as measured by the antifeedant
index (0 is no activity, 100 is
complete feeding deterrence).
No
Compound
Index
(6 h)
94
35
77
69
4
100
69
100
93
Level of
significance
***
Index
(24 h)
54
26
35
Level of
significance
1
2
3
4
5
6
7
8
9
Methyl 2-hydroxy-6-methoxybenzoate
Methyl 3-hydroxy-5-methoxybenzoate
Methyl 3-hydroxy-2-methoxybenzoate
Methyl 4-hydroxy-2-methoxybenzoate
Methyl 5-hydroxy-2-methoxybenzoate
Methyl 2-hydroxy-4-methoxybenzoate
Methyl 3-hydroxy-4-methoxybenzoate
Methyl 2-hydroxy-3-methoxybenzoate
Methyl 2-hydroxy-5-methoxybenzoate
Methyl 4-hydroxy-3-methoxybenzoate
***
***
***
ns
***
***
***
ns
***
***
***
***
4
−3
ns
∗ = p < 0.05,
∗∗ = p < 0.01,
52
***
***
***
***
*
∗∗∗ = p < 0.001 (Fisher exact test
of a 2×2 table).
32
85
56
22
10
56
***
Scheme 4. Structure – activity relationships (decreasing ac-
tivity from left to right).
of methanol can be explained by a rearrangement be-
tween the methylcarboxylate moiety and a hydroxyl
hydrogen in ortho-position [7].
Bioassays were performed with all esters in order
to determine their possible antifeedant effect against
the pine weevil. Eight of the ten compounds showed
antifeedant activity after 24 h exposure to pine weevils
in the bioassay (Table 1). Only compounds 4 and 5 did
not inhibit feeding over the 24 h period, although 4
showed activity after 6 h. The most potent antifeedant
among these compounds was 8. It was closely followed
in activity by compounds 9, 1, and 6, and thereafter 3
and 7. Compound 2 and, particularly, 10 had only a
weak effect.
Apparently, isomers with a hydroxy group in the
ortho position have a stronger antifeedant effect
(Scheme 4). The most potent compound (8) gave a
somewhat higher index value after 24 h than shown by
the strongest antifeedant compound (ethyl cinnamate)
recently isolated from bark of Pinus contorta [2].
Scheme 3. Reaction conditions: (i) MeOH, H2SO4, reflux;
◦
(ii) trimethylacetyl chloride, pyridine, CH2Cl2, −10 C to
RT; (iii) MeI, K2CO3, DMF, 35 ◦C; (iv) MeOH, K2CO3, RT.
Due to the steric hindrance between the bulky pro-
t
tecting group, Bu, and the ester moiety, acylation
was predominant at hydroxyl groups meta and para
to the carbomethoxy group and not with the hydroxyl
group ortho. O–methylation of the hydroxyl group or-
tho to the carbomethoxy group gave the compounds
18a, 18b or 18c. Then, the hydroxyl groups meta or
para to the carbomethoxy group were deprotected us-
ing MeOH/K2CO3, yielding the desired methyl ben-
zoates 3, 4 or 5.
In conclusion, the synthesis of all non-commercial
methyl hydroxy-methoxybenzoate was presented.
Starting from the benzoic esters 11 and 13, the methyl
esters 1 and 2 were synthesized in two steps. A regio-
selective protection was the critical step in the synthe-
ses of the other methyl benzoates 3, 4 and 5.
Conclusion
Starting from commercially available hydroxy-
methoxybenzoic acids, all non-commercial methyl
hydroxy-methoxybenzoates were synthesized. Among
the methyl hydroxy-methoxybenzoic esters tested in
The spectroscopic data of the commercially avail- the bioassay, methyl 2-hydroxy-3-methoxybenzoate
able methyl benzoates 6, 7, 8, 9 and 10 were also had the strongest antifeedant effect on adult pine wee-
recorded. Interestingly, we noted that the mass spec- vils. A comparison with previously discovered an-
tra of all 2-hydroxy-isomers have a strong m/z 150 i.e. tifeedants indicates that methyl 2-hydroxy-3-methoxy-
loss of methanol (32), while all other isomers have benzoate has potential for use in practical protection
a strong 151 fragment. The mechanism for the loss of conifer transplants. Further synthesis and bioassays
Brought to you by | Purdue University Libraries
Authenticated
Download Date | 10/19/15 8:48 AM

832
S. Legrand et al. · Hydroxy-Methoxybenzoic Methyl Esters
are needed to predict the optimal structure for maxi- cyclohexane–ethyl acetate (EtOAc) (2:3) and cyclohexane–
EtOAc (4:1) as eluents. Compound 1 was isolated as a white
mal antifeedant activity. More comparisons of similar
compounds are also needed before structure – activity
patterns can be properly discussed.
◦
solid (55 mg, 20%). M.p. 50 C. – 1H NMR (250 MHz,
DMSO–d6): δ = 3.71 (s, 3 H, OMe), 3.73 (s, 3 H, COOMe),
6.47 – 6.51 (d, 2 H, 2×Har), 7.14 – 7.21 (t, 1 H, Har), 9.98
1
(bs, 1 H, OH). – 13C{ H} NMR (62.9 MHz, DMSO–d6):
Experimental Section
δ = 51.64 (COOMe), 55.54 (OMe) 101.84, 106.59, 108.38,
130.87, 155.32, 157.03, 166.52 (all Car and C=O). – MS:
m/z(%) = 182 (38) [M+], 150 (100), 136 (5.7), 122 (31),
107 (55), 93 (2.9), 79 (5.3), 65 (5.3), 51 (4.3), 39 (4.3).
Methyl 3,5-dihydroxybenzoate (14): 3,5-Dihydroxybenz-
oic acid (13) (1.00 g, 6.49 mmol) was dissolved in MeOH
(40 ml) and some drops of H2SO4 were slowly added to
the reaction mixture, which was stirred at reflux temperature.
The reaction was monitored by TLC. When the reaction was
finished, the solvent was evaporated and the crude product
was dissolved in EtOAc and washed twice with brine. The
organic layer was dried over MgSO4 and the solvent was
evaporated to give 14 as a white powder (871 mg, 80%). M.p.
Synthesis: General synthetic methodology
Melting points were determined on a Bu¨chi 510 in-
strument and were not corrected. Preparative chromatogra-
phy [8] and flash chromatography were done on silica gel
(Merck 60). NMR spectra were recorded on spectrometers
Bruker AC 250 (250 MHz for 1H and 63 MHz for 13C) and
Bruker AMX 500 (500 MHz for 1H and 125 MHz for 13C).
CDCl3 and DMSO-d6 were used as solvents and the signals
of the solvents served as internal standards. Chemical shifts
were expressed in ppm, followed by multiplicity (s, singlet;
t, triplet; d, doublet; m, multiplet; b, broad) and number of
protons. Mass spectra of positive ions obtained by electron
impact (EI, 70 eV) were measured on Hewlett-Packard or
Varian Saturn ws GC-MS instruments.
1
165 – 168 ◦C. – H NMR (250 MHz, DMSO–d6): δ = 3.78
(s, 3 H, COOMe), 6.43 (m, 1 H, Har), 6.80 (m, 2 H, 2×Har),
1
9.64 (s, 2 H, OH). – 13C{ H} NMR (62.9 MHz, DMSO–
Dimethylformamide (DMF) was distilled under N2 before
use. Pyridine and CH2Cl2 were dried over 4 A molecular
sieves. The starting materials employed were purchased from
commercial suppliers and were used without further purifica-
tion.
˚
d6): δ = 51.85 (COOMe), 106.95 (2×Car), 107.04, 131.16
(all Car), 158.41 (2×Car), 166.12 (C=O). – MS: m/z = 168
[M+], 137 (100%), 109, 95, 81, 69, 53, 44.
Methyl 3-hydroxy-5-methoxybenzoate (2): Methyl 3,5-di-
hydroxybenzoate (14) (300 mg, 1.78 mmol) was dissolved
in MeOH, K2CO3 (296 mg, 2.14 mmol) was added and the
reaction mixture was stirred for a couple of minutes. MeI
(0.11 ml, 1.78 mmol) was then added and the mixture was
stirred at room temperature overnight. Silica gel was then
added and the solvent was evaporated. After drying, the im-
pregnated silica gel was put on top of a chromatography col-
umn and subjected to medium pressure liquid chromatog-
raphy (MPLC, cyclohexane:EtOAc 70:30) ◦to give 2 as a
white powder (70 mg, 21%). M.p. 82 – 84 C. – 1H NMR
(250 MHz, DMSO–d6): δ = 3.75 (s, 3 H, COOMe), 3.82 (s,
3 H, OMe), 6.58 (s, 1 H, Har), 6.91 (s, 1 H, Har), 6.97 (s,
Methyl 2,6-dihydroxybenzoate (12): 2,6-Dihydroxybenz-
oic acid (11) (1.00 g, 6.49 mmol) was dissolved in MeOH
(10 ml) and CH2Cl2 (65 ml) was added to the reac-
tion mixture. DCC (1,3-dicyclohexylcarbodiimide) (1.49 g,
7.14 mmol) and DMAP (4-dimethylaminopyridine) (0.158 g,
1.30 mmol) were added and the reaction mixture was stirred
at room temperature (RT) for 72 h. The white precipitate
was then removed by filtration and the solvents were evap-
orated. The crude product was purified by flash chromatog-
raphy on silica gel, using cyclohexane–EtOAc (3:2) as elu-
ent, to give 12 (277 mg, 25%) as a white solid. M.p. 58 −
60 ◦C. – 1H NMR (250 MHz, DMSO–d6): δ = 3.78 (s,
3 H, COOMe), 6.32 − 6.35 (d, 2 H, 2×Har) 7.05 − 7.12 (t,
1
1 H, Har), 9.87 (s, 1 H, OH). – 13C{ H} NMR (62.9 MHz,
1
1 H, Har), 9.94 (bs, 2 H, OH). – 13C{ H} NMR (62.9 MHz,
DMSO–d6): δ = 52.04 (COOMe), 55.15 (OMe), 104.99,
105.99, 108.54, 131.35, 158.53, 160.35, 165.95 (all Car and
C=O). – MS: m/z(%) = 182 (93) [M+], 167 (1), 151 (100),
136 (2.9), 123 (34), 108 (22), 93 (8.6), 79 (3.3), 69 (16),
63 (4.8), 51 (4.8), 44 (9), 39 (3.8).
DMSO–d6): δ = 51.76 (COOMe), 106.60 (2×Car), 106.88,
132.27, 157.22, 157.24, 168.17 (all Car and C=O). – MS:
m/z = 168 [M+], 153, 136 (100%), 108, 96, 80, 69, 63, 52,
44, 39.
Methyl 2-hydroxy-6-methoxybenzoate (1): Methyl 2,6-di-
Methyl 2,3-dihydroxybenzoate (16a): Prepared by the pro-
hydroxybenzoate (12) (260 mg, 1.55 mmol) was dissolved cedure used for compound 14 but with 2,3-dihydroxybenzoic
in DMF (2 ml) and K2CO3 (256 mg, 1.86 mmol) was added acid (15a) (1.50 g, 9.8 mmol) as starting material. 16a was
in 2 portions, followed by MeI (0.12 ml, 1.94 mmol). The isolated as a slightly brown solid (1.27 g, 77%). M.p. 68 –
resulting suspension was vigorously stirred at 35 ◦C for 71 ◦C. – 1H NMR (250 MHz, DMSO–d6): δ = 3.88 (s, 3 H,
3 h. The reaction mixture was then cooled to room tem- COOMe), 6.75 (m, 1 H, Har), 7.01 (m, 1 H, Har), 7.22 (m,
1
perature, the solid was removed by filtration and the sol- 1 H, Har), 9.44 (s, 1 H, OH), 10.41 (s, 1 H, OH). – 13C{ H}
vent was evaporated to give a brown oil. The crude oil NMR (62.9 MHz, DMSO–d6): δ = 52.30 (COOMe), 112.96,
was purified by two flash chromatography procedures using 118.81, 119.42, 120.58, 145.97, 149.26, 169.71 (all Car and
Brought to you by | Purdue University Libraries
Authenticated
Download Date | 10/19/15 8:48 AM

S. Legrand et al. · Hydroxy-Methoxybenzoic Methyl Esters
833
C=O). MS: m/z = 168 [M+], 153, 136 (100%), 119, 108, 38.52 (COOMe), 52.32 (Cq), 110.32, 110.83, 113.20, 131.19,
91, 80, 63, 52, 44, 39.
155.91, 160.93, 168.36, 175.57 (all Car and 2×C=O). – MS:
m/z = 252 [M+], 221, 168, 136 (100%), 108, 95, 85, 69,
57, 41.
Methyl 2,4-dihydroxybenzoate (16b): Prepared by the
same procedure as compound 14 but with 2,4-dihydroxy-
benzoic acid 15b (5.00 g, 32.44 mmol) as starting mate-
Methyl 2-hydroxy-5-pivaloyloxybenzoate (17c): Prepared
rial. The crude product was purified by flash chromatogra- by the procedure used for compound 17a but with the methyl
phy on silica gel using cyclohexane–EtOAc (80:20) as elu- benzoate 16c (650 mg, 3.87 mmol) as starting material. Com-
ent. A white solid 16b (1.43 g, 26%) was obtained. M.p. pound 17c was isolated as a white solid (98 mg, 10%). –
1
115 – 118 ◦C. – H NMR (250 MHz, DMSO–d6): δ = 3.83 1H NMR (250 MHz, DMSO–d6): δ = 1.28 (s, 9 H, 3×Me),
(s, 3 H, COOMe), 6.29 – 6.30 (d, 1 H, Har), 6.34 – 6.38 (dd, 3.88 (s, 3 H, COOMe), 6.99 – 7.03 (d, 1 H, Har), 7.25 – 7.29
1 H, Har), 7.61 – 7.65 (d, 1 H, Har), 10.46 (s, 1 H, OH), (m, 1 H, Har), 7.29 – 7.43 (m, 1 H, Har), 10.50 (bs, 1 H,
1
10.71 (s, 1 H, OH). – 13C{ H} NMR (62.9 MHz, DMSO– OH). – MS: m/z = 252 [M+], 221, 205, 193, 177, 168,
d6): δ = 51.90 (COOMe), 102.36, 103.86, 108.24, 131.50, 136 (100%), 108, 85, 77, 69, 57, 50, 41.
162.58, 164.11, 169.46 (all Car and C=O). – MS: m/z = 168
Methyl 2-methoxy-3-pivaloyloxybenzoate (18a): The di-
ester 17a (400 mg, 1.59 mmol) was dissolved in dry DMF
[M+], 136 (100%), 125, 108, 95, 80, 69, 63, 53, 44, 39.
Methyl 2,5-dihydroxybenzoate (16c): Prepared by the pro- (2 ml) and K2CO3 (242 mg, 1.90 mmol) was added in 2 por-
cedure used for compound 14 but with 2,5-dihydroxybenzoic tions, followed by MeI (0.128 ml, 2.06 mmol). The resulting
acid 15c (3.00 g, 19.4 mmol) as starting material. 16c suspension was stirred vigorously at 35 ◦C for 90 min. The
was isolated as a white solid (0.78 g, 24%). M.p. 73 – reaction mixture was then cooled to RT. The solid was re-
76 ◦C. – 1H NMR (250 MHz, DMSO–d6): δ = 3.87 (s, moved by filtration and the solvent was evaporated to give an
3 H, COOMe), 6.76 – 6.83 (m, 1 H, Har), 6.93 – 6.99 (m, oil. The solid was dissolved in water (10 ml) and added to the
1 H, Har), 7.14 (m, 1 H, Har), 9.18 (s (apparent d), 1 H, oil. The water phase was extracted with Et2O (3×10 ml). The
1
OH), 9.85 (bs, 1 H, OH). – 13C{ H} NMR (62.9 MHz, combined organic layers were washed with water and brine.
DMSO–d6): δ = 52.32 (COOMe), 114.01, 117.63, 123.74, The organic layer was then dried over MgSO4 and the sol-
149.47, 153.02, 153.97, 171.61 (all Car and C=O). – MS: vent was evaporated to give 18a as a pale yellow oil (288 mg,
1
m/z = 168 [M+], 136, 108, 80, 69, 53, 44.
70%). – H NMR (250 MHz, DMSO–d6): δ = 1.31 − 1.33
(s (app. d), 9 H, 3×CH3), 3.72 (s, 3 H, OMe), 3.85 (s, 3 H,
COOMe), 7.21 – 7.39 (m, 2 H, 2×Har),7.60 – 7.63 (m, 1 H,
Methyl 2-hydroxy-3-pivaloyloxybenzoate (17a): Methyl
benzoate 16a (800 mg, 4.76 mmol) was dissolved in CH2Cl2
(8.4 ml) under inert atmosphere and pyridine (2.6 ml) was
added to the reaction mixture. The reaction mixture was then
cooled to −10 ◦C and a solution of pivaloyl chloride (642 mg,
5.30 mmol) in CH2Cl2 (0.7 ml) was added drop wise to the
reaction mixture, which was allowed to reach RT. After stir-
ring for 48 h, the solvent was evaporated and the crude crys-
1
Har). – 13C{ H} NMR (62.9 MHz, DMSO–d6): δ = 26.62
(4×Me), 38.37 (COOMe), 52.19 (Cq), 61.95 (OMe), 124.02,
125.83, 127.35, 127.95, 144.54, 151.01, 165.21, 175.70 (all
Car and 2×C=O). – MS: m/z = 266 [M+], 235, 219, 182,
164, 150, 136, 121, 107, 93, 85, 77, 65, 57 (100%), 41.
Methyl 2-methoxy-4-pivaloyloxybenzoate (18b): Produc-
tals were purified by two consecutive flash chromatography ed by the procedure employed for compound 18a but with the
treatments using cyclohexane-EtOAc (7:3) and cyclohexane- diester 17b (200 mg, 0.793 mmol) as starting material. Com-
Et2O (6:1) as eluents. This procedure yielded 17a as a pound 18b was isolated as a colourless oil (160 mg, 76%). –
◦
white solid (803 mg, 67%). M.p. 64 – 67 C. – 1H NMR 1H NMR (250 MHz, DMSO–d6): δ = 1.30 (s, 9 H, 3×Me),
(250 MHz, DMSO–d6): δ = 1.31 (s, 9 H, 3×Me), 3.77 – 3.91 3.78 (s, 3 H, COOMe), 3.82 (s, 3 H, OMe), 6.74 – 6.78 (m,
(s (app. d), 3 H, COOMe), 6.97 – 7.71 (m, 3 H, 3×Har), 10.52 1 H, Har), 6.91 – 6.92 (m, 1 H, Har), 7.70 – 7.73 (m, 1 H,
1
1
(bs, 1 H, OH). – 13C{ H} NMR (62.9 MHz, DMSO–d6): Har). – 13C{ H} NMR (62.9 MHz, DMSO–d6): δ = 26.56
δ = 26.74 (4×Me), 38.38 (COOMe), 52.61 (Cq), 118.83, (4×Me), 38.52 (COOMe), 51.72 (Cq), 56.06 (OMe), 106.45,
120.49, 125.86, 126.97, 128.62, 139.06, 152.10, 168.96 (all 113.35, 117.16, 131.70, 154.65, 159.35, 165.31, 175.71 (all
Car and 2×C=O), 168.96 (C=O). – MS: m/z = 252 [M+], Car and 2×C=O). – MS: m/z = 266 [M+], 235, 223, 182,
168, 136 (100%), 107, 85, 69, 57, 41.
165, 151, 136, 122, 107, 93, 85, 77, 65, 57 (100%), 41.
Methyl 2-hydroxy-4-pivaloyloxybenzoate (17b): Prepared
Methyl 2-methoxy-5-pivaloyloxybenzoate (18c): Synthe-
by the procedure used for compound 17a but with the methyl sized by the procedure used for compound 18a but with the
benzoate 15b (500 mg, 2.97 mmol) as starting material. diester 17c (100 mg, 0.39 mmol) as starting material. Com-
Compound 17b was isolated as a white solid (245 mg, pound 18c was isolated as a colourless oil (74 mg, 70%). –
33%). M.p. 71 – 73 ◦C. – 1H NMR (250 MHz, DMSO– 1H NMR (250 MHz, DMSO–d6): δ = 1.28 (s, 9 H, 3×Me),
d6): δ = 1.30 (s, 9 H, 3×Me), 3.89 (s, 3 H, COOMe), 3.78 (s, 3 H, COOMe), 3.82 (s, 3 H, OMe), 7.16 – 7.36 (m,
1
6.69 – 6.77 (t, 2 H, 2×Har), 7.80 – 7.85 (d, 1 H, Har). – 3 H, 3×Har). – 13C{ H} NMR (62.9 MHz, DMSO–d6): δ =
1
13C{ H} NMR (62.9 MHz, DMSO–d6): δ = 26.50 (4×Me), 26.65 (4×Me), 38.52 (COOMe), 51.97 (Cq), 56.14 (OMe),
Brought to you by | Purdue University Libraries
Authenticated
Download Date | 10/19/15 8:48 AM

834
S. Legrand et al. · Hydroxy-Methoxybenzoic Methyl Esters
113.42, 120.28, 123.45, 126.43, 143.12, 155.66, 165.17, 2 H, 2×Har), 7.71 – 7.73 (d, 1 H, Har), 10.78 (bs, 1 H,
1
176.45 (all Car and 2×C=O). – MS: m/z = 266 [M+], 235, OH). – 13C{ H} NMR (125.76 MHz, DMSO–d6): δ = 53.06
182 (100%), 167, 149, 135, 121, 107, 93, 85, 77, 65, 57, 41.
Methyl 3-hydroxy-2-methoxybenzoate (3): The diester 18a
(98 mg, 0.37 mmol) was dissolved in MeOH (3.7 ml) and
K2CO3 (0.108 g, 0.78 mmol) was added to the reaction
mixture that was stirred at room temperature for 3 h. Then
the liquid was decanted from the solid residue and the sol-
vent was evaporated to give crude white crystals. The pre-
vious solid residue was dissolved in water (3 ml) and HCl
(37%) was added until pH=2. Then the aqueous solution
was added to the crude white crystals and the mixture was
extracted with Et2O (3×5 ml). The combined organic lay-
ers were washed (water, brine) and dried (MgSO4). The sol-
vent was evaporated to give 3 as a colourless oil (34 mg,
50%). – 1H NMR (250 MHz, DMSO–d6): δ = 3.74 (s, 3 H,
COOMe), 3.80 (s, 3 H, OMe), 7.03 (m, 3 H, 3×Har), 9.64
(COOMe), 56.49 (OMe), 101.84, 106.15, 108.32, 132.21,
163.49, 166.11, 170.25 (all Car and C=O). – MS: m/z(%) =
182 (40) [M+], 168 (2), 150 (100), 139 (3.5), 122 (57),
107 (28), 95 (10), 79 (18), 63 (5), 51 (7.5).
Methyl 3-hydroxy-4-methoxybenzoate (7): M.p. 64 –
◦
67 C. – 1H NMR (500.14 MHz, DMSO–d6): δ = 3.79 (s,
3 H, COOMe), 3.84 (s, 3 H, OMe), 7.01 – 7.04 (d, 1 H, Har),
7.37 – 7.40 (d, 1 H, Har), 7.43 – 7.47 (dd, 1 H, Har), 9.48
1
(bs, 1 H, OH). – 13C{ H} NMR (125.76 MHz, DMSO–
d6): δ = 52.62 (COOMe), 56.47 (OMe), 112.28, 116.56,
122.35, 122.74, 147.13, 152.78, 166.92 (all Car and C=O). –
MS: m/z(%) = 182 (54) [M+], 167 (5), 151 (100), 139 (4),
123 (13), 108 (7.5), 95 (2), 79 (6), 65 (6), 51 (7), 39 (2.5).
Methyl 2-hydroxy-3-methoxybenzoate (8): M.p. 61.5 –
◦
62.5 C. – 1H NMR (500.14 MHz, DMSO–d6): δ = 3.81
1
(bs, 1 H, OH). – 13C{ H} NMR (62.9 MHz, DMSO–d6):
(s, 3 H, COOMe), 3.90 (s, 3 H, OMe), 6.88 – 6.90 (t, 1 H,
Har), 7.22 – 7.24 (d, 1 H, Har), 7.35 – 7.36 (d, 1 H, Har), 10.50
δ = 51.82 (COOMe), 60.51 (OMe), 119.87, 119.97, 123.79,
125.80, 146.62, 150.83, 166.20 (all Car and C=O). – MS:
m/z(%) = 182 (81) [M+], 164 (24), 151 (80), 136 (43),
121 (100), 107 (62), 93 (12), 79 (12), 65 (16), 59 (2.4),
51 (14), 45 (3.8), 39 (6.7).
1
(bs, 1 H, OH). – 13C{ H} NMR (125.76 MHz, DMSO–
d6): δ = 53.40 (COOMe), 56.78 (OMe), 113.89, 117.89,
119.69, 121.63, 149.13, 151.37, 170.50 (all Car and C=O). –
MS: m/z(%) = 182 (58) [M+], 167 (2), 150 (65), 136 (7),
122 (100), 107 (28), 92 (18), 79 (13), 65 (9), 53 (11), 39 (5).
Methyl 4-hydroxy-2-methoxybenzoate (4): Prepared by
the procedure used for compound 3 with the diester 18b
(160 mg, 0.60 mmol) as starting material. Compound 4
was isolated as a white solid (51 mg, 46%). M.p. 130 –
135 ◦C. – 1H NMR (250 MHz, DMSO–d6): δ = 3.70
(s, 3 H, COOMe), 3.75 (s, 3 H, OMe), 6.38 – 6.46 (m,
2 H, 2×Har), 7.58 – 7.62 (m, 1 H, Har), 10.36 (bs, 1 H,
Methyl 2-hydroxy-5-methoxybenzoate (9): B.p. 235 –
240 ◦C. – 1H NMR (500.14 MHz, DMSO–d6): δ = 3.72 (s,
3 H, COOMe), 3.89 (s, 3 H, OMe), 6.91 – 6.93 (dd, 1 H, Har),
7.13 – 7.16 (dd, 1 H, Har), 7.21 – 7.22 (d, 1 H, Har), 10.09
1
(bs, 1 H, OH). – 13C{ H} NMR (125.76 MHz, DMSO–d6):
δ = 53.30 (COOMe), 56.41 (OMe), 112.97, 113.43, 119.36,
124.26, 152.56, 155.23, 169.90 (all Car and C=O). – MS:
m/z(%) = 182 (43) [M+], 167 (2.5), 150 (100), 135 (15),
122 (20), 107 (30), 93 (7.5), 79 (27), 65 (5), 51 (10), 39 (2.5).
1
OH). – 13C{ H} NMR (62.9 MHz, DMSO–d6): δ = 51.10
(COOMe), 55.38 (OMe), 99.45, 107.10, 109.85, 133.14,
160.84, 162.62, 165.31 (all Car and C=O). – MS: m/z(%) =
182 (32) [M+], 151 (100), 136 (5.2), 121 (12), 108 (12),
93 (5.7), 65 (5.7), 53 (5.3), 44 (0.4), 39 (5.7).
Methyl 4-hydroxy-3-methoxybenzoate (10): M.p. 69 –
◦
70 C. – 1H NMR (500.14 MHz, DMSO–d6): δ = 3.80 (s,
Methyl 5-hydroxy-2-methoxybenzoate (5): Prepared by
the procedure used for compound 3 but with the diester 18c
(288 mg, 1.08 mmol) as starting material. Purification of the
crude product by flash chromatography on silica gel using
cyclohexane–EtOAc (3:1) as eluent yielded 5 as a slightly
yellow oil (93 mg, 50%). – 1H NMR (250 MHz, CDCl3):
δ = 3.83 (s (app. d), 3 H, COOMe), 3.88 (s (app. d), 3 H,
OMe), 6.83 – 6.98 (m, 2 H, 2×Har), 7.24 – 7.34 (m, 1 H,
3 H, COOMe), 3.82 (s, 3 H, OMe), 6.86 – 6.88 (d, 1 H, Har),
7.44 – 7.45 (d, 1 H, Har), 7.46 – 7.48 (dd, 1 H, Har), 9.96
1
(bs, 1 H, OH). – 13C{ H} NMR (125.76 MHz, DMSO–
d6): δ = 52.58 (COOMe), 56.47 (OMe), 113.34, 116.06,
121.31, 124.28, 148.22, 152.38, 166.93 (all Car and C=O). –
MS: m/z(%) = 182 (55) [M+], 167 (5), 151 (100), 140 (5),
124 (11), 108 (6), 93 (2), 79 (5), 65 (5), 51 (6), 39 (2.5).
1
Har). – 13C{ H} NMR (62.9 MHz, CDCl3): δ = 52.25
Bioassay
(COOMe), 56.63 (OMe), 113.89, 118.12, 120.71, 122.48,
149.22, 153.35, 166.86 (all Car and C=O). – MS: m/z(%) =
182 (75) [M+], 167 (6.7), 151 (100), 136 (17), 121 (15),
108 (21), 93 (20), 80 (9), 65 (18), 52 (15), 44 (5.7).
Characterization of commercially available isomers, all
purchased from Aldrich.
The various esters were tested for antifeedant effect on
the pine weevil Hylobius abietis (L.) (Coleoptera, Cur-
culionidae). For each test, 40 pine weevils (20 females + 20
males) were used. They were placed in separate Petri dishes
provided with a pine twig prepared with delimited treatment
Methyl 2-hydroxy-4-methoxybenzoate (6): M.p. 50 – and control areas. These pine twigs were enveloped in
53 ◦C. – 1H NMR (500.14 MHz, DMSO–d6): δ = 3.81 aluminium foil and two holes with a diameter of 5 mm and
(s, 3 H, COOMe), 3.87 (s, 3 H, OMe), 6.52 – 6.54 (m, separated by 25 mm were punched in the foil with metal
Brought to you by | Purdue University Libraries
Authenticated
Download Date | 10/19/15 8:48 AM

S. Legrand et al. · Hydroxy-Methoxybenzoic Methyl Esters
835
rings. After removal of the aluminium foil inside the rings, T is the number of treated surfaces with feeding marks. It was
one of the two surfaces exposed was treated with 100 µl tested if there was a statistic significant difference between
of a 50 mM methanol solution of the compound that was treatment and control with a Fisher exact test of a 2×2 table.
tested, and the other surface was treated with the same
amount of methanol alone (control). The following day, after
the solvent had evaporated, the metal rings were removed
and the test started. After 6 and 24 hours it was recorded
whether the pine weevil had started to feed on the treated and
untreated surfaces. The antifeedant effect was expressed by
means of the following index: (C-T)×100/(C+T), wherein
C is the number of control surfaces with feeding marks and
Acknowledgements
This work has been supported financially by The Univer-
sity of Kalmar, by Robigus AB and by the Swedish Hylobius
Research Program. Help from Professors Roland Isaksson
and Ian Nicholls (both at the University of Kalmar) in the
form of discussions is gratefully acknowledged.
¨
M. T. Guidotti, R. A. Halpin, D. W. Hobbs, C. F. Hom-
nick, M. S. Kuo, E. V. Lis, D. J. Mathre, S. R. Michel-
son, J. M. Pawluczyk, D. J. Pettibone, D. R. Reiss,
S. Vickers, P. D. Williams, C. J. Woyden, J. Med.
Chem. 41, 2146 (1998).
[1] G. Orlander, U. Nilsson, Scand. J. For. Res. 14, 341
(1999).
[2] K. Bratt, K. Sunnerheim, H. Nordenhem, G. Nordlan-
der, B. La˚ngstro¨m, J. Chem. Ecol. 27, 2253 (2001).
[3] G. Nordlander, H. Nordenhem, A.-K. Borg-Karlson,
R. Unelius, Swedish and PCT Patent Application WO
0056152 A1, 2000.
[5] L. M. Harwood, J. Chem. Soc., Chem. Commun. 9, 530
(1983).
[6] J. Dornhagen, H.-D. Scharf, Tetrahedron 1, 173 (1985).
[7] F. W. McLafferty, Interpretation of Mass Spectra, 3rd
Ed. University Science Books, Mills Valley California
(1980).
[8] P. Baeckstro¨m, K. Stridh, L. Li, T. Norin, Acta Chem.
Scand. B41, 442 (1987).
[4] a) For 1 see: S. E. Maier, S. Ku¨hnert, Org. Lett. 4,
643 (2002); b) for 2 see: T. K. Chakrabotry, G. Venkat
Reddy, J. Org. Chem. 57, 5462 (1992); c) for 3 see:
R. S. Coleman, E. B. Grant, J. Am. Chem. Soc. 117,
10889 (1995). I. Churcher, D. Hallet, P. Magnus,
Tetrahedron 55, 1597 (1999); d) for 4 see: M. I. Bell,
J. M. Erb, R. M. Freidinger, S. N. Gallicchio, J. P. Guare,
Brought to you by | Purdue University Libraries
Authenticated
Download Date | 10/19/15 8:48 AM