Dillapiole as Antileishmanial Agent: Discovery, Cytotoxic Activity and Preliminary SAR Studies of Dillapiole Analogues

In this paper, the isolation of dillapiole (1) from Piper aduncum was reported as well as the semi-synthesis of two phenylpropanoid derivatives [di-hydrodillapiole (2), isodillapiole (3)], via reduction and isomerization reactions. Also, the compounds' molecular properties (structural, electronic, hydrophobic, and steric) were calculated and investigated to establish some preliminary structureactivity relationships (SAR). Compounds were evaluated for in vitro antileishmanial activity and cytotoxic effects on fibroblast cells. Compound 1 presented inhibitory activity against Leishmania amazonensis (IC50?=?69.3 mu M) and Leishmania brasiliensis (IC50?=?59.4 mu M) and induced cytotoxic effects on fibroblast cells mainly in high concentrations. Compounds 2 (IC50?=?99.9 mu M for L. amazonensis and IC50?=?90.5 mu M for L. braziliensis) and 3 (IC50?=?122.9 mu M for L. amazonensis and IC50?=?109.8 mu M for L. brasiliensis) were less active than dillapiole (1). Regarding the molecular properties, the conformational arrangement of the side chain, electronic features, and the hydrophilic/hydrophobic balance seem to be relevant for explaining the antileishmanial activity of dillapiole and its analogues.

Larvicidal activity of Piper tuberculatum on Spodoptera frugiperda (Lepidoptera: Noctuidae) under laboratory conditions

The larvicidal activity of the neotropical "matico" Piper tuberculatum was evaluated. The secondary compounds were extracted of leaves, stems and mature spikes with fruits and seeds from wild plants and in vitro plants of Piper tuberculatum. The acute toxicities to the fall armyworm, Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae), of extracts of spikes with fruits and seeds and in vitro plants of P tuberculatum were evaluated by means of contact bioassays. Only CH2Cl2:MeOH (2:1) and EtOH extracts of mature spikes and CH2C12:MeOH (2:1) extract from in vitro plants showed significant levels of larval mortality. The CH2Cl2:MeOH (2:1) and EtOH extracts of mature spikes caused 90% mortality when doses of 0.1850 mg/mu L were applied to the S. frugiperda in 24 and 48 h of exposure, respectively. The CH2Cl2:MeOH (2:1) extract from in vitro plants caused 95% mortality when doses of 0.1850 mg/mu L were too applied in 48 h of exposure. The mature spikes test best results were: LD50 0.001 mg/mu L with EtOH and 0.007 mg/mu L with CH2Cl2:MeOH (2:1) and LD90 0.027 mg/mu L with EtOH and 0.103 mg/mu L with CH2Cl2:MeOH (2:1); and, in the case of in vitro plants, only CH2Cl2:MeOH (2:1) extract was: LD50 0.003 mg/mu L and LD90 0.060 mg/mu L. The potential value of extracts derived from P. tuberculatum as efficient insecticides against S.frugiperda is discussed.

Non-substituted B-ring flavonoids and an indole alkaloid from Piper aleyreanum (Piperaceae)

Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)

Biotransformation of 4-nerolidylcatechol by Heraclides brasiliensis (Lepidoptera: Papilionidae) reduces the toxicity of Piper umbellata (Piperaceae)

Heraclides brasiliensis (Lepidoptera: Papilionidae) larvae feed preferably on Piperaceae, foraging successfully on leaf tissues even though species of this contain high levels of secondary metabolites such as amides and lignans, associated with diverse biological activities including insecticidal properties. Studies examining the metabolism of chemical constituents in Piperaceae by insects are rare. In this study, we characterized the metabolites of 4-nerolidylcatechol (4-NC), the major constituent of Piper umbellata (Piperaceae), and E-2,3-dihydro-3-(3,4-dihydroxyphenyl)farnesoic acid, compounds from fecal material of H. brasiliensis larvae fed a diet containing only P. umbellata leaves. The biotransformed product was also detected in larval and pupal tissues. Moreover, we observed deactivation of the toxicity of P. umbellata leaves against brine shrimp after their metabolism in H. brasiliensis larvae from a LC50 of 523.3 to 3,460.7 mu g/mL. This deactivation is closely associated with the biotransformation of 4-NC to E-2,3-dihydro-3-(3,4-dihydroxyphenyl)farnesoic acid, which showed LC50 of 8.0 and >1,000 mu g/mL, respectively.

Leishmanicidal activity of an alkenylphenol from Piper malacophyllum is related to plasma membrane disruption

Leishmaniasis and Chagas disease are parasitic protozoan infections that affect the poorest population in the world, causing high mortality and morbidity. As a result of highly toxic and long-duration treatments, novel, safe and more efficacious drugs are essential. In this work, the methanol (MeOH) extract from the leaves of Piper malacophyllum (Piperaceae) was fractioned to afford one alkenylphenol, which was characterized as 4-[(3'E)-decenyl]phenol (gibbilimbol B) by spectroscopic methods. Anti-protozoan in vitro assays demonstrated for the first time that Leishmania (L.) infantum chagasi was susceptible to gibbilimbol B. with an in vitro EC50 of 23 mu g/mL against axenic promastigotes and an EC50 of 22 mu g/mL against intracellular amastigotes. Gibbilimbol B was also tested for anti-trypanosomal activity (Trypanosoma cruzi) and showed an EC50 value of 17 mu g/mL against trypomastigotes. To evaluate the cytotoxic parameters, this alkenylphenol was tested in vitro against NCTC cells, showing a CC50 of 59 mu g/mL and absent hemolytic activity at the highest concentration of 75 mu g/mL. Using the fluorescent probe SYTOX Green suggested that the alkenylphenol disrupted the Leishmania plasma membrane upon initial incubation. Further drug design studies aiming at derivatives could be a promising tool for the development of new therapeutic agents for leishmaniasis and Chagas disease. (C) 2012 Elsevier Inc. All rights reserved.

Secondary metabolites isolated from Piper chimonantifolium and their antifungal activity

In this study, the CH2Cl2 extract from leaves of Piper chimonantifolium was subjected to several chromatographic separation procedures to afford one chromene (gaudichaudianic acid) as a major compound as well as two flavonoids (dihydrooroxylin and pinocembrin) and three steroids (sitosterol, sitosteryl palmitate and stigmasterol). The structures of all determined compounds were characterised by spectrometric analysis, mainly mass spectrometry and NMR, as well as their optical properties. This article describes the first phytochemical study of the leaves of P. chimonantifolium and an evaluation of the antifungal activity of its major compounds.

Pro-inflammatory effects of the aqueous extract of Echinometra lucunter sea urchin spines

The sea urchin, Echinometra lucunter, can be found along the Western Central Atlantic shores. In Brazil, it is responsible by circa 50% of the accidents caused by marine animals. The symptoms usually surpass trauma and may be pathologically varied and last differently, ranging from spontaneous healing in a few days, to painful consequences lasting for weeks. In this work, we have mimicked the sea urchin accident by administering an aqueous extract of the spine into mice and rats and evaluated the pathophysiological developments. Our data clearly indicate that the sea urchin accident is indeed a pro-inflammatory event, triggered by toxins present in the spine that can cause edema and alteration in the leukocyte-endothelial interaction. Moreover, the spine extract was shown to exhibit a hyperalgesic effect. The extract is rich in proteins, as observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but also contains other molecules that can be analyzed by reversed phase high-performance liquid chromatography. Altogether, these effects corroborate that an E. lucunter encounter is an accident and not an incident, as frequently reported by the victims.

Secondary Metabolites from the Phloem of Piper solmsianum (Piperaceae) in the Honeydew of Edessa meditabunda

Introduction The phytochemistry of species of the genus Piper has been studied extensively, including Piper solmsianum. However, no studies have addressed the phytochemistry of the sap content of Piper species. Objective To evaluate the transferring of secondary compounds from the saps of P. solmsianum to the honeydew of Edessa meditabunda. Methodology The honeydew of E. meditabunda and saps of P. solmsianum were analysed by GC-MS, H-1-NMR and LC-MS. Results The lignan (-)-grandisin and the phenylpropanoid (E)-isoelemicin were detected in both saps of P. solmsianum and honeydew of E. meditabunda. Conclusion Analysis of honeydew secreted by the sap-sucking insect E. meditabunda indicated that (-)-grandisin and (E)-isoelemicin are absorbed from the phloem of Piper solmsianum. Copyright (c) 2012 John Wiley & Sons, Ltd.

The effects of conjugated estrogen, raloxifene and soy extract on collagen in rat bones

Objective To evaluate the action of conjugated equine estrogen, raloxifene and isolated or combined genistein-rich soy extracts on collagen fibers in the bones of oophorectomized rats. Materials and methods Seventy female rats received testosterone propionate (0.1 mu g/g) on the 9th day after birth. At 6 months of age, the rats were administered the vehicle (propylene glycol, 0.5 ml/day), and ten of the rats were randomly chosen to comprise the non-oophorectomized control group (GI). The other 60 rats were ovariectomized and randomized into six groups of ten as follows: GII, vehicle; GIII, conjugated equine estrogen (CEE), 50 mu g/kg/day; GIV, raloxifene (RAL), 0.75 mg/kg/day; GV, genistein-rich soy extract (GSE), 300 mg/kg/day; GVI, CEE + GSE, 50 mu g/kg/day + 300 mg/kg/day; and GVII, CEE + RAL, 50 mu g/kg/day + 0.75 mg/kg/day. Three months after surgery, the drugs were administered for 60 consecutive days. All rats were euthanized, and their left tibiae were removed for histological routine. The histological sections were stained with hematoxylin-eosin, and picrosirius for evaluating bone microarchitecture. Types I and II collagen fibers were analyzed by immunofluorescence. Data analysis was carried out with ANOVA and Tukey's test. Results Collagen reduction was significant in the GIII animals when compared to the other groups (p < 0.05). There was no significant difference in the thickness of collagen fibers among the groups. There was a greater quantity of type III collagen in GVI than in the other groups. Conclusion Our data indicate that conjugated equine estrogen improves bone quality because it increases the quantity of type I collagen while reducing the quantity of thin collagen fibers. In addition, the combination of CEE and raloxifene or genistein-rich soy extract is not as efficient as CEE itself to improve bone quality.

Increased Antioxidant Content in Juice Enriched with Dried Extract of Pomegranate (Punica granatum) Peel

Antioxidants are compounds responsible for free radical scavenging in the body. They protect the organism from oxidative modification of cells and tissues. These modifications have been associated with degenerative diseases, atherosclerosis and carcinogenesis. Punica granatum displays high antioxidant potential due to the presence of phenolic compounds, which are capable of disease prevention. The present study showed the highest antioxidant activity in pomegranate peel than in seeds and pulp. Based on these results, pomegranate peel was used to produce dried extract that was added to commercial tomato juice and orange juice with strawberries. Analysis to determine the content of phenolic compounds and antioxidant activity was performed on pomegranate pulp, seeds and peel and in juices enriched with dried extract of pomegranate peel. The dried extract was responsible for a significant increase in antioxidant activity of the juices, proportional to the concentrations added. However, although both flavors of enriched juices displayed high antioxidant levels, the samples with higher dried extract concentrations received the lowest scores from sensory analysis participants due to the characteristic astringent flavor of pomegranate peels. Therefore, to obtain greater acceptance in the consumer market, we concluded that the maximum addition of dried pomegranate peel extract is 0.5% in tomato juice and orange juice with strawberries.

A new HPLC-DAD-MS/MS method for the simultaneous determination of major compounds in the crude extract of Lychnophora salicifolia Mart. (Brazilian arnicao) leaves: Application to chemical variability evaluation

Lychnophora salicifolia Mart., which occurs in the Brazilian Cerrado in the states of Bahia and Minas Gerais as well as in the southeast of the state of Goias, is the most widely distributed and also the most polymorphic species of the genus. This plant is popularly known to have anti-inflammatory and analgesic activities. In this work, we have studied the variation in terms of polar metabolites of ninety-three Lychnophora salicifolia Mart, specimens collected from different regions of the Brazilian Cerrado. Identification of the constituents of this mixture was carried out by analysis of the UV spectra and MS data after chromatographic separation. Twenty substances were identified, including chlorogenic acid derivatives, a flavonoid C-glucoside, and other sesquiterpenes. The analytical method was validated, and the reliability and credibility of the results was ensured for the purposes of this study. The concentration range required for analysis of content variability within the analyzed group of specimens was covered with appropriate values of limits of detection and quantitation, as well as satisfactory precision and recovery. A quantitative variability was observed among specimens collected from the same location, but on average they were similar from a chemical viewpoint. In relation to the study involving specimens from different locations, there were both qualitative and quantitative differences among plants collected from different regions of Brazil. Statistical analysis revealed that there is a correlation between geographical localization and polar metabolites profile for specimens collected from different locations. This is evidence that the pattern of metabolites concentration depends on the geographical distribution of the specimens. (C) 2012 Elsevier B.V. All rights reserved.

PHYSIOLOGICAL CHANGES IN SEEDS AND ANTIOXIDANT METABOLISM OF LETTUCE SEEDLING EXPOSED TO THE ACTION OF Zantedeschia aethiopica Spreng. LEAF EXTRACT

This work aimed to evaluate the influence of different concentrations of Zantedeschia aethiopica Spreng. extract on the physiological performance of the seed and on the response of the antioxidant metabolism of lettuce seedlings. The treatments consisted of leaves extracts from Z. aethiopica at concentrations of 0, 6, 12, 25 and 50%. Germination, first germination count, germination speed and index, length of shoot and radicle, seedling total dry mass, chlorophyll content, activity of superoxide dismutase, catalase and ascorbarte peroxidase enzymes, lipid peroxidation, hydrogen peroxide quantification and seedling emergence, length of organs, and total dry mass of seedlings were evaluated. The percentage of germination, the length of the shoot and radicle of seedlings and the total dry mass of seedlings grown in the greenhouse were reduced as the concentration of the extract increased. There were increases of electrical conductivity, of superoxide dismutase, catalase and ascorbate peroxidadase enzymes and the amount of hydrogen peroxide and lipid peroxidation in seedlings with increasing extract concentration. The extract reduced the physiological quality of lettuce seeds and induced an increased production of hydrogen peroxide in seedlings, which increased the activity of antioxidant enzymes that were not effective in tissue detoxification, resulting in cellular damage and increased numbers of abnormal seedlings.

Processing of Apeiba tibourbou Aubl. Extract Via Spray Drying

The effects of drying air inlet temperature (IT) and concentration of Aerosil 200 (C-A) on several properties of spray-dried Apeiba tibourbou extracts were investigated following a 3(2) full factorial design. Powder recovery varied from 9.83 to 46.95% and dried products showed moisture contents below 7%. Although the spray-dried products lost some of their polyphenols, they still present excellent antioxidant activity, opening perspectives for its use to medicinal purpose. C-A exerted a key role on the properties of spray-dried extracts, while IT did not present a significative influence. Aerosil (R) 200 proved to be an interesting alternative as an excipient for the drying of the herbal extract, even at intermediate concentrations such as 15%. The best combination of conditions to use for obtaining dry A. tibourbou extracts with adequate physicochemical and functional properties involves an IT of 100 degrees C and a C-A of 15%.

Cytotoxicity evaluation of Curcuma zedoaria (Christm.) Roscoe fluid extract used in oral hygiene products

Objective. This in vitro study evaluated the cytotoxic effects of the Curcuma zedoaria (Christm.) Roscoe (popular name: zedoary) fluid extract, as used in preparations for oral hygiene, mostly for anti-septic purposes. Materials and methods. The cell viability and cell growth were assessed by Trypan blue dye exclusion assay using the LMF cell line derived from oral mucosa. Cell viability (short-term assay) was measured 0, 6, 12 and 24 h after contact with the fluid extract. Cell growth (long-term assay) was analyzed in 1, 3, 5 and 7 days. The experimental groups were those testing the fluid extract obtained from the zedoary rhizome and the extractor liquid (ethanol 70 degrees GL) in the concentrations of 0.01-0.0001% v/v. Fresh DMEM were used in the control cultures. Results. Short-term assay-all studied cultures maintained stable cell viability; Long-term assay-there was progressive cell growth in all studied cultures. Conclusion. According to the results, the zedoary fluid extract presents low cytotoxicity and probably can be used in the oral hygiene products.

Biochemical Analysis of a Castor Bean Leaf Extract and its Insecticidal Effects Against Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae)

Plant extracts represent a great source of molecules, with insecticidal activity, which are used for pest control in several crop production systems. This work aimed to evaluate the toxicity of an aqueous extract of leaves of castor bean against larvae of Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) in search for different classes of molecules with insecticidal activities by using in vitro assays. The effects of the castor bean leaf extract on the food utilization, development, and survival of S. frugiperda larvae was evaluated by feeding the larvae an artificial diet supplemented with different concentrations of the extract (0%, 1%, 2.5%, 5%, and 10% w/v). The effects observed were dose-dependent, and the highest concentration evaluated (10% w/v) was the one the most affected food utilization by altering the nutritional indices, as well as larval weight gain, development time, and survivorship. In vitro assays to detect saponins, lectins, and trypsin inhibitors in the castor bean leaf extract were performed, but only trypsin inhibitors were detected. No preference for the diet source was detected in S. frugiperda by feeding the larvae in choice experiments with diets containing different concentrations of the castor bean extract tested. The data obtained indicate the existence of a potential molecule in the tested extract of castor bean to be used as an alternative insecticide to be integrated in the management of S. frugiperda.