Identificación y determinación estructural de un sesquiterpeno de las hojas de Tessaria integrifolia Ruiz & Pav. y evaluación de su actividad leishmanicida / Identification and structural determination of a sesquiterpene of Tessaria integrifolia Ruiz & Pav. leaves and evaluation of its leishmanicidal activity

RESUMEN Objetivos. Identificar y determinar la estructura del fitoconstituyente de hojas de Tessaria integrifolia Ruiz & Pav con efecto leishmanicida. Materiales y métodos. Se preparó un extracto fluido de hojas de Tessaria integrifolia Ruiz & Pav. que fue concentrado a extracto blando y se utilizó para evaluar la actividad leishmanicida en Mesocricetus auratus con leishmaniasis experimental, administrando vía intramuscular la dosis de 250 mg/kg del extracto blando por 15 días. El extracto fue fraccionado en columna cromatográfica de 45 cm, con un diámetro de 2,5 cm que contiene silicagel G-60, de 70-230 mesh (Sigma-Aldrich®), las nueve fracciones obtenidas fueron evaluadas sobre macrófagos infectados con Leishmania sp para determinar la fracción activa y aislar el compuesto activo, mediante separación, purificación y cristalización, siendo analizado por resonancia magnética nuclear (RMN) de 1H, 13C, y cromatografía líquida acoplada a espectroscopía de masas (LC/MS). Resultados. El extracto fluido de las hojas de Tessaria integrifolia Ruiz & Pav. presenta actividad leishmanicida en Mesocricetus auratus y la fracción F8 es activa sobre macrófagos infectados a dosis de 14 µg/mL. Se elucidó en esa fracción un sesquiterpeno tipo eudesmano ((4aS, 5R,6R,8aR)-6-hidroxi-5,8a-dimetil-3-(1-metiletiliden) octahidronaftalen-2(1H)-ona), según análisis de RMN 1H, 13C, y LC/MS. Conclusiones. El extracto fluido de hojas de Tessaria integrifolia Ruiz & Pav. presenta actividad leishmanicida sobre Mesocricetus auratus. La fracción F8 presenta actividad leishmanicida sobre macrófagos infectados a dosis de 14 µg/mL. Se elucidó en la fracción activa un sesquiterpeno tipo eudesmano según los análisis de RMN 1H, 13C, y LC/MS.

ABSTRACT Objective. To identify and determine the phytoconstituent structure of Tessaria integrifolia Ruiz & Pav. leaves with leishmanicidal activity. Materials and Methods. Fluid extract of leaves was prepared, concentrated to soft extract, and used to evaluate leishmanicidal activity in Mesocricetus auratus with experimental leishmaniasis, at the dose of 250 mg/kg of soft extract by intramuscular route for 15 days. Extract was fractionated in 45 cm column chromatography with a 2.5 cm diameter, containing G-60 silica gel, and 70-230 mesh (Sigma-Aldrich®). Nine fractions were obtained and assessed on macrophages infected with Leishmania sp to determine the active fraction and isolate the active compound, by separation, purification, and crystallization, analyzed by nuclear magnetic resonance (NMR) of 1H, 13C, and liquid chromatography coupled to mass spectroscopy (LC/ MS). Results. Fluid extract from the leaves of T. integrifolia presents leishmanicidal activity in M. auratus. Fraction F8 is active on infected macrophages at a dose of 14 μg/mL. An eudesman type sesquiterpene ((4aS, 5R, 6R, 8aR) -6-hydroxy-5, 8a-dimethyl-3- (1-methylethylidene) octahydronaphthalen-2 (1H) -one) was identified, by RMN 1 H, 13C, and LC / MS analysis. Conclusions. Fluid extract of leaves of Tessaria integrifolia Ruiz & Pav. presents leishmanicidal activity on Mesocricetus auratus with experimental leishmaniasis. Fraction F8 presents leishmanicidal activity on infected macrophages at a dose of 14 μg/mL. An eudesman type sesquiterpene was identified, according to 1 H, 13C, and LC / MS NMR analysis.

Triterpenes and Beta-agarofurane sesquiterpenes from tissue culture ofMaytenus boaria / Triterpenos y Beta-agarofuranos sesquiterpenos desde cultivo de tejidos de Maytenus boaria

Leaf explants of Maytenus boaria were induced towards callus tissue culture with different mixture of cytokinins and auxins. MeOH extract of callus was partitioned with AcOEt and water, and through repeated chromatography procedures were isolated and identified, four triterpenes and three beta-agarofuran sesquiterpenes.

Explantes de hojas de Maytenus boaria fueron inducidos a formar callos mediante diferentes mezclas de citoquininas y auxinas. Un extracto metanólico de los callos fue fraccionado con AcOEt y agua, y mediante repetidas cromatografías fueron aislados e identificados siete compuestos, cuatro triterpenos y 3 sesquiterpenos del tipo agarofurano.

Induction of apoptosis by zerumbone isolated from Zingiber zerumbet (L.) Smith in protozoan parasite Leishmania donovani due to oxidative stress

Abstract In the present context of emergence of resistance aligned with the conventional anti-leishmanial drugs and occasional treatment failure compelled us to continue the search for replaceable therapeutic leads against Leishmaniainfection. Various ginger spices of the Zingiberaceae family are widely used as spices, flavouring agents, and medicines in Southeast Asia because of their unique flavour as well as due to their medicinal properties. Zerumbone, a natural component of Zingiber zerumbet (L.) Smith, has been studied for its pharmacological potential as antiulcer, antioxidant, anticancer, and antimicrobial. In this study, we have shown that zerumbone could induce ROS mediated apoptosis in Leishmania donovani promastigotes and also found effective in reducing intracellular amastigotes in infected-macrophages. We emphasized the potential of zerumbone to be employed in the development of new therapeutic drugs against L. donovaniinfection and provided the basis for future research on the application of transitional medicinal plants.

Antifungal activity of extracts from Piper aduncum leaves prepared by different solvents and extraction techniques against dermatophytes Trichophyton rubrum and Trichophyton interdigitale

The effects of different solvents and extraction techniques upon the phytochemical profile and anti-Trichophyton activity of extracts from Piper aduncum leaves were evaluated. Extract done by maceration method with ethanol has higher content of sesquiterpenes and antifungal activity. This extract may be useful as an alternative treatment for dermatophytosis.

Sedative and anesthetic activities of the essential oils of Hyptis mutabilis (Rich.) Briq. and their isolated components in silver catfish (Rhamdia quelen)

This study evaluated the sedative and anesthetic effects of the essential oils (EO) of Hyptis mutabilis (Rich.) Briq. and their isolated components on silver catfish (Rhamdia quelen). Quantitative chemical differences between the EOs obtained from leaves and inflorescences were verified, and a new chemotype rich in globulol was described. Although there were no significant differences in the time of induction for sedation and anesthesia between the EOs, only the leaf EO at 344 mg/L anesthetized all fish without side effects. Fractionation of the leaf EO was carried out by column chromatography. The isolated compounds [(+)-1-terpinen-4-ol and (-)-globulol] showed different activity from that detected for the leaf EO in proportional concentrations and similar sedation to a eugenol control at 10 mg/L. However, fish exposed to 1-terpinen-4-ol (3 and 10 mg/L) did not remain sedated for 30 min. Anesthesia was obtained with 83-190 mg/L globulol, but animals showed loss of mucus during induction and mortality at these concentrations. Synergism of the depressor effects was detected with the association of globulol and benzodiazepine (BDZ), compared with either drug alone. Fish exposed to BDZ or globulol+BDZ association showed faster recovery from anesthesia in water containing flumazenil, but the same did not occur with globulol. In conclusion, the use of globulol in aquaculture procedures should be considered only at sedative concentrations of 10 and 20 mg/L, and its mechanism of action seems not to involve the GABAA-BDZ system.

Biological activity of a furanoeremophilane isolated from Senecio filaginoides var. filaginoides / Actividad biológica de un furanoeremofilano aislado de Senecio filaginoides var. filaginoides

Compound, 10H-9-oxofuranoeremophilane was isolated from Senecio filaginoides var. filaginoides, and its structure established on the basis of spectral analysis and and comparison with literature data. Antimicrobial activity was demonstrated against Candida spp but not against bacteria (Staphylococcus aureus, Escherichia coli and Pseudomona aeruginosa). Besides, the compound showed antifungal activity and effect on mycelial growth of the phytopathogenic fungus Botrytis cinerea. and comparison with literature data.

Un compuesto, 10H-9-oxofuranoeremofilano fue aislado de Senecio filaginoides var. filaginoides, esta estructura fue establecida sobre la base del análisis espectral y comparación con datos bibliográficos. La actividad antimicrobiana fue demostrada contra Candida spp pero no contra las bacterias (S.aureus, E.coli y P. aeruginosa). Además el compuesto mostró actividad antifúngica y efectos sobre el crecimiento micelial del hongo fitopatógeno Botrytis cinerea.

In vitro activity of hypnophilin from Lentinus strigosus: a potential prototype for Chagas disease and leishmaniasis chemotherapy

Hypnophilin and panepoxydone, terpenoids isolated from Lentinus strigosus, have significant inhibitory activity onTrypanosoma cruzi trypanothione reductase (TR). Although they have similar TR inhibitory activity at 10 μg/mL (40.3 μM and 47.6 μM for hypnophilin and panepoxydone, respectively; ~100 percent), hypnophilin has a slightly greater inhibitory activity (~71 percent) on T. cruzi amastigote (AMA) growth in vitro as well as on in vitro phytohemagglutinin (PHA)-induced peripheral blood mononuclear (PBMC) proliferation (~70 percent) compared to panepoxydone (69 percent AMA inhibition and 91 percent PBMC inhibition). Hypnophilin and panepoxydone at 1.25 μg/mL had 67 percent inhibitory activity onLeishmania (Leishmania) amazonensis amastigote-like (AMA-like) growth in vitro. The panepoxydone activity was accompanied by a significant inhibitory effect on PHA-induced PBMC proliferation, suggesting a cytotoxic action. Moreover, incubation of human PBMC with panepoxydone reduced the percentage of CD16+ and CD14+ cells and down-regulated CD19+, CD4+ and CD8+ cells, while hypnophilin did not alter any of the phenotypes analyzed. These data indicate that hypnophilin may be considered to be a prototype for the design of drugs for the chemotherapy of diseases caused by Trypanosomatidae.

Lactonas sesquiterpénicas de las plantas Viguiera sylvatica y Decachaeta thieleana (Asteraceae) modulan la producción de óxido nítrico y la fagocitosis de macrófagos RAW

Sesquiterpene lactones of the plants Viguiera sylvatica and Decachaeta thieleana (Asteraceae) modulate nitric oxide production and phagocytosis of macrophages cell line RAW. Different species of the Asteraceae family are a potential source of sesquiterpene lactones with anti-inflammatory properties. Macrophages play a central role in the regulation of immune responses. In the present study, the in vitro effect of two sesquiterpene lactones, a millerenolide and a thieleanin, was assessed by measuring the production of nitric oxide (NO) by cell line RAW (murine macrophages) using the Griess reagent. Additionally, the effect of these sesquiterpene lactones on phagocytic capacity of latex particles and the reduction of nitroblue tetrazolium (NBT) were evaluated microscopically. Treatment of macrophages with >2.5 µg/ml of both sesquiterpene lactones, reduced the production of NO. A decreased number of macrophages able to reduce NBT were observed when these cells were treated with 3 µg/ml of millerenolide or 7.5 µg/ml of thieleanin. Treatment of macrophages with 4 µg/ml of millerenolide or 7.5 µg/ml of thieleanin, reduced the phagocytic capacity of macrophages. Cytotoxic effects on the macrophages were only observed when the concentration was increased to 8 µg/ml of millerenolide or 25 µg/ml of thieleanin. Our results suggest that these sesquiterpene lactones could be useful compounds in the elaboration of anti-inflammatory drugs. Rev. Biol. Trop. 56 (3): 1063-1073. Epub 2008 September 30.

Las plantas de la familia Asteraceae son una fuente potencial de lactonas sesquiterpénicas con propiedades antiinflamatorias. Los macrófagos son células que desempeñan una función central en la regulación de la respuesta inmune. Este estudio evaluó el efecto in vitro de dos lactonas sesquiterpénicas, un millerenólido y thieleanina, sobre la producción de óxido nítrico (NO) en una línea celular de macrófagos de ratón (RAW), utilizando el reactivo de Griess. Además, se estudió el efecto sobre la capacidad fagocítica de RAW, evaluando al microscopio la fagocitosis de partículas inertes de látex y la reducción de nitroazul de tetrazolio (NBT). Se observó que los macrófagos tratados con lactona sesquiterpénica (>2.5 µg/ml) disminuyeron la producción de NO. Además, se observó una disminución de la cantidad de macrófagos capaces de reducir NBT, después que fueron tratados con millerenólido (3 µg/ml) o thieleanina (7.5 µg/ ml). Por otro lado, la exposición a 4 µg/ml de millerenólido ó 7.5 µg/ml de thieleanina redujo la cantidad promedio de partículas de látex fagocitadas. No se observaron diferencias entre tratamientos y control en cuanto al porcentaje de células fagocíticas. Sólo se observaron efectos citotóxicos sobre los macrófagos, cuando la concentración de millerenólido se incrementó a 8 µg/ml o la de thieleanina se aumentó a 25 µg/ml. Estos resultados sugieren que el millerenólido y la thieleanina podrían ser compuestos útiles en la elaboración de drogas antiinflamatorias.

Bioactivity of the extract and isolation of lignansand a sesquiterpene from the aerial parts of centaurea pamphlica[Asteraceae]

Centaurea pamphylica Boiss. and Heldr. [Family: Asteraceae], commonly known as "pamphylia daisy", is a Turkish endemic species of the genus Centaurea that comprises ca. 500 species, many of which have been used as traditional medicines. The n-hexane, dichloromethane [DCM] and methanol [MeOH] extracts of the aerial parts of C. pamphylica were assessed for antioxidant activity and general toxicity using, respectively, the 2, 2-diphenyl-1-picrylhydrazyl [DPPH], and the brine shrimp lethality assays. The reversed-phase preparative HPLC and PTLC were used to isolate compounds from the extracts. The structures of these compounds [1-4] were elucidated by spectroscopic means, and also by direct comparison with the respective published data. Both the DCM and the MeOH extract showed significant levels of antioxidant activities with the RC[50] values of 72.6 x 10[-2] and 47.3 x 10[-2] mg/mL, respectively. The MeOH extract exhibited low levels of toxicity towards brine shrimps [LD[50] = 125.0 x 10[-2] mg/mL]. Three major bioactive components of the MeOH extract were matairesinoside [1], arctiin [2] and matairesinol [3]. An eudesmane-type sesquiterpene, pterodontriol [4], was also isolated from the DCM extract. Since reactive oxygen species are important contributors to various ailments, the antioxidant properties of the extracts as well as the isolated compounds may be of medicinal significance. This is the first report on the occurrence of 1-4 in C. pamphylica, and 4 in the genus Centaurea

Laboratory evaluation of sensitivity of three keratinophilic fungi to some vicolides.

Four vicolides (sesquiterpenoides) isolated from Vicoa indica were evaluated against three keratinophilic fungi, viz., Microsporum gypseum, Chrysosporium tropicum, and Trichophyton terrestris. All the test fungi were found to be sensitive to vicolides. Vicolides A and C showed the maximum efficacy while B and D exhibited moderate activity. The Minimum Inhibitory Concentration (MIC) was observed in the range of 15.62-125 micrograms. The most sensitive fungus tested was C. tropicum followed by T. terrestris and M. gypseum.

Aislamiento y elucidación estructural de lactonas sesquiterpenicas de ambrosia arborescens mill (asteraceae) / Isolation and structural elucidation of sesquiterpenics lactonesof ambrosia arborescens mill (asteraceae)

La especie Ambrosia arborescens mill de la familia asteracea, contiene cuatro lactonas sesquiterpénicas. Fueron identificadas dos lactonas: la damsina, comprobada mediante cromatografía en capa fina y punto de fusión, y que la coronopulínea cuya estructura fue determinada mediante sus constantes físicas, espectroscópicas, uv, Ir y formación de derivados, una tercera lactona sesquiterpénica cristalizable y una cuarta, que sólo se llegó a aislar; no pudieron ser identificadas por la proporción en que se encontraban presentes en la muestra.

Sesquiterpene lactones from Vernonia

Some informations about the sesquiterpene lactones isolated from Brazilian species of Vernonia are described, as well the results of tests developed with such compounds with respect to their anti-feedant, molluscicide, antimicrobial and analgesic properties.