Antinociceptive and anti-inflammatory activity of the siaresinolic acid, a triterpene isolated from the leaves of Sabicea grisea Cham. & Schltdl. var. grisea.

In the present study, siaresinolic acid (siaresinol, SA) was isolated from the leaves of Sabicea grisea and studied to evaluate its antinociceptive and anti-inflammatory activity. The antinociceptive effect of SA was investigated in mice using different animal models to study pain. In the acetic acid-induced writhing test, intraperitoneal (i.p.) injection of SA (0.1, 1, and 10 mg/kg, i.p.) 1 h before a pain stimulus significantly reduced the nociceptive response (by 42.3, 68.2, and 70.9 %, respectively). Pretreatment with glibenclamide, but not with yohimbine, metoclopramide, ketanserin, or naloxone, restored the antinociceptive effect induced by SA in the writhing test, suggesting that the K(+)ATP channel pathway might be involved in its mechanism of action. In the formalin test, SA (1 mg/kg, i.p.) decreased licking time in the second phase only, thereby indicating an anti-inflammatory effect. In the hot plate test, there was no significant difference in nociceptive behavior. In the rota-rod test, it was verified that a high dose of SA (10 mg/kg, i.p.) did not affect the locomotor activity of mice. In the pleurisy model, induced by carrageenan, treatment with SA inhibited important events involved in inflammatory responses, namely leukocyte influx, plasma leakage, and increased inflammatory mediators (TNF-α, IL-1ß, and chemokine CXCL1), in the pleural exudate. Additionally, SA itself was not cytotoxic when evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in macrophages cultured for 24 h at concentrations ranging from 1 to 200 µg/mL. These results suggest, for the first time, that SA attenuates nociceptive behavior through mechanisms involving receptors for ATP-dependent potassium channels, in addition to suppressing acute inflammatory responses.

Isolation of a dihydrobenzofuran lignan, icariside E4, with an antinociceptive effect from Tabebuia roseo-alba (Ridley) Sandwith (Bignoniaceae) bark.

The antinociceptive activity of icariside E4, a dihydrobenzofuran-type lignan isolated from Tabebuia roseo-alba (Ridley) Sandwith (Bignoniaceae) bark, was evaluated in mice by using chemical and thermal models of nociception. Intraperitoneal (i.p.) administration of crude T. roseo-alba bark extract and its methanol fraction inhibited acetic acid-induced abdominal constriction in mice. Furthermore, i.p. administration of 0.1, 1, and 10 mg/kg of icariside E4 reduced the number of writhes evoked by acetic acid injection by 46.9, 82.3, and 66.6%, respectively. Icariside E4 administration had no effect in the first phase of the formalin test, but it reduced nociceptive behavior in the second phase as indicated by a reduction in the licking time. Icariside E4 did not modify thermal nociception in the hot-plate test model, suggesting that it had a peripheral antinociceptive action. The antinociceptive effect of icariside E4 in the writhing test was reversed by pre-administration of glibenclamide, but not of naloxone, atropine, yohimbine, or haloperidol. Together, these results indicated that the antinociceptive activity of icariside E4 from T. roseo-alba in models of chemical pain occurred through ATP-sensitive K(+) channel-dependent mechanisms.

Larvicidal activities and chemical composition of essential oils from Piper klotzschianum (Kunth) C. DC. (Piperaceae).

BACKGROUND: Volatile oils from fresh roots, stems, leaves and seeds of Piper klotzschianum (Piperaceae) were obtained by hydrodistillation and analysed by GC-FID and GC-MS. In total, 25 components, representing more than 95% of the examined oils, were identified. The essential oils were evaluated against Artemia salina Leach nauplii and fourth-instar Aedes aegypti larvae. RESULTS: The major chemical constituents that were identified from various parts of this plant were 1-butyl-3,4-methylenedioxybenzene and 2,4,5-trimethoxy-1-propenylbenzene in the root, 1-butyl-3,4-methylenedioxybenzene in the stems and leaves and 1-butyl-3,4-methylenedioxybenzene, limonene and α-phellandrene in the seeds. The biological activities of these essential oils generally exhibited high toxicity against A. salina, with LC50 values that ranged from 7.06 to 15.43 µg mL(-1), and significant larvicidal activity against fourth-instar A. aegypti larvae was observed in the essential oils from the seeds (LC50 of 13.27 µg mL(-1)) and roots (LC50 of 10.0 µg mL(-1)) of the plant. CONCLUSION: The present study indicates that both essential oil of P. klotzsdhianum and the isolate 1-butyl-3,4-methylenedioxybenzene are potential resources for A. aegypti larva control. This is the first report of the biological activities of the oil and isolated compound.

Chemical constituents of Rourea doniana

The chromatography fractionation of the hexane, chloroform and ethyl acetate extracts from the leaves and stems of Rourea doniana Baker, Connaraceae, resulted in the isolation of five triterpenes (lupeol, lupenone, α-amyrenone, β-amyrenone, and taraxerol), a flavonol (7,4'-dimethylkaempferol), a coumarin (scopoletin) and four phytosteroids (β-sitosterol, stigmasterol, β-sitosteryl-3-O-β- D-glucopyranoside and stigmasteryl-3-O-β-D-glucopyranoside). All compounds are being for the first time in this species and all triterpenes and the flavonol are being described for the first time in the family Connaraceae. These compounds were identified on basis of their IR and NMR (¹H, 13C, DEPT, HSQC, HMBC, and NOESY) spectral data and by comparison with literature data.

Antinociceptive and anti-inflammatory effects of octacosanol from the leaves of Sabicea grisea var. grisea in mice.

Sabicea species are used in the Amazon for treatment of fever and malaria, which suggests that its chemical constituents may have some effect on pain and inflammation. Phytochemical analysis of the hexane fraction obtained from the crude ethanol extract from Sabicea grisea var. grisea Cham. & Schltdl (Rubiaceae), an endemic plant in Brazil, resulted in the isolation of octacosanol. This study investigated the antinociceptive and anti-inflammatory effects of the octacosanol in different experimental models. The crude ethanolic extract and hexane fraction obtained from the leaves of S. grisea produced an inhibition of acetic acid-induced pain. Moreover, octacosanol isolated from the hexane fraction produced a significant inhibition of pain response elicited by acetic acid. Pre-treatment with yohimbine, an alpha 2-adrenergic receptor antagonist, notably reversed the antinociceptive activity induced by octacosanol in the abdominal constriction test. Furthermore, mice treated with octacosanol did not exhibit any behavioral alteration during the hot plate and rota-rod tests, indicating non-participation of the supraspinal components in the modulation of pain by octacosanol with no motor abnormality. In the formalin test, octacosanol did not inhibit the licking time in first phase (neurogenic pain), but significantly inhibited the licking time in second phase (inflammatory pain) of mice. The anti-inflammatory effect of octacosanol was evaluated using carrageenan-induced pleurisy. The octacosanol significantly reduced the total leukocyte count and neutrophils influx, as well as TNF-α levels in the carrageenan-induced pleurisy. This study revealed that the mechanism responsible for the antinociceptive and anti-inflammatory effects of the octacosanol appears to be partly associated with an inhibition of alpha 2-adrenergic transmission and an inhibition of pathways dependent on pro-inflammatory cytokines. Finally, these results demonstrated that the octacosanol from the leaves of S. grisea possesses antinociceptive and anti-inflammatory activities, which could be of relevance for the pharmacological control of pain and inflammatory processes.

Modulation of inflammatory processes by leaves extract from Clusia nemorosa both in vitro and in vivo animal models.

The present study was carried out to investigate the anti-inflammatory effect of the hexane extract of the leaves from Clusia nemorosa G. Mey, called HECn, using carrageenan-induced mice pleurisy and cotton pellet-induced mice granuloma. Additionally, the ability of HECn to affect both neutrophil migration as viability was investigated by use of the Boyden chamber assay and flow cytometry, respectively. The HECn significantly inhibited exudation, total leukocytes and neutrophils influx, as well as TNFα levels in carrageenan-induced pleurisy. However, the extract not suppressed the granulomatous tissue formation in the cotton pellet-induced granuloma test. Experiments performed in vitro revealed that HECn on human neutrophils inhibited a dose-dependent manner the CXCL1-induced neutrophil chemotaxis. Furthermore, HECn also inhibited the chemoattraction of human neutrophils induced by formyl-methionyl-leucyl-phenylalanine (fMLP), leukotriene B4 (LTB4) and platelet activating factor (PAF) in a Boyden chamber. However, this same treatment not was able to induce apoptosis. The results obtained in this study showed that the extract from leaves of C. nemorosa possess a potent inhibitory activity in acute model of inflammation, being the effects mediated, in part, by inhibition of neutrophil responsiveness. These results indicate that C. nemorosa could be a good source for anti-inflammatory compounds.

A new iridoid from Guettarda grazielae M.R.V. Barbosa (Rubiaceae).

Chromatographic fractionation of the chloroform extracts from the stem bark and stems of Guettarda grazielae resulted in the isolation of a new iridoid (guettardodiol, 1) and the secoiridoid sarracenin (2), described for the first time in this genus. The structural elucidation of these compounds was based on spectroscopic analyses (IR, MS as well as 1-D and 2-D NMR experiments).

Determination of the phenolic content and antioxidant potential of crude extracts and isolated compounds from leaves of Cordia multispicata and Tournefortia bicolor.

In this work, the total phenolic content and antioxidant activity of extracts and four flavonoids isolated from leaves of two Boraginaceae species (Cordia multispicata Cham. and Tournefortia bicolor Sw.) were evaluated using Folin-Ciocalteu reagent, DPPH free radical scavenging and inhibition of peroxidation of linoleic acid by FTC method. For comparison, ascorbic acid, alpha-tocopherol and BHT were used. In general, extracts from T. bicolor (68.8 +/- 0.001 to > 1000 mg/g) showed higher phenolic content than C. multispicata (66.1 +/- 0.009 to 231 +/- 0.07 mg/g), and also scavenged radicals (IC(50) 12.8 +/- 2.5 to 437 +/- 3.5 mg/L) and inhibited lipid peroxide formation (IC(50) 51.2 +/- 2.29 to 89 +/- 0.59 mg/L). For these extracts a good correlation between the phenolic content and antioxidant activity was observed, suggesting that T. bicolor is richer in phenolic compounds and that it could serve as a new source of natural antioxidants or nutraceuticals with potential applications. Chromatographic procedures monitored by antioxidant assays afforded seven compounds, which were identified by spectral analyses (IR, MS and 1D and 2D NMR) and comparison with reported data as being trans-phytol (1), taraxerol (2), 3,7,4'-trimethoxyflavone (3), 5,3'-dihydroxy-3,7,4'-trimethoxyflavone (4), quercetin (5), tiliroside (6), and rutin (7). Compounds (4-7) were also evaluated and were effective as DPPH quenching (IC(50) 7.7 +/- 3.6 to 79.3 +/- 3.4 mg/L) and as inhibition of lipid peroxidation (IC(50) 80.1 +/- 0.98 to 88.7 +/- 3.62 mg/L). This is the first report on the total phenolic content, radical-scavenging and antioxidant activities of these species.

Larvicidal activity of 94 extracts from ten plant species of northeastern of Brazil against Aedes aegypti L. (Diptera: Culicidae).

In an effort to find effective and affordable ways to control of Aedes aegypti L. (Diptera: Culicidae), the larvicidal activities of 94 extracts from ten plant species belonging to eight families [Guettarda grazielae and Spermacoce verticillata (Rubiaceae), Coccoloba mollis and Triplaris americana (Polygonaceae), Eschweilera ovata (Lecytidaceae), Merremia aegyptia (Convolvulaceae), Ouratea nitida (Ochnnaceae), Protium heptaphyllum (Burseraceae), Rourea doniana (Connaraceae), and Tovomita brevistaminea (Guttiferae)], widely found in the northeast of Brazil, were initially screened at 250 microg/mL. Nineteen of the 94 extracts from six plant species (C. mollis, G. grazielae, M. aegyptia, R. doniana, S. verticillata, and T. americana) showed significant activity (>75% mortality) against the fourth instar larvae. Some of these extracts showed 100% mosquito larvae mortality, with LD(50) values lower 200 microg/mL. When compared to synthetic insecticide Temephos (LD(50) 8.7 microg/mL), two of them, hexane (LD(50) 12.1 microg/mL) and ethyl acetate (LD(50) 51.6 microg/mL) extracts from stems of R. doniana and G. grazielae, respectively, were the most effective. The isolation and purification of these extracts are in progress and evaluation of these compounds will be needed to identify the active component.

Studies on the antimicrobial activity and brine shrimp toxicity of Zeyheria tuberculosa (Vell.) Bur. (Bignoniaceae) extracts and their main constituents.

BACKGROUND: Due to the indiscriminate use of antimicrobial drugs, the emergence of human pathogenic microorganisms resistant to major classes of antibiotics has been increased and has caused many clinical problems in the treatment of infectious diseases. Thus, the aim of this study was to evaluate for the first time the in vitro antimicrobial activity and brine shrimp lethality of extracts and isolated compounds from Zeyheria tuberculosa (Vell.) Bur., a species used in Brazilian folk medicine for treatment of cancer and skin diseases. METHODS: Using the disc diffusion method, bioautography assay and brine shrimp toxicity test (Artemia salina Leach), we studied the antimicrobial activity and lethality of extracts and isolated compounds against three microorganisms strains, including Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa) bacteria and yeasts (Candida albicans). RESULTS: In this study, the extracts inhibited S. aureus (8.0 +/- 0.0 to 14.0 +/- 0.0 mm) and C. albicans (15.3 +/- 0.68 to 25.6 +/- 0.4 mm) growth. In the brine shrimp test, only two of them showed toxic effects (LC50 29.55 to 398.05 microg/mL) and some extracts were non-toxic or showed weak lethality (LC50 705.02 to > 1000 microg/mL). From these extracts, four flavones [5,6,7,8-tetramethoxyflavone (1), 5,6,7-trimethoxyflavone (2), 4'-hydroxy-5,6,7,8-tetramethoxyflavone (3), and 4'-hydroxy-5,6,7-trimethoxyflavone (4)] were isolated through bioassay-guided fractionation and identified based on the 1D and 2D NMR spectral data. By bioautography assays, compounds 1 [S. aureus (16.0 +/- 0.0 mm) and C. albicans (20.0 +/- 0.0 mm)] and 3 [S. aureus (10.3 +/- 0.6 mm) and C. albicans (19.7 +/- 0.6 mm)] inhibited both microorganisms while 2 inhibited only S. aureus (11.7 +/- 0.6 mm). Compound 4 did not restrain the growth of any tested microorganism. CONCLUSION: Our results showed that extracts and isolated flavones from Z. tuberculosa may be particularly useful against two pathogenic microorganisms, S. aureus and C. albicans. These results may justify the popular use this species since some fractions tested had antimicrobial activity and others showed significant toxic effects on brine shrimps. However, in order to evaluate possible clinical application in therapy of infectious diseases, further studies about the safety and toxicity of isolated compounds are needed.

Triterpenos de Guettarda grazielae M.R.V. Barbosa (Rubiaceae) / Triterpenes from Guettarda grazielae M.R.V. Barbosa (Rubiaceae)

O presente trabalho descreve o isolamento e a identificação estrutural de seis triterpenos (acetato de ±-amirina, cicloartenona, 3²,19±,23-triidroxi-urs-12-eno e os ácidos ursólico, 3-²-O-²-D-glicopiranosilquinóvico e 3²,6²,19±,23-tetraidroxi-urs-12-en-28-óico), além de um fitoesteróide (sitostenona) de extratos das folhas, cascas do caule e caule de Guettarda grazielae M.R.V. Barbosa (Rubiaceae). Os compostos isolados, exceto o ácido 3-²-O-²-D-glicopiranosilquinóvico, estão sendo descritos pela primeira vez no gênero Guettarda e foram identificados com base na análise de dados espectrais de RMN, incluindo APT e DEPT, bem como pela comparação com dados da literatura.

This work describes the isolation and structural identification of six triterpenes (a-amyrin acetate, cycloartenone,3²,19±,23-trihydroxyurs-12-ene,andursolic,3-²-O-²-D-glucopyranosylquinovic, 3²,6²,19±,23-tetrahydroxyurs-12-en-28-oic acids), beside a phytosteroid (sitostenone) from leaves, stem barks and stems of Guettarda grazielae (Rubiaceae). All isolated compounds, except 3-²-O-²-D-glucopyranosylquinovic acid, are being described for the first time in the genus Guettarda and were identified on basis of the spectral NMR data, including APT and DEPT, as well as by comparison with data of the literature.

Constituintes químicos das folhas e do caule de Coccoloba mollisCasaretto (Polygonaceae) / Chemical constituents from leaves and stem of Coccoloba mollis Casaretto (Polygonaceae)

O estudo químico das frações neutra em hexano das folhas e em diclorometano do caule de Coccoloba mollis resultou no isolamento de um triterpeno pentacíclico (simiarenol), que pela primeira vez ocorre em Polygonaceae, dois fitoesteróides (sitostenona e sitosterol), um diterpeno (trans-fitol) e de um benzenóide (ácido vanílico) que está sendo descrito pela vez neste gênero. A identificação estrutural destes compostos foi feita com base na análise dos dados espectrais (IV, EM e RMN, incluindo DEPT e HMQC), bem como pela comparação com dados descritos na literatura.

This work describes the isolation of a pentacyclic triterpene (simiarenol), described for the first time in this family, two phytosteroids (sitostenone and sitosterol), a diterpene (trans-phytol) and a benzenoid (vanillic acid), described for the first time in this genus, from leaves and stem of Coccoloba mollis. These compounds were identified on basis of their spectral data (IR, MS and NMR, including DEPT and HMQC), as well as by comparison with literature data.

Terpenóides e avaliação do potencial antimalárico, larvicida, anti-radicalar e anticolinesterásico de Pouteria venosa (Sapotaceae) / Terpenoids and evaluation of the antimalarial, larvicidal, anti-radicalar and anticholinesterase potential of Pouteria venosa (Sapotaceae)

O presente trabalho descreve o isolamento de quatro triterpenos (taraxerol, ácido ursólico, ácido 3b,19a,23-triidroxiurs-12-en-28-óico e ácido 2a,3a,19a,23-tetraidroxiurs-12-en-28-óico) e um fitoesteróide (espinasterol), bem como a avaliação do potencial antimalárico (cepa NK-65 do Plasmodium berghei), larvicida (larvas do 4º instar do Aedes aegypti), anti-radicalar (2,2-difenil-1-picril-hidrazila, DPPH) e anticolinesterásico de extratos das folhas, cascas do caule e caule de Pouteria venosa (Mart.) Baehni. Todos os compostos isolados estão sendo descritos pela primeira vez nesta espécie e foram identificados com base na análise de dados espectrais (IV e RMN, incluindo APT e DEPT), bem como pela comparação com dados descritos na literatura.

This work describes the isolation of four triterpenes (taraxerol, ursolic acid, 3b,19a,23-trihydroxyurs-12-en-28-oic acid and 2a,3a,19a,23-tetrahydroxyurs-12-en-28-oic acid) and a phytosteroid (spinasterol), as well as a preliminary evaluation of antimalarial (NK-65 strains of Plasmodium berghei), larvicidal (4th instar of Aedes aegypti), anti-radicalar (2,2-diphenyl-1-pycryl-hydrazyl, DPPH) and anticholinesterase activities of Pouteria venosa (Mart.) Baehni extracts from leaves, stem barks and stems. All isolated compounds are being described for the first time in this species and were identified on basis of the spectral data (IR and NMR, including APT, DEPT), as well as by comparison with literature data.

Constituintes químicos das folhas e caule de Croton sellowii (Euphorbiaceae) / Chemical constituents from the leaves and stems of Croton sellowii (Euphorbiaceae)

O presente trabalho descreve o isolamento de terpenóides (sitosterol, estigmasterol, cicloart-25-eno-3b,24b-diol, cicloart-25-eno-3b,24a-diol e ácido betulônico), flavonóides (artemetina, crisosplenetina, casticina, penduletina e tilirosídeo) e fenilpropanóide (treo-7-etoxi-8-hidroxi-diidroanol) nas folhas e caule de Croton sellowii Baill. Todos os compostos isolados estão sendo descritos pela primeira vez nesta espécie e foram identificados com base na análise de dados espectrais (IV, UV e RMN, incluindo APT, DEPT, COSY, NOESY, HMQC e HMBC), bem como pela comparação com dados descritos na literatura.

This work describes the isolation of terpenoids (sitosterol, stigmasterol, cycloart-25-ene-3b,24b-diol, cycloart-25-ene-3b,24a-diol and betulonic acid), flavonoids (artemetin, crysosplenetin, casticin, penduletin and tiliroside) and phenylpropanoid (threo-7-ethoxy-8-hydroxy-dihydroanol) from the leaves and stems of Croton sellowii Baill. All isolated compounds are being described for the first time in this species and were identified on basis of the spectral data (IR, UV, and NMR, including APT, DEPT, COSY, NOESY, HMQC and HMBC), and by comparison with literature data.

Constituintes químicos e avaliação preliminar in vivo da atividade antimalárica de Ouratea nitida Aubl (Ochnaceae) / Chemical constituents and in vivo preliminary evaluation of antimalarial activity of Ouratea nitida Aubl. (Ochnaceae)

O presente trabalho descreve o isolamento de triterpenos (ácido 3,4-seco-friedelan-3-óico, friedelina e b-amirina) e outros compostos (éster etílico do ácido p-hidroxibenzóico e tetracosano), bem como a avaliação preliminar in vivo da atividade antimalárica de extratos das folhas de Ouratea nitida Aubl. Análise qualitativa através de CG-EM de uma fração apolar do extrato em hexano também foi efetuada. Ésteres metílicos e etílicos dos ácidos laúrico, mirístico, palmítico, esteárico e oléico, metílicos dos ácidos pentadecanóico, heptadecanóico, araquidônico, behênico e lignocérico e o etílico do ácido linoléico foram os componentes majoritários; enquanto que o estearato de n-butila, o tetracosano e a 6,10,14-trimetil-2-pentadecanona foram os minoritários. Os compostos isolados foram identificados com base na análise dos dados espectrais (IV, EM e RMN, incluindo DEPT) e estão sendo descritos pela primeira vez nesta espécie.

This work describes the isolation of triterpenes (3,4-seco-friedelan-3-oic acid, friedelin, and b-amyrin) and other compounds (p-hydroxybenzoic acid ethyl ester and tetracosane) as well as a preliminary evaluation of in vivo antimalarial activity of the extracts from the leaves of Ouratea nitida Aubl. Qualitative analysis by GC-MS of an apolar fraction from the hexane extract was also carried out. Methyl and ethyl esters of lauric, myristic, palmitic, stearic and oleic acids, methyl ester of pentadecanoic, heptadecanoic, arachidonic, beenic and lignoceric acids, and ethyl ester of linoleic acid were found to be the main constituents while n-butyl stearate, tetracosane and 6,10,14-trimetthyl-2-pentadecanone were the minor. All isolated compounds were identified on basis of the spectral data (IR, MS and NMR, including DEPT) and are being described for the first time in this specie.