Antibacterial potencial of 12 Lichen species.

Resistant bacterial infections are a major public health problem worldwide, which entails the need to search for new therapeutic agents. In this context, lichens stand out, provided that they are producers of structurally diverse compounds that have attractive biological properties, including antimicrobial activity. Thus, extracts of 12 lichen species were prepared and their potential to inhibit the growth of 5 bacterial strains was evaluated in this work. The chemical compositions of these extracts were examined using TLC and microcrystallization, being the identity of the active compounds in each extract attributed based on the bioautography technique. The most active extracts (and their identified active compounds) were from Cladonia borealis (usnic, barbatic and 4-O-demethylbarbatic acids), Cladina confusa (usnic and perlatolic acids), Stereocaulom ramulosum (atranorin, perlatolic and anziaic acids) and Canoparmelia cryptochlorophaea (cryptochlorophaeic and caperatic acids), with MICs ranging from 7.8 to 31.25 µg/mL, including for resistant clinical strains. MIC values ​​were also obtained for substances isolated from lichens for comparison purposes. A group of four extracts containing usnic acid was analyzed by 1H NMR in order to correlate relative proportion of major metabolites and extracts activity. The less active extracts in this group, in fact, presented low proportion of usnic acid.

Molecular modeling and cytotoxicity of diffractaic acid: HP-ß-CD inclusion complex encapsulated in microspheres.

In this pioneer study, 2-hydroxypropyl-ß-cyclodextrin (HP-ß-CD) was used to improve the solubility of the diffractaic acid (DA) via inclusion complex (DA:HP-ß-CD). Subsequently, DA:HP-ß-CD was incorporated into poly-ε-caprolactone (PCL) microspheres (DA:HP-ß-CD-MS). Microspheres containing DA (DA-MS) or DA:HP-ß-CD (DA:HP-ß-CD-MS) were prepared using the multiple W/O/W emulsion-solvent evaporation technique. The phase-solubility diagram of DA in HP-ß-CD (10-50mM) showed an AL type curve with a stability constant K1:1=821M-1. 1H NMR, FTIR, X-ray diffraction and thermal analysis showed changes in the molecular environment of DA in DA:HP-ß-CD. The molecular modeling approach suggests a guest-host complex formation between the carboxylic moiety of both DA and the host (HP-ß-CD). The mean particle size of the microspheres were ∅DA-MS=5.23±1.65µm and ∅DA:HP-ß-CD-MS=4.11±1.39µm, respectively. The zeta potential values of the microspheres were ζDA-MS=-7.85±0.32mV and ζDA:HP-ß-CD-MS=-6.93±0.46mV. Moreover, the encapsulation of DA:HP-ß-CD into microspheres resulted in a more slower release (k2=0.042±0.001; r2=0.996) when compared with DA-MS (k2=0.183±0.005; r2=0.996). The encapsulation of DA or DA:HP-ß-CD into microspheres reduced the cytotoxicity of DA (IC50=43.29µM) against Vero cells (IC50 of DA-MS=108.48µM and IC50 of DA:HP-ß-CD-MS=142.63µM).

Influência da variação sazonal nos teores de flavonóides e atividade antioxidante das folhas de Campomanesia adamantium (Cambess.) O. Berg, Myrtaceae / The Influence of seasonal variation in levels of flavonoids and antioxidant activity of the leaves of Campomanesia adamantium

Campomanesia adamantium (Cambess.) O. Berg, Myrtaceae, é uma espécie nativa. As folhas são utilizadas na medicina popular. Os extratos hexânico e acetato de etila das folhas mostraram pouca variação na sua composição química em diferentes estações do ano associado com o desenvolvimento do vegetal, enquanto o extrato etanólico mostrou alteração significativa em relação à composição química. Todos os extratos etanólicos mostraram alta atividade antioxidante frente ao método DPPH e de moderada a alta para β-caroteno/ácido linoléico.

Campomanesia adamantium (Cambess.) O. Berg, Myrtaceae, is a native species. The extracts of ethyl acetate and hexane showed little variation in their chemical composition at different seasons of the year associated with the development of the plant, while the ethanol extract showed significant change in relationship to chemical composition. All ethanol extracts showed high antioxidant activity against DPPH method and from moderate to high antioxidant activity for β-carotene/linoleic acid.

Lichen metabolites modulate hydrogen peroxide and nitric oxide in mouse macrophages.

The activities of perlatolic acid (1), atranorin (2), and lecanoric acid (3) and their derivatives, such as orsellinates and beta-methyl orsellinates obtained by alcoholysis, were assessed for stimulation of the release of hydrogen peroxide and nitric oxide in cultures of peritoneal macrophage cells from mice. The hydrogen peroxide production was estimated by oxidation of phenol red, while the Griess reagent was used to determine the nitric oxide production. 1 and 4-methoxy-ethyl orsellinate (XVII) were the compounds that induced the greatest release of H2O2, whereas n-pentyl orsellinate (IV), iso-propyl orsellinate (V), sec-butyl orsellinate (VI), and XVII induced a small release of NO. These results indicate that lichen products and their derivatives have potential immune-modulating activities.

Immunostimulatory effects of the phenolic compounds from lichens on nitric oxide and hydrogen peroxide production / Efeito imunoestimulante de compostos fenólicos de líquens na produção de peróxido de hidrogênio e óxido nítrico

The effects of isolated compounds from Brazilian lichens and their derivatives on H2O2 and NO production were studied using murine macrophages as a part of an attempt to understand their possible immunomodulatory properties. The compound cytotoxicity was studied using MTT assay. Macrophage stimulation was evaluated by the determination of NO (Griess assay) and H2O2 (horseradish peroxidase/phenol red) in supernatants of peritoneal macrophage cultures of Swiss mice. This research demonstrated stimulatory activities of some phenolic compounds isolated from lichens and their derivatives on H2O2 and NO production. Structure-activity relationships suggest several synthetic directions for further improvement of immunological activity.

Os efeitos dos compostos isolados de líquens brasileiros e seus derivados na produção de NO e H2O2 foram estudados utilizando macrófagos murinos na tentativa de desvendar suas possíveis propriedades imunomodulatórias. A citotoxicidade dos compostos foi estudada utilizando o ensaio de MTT. A estimulação dos macrófagos foi avaliada através da determinação de NO (metodologia de Griess) e H2O2 (peroxidase de raíz forte/vermelho de fenol) no sobrenadante de culturas de macrófagos peritoneais de camundongos Swiss. Este estudo demonstrou atividade estimulante de alguns compostos fenólicos e seus derivados na produção de NO e H2O2. A relação estrutura atividade sugere inúmeras direções sintéticas para futuros melhoramentos da atividade imunológica.

Activities of 2,4-dihydroxy-6-n-pentylbenzoic acid derivatives.

Esters of 2-hydroxy-4-methoxy-6-n-pentylbenzoic acid (2-8) (methyl, ethyl, butyl, pentyl, isopropyl, sec-butyl and benzyl), olivetol (9), methyl, ethyl, butyl perlatolates (10-12), 2,4-dihydroxy-6-n-pentylbenzoic acid (15), and methyl and ethyl esters of (15) were prepared through structural modifications of perlatolic acid (1) with the aim to detect new antifungal and antibacterial substances and also to evaluate the toxicity by the brine shrimp lethality assay against Artemia salina. The antifungal assays were carried out against the fungus Cladosporium sphaerospermum through the bioautography method, and methyl 2,4-dihydroxy-6-n-pentylbenzoate (13) showed the highest antifungal activity (2.5 yg). Olivetol (9) and 2,4-dihydroxy-6-n-pentylbenzoic acid (15) are also potent inhibitors of the growth of the fungus (5.0 microg). Except for methyl (10), the ethyl (11) and butyl (12) perlatolates were less active than perlatolic acid (1). The activities presented by methyl (2) and ethyl (3) 2-hydroxy-4-methoxy-6-n-pentylbenzoates and methyl (13) and ethyl (14) 2,4-dihydroxy-6-n-pentylbenzo-ates suggest that compounds with a free hydroxy group in the aromatic ring (C-4) have a more pronounced effect against C. sphaerospermum. Antibacterial activities were tested by the disc diffusion method using pathogenic strains of S. aureus and E. coli. The compounds were weakly active with inhibition zones between 9-15 mm. The 2-hydroxy-4-methoxy-6-n-pentylbenzoic esters 2-8 and alkyl perlatolates 10-12 were selective against E. coli. Perlatolic acid (1) and methyl 2-hydroxy-4-methoxy-6-n-pentylbenzoate (2) were the most active with LD50 values of 24.1 microM and 27.2 microM, respectively. The other compounds were not toxic to Artemia salina larvae.

Cytotoxic activity of orsellinates.

The series of 2,4-dihydroxy-6-methylbenzoates 2-10 (methyl to hexyl orsellinates) prepared by alcoholysis of lecanoric acid (1)--a natural product from the lichen Parmotrema tinctorum (Nyl.) Hale - was submitted to the brine shrimp lethality test (BST), which was also performed for 2,4-dihydroxy-6-methylbenzoic acid (11) (orsellinic acid) and the derivative ethyl-2-hydroxy-4-methoxy-6-methylbenzoate (12) (4-methoxy-ethyl orsellinate), in order to detect new substances with probable antineoplasic activity. Results showed that chain elongation--increase in lipophilicity (log P)--causes a rise in the cytotoxic activity of orsellinates. Hexyl orsellinate (7) showed the highest cytotoxic activity (LC50 = 31 microM). A correlation between lipophilicity (log P) and cytotoxic activity (log 1/LC50) is presented. Compounds with ramified chains--iso-propyl, sec-butyl and tert-butyl orsellinates (8-10)--were less active than those with the correspondent linear chain. The activities presented by 4-methoxy-ethyl orsellinate (12) and ethyl orsellinate (3) suggest that the hydroxy group at the C-4 position causes effect in the cytotoxic activity of orsellinates against Artemia salina.

Simultaneous determination of furanocoumarins in infusions and decoctions from "Carapiá" (dorstenia species) by high-performance liquid chromatography.

The use of furanocoumarins, which are photosensitizing compounds, combined with exposure to UV-A radiation is a common treatment for vitiligo, psoriasis, and a number of other skin diseases. Although furanocoumarins plus UV-A treatment is highly effective, several studies have shown that exposure to high doses increases the risk to development of cutaneus carcinoma. Several Dorstenia species are used in folk medicine, mainly against skin diseases, because of the presence of biologically active compounds. We present here analysis of the chemical composition of furanocoumarins from infusion and decoction of "Carapiá" (Dorstenia species), which is used in Brazil against several diseases. We have employed high-performance liquid chromatography (HPLC) procedures for the quantitative determination of psoralen, bergapten, and isopimpinellin. The contents of furanocoumarins revealed an insignificant difference between infusion and decoction. Dorstenia tubicina and D. asaroides contained psoralen and bergapten only in the rhizomes, whereas D. vitifolia shows solely isopimpinellin in both rhizomes and aerial parts.

Simple and rapid determination of psoralens in topic solutions using liquid chromatography.

A method for sample preparation and high-performance liquid chromatography detection and quantification has been developed for the routine analysis of psoralen and bergapten, photosensitizing compounds, in topic solutions employed in Brazil for treatment of vitiligo and psoriasis. The linearity, accuracy, the inter- and intra-day precision of the procedure were evaluated. The calibration curve for psoralen and bergapten were linear in the range 1.0-20.0 microg ml(-1). The best recoveries of the psoralens in the topic solutions analysed were 96.3-98.2%. The percentage coefficient of variation of the quantitative analysis of the psoralens in the product analyses was within 5%.

Bioactive derivatives obtained from lecanoric acid, a constituent of the lichen Parmotrema tinctorum (Nyl. ) Hale (Parmeliaceae)

With the aim of obtaining new compounds with potential antifungal activity, lecanoric acid, a chemical constituent of the lichen Parmotrema tinctorum and its derivatives prepared from structural modification were tested against the fungus Cladosporium sphaerospermum, by employing the bioautographic method. Activity of the derivatives ranged from 10- to 1-ìg concentrations. Results demonstrated this series of compounds to have potent fungitoxic activity.