Synthesis and Cytotoxic Activity of Friedelinyl Esters.

Commonly isolated from plants of Celastraceae family, pentacyclic triterpenoids have a broad spectrum of biological activities, such as antitumor, anti-inflammatory, antinociceptive properties, among others. Structural modifications in these triterpenoids can enhance their biological activity, as well as their selectivity, while improving their physicochemical and pharmacokinetic aspects. In this study, eight novel esters were synthesized: four derivatives of 3α-friedelinol (friedelan-3α-yl p-bromobenzoate (1a); friedelan-3α-yl naproxenate (1b); friedelan-3α-yl pent-4-ynoate (1c); friedelan-3α-yl undec-10-ynoate (1d)) and four derivatives of 3ß-friedelinol (friedelan-3ß-yl p-bromobenzoate (2a); friedelan-3ß-yl naproxenate (2b); friedelan-3ß-yl pent-4-ynoate (2c); friedelan-3ß-yl undec-10-ynoate (2d)). Overall, 3α-friedelinol showed greater reactivity when compared to the ß-epimer. The esters 1b-d and 2b-c were tested for antileukemic activity against THP-1 and K-562 cells but showed low cytotoxicity for both cell lines. The most active against THP-1 cells was friedelan-3ß-yl naproxenate (2b, IC50=266±6 µM), and the most active against K-562 cells was friedelan-3α-yl pent-4-ynoate (1c, IC50=267±5 µM).

Identification and Synthesis of a Macrolide as an Anti-aphrodisiac Pheromone from Males of Heliconius erato phyllis.

Investigation of the contents of the scent glands of the heliconiine butterfly Heliconius erato phyllis via gas chromatography/electroantennography revealed an unprecedented active compound. The males transfer this compound to females during mating. The structure of (2R,6E,10R)-2,6-dimethyl-6-undecen-10-olide, a derivative of geranylacetone, was proposed on the basis of infrared and mass spectrometry spectra and microderivatization and confirmed by racemic and stereoselective syntheses. Bioassays with the synthetic macrolide showed the repellency of this compound, termed phyllisolide, when applied to scent glands of females, identifying it as an anti-aphrodisiac pheromone.

The dear enemy effect drives conspecific aggressiveness in an Azteca-Cecropia system.

Territoriality is costly, and the accurate identification of intruders and the decision to perform aggressive responses are key behavioral traits in social animals. We studied aggression among individuals belonging to close and distant nests of the plant-ant Azteca muelleri, which lives in stems of the pioneer tree Cecropia glaziovii. More specifically, we aim to investigate if the DE (dear-enemy effect-less aggression towards neighbors than strangers) or NN (nasty-neighbor effect-less aggression to strangers than neighbors) effects or even none of them apply for this iconic Azteca-Cecropia system. We further checked if ant aggression towards conspecifics is related to cuticular hydrocarbon profiles (CHCs), which provide chemical cues for nestmate recognition. Therefore, we sampled 46 nests of A. muelleri in three Brazilian Atlantic forest fragments and performed behavioral trials within and between sites. Consistently with the DE effect, we found higher aggression levels in 'between sites' versus 'within sites' treatments as well as a positive effect of spatial distance on ant aggressiveness. We found no effect of the overall dissimilarities on CHC blend on ant aggressiveness, but of one CHC class, the methylated alkanes. Overall, we provide key insights on nest-mate recognition in obligatory ant-plant mutualisms.

Tergal Gland Secretion of the Rove Beetle Aleochara pseudochrysorrhoa (Staphylinidae: Aleocharinae): Chemical Composition and Biological Roles.

Aleochara pseudochrysorrhoa has a glandular complex known as the tergal gland. Generally, the tergal gland secretion (TGS) has been described to have defensive function, but some reports point to a possible secondary function of this complex. For example, the TGS of the related species A. curtula has been demonstrated to possess an important role in intraspecies communication. In this work, we describe the chemical composition of the TGS of A. pseudochrysorrhoa males and females. Eleven compounds were identified based on GC/MS and GC-FT-IR analyses, retention indexes and derivatization products. Furthermore, a brief study regarding the biological function of the TGS in mating behavior is provided, in which the stimulation of male grasping response reaction by female TGS proved to be dependent on concentration.

Chemodiversity of Essential Oils from Nine Species of Celastraceae.

Extracts and compounds obtained from several species of Celastraceae family are reported as potential sources of drugs due to their diverse pharmacological properties. Nevertheless, essential oil composition from these species is still little known. This work aimed at the analysis of essential oils obtained from different Brazilian Celastraceae species. A total of seventeen oils were obtained using hydrodistillation process and analyzed by gas chromatography/mass spectrometry (GC/MS). Principal component analysis (PCA) allowed the identification of a chemical composition pattern among the analyzed essential oils. Some compounds were more frequent among Celastraceae species, such as cis- and trans-linalool oxide (14/17 oil samples), nerylacetone (13/17), linalool (11/17), ß-ionone (10/17), α-ionone (9/17), nerolidol (10/17), decanal (10/17), and dodecanoic acid (10/17). These results contribute to the chemophenetics of Celastraceae species.

Aggregation Pheromone of the Bearded Weevil, Rhinostomus barbirostris (Coleoptera: Curculionidae): Identification, Synthesis, Absolute Configuration and Bioactivity.

The bearded weevil, Rhinostomus barbirostris (Coleoptera: Curculionidae: Dryophthorinae), attacks coconut trees, oil palms and other species of Arecaceae. Besides direct damage, R. barbirostris may be a vector of diseases in coconut and oil palms, such as stem bleeding (resinosis) and red ring disease. Currently, the only method to control this weevil is by visual observation of damage and removal of infected plants. Semiochemical-based trapping could improve the effectiveness of monitoring and management of R. barbirostris. In comparisons of volatiles released by R. barbirostris males and females by gas chromatography (GC) two male-specific compounds were observed. GC-mass spectrometry (MS) and GC-Fourier transform-infrared (FTIR) analyses of the natural compounds suggested these were diastereoisomers of 5-hydroxy-4-methylheptan-3-one, also known as sitophilure, a pheromone component of other dryophthorine species. Synthesis of the mixture of all four stereoisomers of sitophilure was performed in two steps, and the chemical structures were confirmed by comparing GC retention times and MS and FTIR spectra of natural and synthetic compounds. The absolute configurations of the two male-specific compounds were elucidated by enantioselective GC; the major component was the (4S,5R)-isomer, and the minor component (4S,5S)-sitophilure. In analyses by GC-electroantennography (EAG) the antennae of male and female R. barbirostris only responded to the (4S,5R)-isomer of the synthetic sitophilure. The stereoisomeric mixture of sitophilure was attractive to both sexes of R. barbirostris in laboratory experiments in the presence of sugar cane volatiles, and a similar result was obtained in a preliminary field trapping test.

Moth sex pheromone receptors and deceitful parapheromones.

The insect's olfactory system is so selective that male moths, for example, can discriminate female-produced sex pheromones from compounds with minimal structural modifications. Yet, there is an exception for this "lock-and-key" tight selectivity. Formate analogs can be used as replacement for less chemically stable, long-chain aldehyde pheromones, because male moths respond physiologically and behaviorally to these parapheromones. However, it remained hitherto unknown how formate analogs interact with aldehyde-sensitive odorant receptors (ORs). Neuronal responses to semiochemicals were investigated with single sensillum recordings. Odorant receptors (ORs) were cloned using degenerate primers, and tested with the Xenopus oocyte expression system. Quality, relative quantity, and purity of samples were evaluated by gas chromatography and gas chromatography-mass spectrometry. We identified olfactory receptor neurons (ORNs) housed in trichoid sensilla on the antennae of male navel orangeworm that responded equally to the main constituent of the sex pheromone, (11Z,13Z)-hexadecadienal (Z11Z13-16Ald), and its formate analog, (9Z,11Z)-tetradecen-1-yl formate (Z9Z11-14OFor). We cloned an odorant receptor co-receptor (Orco) and aldehyde-sensitive ORs from the navel orangeworm, one of which (AtraOR1) was expressed specifically in male antennae. AtraOR1•AtraOrco-expressing oocytes responded mainly to Z11Z13-16Ald, with moderate sensitivity to another component of the sex pheromone, (11Z,13Z)-hexadecadien-1-ol. Surprisingly, this receptor was more sensitive to the related formate than to the natural sex pheromone. A pheromone receptor from Heliothis virescens, HR13 ( = HvirOR13) showed a similar profile, with stronger responses elicited by a formate analog than to the natural sex pheromone, (11Z)-hexadecenal thus suggesting this might be a common feature of moth pheromone receptors.

Male-produced sex pheromone of the stink bug Edessa meditabunda.

Edessa meditabunda is a secondary pest within the piercing-sucking stink bug complex that attacks soybean crops in Brazil. The behavioral responses of males and females to aeration extracts from conspecifics suggested the presence of a male-produced sex pheromone. Gas chromatographic (GC) analysis of male and female aeration extracts revealed the presence of two male-specific compounds in a ratio of 92:8. Gas chromatographic -electroantennographic detection (GC-EAD) assays indicated that the major component is bioactive for females, supporting the behavioral data. Analysis of the mass and infrared spectra of the male-specific compounds suggested that they were both methyl-branched long-chain methyl esters. On the basis of the mass spectra of the respective hydrocarbons obtained by micro derivatizations, the structures of these methyl esters were proposed to be methyl 4,8,12-trimethylpentadecanoate (major) and methyl 4,8,12-trimethyltetradecanoate (minor). An 11 step synthetic route that was based on a sequence of Grignard reactions, starting from cyclopropyl methyl ketone, was developed to obtain synthetic standards with a 7.9 % overall yield for the major compound and a 9.9 % yield for the minor. The synthetic standards co-eluted with the natural pheromones on three different GC stationary phases. Y-tube olfactometer assays showed that the synthetic standards, including the major compound alone and a mixture of the major and minor compounds in the proportion found in natural extracts, were strongly attractive to females.

Male-produced sex pheromone of the cerambycid beetle Hedypathes betulinus: chemical identification and biological activity.

We identified, synthesized, determined the diel periodicity of release, and tested the bioactivity of components of the male-produced sex pheromone of Hedypathes betulinus (Coleoptera: Cerambycidae: Lamiinae). Gas chromatographic-mass spectrometric analysis of headspace volatiles from adult beetles showed three male-specific compounds, which were identified as (E)-6,10-dimethyl-5,9-undecadien-2-yl acetate (major component), (E)-6,10-dimethyl-5,9-undecadien-2-one (geranylacetone), and (E)-6,10-dimethyl-5,9-undecadien-2-ol. Release of these chemicals was dependent on time of the photoperiod and presence of the host plant. Pheromone release took place primarily during the photophase, with maximum release occurring between 4 and 6 hr after the onset of photophase. The amount of pheromone released by males was much greater when they were in the presence of their host plant than when they were not. In Y-tube olfactometer tests, a ternary mixture of the compounds was attractive to female beetles, although the individual compounds were not attractive by themselves. Addition of volatiles from the host plant greatly increased the attractiveness of the ternary pheromone mixture and of the major pheromone component alone.

Pheromone binding to general odorant-binding proteins from the navel orangeworm.

General odorant-binding proteins (GOBPs) of moths are postulated to be involved in the reception of semiochemicals other than sex pheromones, the so-called "general odorants." We have expressed two GOBPs, AtraGOBP1 and AtraGOBP2, which were previously isolated from the antennae of the navel orangeworm, Amyelois transitella. Surprisingly, these two proteins did not bind compounds that are known to attract adult moths, particularly females. The proper folding and functionality of the recombinant proteins was inferred from circular dichroism analysis and demonstration that both GOBPs bound nonanal in a pH-dependent manner. EAG experiments demonstrated that female attractants (1-phenylethanol, propionic acid phenyl ester, and isobutyric acid phenyl ester) are detected with high sensitivity by the antennae of day-0 to day-4 adult females, with response declining in older moths. The same age-dependence was shown for male antennae responding to constituents of the sex pheromone. Interestingly, AtraGOBP2 bound the major constituent of the sex pheromone, Z11Z13-16Ald, with affinity comparable to that shown by a pheromone-binding protein, AtraPBP1. The related alcohol bound to AtraPBP1 with higher affinity than to AtraGOBP2. AtraGOBP1 bound both ligands with low but nearly the same affinity.

Sex pheromone of Lonomia obliqua: daily rhythm of production, identification, and synthesis.

The sex pheromone of Lonomia obliqua Walker (Lepidoptera: Saturniidae) was studied in the laboratory. All female calling occurred during the scotophase. Most females (70.6%) called first within 24 hr of eclosion. Calling varied with age of female, with older (5- to 6-day-old) females calling earlier in the scotophase and for longer durations than younger (0- to 1-day-old) females. The sex pheromone gland of 1- to 3-day-old virgin females was extracted during the calling peak. A Y-olfactometer bioassay showed significant attraction of males to a filter paper containing the female gland extract. Gas chromatographic-electroantennogram detection (GC-EAD) analysis of the extract indicated the presence of at least two possible pheromone components. Gas chromatographic-mass spectrometric analysis of the major GC-EAD-active peak indicated a hexadecenyl acetate; chemical derivatization indicated delta11 unsaturation. Synthetic samples of (E)- and (Z)-11-hexadecenyl acetate were obtained by coupling 10-bromo-1-decanol and 1-hexyne, utilizing lithium chemistry. The comparison of the retention time of dimethyl disulfide derivatives of the natural compound, to those of synthetic chemicals, confirmed the natural compound as (E)-11-hexadecenyl acetate. The minor component was identified as the related alcohol, (E)-11-hexadecenol. The ratio of the two components in female extract was 100:35. Preliminary tests of males in a Y-olfactometer showed that their response to a mixture of the two compounds was not significantly different from that to gland extract.