Synthesis and insecticidal activity of lactones derived from furan-2(5H)-one

Article abstract of DOI:10.5935/0103-5053.20150217

Ten 4,7-methanoisobenzofuran-1(3H)-ones were synthesized and their insecticidal activities against the insect pest Diaphania hyalinata were evaluated. The most active substances have been selected from the initial screening to find the dose to kill 50 (LD₅₀) and 90percent (LD₉₀) of the melonworm. Time-mortality curves of the three selected formulations at the LD₉₀ concentration have been made against D. hyalinata. From the time-mortality curves we found that the formulation (3aR,4R,5S,6S,7S,7aS)- and (3aS,4S,5R,6R,7R,7aR)-5,6-dichlorohexahydro-4,7-methanoisobenzofuran-1(3H)-one + (3aR,4R,5R,6R,7S,7aS)- and (3aS,4S,5S,6S,7R,7aR)-5,6-dichlorohexahydro-4,7-methanoisobenzofuran-1(3H)-one has killed 50percent of the melonworm after 2 h, presenting the best knockdown effect. Bioassays against Solenopsis saevissima and Tetragonisca angustula were carried out for the lactones selected in the initial screening against D. hyalinata. The formulation (3aS,4R,5S,6S,7S,7aR)- and (3aR,4S,5R,6R,7R,7aS)-5,6-dibromohexahydro-4,7-methanoisobenzofuran-1(3H)-one + (3aS,4R,5R,6R,7S,7aR)- and (3aR,4S,5S,6S,7R,7aS)-5,6-dibromohexahydro-4,7-methanoisobenzofuran-1(3H)-one has killed 31.25 and 68.30percent of the pest natural enemy and the pollinator bee, respectively. At the same concentration this formulation killed 90percent of D. hyalinata. The selectivity in favor of the non-target organisms has rendered this formulation a position as a promising agrochemical.

Full text of DOI:10.5935/0103-5053.20150217

http://dx.doi.org/10.5935/0103-5053.20150217
J. Braz. Chem. Soc., Vol. 26, No. 11, 2279-2289, 2015.
Printed in Brazil - ©2015 Sociedade Brasileira de Química
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Article
A
Synthesis and Insecticidal Activity of Lactones Derived from Furan-2(5H)-one
Milena G. Teixeira,a Elson S. Alvarenga,*,a Mirian F. Pimentelb and
Marcelo C. Picançob
aDepartamento de Quimica and bDepartamento de Entomologia,
Universidade Federal de Viçosa, 36570-900 Viçosa-MG, Brazil
Ten 4,7-methanoisobenzofuran-1(3H)-ones were synthesized and their insecticidal activities
against the insect pest Diaphania hyalinata were evaluated. The most active substances have
been selected from the initial screening to find the dose to kill 50 (LD50) and 90% (LD90) of the
melonworm. Time-mortality curves of the three selected formulations at the LD90 concentration
have been made against D. hyalinata. From the time-mortality curves we found that the
formulation (3aR,4R,5S,6S,7S,7aS)- and (3aS,4S,5R,6R,7R,7aR)-5,6-dichlorohexahydro-4,7-
methanoisobenzofuran-1(3H)-one + (3aR,4R,5R,6R,7S,7aS)- and (3aS,4S,5S,6S,7R,7aR)-5,6-
dichlorohexahydro-4,7-methanoisobenzofuran-1(3H)-one has killed 50% of the melonworm after
2 h, presenting the best knockdown effect. Bioassays against Solenopsis saevissima and Tetragonisca
angustula were carried out for the lactones selected in the initial screening against D. hyalinata.
The formulation (3aS,4R,5S,6S,7S,7aR)- and (3aR,4S,5R,6R,7R,7aS)-5,6-dibromohexahydro-
4,7-methanoisobenzofuran-1(3H)-one + (3aS,4R,5R,6R,7S,7aR)- and (3aR,4S,5S,6S,7R,7aS)-5,6-
dibromohexahydro-4,7-methanoisobenzofuran-1(3H)-one has killed 31.25 and 68.30% of the pest
natural enemy and the pollinator bee, respectively. At the same concentration this formulation
killed 90% of D. hyalinata. The selectivity in favor of the non-target organisms has rendered this
formulation a position as a promising agrochemical.
Keywords: Diaphania hyalinata, Tetragonisca angustula, Solenopsis saevissima, insecticide
Introduction
ecosystems. However, an alarming fact is that during the
last decades there were losses in the number of colonies in
locations around the world.7 Multiple environmental factors
can potentially contribute to these losses, including biotic
ones like pathogens, parasites, availability of resources
due to habitat fragmentation and loss; and abiotic ones
like climate change and pollutants.8-10 Although the causes
of the loss of pollinators are still under study, which
currently go through a screening process, extensive use of
chemical pesticides against insect pests for crop protection
may have contributed to the decline of pollinators.11 One
hypothesis for the disappearance of bees relates to the use of
neonicotinoid insecticides.12-17 Concern for bees is justified
by the fact that they play a vital role in agriculture.18 Nearly
35% of the world’s food crops depend on pollination mostly
by insects,19 especially bees and butterflies. The pollinator
insects can increase genetic biodiversity in the cultivated
species by the cross pollination leading to better fruits,
seeds and plants.20
The melonworm Diaphania hyalinata (Lepidoptera:
Crambidae) is an important pest of plants of the family
Curcubitaceae and occurs in theAmericas.1,2 The leaves are
first attacked by these insects, limiting plant photosynthesis,
therefore, growth and development. After this the stems
and shoots are damaged leading to the plant death. When
attacked by the caterpillar, fruits become unfit for human
consumption.3
Solenopsis saevissima, an ant widely distributed in South
America, is among the natural enemies of D. hyalinata.
Worker individuals of this species are aggressive and have
a painful sting.4 Predation by S. saevissima contributes
significantly to the biological control of agricultural crops.5
Species of the genus Solenopsis have been considered
important predators contributing to insect pest reduction
in the tropics and subtropics.6
Bees are another important non-target group of
organisms that contribute to the biodiversity of the
In recent years, several species are rapidly going into
extinction because the insecticides have been planned to
be very effective against pests in general, but with great
*e-mail: elson@ufv.br

2280
Synthesis and Insecticidal Activity of Lactones Derived from Furan-2(5H)-one
J. Braz. Chem. Soc.
potential to affect beneficial organisms.21 This concept
has fueled concerns about the ecological consequences of
biodiversity loss, leading to a flurry of studies examining
how changes in diversity affect the ecosystems, particularly
those that provide goods and services that humans depend
upon.22,23
Pest control should be based on equilibrium between the
use of chemical agents and survival of the natural enemies.24
Therefore, to avoid the possible interruption of the natural
control of pests it is important to evaluate the effects of
pesticides against the beneficial organisms.25
The idea that beneficial organisms and insecticides
were incompatible has been modified in recent years. The
outdated broad-spectrum insecticides have been superseded
by more modern agrochemicals like spinosads,26 and novel
pyrethroids that present selectivity in favor of the beneficial
organisms and a desirable ecotoxicology.27 The novel
pyrethroids have been evaluated for their toxicological
effect against target insects and adverse activity on natural
enemies.28,29
compounds display insecticidal,41,42 nematicidal,43 and
acaricidal44 activities. These plant-based agrochemicals
have presented themselves as an important tool to combat
the evolution of resistance to pesticides.45
Therefore, the aim of this work was to investigate the
toxicity of compounds of the class of phthalides against
D. hyalinata and non-target organisms: bee pollinator
(T. angustula) and the pest natural enemy (S. saevissima).
The insecticidal activities of these compounds have been
evaluated against the important pest of Curcubitaceae
from which three of them have shown stunning insecticidal
activities. Besides that we have been involved in functional
group modification and/or stereochemistry aiming for novel
compounds which could play an important role with respect
to the biological activity.
Results and Discussion
Synthesis
The conservation of beneficial organisms is an essential
component in integrated pest management.22,23,30 The most
frequent source of mortality for phytophagous arthropods
in ecosystems is attack by natural enemies.5
Cycloaddition reaction of furan-2(5H)-one 1 and
cyclopentadiene has been employed in first stage of our
work for the synthesis of 4,7-methanoisobenzofuran-
1(3H)-ones. The synthesis of the endo 2 and exo 3 adducts
using this methodology is represented in Scheme 1. This
reaction has been carried out with regio- and stereoselective
control but without asymmetric induction. Therefore, to
simplify we have opted to use only one of the enantiomers
to represent each adduct.
The most distinct signals of adduct 2 in the 13C nuclear
magnetic resonance (NMR) spectrum were the carbonyl
and the alkene carbons at d 178.0, 134.3, and 136.7,
respectively. The double bond is confirmed by the multiplet
at 6.25-6.33 ppm in the 1H NMR spectrum. These distinct
signals have also been observed in the 13C NMR of adduct 3
but at slightly different chemical shifts (d 177.6, 137.3, and
137.6). The olefinic protons H5 and H6 of 3 are displayed
as doublet of doublets at d 6.18 and 6.23, respectively.
Long distance detection experiments in the NMR (nuclear
Overhauser effect (NOE)) have enabled us to differentiate
these isomers. Irradiation of the signal of H8 of adduct 2
has resulted in 1.32 and 1.87% increase in the signals of
The synthesis of new lactones has been inspired by
small molecules from natural products that have been
the mainstay of research in organic chemistry since its
initial development.31 Within this context, phthalides or
isobenzofuranones, natural volatile compounds present
in apiaceous plants have attracted the attention of many
research groups.32 This class of compound has presented
a wide spectrum of activities such as vasodilatation,33
analgesic,34 anti-inflammatory,35,36 and protection effect
against neuronal impairment induced by deprival of oxygen
and glucose.37
Thefocusontheuseofnaturalproductsforagrochemicals
is widely used, because the resulting secondary metabolites
of plant interaction with the environment during its
evolution play an important function in plant protection
against herbivores.38-40
Phthalide derivatives possess potential as agrochemicals
because recent research indicates that a large number of
O
O
H
H
7
1
O
6
O
7a
3a
O
8
+
O
+
5
o
100 C
3
4
H
H
1
exo-3
2
endo-
:
1
3.2
Proportion
Scheme 1. Preparation of adducts 2 and 3 using Diels-Alder reaction.

Vol. 26, No. 11, 2015
Teixeira et al.
2281
H7a and H3a, respectively. However, irradiation of H8 of
adduct 3 has resulted in 0.85% NOE in the signal of H3’
which is close in space to H8.
correlation spectroscopy (COSY), and nuclear Overhauser
effect spectroscopy (NOESY).
In the next stage, the double bond of lactones 2 and
3 was oxidized with meta-chloroperbenzoic acid to give
only epoxides 4 (98% yield) and 5 (83% yield) with
stereospecific control (Scheme 2). In the 1H NMR of 4 and
5, H5 and H6 (d ca. 3.2 ppm) were chemically shielded
compared to H5 and H6 (d ca. 6 ppm) at the double bond
in compounds 2 and 3, which is a strong evidence of the
epoxide formation. The oxygen at the exo position in
epoxides 4 and 5 was established by NOE experiments.
Irradiation of the signal of H5/H6 of epoxide 4 has resulted
in 1.17% increase in the signal of H3’. However, irradiation
of the signal of H5 of epoxide 5 has led to 0.9% increase
of H3a. To reinforce the exo position of the oxygen in the
epoxide 5 we have irradiated the signal of H6 resulting in
2.03% increase of H7a.
Following the idea of preparing novel isobenzofuranone
derivatives we have also performed hydrogenation,
chlorination, and bromination reactions of these adducts
(Scheme 2). Bromination and chlorination of unsaturated
lactone 3 have proceeded stereoselectively to give only
the trans-1,2-dihalogenated products, as expected. The
structures of all compounds have been established by
using the spectrometric methods, including in these
methods the bidimensional NMR experiments such
as heteronuclear single quantum coherence (HSQC),
Insecticidal activity
In this study, we have assessed the efficacy of
compounds 1-7, 9a, and 9b against second-instar larvae of
D. hyalinata. Furthermore, the insecticidal activities of the
mixtures of 8a/8b (2:1 ratio), and 9a/9b (1:1 ratio) have
also been evaluated. Compounds 8a and 8b have not been
evaluated separately because only a small quantity of 8b
was isolated and purified (57.5 mg).
There was a significant difference in the mortality
of larvae of D. hyalinata as a function of the treatments
(analysis of variance (ANOVA), F14.75 = 48.24, p < 0.001).
The mortality of D. hyalinata as a result of the treatments
is presented in Figure 1 as a histogram with standard
deviation bars and assignment letters according to the Scott-
Knott grouping analysis test at p < 0.05. The compounds
1, 9a, 9a + 9b, 9b, and 8a + 8b have presented significant
insecticidal activities killing more than the negative control.
The lactones 9a + 9b, 9b, and 8a + 8b at the concentration
of 45.2 µmol g-1 insect were the most potent, killing 84.8,
91.3 and 96.3% of D. hyalinata, respectively. These
compounds were more active than the commercial piperine,
which has presented 64.5% mortality.46 Compounds 1 and
9a have shown moderate activities of 23.1 and 40.0%,
respectively.
O
O
O
O
H
H
H
H
(a)
(a)
O
O
O
O
O
O
H
H
H
H
3
4
(b)
5
2
(d)
O
(c)
(b)
H
Br
O
O
H
H
O
O
Cl
H
Br
O
O
H
9a
O
Cl
H
H
8a
+
7
O
H
H
6
Br
+
O
H
O
Cl
Br
O
H
9b
Cl
H
8b
Scheme 2. Reagents: (a) meta-chloroperbenzoic acid (MCPBA), CH2Cl2; (b) H2, Pd/C (10%), EtOH; (c) Cl2, CH2Cl2; (d) Br2, CH2Cl2.

2282
Synthesis and Insecticidal Activity of Lactones Derived from Furan-2(5H)-one
J. Braz. Chem. Soc.
separation of the chemicals 9a and 9b is not necessary
considering only the difference in insecticidal activity.
The most active lactones 8a + 8b, 9a + 9b, and 9b were
selected for a more thorough investigation after the general
insecticidal bioassay. This selection is in line with the
Brazilian Health SurveillanceAgency (ANVISA) for tests
of efficacy on pest control products, which recommends
that only values between 90 10% mortality should be
considered satisfactory.61
The dose-mortality results from the lactones application
in larvae of D. hyalinata have shown low chi-square test
(χ2) and high p values (< 2.39 and > 0.05, respectively)
indicating the suitability of the probit model for fitting the
dose-response curves. Curves with acceptance probability
of the null hypothesis by χ2 greater than 0.05 were accepted.
The doses to kill 50 (LD50) and 90% (LD90) of the insects
were obtained from these curves (Figure 2).
Figure 1. Mortality (mean standard error) of second-instar larvae of
D. hyalinata. Columns followed by the same letter are not significantly
different by the Scott-Knott grouping analysis test at p > 0.05.
Piperine was used as the positive control because the
insecticidal activity of this substance has been extensively
described in the literature.47-51 Furthermore, extracts of
black pepper Piper nigrum L. (Piperaceae), whose main
constituent is piperine are toxic to Lepidoptera insects,
including Ascia monuste orseis Latreile (Lepidoptera:
Pieridae)52 and Spodoptera frugiperda Smith (Lepidoptera:
Noctuidae).53
The isolated isomers 9a and 9b, in the concentration
of 45.2 µmol g-1 insect and a period of 48 h, have caused
40.0 and 91.3% mortalities of the insects, respectively.
According to Ahern and Whitney,54 many secondary
metabolite classes are responsible for the plant resistance
against herbivore. Therefore, totally different compounds
could play an important defense mechanism for the plants.
To the best of our knowledge we have not found many
references describing the ecological effects played by
modifying the stereochemistry of the active ingredients.55-59
In some cases, individual isomers have identical
bioactivities, but in other cases different isomers can
drastically differ in bioactivity.60
A significant difference in insecticide activity has been
observed for stereoisomers 9a and 9b. The isomer 9b is
the most active killing 91.3% of the insect pest. However
the formulation 9a + 9b at the ratio of 1:1 has killed 84.8%
of the pest, which does not differ significantly from that
obtained for the pure isomer 9a. Therefore, the laborious
Figure 2. Dose-mortality curves of the compounds 8a + 8b, 9a + 9b, and
9b against D. hyalinata caterpillars.
According to Table 1, the formulation 8a + 8b exhibited
the highest toxicity to D. hyalinata, with lethal doses of
20.58 (LD50) and 40.37 (LD90) µmol g-1 insect. However,
lactones 9a + 9b and 9b have shown similar insecticidal
activities.
The curves with the activity rates for the most promising
chemicals at the LD90 concentration against D. hyalinata are
presented in Figure 3. The survival analysis of D. hyalinata
exposed to lactones indicated significant differences among
2
treatments (log-rank test, χ = 190.28, df = 3, p < 0.001).
Table 1. Toxicity of the chemicals 8a + 8b, 9a + 9b and 9b against second instar D. hyalinata caterpillars 48 h after application
2
Lactones
8a + 8b
9a + 9b
9b
LD50a / (µmol g-1 insect)
20.58 (19.37-21.86)
21.49 (19.95-23.08)
21.40 (19.97-22.86)
LD90a / (µmol g-1 insect)
32.03 (29.57-35.39)
33.90 (31.10-37.67)
36.73 (33.55-41.21)
y
χ
df
4
p
–0.75 + 4.38x
–0.66 + 4.25x
0.24 + 3.58x
2.39
1.88
0.72
0.67
0.60
0.98
3
5
aThe numbers in parentheses are the confidence intervals. LD50 and LD90: lethal dose to kill 50 and 90% of the insects, respectively; y: curve equation;
2
χ : chi-square test; df: degree of freedom; p: probability.

Vol. 26, No. 11, 2015
Teixeira et al.
2283
The chemicals 8a + 8b, 9a + 9b and 9b have killed 50%
of the population of the pest in intervals of 2, 4 and 3 h,
respectively. The rate of activity of the insecticide is very
important to control the outbreak of pests in the agriculture.
The fast population growth of pests should be controlled
to avoid great damages to the crops. The formulation
8a + 8b has presented the best time mortality result, which
is adequate to control the outbreak of pests because it has
killed 50% of the population of D. hyalinata in the shortest
period (2 h).
9a + 9b (t10 = 4.59, p < 0.001) were selective in favor of
S. saevissima and T. angustula (more toxic to the pest than
to the natural enemy).
Besides the efficiency against insect pests, novel
agrochemical agents should preferably provide selectivity
to non-target species, especially predators and pollinators.
This is important because natural enemies can aid in the
control of the population of pests in agricultural crops.62
Moreover, the conservation and enhancement of natural
enemy activity in agroecosystems is one of the key elements
of sustainable agricultural production.63-67
However, the natural enemies alone are not able to
prevent any damage caused by the pest.24 Because of this,
finding a balance between the use of chemical insecticides
and the survival of natural enemies is a priority of integrated
pest management.
Furthermore, pollination is essential to ensure the
reproduction of most plants.68 Thus pollinator should
be retained, so that the biodiversity of the ecosystem is
maintained.69,70
The International Organization for Biological Control
(IOBC) uses a standardized classification for the impact of
pesticides on natural enemies for experiments in laboratory
which consists of four categories: harmless (< 30%
effect), slightly harmful (30-79% effect), moderately
harmful (80-99% effect), and harmful (> 99% effect).71
The formulation 9a + 9b has killed 31.25 and 68.30% of
S. saevissima and T. angustula, respectively. However,
the isolated compound 9b has killed 46.99 and 65.33% of
S. saevissima and T. angustula, respectively. The selectivity
of these lactones in favor of S. saevissima and T. angustula
surpasses the negative impact of the outdated broad-spectrum
insecticides which kill the insects without selectivity.
The selectivity provided by lactones (9a + 9b and 9b) to
S. saevissima and T. angustula suggests that the use of these
compounds to control D. hyalinata presents a low risk to
these non-target insects according to the IOBC classification.
To sum up we have found that a number of structural
features have influenced the insecticidal activity, particularly
the presence of halogens and the cis-relationship between
the lactone ring and the methylene bridge. The most active
compounds were the exo adducts and the ones containing
halogens which is in agreement with literature reports
where various compounds with biological activities such
as fungicide, herbicide, antibiotic, and insecticide have
been described.72-74
Figure 3. Survival curves of D. hyalinata subjected to the application of
the chemicals 8a + 8b, 9a + 9b, and 9b at the LD90 concentration.
The insecticides have been planned to be very effective
against pests in general, but with great potential to affect
beneficial organisms. Thus we have evaluated the toxicity of
eleven chemical formulations against the pest D. hyalinata,
and the three most promising formulations have been tested
against the beneficial bee pollinator and the natural enemy
fire ant.
The formulations 8a + 8b, 9a + 9b, and 9b have been
evaluated against the beneficial organisms S. saevissima
and T. angustula (Table 2). The results from this study
showed that lactones 9b (t10 = 6.35, p < 0.001) and
Table 2. Mortalities of the pest natural enemy S. saevissima and the
pollinator T. angustula 48 h after application of the formulations 8a + 8b,
9a + 9b, and 9b at 40.37, 43.03, and 48.74 mmol g-1 of insect, respectively
Average mortality standard error / %
Lactones
S. saevissima
91.37 4.24
31.25 12.81a
46.99 6.79a
T. angustula
82.61 5.74
68.30 7.03a
65.22 6.59a
8a + 8b
9a + 9b
9b
Conclusions
aSignificantly different from the mortality caused by these chemicals to
D. hyalinata by t-test at p < 0.05.
In summary, we have synthesized and determined the
insecticidal activity of ten lactones. The agrochemicals

2284
Synthesis and Insecticidal Activity of Lactones Derived from Furan-2(5H)-one
J. Braz. Chem. Soc.
9a +9b, 9b, and 8a +8b at the concentration of 45.2 µmol g-1
insect were the most active against D. hyalinata killing 84.8,
91.3 and 96.3% of the worms, respectively. The activities
presented by these lactones were better than the commercial
positive control. The lactones 9a + 9b and 9b were selective
in favor of non-target organisms. Therefore, these lactones
are promising as potential novel agrochemicals for the
integrated pest management.
Data for compound 2
White solid; TLC Rf = 0.44 (hexane:ethyl acetate 2:1
v/v); m.p. 119.8-120.6 °C; IR (film) νmax / cm-1 3065,
2097, 2873, 1765, 1481, 1381, 1345, 1185, 1001; 1H NMR
(300 MHz, CDCl3) d 1.46 (d, 1H, J 8.6 Hz, H8), 1.64 (d,
1H, J 8.6 Hz, H8’), 3.04-3.15 (m, 2H, H3a and H4), 3.25
(dd, 1H, J 9.2, 4.6 Hz, H7a), 3.30-3.36 (m, 1H, H7), 3.79
(dd, 1H, J 9.7, 3.1 Hz, H3’), 4.28 (dd, 1H, J 9.7, 8.3 Hz,
H3), 6.25-6.33 (m, 2H, H5 and H6); 13C NMR (75 MHz,
CDCl3) d 40.1 (C3a), 45.6 (C7), 46.0 (C4), 47.4 (C7a),
51.7 (C8), 70.2 (C3), 134.3 (C5), 136.7 (C6), 178.0 (C1);
MS m/z (%) 91 (7), 85 (11), 66 (100).
Experimental
General
Reagents and solvents were purified, when necessary,
according to the procedures described by Perrin and
Armarego.75 The reactions were followed by visualizing
the thin layer chromatography (TLC) plates coated with
silica-gel in an ultraviolet chamber at 254 nm.76 The
furan-2(5H)-one was obtained as previously described.77
The cyclopentadiene was obtained by distillation of
dicyclopentadiene commercially available (Sigma-
Aldrich) before being used in the Diels-Alder reaction.
The 1H, 13C NMR, COSY, HSQC, heteronuclear correlation
(HETCOR), heteronuclear multiple-bond correlation
(HMBC), NOE difference (NOEDIFF), and NOESY
spectra were recorded on aVarian Mercury 300 instrument
(300 MHz), using deuterated chloroform as solvent.
Infrared (IR) spectra were recorded on a Varian 660-IR,
equipped with GladiATR scanning from 4000 to 500 cm-1.
Mass spectra (MS) were recorded on a Shimadzu GCMS-
QP5050A instrument under electron impact (70 eV)
conditions. Melting points are uncorrected and were
obtained using MQAPF-301 melting point apparatus
(Microquímica). Column chromatography was performed
over silica gel (60-230 mesh).
Data for compound 3
Colorless oil; TLC Rf = 0.56 (hexane:ethyl acetate 2:1
v/v); IR (film) νmax / cm-1 3061, 2974, 2909, 1761, 1485,
1381, 1185, 1005; 1H NMR (300 MHz, CDCl3) d 1.47 (dt,
1H, J 9.8, 1.7 Hz, H8), 1.55 (dquint, 1H, J 9.8, 1.7 Hz,
H8’), 2.51-2.60 (m, 1H, H3a), 2.65 (dt, 1H, J 8.5, 1.7 Hz,
H7a), 2.90 (sl, 1H, H4), 3.27 (sl, 1H, H7), 3.96 (dd, 1H,
J 9.8, 3.5 Hz, H3’), 4.48 (dd, 1H, J 9.8, 8.7 Hz, H3), 6.18
(dd, 1H, J 5.7, 3.0 Hz, H5) 6.23 (dd, 1H, J 5.7, 3.0 Hz,
H6); 13C NMR (75 MHz, CDCl3) d 41.7 (C3a), 43.1 (C8),
46.2 (C7), 47.8 (C7a), 48.0 (C4), 71.7 (C3), 137.3 (C6),
137.6 (C5), 177.6 (C1); MS m/z (%) 150 ([M]+, 1), 91 (7),
85 (10), 66 (100).
(3aR,4R,5R,6S,7S,7aR)- and (3aS,4S,5S,6R,7R,7aS)-5,6-
epoxyhexahydro-4,7-methanoisobenzofuran-1(3H)-one (4)
meta-Chloroperbenzoic acid (MCPBA) (2.6366 g of
MCPBA 70%, 15.2 mmol) was added portionwise to a round-
bottom flask containing phthalide 2 (1.1386 g, 7.59 mmol)
and dichloromethane (100 mL). The reaction mixture was
stirred for 15 h and quenched with aqueous Na2SO3 solution
(20%, 100 mL). The mixture was extracted with diethyl
ether (3 × 100 mL), and the combined organic layers were
washed with aqueous NaHCO3 solution (10%, 100 mL)
and brine until neutral. The organic layer was dried with
anhydrous MgSO4 and filtered.After evaporation of solvent,
the product mixture was purified by column chromatography
with silica-gel using hexane:ethyl acetate 1:1 as eluent to
give 1.238 g of 4 (98% yield): white solid; TLC Rf = 0.36
(hexane:ethyl acetate 1:1 v/v); m.p. 120.9-121.7 °C; IR (film)
Synthesis of lactones
(3aR,4S,7R,7aS)- and (3aS,4R,7S,7aR)-3a,4,7,7a-
tetrahydro-4,7-methanoisobenzofuran-1(3H)-one (2)
and (3aS,4S,7R,7aR)- and (3aR,4R,7S,7aS)-3a,4,7,7a-
tetrahydro-4,7-methanoisobenzo-furan-1(3H)-one (3)
Furan-2(5H)-one 1 (1.0012 g, 11.9 mmol) and
cyclopentadiene (8.1 g, 0.12 mol) have been added to
a sealed tube. The resulting reaction mixture has been
magnetically stirred and heated at 100 °C for 72 h. The
excess of cyclopentadiene has been evaporated, and the
crude product was purified by column chromatography
(eluent: hexane:ethyl acetate 2:1 v/v) to give 1.139 g (66%
yield) of 2 and 0.363 g (18% yield) of 3.
ν
max / cm-1 2997, 2961, 1761, 1381, 1193, 1001, 849; 1H NMR
(300 MHz, CDCl3) d0.96 (d, 1H, J10.2 Hz, H8), 1.56 (d, 1H,
J 10.2 Hz, H8’), 2.70-2.74 (m, 1H, H4), 2.95-2.99 (m, 1H,
H7), 2.99-3.06 (m, 1H, H3a), 3.11 (dd, 1H, J 10.3, 5.0 Hz,
H7a), 3.32-3.36 (m, 2H, H5 and H6), 4.26-4.41 (m, 2H, H3
and H3’); 13C NMR (75 MHz, CDCl3) d 29.3 (C8), 39.2 (C7),
39.9 (C4), 41.3 (C3a), 45.5 (C7a), 47.3 (C5)*, 48.5 (C6)*,
67.6 (C3), 176.7 (C1); MS m/z (%) 138 (11), 110 (13), 109

Vol. 26, No. 11, 2015
Teixeira et al.
2285
(20), 92 (17), 91 (25), 82 (42), 81 (100), 80 (11), 79 (49), 78
(19), 77 (45), 70 (27), 69 (57), 66 (17), 65 (14), 55 (17), 54
(27), 53 (29), 51 (21), 41 (22), 40 (12). *These assignments
could be reversed.
1:1 v/v); m.p. 112.9-113.4 °C; IR (film) νmax / cm-1 2969,
1769, 1385, 1189, 1001, 853; 1H NMR (300 MHz, CDCl3)
d 1.15-1.38 (m, 4H, H5, H6, H8 and H8’), 1.45-1.67 (m, 2H,
H5’ and H6’), 2.24-2.28 (m, 1H, H4), 2.42 (tdd, 1H, J 8.7,
3.6, 1.3 Hz, H3a), 2.51 (dt, 1H, J 8.7, 1.3 Hz, H7a), 2.65-2.69
(1H, m, H7), 3.89 (dd, 1H, J 9.5, 3.8 Hz, H3’), 4.43 (dd,
1H, J 9.5, 9.0 Hz, H3); 13C NMR (75 MHz, CDCl3) d 27.1
(C5)*, 27.8 (C6)*, 33.4 (C8), 40.5 (C7), 41.7 (C3a), 42.6
(C4), 48.0 (C7a), 72.9 (C3), 179.6 (C1); MS m/z (%) 152
([M]+, 14), 93 (19), 91 (11), 85 (10), 81 (10), 80 (100), 79
(72), 78 (11), 77 (20), 68 (17), 67 (39), 66 (86), 64 (15), 54
(14), 53 (13), 51 (10), 41 (27), 40 (10). *These assignments
could be reversed.
(3aS,4R,5R,6S,7S,7aS)- and (3aR,4S,5S,6R,7R,7aR)-5,6-
epoxyhexahydro-4,7-methanoisobenzofuran-1(3H)-one (5)
Epoxide 5 was obtained (137.5 mg) in 83% yield
by reaction of lactone 3 (150 mg, 1.0 mmol) with 70%
MCPBA (346.8 mg, 2.0 mmol), by following the same
procedure described for lactone 4: white solid; TLC Rf =
0.30 (hexane:ethyl acetate 1:1 v/v); m.p. 112.9-113.4 °C;
IR (film) νmax / cm-1 2969, 1769, 1385, 1189, 1001, 853;
1H NMR (300 MHz, CDCl3) d 0.87 (dm, 1H, J 11.2 Hz,
H8), 1.42 (dquint, 1H, J 11.2, 1.9 Hz, H8’), 2.57 (sl, 1H,
H4), 2.61 (ddt, 1H, J 8.9, 3.6, 1.9 Hz, H3a), 2.65-2.70 (m,
1H, H7a), 3.94 (sl, 1H, H7), 3.16 (d, 1H, J 4.4 Hz, H5),
3.26 (d, 1H, J 4.4 Hz, H6) 3.97 (dd, 1H, J 9.7, 3.6 Hz,
H3’), 4.49 (dd, 1H, J 9.7, 8.9 Hz, H3); 13C NMR (75 MHz,
CDCl3) d 21.3 (C8), 39.1 (C3a), 41.6 (C7), 43.2 (C4), 45.6
(C7a), 50.5 (C5), 50.9 (C6), 71.4 (C3), 177.3 (C1); MS m/z
(%) 93 (21), 91 (27), 82 (92), 81 (100), 79 (34), 77 (43),
66 (12), 65 (11), 54 (22), 53 (21), 51 (18), 41 (15), 40 (9).
(3aR,4R,5S,6S,7S,7aS)- and (3aS,4S,5R,6R,7R,7aR)-
5,6-dichlorohexahydro-4,7-methanoisobenzofuran-
1(3H)-one (8a) and (3aR,4R,5R,6R,7S,7aS)- and
(3aS,4S,5S,6S,7R,7aR)-5,6-dichlorohexahydro-4,7-
methanoisobenzofuran-1(3H)-one (8b)
Chlorine was bubbled for 20 min into a solution
of lactone 3 (537.1 mg, 3.58 mmol) dissolved in
dichloromethane (20 mL). The chlorine was produced by
adding concentrated HCl to a two-neck round-bottom flask
containing MnO2 under vigorous stirring. The progress of
the reaction was checked by TLC, and the mixture was
stirred until all the substrate was fully consumed. The
solvent was evaporated and the pure product was obtained
after column chromatography packed with silica-gel using
hexane:diethyl ether 1:1 as eluent to give 0.2761 g (35%
yield) of 8a and 0.0575 g (7% yield) of 8b.
(3aR,4R,7S,7aS)- and (3aS,4S,7R,7aR)-hexahydro-4,7-
methanoisobenzofuran-1(3H)-one (6)
To a 25 mL round-bottom flask were added compound 2
(0.1024 g, 0.68 mmol), ethanol (10 mL) and Pd/C 10%
(7.3 mg). The resulting mixture was stirred under hydrogen
atmosphere for 1 h.After the solvent evaporation the residue
was purified by column chromatography with silica-gel using
hexane:ethyl acetate 1:1 as eluent to give 73.1 mg of 6 (70%
yield): white solid; TLC Rf = 0.63 (hexane:ethyl acetate 1:1
v/v); m.p. 64.8-65.6 °C; IR (film) νmax / cm-1 2961, 2881,
Data for compound 8a
Colorless oil; TLC Rf = 0.37 (hexane:ether 1:1 v/v);
IR (film) νmax / cm-1 2981, 2913, 1769, 1485, 1385, 1273,
1189, 1052, 1009, 945, 812, 776, 689, 641, 480; 1H NMR
(300 MHz, CDCl3) d 1.55-1.64 (m, 1H, H8), 2.00 (dquint,
1H, J 12.1, 1.6 Hz, H8’), 2.53-2.58 (m, 1H, H4), 2.70 (d,
1H, J 8.9 Hz, H7a), 2.86 (sl, 1H, H7), 3.20 (tdd, 1H, J 8.9,
3.7, 1.6 Hz, H3a), 3.74 (t, 1H, J 3.0 Hz, H6), 3.97 (dd, 1H,
J 9.7, 3.7 Hz, H3’), 4.24 (dd, 1H, J 4.0, 3.0 Hz, H5), 4.56
(t, 1H, J 9.7 Hz, H3); 13C NMR (75 MHz, CDCl3) d 30.4
(C8), 33.6 (C3a), 45.6 (C7a), 49.3 (C4), 49.6 (C7), 66.5
(C6), 67.4 (C5), 71.6 (C3), 177.3 (C1); MS m/z (%) 224
([M+4]+, 0.22), 222 ([M+2]+, 1.21), 220 ([M]+, 1.70), 141
(12), 105 (29), 91 (25), 80 (100), 79 (52), 78 (11), 77 (23),
66 (38), 65 (42), 63 (19), 53 (17), 52 (15), 51 (44), 50 (15),
49 (10), 41 (15), 40 (13).
1
1761, 1485, 1381, 1172, 1069, 997; H NMR (300 MHz,
CDCl3) d 1.50-1.63 (m, 6H, H5, H6, H8 and H8’), 2.34-2.40
(m, 1H, H4), 2.64-2.69 (m, 1H, H7), 2.80-3.92 (m, 1H, H3a),
2.98 (dd, 1H, J 11.2, 5.7 Hz, H7a) 4.21-4.34 (m, 2H, H3 and
H3’); 13C NMR (75 MHz, CDCl3) d 21.5 (C5)*, 25.4 (C6)*,
39.8 (C7), 40.3 (C4), 41.9 (C8/C3a), 46.7 (C7a), 68.4 (C3),
178.8 (C1); MS m/z (%) 93 (13), 91 (12), 85 (40), 80 (33),
79 (31), 77 (15), 68 (12), 67 (32), 66 (100), 65 (10), 53 (9),
41 (19). *These assignments could be reversed.
(3aS,4R,7S,7aR)- and (3aR,4S,7R,7aS)-hexahydro-4,7-
methanoisobenzofuran-1(3H)-one (7)
Product 7 was obtained (197.8 mg, 1.30 mmol) in 92%
yield by reaction of lactone 3 (0.2132 mg, 1.42 mmol)
and Pd/C 10% (14.2 mg) under hydrogen atmosphere, by
using the same procedure described for the synthesis of
lactone 6: white solid; TLC Rf = 0.30 (hexane:ethyl acetate
Data for compound 8b
Colorless oil; TLC Rf = 0.31 (hexane:ether 1:1 v/v);
IR (film) νmax / cm-1 2975, 2911, 1769, 1485, 1385, 1189,

2286
Synthesis and Insecticidal Activity of Lactones Derived from Furan-2(5H)-one
J. Braz. Chem. Soc.
1
1048, 1009, 821, 785, 653, 493; H NMR (300 MHz,
CDCl3) d 1.57-1.66 (m, 1H, H8), 2.03 (dquint, 1H, J 12.1,
1.7 Hz, H8’), 2.49 (sl, 1H, H4), 2.61 (tdd, 1H, J 9.0, 3.7,
1.7 Hz, H3a), 2.93-2.97 (m, 1H, H7), 3.27 (dt, 1H, J 9.0,
1.7 Hz, H7a), 3.70 (t, 1H, J 2.7 Hz, H5), 3.97 (dd, 1H,
J 9.9, 3.7 Hz, H3’), 4.30 (dd, 1H, J 4.1, 2.7 Hz, H6), 4.54
1.8 Hz, H8’), 2.55 (sl, 1H, H4), 2.63 (tdd, 1H, J 8.7, 3.6,
1.8 Hz, H3a), 2.92-2.96 (m, 1H, H7), 3.31 (ddd, 1H, J 8.7,
1.8, 1.8 Hz, H7a), 3.85 (t, 1H, J 2.9 Hz, H5), 3.97 (dd,
1H, J 10.0, 3.6 Hz, H3’), 4.47-4.56 (m, 2H, H3 and H6);
13C NMR (75 MHz, CDCl3) d 30.8 (C8), 40.0 (C3a), 43.0
(C7a), 48.3 (C7), 52.6 (C4), 55.9 (C5), 57.6 (C6), 71.6 (C3),
178.6 (C1); MS m/z (%) 231 (40), 229 (43), 106 (29), 105
(100), 91 (60), 80 (16), 79 (45), 78 (21), 77 (34), 66 (79),
65 (73), 63 (31), 55 (14), 53 (23), 52 (24), 51 (68), 50 (28),
41 (26), 40 (30).
13
(dd, 1H, J 9.9, 9.2 Hz, H3); C NMR (75 MHz, CDCl3)
d 30.5 (C8), 39.3 (C3a), 47.5 (C7a), 47.6 (C7), 52.1 (C4),
66.3 (C5), 67.2 (C6), 71.5 (C3), 178.6 (C1); MS m/z (%)
224 ([M+4]+, 0.31), 222 ([M+2]+, 1.88), 220 ([M]+, 2.87),
141 (13), 105 (50), 91 (40), 80 (100), 79 (72), 78 (16), 77
(31), 75 (12), 66 (66), 65 (65), 63 (31), 53 (25), 52 (22),
51 (74), 50 (27), 49 (20), 41 (23), 40 (24).
Biological assays
Biological assays were conducted with second-instar
larvae of D. hyalinata and adults of S. saevissima and
T. angustula. Larvae of D. hyalinata were obtained from
a laboratory rearing and adults of S. saevissima and
T. angustula were collected from nests located at the
University campus.
(3aS,4R,5S,6S,7S,7aR)- and (3aR,4S,5R,6R,7R,7aS)-
5,6-dibromohexahydro-4,7-methanoisobenzofuran-
1(3H)-one (9a) and (3aS,4R,5R,6R,7S,7aR)- and
(3aR,4S,5S,6S,7R,7aS)-5,6-dibromohexahydro-4,7-
methanoisobenzofuran-1(3H)-one (9b)
The phthalide 3 (420.3 mg, 2.80 mmol) was dissolved
in dichloromethane (20 mL) in a round-bottom flask
with a magnetic stirring bar. Then a solution of bromine
in dichloromethane was added dropwise using a Pasteur
pipette. The reaction mixture was stirred until the substrate
was fully consumed. When the reaction was completed
by TLC (1 h), the mixture was concentrated. The product
mixture was purified by column chromatography with
silica-gel using hexane:diethyl ether 1:1 to give 0.1952 g
(23% yield) of 9a and 0.1858 g (21% yield) of 9b.
Screening bioassay for D. hyalinata
The experimental design was completely randomized
with six replications. Each experimental unit consisted of a
glass petri dish (9.5 cm × 2.0 cm) containing ten insects. The
average weights of the insects were obtained by measuring,
on an analytical balance, the mass of ten insects and taking the
average. Bioassays were conducted by topical application.A
10 μL Hamilton micro syringe was employed to apply, on the
thoracic tergite of each individual insect, 0.5 μL of a solution
of the test compound, dissolved in acetone, corresponding to
a concentration of 45.2 μmol g-1 insect. In a negative control
experiment, carried out under the same conditions, 0.5 μL
of acetone was applied on each insect.After the application,
the insects were kept in individual petri dishes containing
discs of chayote leaf as food. The petri dishes were placed
in an incubator at 25 0.5 oC and 75 5% relative humidity
with a photoperiod of 12 h. The mortality counts were made
after 6, 12, 24 and 48 h of treatment. Mortalities included
dead individuals as well as those without movements. The
insecticidal activity of piperine (Sigma-Aldrich) was taken
as positive control.
Data for compound 9a
Colorless oil; TLC Rf = 0.31 (hexane:diethyl ether 1:1
v/v); IR (film) νmax / cm-1 2976, 2917, 1761, 1475, 1457,
1371, 1180, 1052, 996, 754, 677, 605, 543, 466; 1H NMR
(300 MHz, CDCl3) d 1.56-1.64 (m, 1H, H8), 2.08 (d, 1H,
J 12.1 Hz, H8’), 2.55 (d, 1H, J 3.7 Hz, H4), 2.72 (d, 1H,
J 8.9 Hz, H7a), 2.92 (s, 1H, H7), 3.22 (tdd, 1H, J 8.9, 3.7,
1.9 Hz, H3a), 3.88 (t, 1H, J 3.0 Hz, H6), 3.96 (dd, 1H,
J 9.9, 3.6 Hz, H3’), 4.43 (t, 1H, J 3.7 Hz, H5), 4.56 (t, 1H,
13
J 9.6 Hz, H3); C NMR (75 MHz, CDCl3) d 30.6 (C8),
35.7 (C3a), 46.2 (C7a), 50.0 (C4/C7), 56.0 (C6), 58.0 (C5),
71.7 (C3), 177.0 (C1); MS m/z (%) 231 (33), 229 (35), 106
(31), 105 (100), 92 (11), 91 (60), 80 (19), 79 (48), 78 (22),
77 (40), 66 (86), 65 (85), 63 (36), 53 (28), 52 (32), 51 (78),
50 (34), 41 (26), 40 (33).
Dose-mortality bioassays for D. hyalinata
The most active lactones against D. hyalinata were
subjected to experiments to obtain dose-mortality curves.
The experimental design was completely randomized with
six replications. Each experimental unit consisted of a glass
petri dish (9.5 cm × 2.0 cm) containing ten insects. Initially,
four doses of each compound were tested to identify the
range of concentrations that would provide mortalities
greater than zero and less than 100%. Once the ranges of
Data for compound 9b
Colorless oil; TLC Rf = 0.29 (hexane:diethyl ether 1:1
v/v); IR (film) νmax / cm-1 2977, 2911, 1761, 1478, 1380,
1
1180, 1053, 1009, 757, 613, 485; H NMR (300 MHz,
CDCl3) d 1.60-1.66 (m, 1H, H8), 2.13 (dquint, 1H, J 12.1,

Vol. 26, No. 11, 2015
Teixeira et al.
2287
concentrations were defined, other doses were tested for
each compound. The number of doses used to obtain the
dose-mortality curves varied from five to seven. Bioassays
were conducted by topical application using the same
procedure described above.
were compared with pest mortality by t-test for independent
samples (p < 0.05).
Supplementary Information
Supplementary data are available free of charge at
http://jbcs.sbq.org.br as PDF file.
Time-mortality bioassays for D. hyalinata
The most active lactones were subjected to experiments
to obtain curves of survival. The experimental design
was completely randomized with twelve replicates. Each
experimental unit consisted of ten insects kept on a glass
Petri dish (9 cm diameter × 2 cm height) covered with
organza. Bioassays were conducted by the same procedure
as that described above. The dose used was equivalent to
LD90 obtained for the three most active lactones. Insect
mortality was observed every 30 min during the initial 12 h
exposure followed by 5 h intervals of observation until the
death of approximately 90% of insects’ populations.
Acknowledgments
We are grateful to FAPEMIG, CNPq, and CAPES for
financial support.
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