Evaluation of Cytotoxicity and Antifungal Activity of Friedelanes from Salacia elliptica Roots.

Plants from Salacia genus are used in traditional medicine for a wide range of diseases. Previous studies reported bioactive pentacyclic triterpenoids from S. elliptica leaves and branches. In this study, the novel pentacyclic triterpenoid 7α,15α-dihydroxyfriedelan-3-one (1) was obtained from the roots of Salacia elliptica, along with seven known compounds: friedelan-3-one (2), friedelan-3ß-ol (3), friedelan-1,3-dione (4), friedelan-3,15-dione (5), 15α-hydroxyfriedelan-3-one (6), 15α,26-dihydroxyfriedelan-3-one (7), and 26-hydroxyfriedelan-3,15-dione (8). Additionally, one steroid, spinasterol (9), was also identified. The chemical structures of all compounds were established through 1 H and 13 C-NMR. Compound 1 was analysed by additional 2D experiments (HMBC, HSQC, COSY, and NOESY) for complete elucidation. Furthermore, the cytotoxicity of compounds 2, 3, 6, 7 and 8 against the A549 lung cancer cells model was evaluated. The flow cytometry analysis revealed a significant cytotoxic activity similar to that exhibited by the triterpenoid lupeol. Additionally, compounds 2, 3, 6, and 7 were tested for in vitro antifungal activity against Candida, Cryptococcus and Sporothrix strains. However, all compounds showed no activity at the tested concentrations.

Antivirulence activity and in vivo efficacy of a thiazole derivative against candidiasis.

Candida albicans is a pathogen equipped with a variety of commensal and virulence traits that help it colonize the microbiota and invade host tissue during infection. In this study, we investigated the potential anticandidal activity of 3-[2-(4-(4-methoxyphenyl)thiazol-2-yl)hydrazino)]butan-1-ol (MT), a thiazolylhydrazone compound synthesized by our group, and identified it as a promising antifungal agent. The activity of MT was evaluated in vitro and in vivo against C. albicans as well as its ability to inhibit virulence factors. For this, the ability of MT to inhibit the adhesion of C. albicans to human buccal epithelial cells and biofilm formation and filamentation was tested. In addition, the potential in vivo activity of MT was evaluated in murine models of oral candidiasis. Our results confirmed the antifungal activity of MT, with a minimal inhibitory concentration range of 0.5-2 µg/mL. Indeed, MT treatment in vitro decreased the expression of C. albicans genes involved in biofilm formation and morphogenesis and encoding hydrolytic enzymes, which was also confirmed through phenotypic observations. In addition, MT promoted a decrease in the colony forming units recovered from the tongues of mice with oral candidiasis. In this work, we present a potent antivirulence compound that shows potential for candidiasis therapy, especially for topical use.

Study and Development of an Anthroposophical Formula Based on Phosphorus and Formica rufa for Onychomycosis´s Treatment.

Onychomycosis is a fungal infection of the nail plate or nail bed that leads to the gradual destruction of the nail. The main difficulties in the treatment of onychomycosis refer to the duration of treatments and their side effects. Thus, it becomes relevant to look for new therapeutic alternatives in the treatment of such common diseases that are efficient without causing the undesirable side effects on the patient's body. In this way, the objective of this study was to develop an anthroposophical formula for the treatment of onychomycosis, based on Phosphorus and Formica rufa, from an extensive bibliographic survey on the functions of these components, evaluating within the principles of Anthroposophy. Considering the set of knowledge and practices on the use of these components, it was possible to arrive at a proposal therapy that can be effective for the treatment of onychomycosis. After an extensive review of several existing patents, it was observed that formulations containing Phosphorus and Formica rufa together have not been described in other studies. Subsequently, our research group published a patent of the anthroposophical formula using these two components, with the number BR1020180750755, which will be efficient to help the recovery of nails, and facilitate normal growth.

Antifungal and antioxidant activity of fatty acid methyl esters from vegetable oils.

Fatty acid methyl esters (FAMEs) were obtained from vegetable oils of soybean, corn and sunflower. The current study was focused on evaluating the antifungal activity of FAMEs mainly against Paracoccidioides spp., as well as testing the interaction of these compounds with commercial antifungal drugs and also their antioxidant potential. FAMEs presented small IC50 values (1.86-9.42 µg/mL). All three FAMEs tested showed antifungal activity against isolates of Paracoccidioides spp. with MIC values ranging from 15.6-500 µg/mL. Sunflower FAMEs exhibited antifungal activity that extended also to other genera, with an MIC of 15.6 µg/mL against Candida glabrata and C. krusei and 31.2 µg/mL against C. parapsilosis. FAMEs exhibited a synergetic effect with itraconazole. The antifungal activity of the FAMEs against isolates of Paracoccidioides spp. is likely due to the presence of methyl linoleate, the major compound present in all three FAMEs. The results obtained indicate the potential of FAMEs as sources for antifungal and antioxidant activity.

Antifungal and antioxidant activity of fatty acid methyl esters from vegetable oils

ABSTRACT Fatty acid methyl esters (FAMEs) were obtained from vegetable oils of soybean, corn and sunflower. The current study was focused on evaluating the antifungal activity of FAMEs mainly against Paracoccidioides spp., as well as testing the interaction of these compounds with commercial antifungal drugs and also their antioxidant potential. FAMEs presented small IC50 values (1.86-9.42 μg/mL). All three FAMEs tested showed antifungal activity against isolates of Paracoccidioides spp. with MIC values ranging from 15.6-500 µg/mL. Sunflower FAMEs exhibited antifungal activity that extended also to other genera, with an MIC of 15.6 μg/mL against Candida glabrata and C. krusei and 31.2 μg/mL against C. parapsilosis. FAMEs exhibited a synergetic effect with itraconazole. The antifungal activity of the FAMEs against isolates of Paracoccidioides spp. is likely due to the presence of methyl linoleate, the major compound present in all three FAMEs. The results obtained indicate the potential of FAMEs as sources for antifungal and antioxidant activity.

The diversity and antimicrobial activity of endophytic fungi associated with medicinal plant Baccharis trimera (Asteraceae) from the Brazilian savannah.

The fungal endophyte community associated with Baccharis trimera, a Brazilian medicinal plant, was characterized and screened for its ability to present antimicrobial activity. By using molecular methods, we identified and classified the endophytic fungi obtained into 25 different taxa from the phyla Ascomycota and Basidiomycota. The most abundant species were closely related to Diaporthe phaseolorum, Pestalotiopsis sp. 1, and Preussia pseudominima. The differences observed in endophytic assemblages from different B. trimera specimens might be associated with their crude extract activities. Plants that had higher α-biodiversity were also those that contributed more to the regional (γ) diversity. All fungal isolates were cultured and their crude extracts screened to examine the antimicrobial activities. Twenty-three extracts (12.8%) displayed antimicrobial activities against at least one target microorganism. Among these extracts, those obtained from Epicoccum sp., Pestalotiopsis sp. 1, Cochliobolus lunatus, and Nigrospora sp. presented the best minimum inhibitory concentration values. Our results show that the endophytic fungal community associated with the medicinal plant B. trimera included few dominant bioactive taxa, which may represent sources of compounds with antifungal activity. Additionally, the discovery of these bioactive fungi in association with B. trimera suggests that Brazilian plants used as folk medicine may shelter a rich fungal diversity as well as taxa able to produce bioactive metabolites with antimicrobial activities.

In vitro antifungal activities of leaf extracts of Lippia alba (Verbenaceae) against clinically important yeast species.

INTRODUCTION: There are few studies reporting the antifungal activities of Lippia alba extracts. METHODS: A broth microdilution assay was used to evaluate the antifungal effects of Lippia alba extracts against seven yeast species of Candida and Cryptococcus. The butanol fraction was investigated by gas chromatography-mass spectrometry. RESULTS: The butanol fraction showed the highest activity against Candida glabrata. The fraction also acted synergistically with itraconazole and fluconazole against C. glabrata. The dominant compounds in the butanol fraction were 2,2,5-trimethyl-3,4-hexanedione, 3,5-dimethyl-4-octanone and hexadecane. CONCLUSIONS: The butanol fraction may be a good candidate in the search for new drugs from natural products with antifungal activity.

Activity of compounds isolated from Baccharis dracunculifolia D.C. (Asteraceae) against Paracoccidioides brasiliensis.

Paracoccidioidomycosis is a prevalent systemic mycosis in Latin America which requires prolonged treatment with highly toxic antifungals. Baccharis dracunculifolia is a medicinal plant in Brazil that is a candidate in the search for new drugs. Fractions of the hexanic extracts were obtained using chromatographic procedures and assessed using an antifungal assay with Paracoccidioides brasiliensis (Pb18), tumor cell lines and amastigote forms of Leishmania, L. amazonensis. Four compounds were isolated, i.e., ursolic acid (1), methyl linolenate (2), caryophyllene oxide (3), and trans-nerolidol (4). Compounds 2, 3 and 4 displayed antifungal activity against four isolates of Paracocci dioides with MIC values ranging from 3.9-250 µg/ml. Only caryophyllene oxide showed differences in the MIC values against Pb18 when the medium was supplemented with ergosterol, which suggested that the compound interacts with ergosterol. Ursolic acid was active in the cytotoxic assays and showed leishmanicidal activity. Scanning electron microscopy demonstrated that compounds 2, 3 and 4 decreased the cell size and produced an irregular cell wall surface on P. brasiliensis cells. The present results showed the biological activities of the isolated compounds and revealed that these compounds may affect the cell surface and growth of P. brasiliensis isolates.

In vitro antifungal activity of fatty acid methyl esters of the seeds of Annona cornifolia A.St.-Hil. (Annonaceae) against pathogenic fungus Paracoccidioides brasiliensis / Atividade antifúngica in vitro dos ésteres metílicos de ácidos graxos das sementes de Annona cornifolia A.St.-Hil. (Annonaceae) sobre o fungo patogênico Paracoccidioides brasiliensis

INTRODUCTION: Fatty acids are abundant in vegetable oils. They are known to have antibacterial and antifungal properties. METHODS: Antifungal susceptibility was evaluated by broth microdilution assay following CLSI (formerly the NCCLS) guidelines against 16 fungal strains of clinical interest. RESULTS: In this work, fatty acid methyl esters (FAME) was able to inhibit 12 clinical strains of the pathogenic fungus Paracoccidioides brasiliensis and were also active in the bioautographic assay against Cladosporium sphaerospermum. CONCLUSIONS: FAME was a more potent antifungal than trimethoprim-sulfamethoxazole against P. brasiliensis under the experimental conditions tested.

INTRODUÇÃO: Os ácidos graxos são abundantes em óleos vegetais. Eles são conhecidos por suas propriedades antibacterianas e antifúngicas. MÉTODOS: A suscetibilidade a antifúngicos foi avaliada pelo ensaio de microdiluição em caldo de acordo com CLSI (anteriormente NCCLS) sobre 16 isolados de interesse clínico. RESULTADOS: Nesse trabalho, os ésteres metílicos de ácidos graxos (FAME) inibiram doze isolados clínicos do fungo patogênico Paracoccidioides brasiliensis, e também foi muito ativo no ensaio de bioautografia sobre o fungo Cladosporium sphaerospermum. CONCLUSÕES: FAME foi um antifúngico mais potente do que sulfametoxazol-trimetoprim contra P. brasiliensis, nas condições utilizadas no presente trabalho.

In vitro antifungal activity of fatty acid methyl esters of the seeds of Annona cornifolia A.St.-Hil. (Annonaceae) against pathogenic fungus Paracoccidioides brasiliensis.

INTRODUCTION: Fatty acids are abundant in vegetable oils. They are known to have antibacterial and antifungal properties. METHODS: Antifungal susceptibility was evaluated by broth microdilution assay following CLSI (formerly the NCCLS) guidelines against 16 fungal strains of clinical interest. RESULTS: In this work, fatty acid methyl esters (FAME) was able to inhibit 12 clinical strains of the pathogenic fungus Paracoccidioides brasiliensis and were also active in the bioautographic assay against Cladosporium sphaerospermum. CONCLUSIONS: FAME was a more potent antifungal than trimethoprim-sulfamethoxazole against P. brasiliensis under the experimental conditions tested.

Antifungal activity of schinol and a new biphenyl compound isolated from Schinus terebinthifolius against the pathogenic fungus Paracoccidioides brasiliensis.

BACKGROUND: The aim of this study was to isolate and identify the antifungal compounds from the extracts of Schinus terebinthifolius (Anacardiaceae) against clinical isolates of the pathogenic fungus Paracoccidioides brasiliensis. METHODS: The hexane and dichlomethane fractions from leaves and stems of S. terebinthifolius were fractionated using several chromatography techniques to afford four compounds. RESULTS: The compounds isolated from S. terebinthifolius were identified as schinol (1), a new biphenyl compound, namely, 4'-ethyl-4-methyl-2,2',6,6'-tetrahydroxy[1,1'-biphenyl]-4,4'-dicarboxylate (2), quercetin (3), and kaempferol (4). Compounds 1 and 2 were active against different strains of P. brasiliensis, showing a minimal inhibitory concentration value against the isolate Pb B339 of 15.6 µg/ml. The isolate Pb 1578 was more sensitive to compound 1 with a MIC value of 7.5 µg/ml. Schinol presented synergistic effect only when combined with itraconazole. The compounds isolated from S. terebinthifolius were not able to inhibit cell wall synthesis or assembly using the sorbitol assay. CONCLUSION: This work reveals for the first time the occurrence of compound 2 and discloses activity of compounds 1 and 2 against several clinical isolates of P. brasiliensis. These results justify further studies to clarify the mechanisms of action of these compounds.

Antifungal activity of extracts of some plants used in Brazilian traditional medicine against the pathogenic fungus Paracoccidioides brasiliensis.

Paracoccidioidomycosis (PCM) is a systemic granulomatous disease caused by Paracoccidioides brasiliensis Almeida (Onygenales) that requires 1-2 years of treatment. In the absence of drug therapy, the disease is usually fatal, highlighting the need for the identification of safer, novel, and more effective antifungal compounds. With this need in mind, several plants employed in Brazilian traditional medicine were assayed on P. brasiliensis and murine macrophages. Extracts were prepared from 10 plant species: Inga spp. Mill. (Leguminosae), Schinus terebinthifolius Raddi (Anacardiaceae), Punica granatum L. (Punicaceae), Alternanthera brasiliana Kuntze (Amaranthaceae), Piper regnellii CDC. (Piperaceae), P. abutiloides Kunth (Piperaceae), Herissantia crispa L. Briz. (Malvaceae), Rubus urticaefolius Poir (Rosaceae), Rumex acetosa L. (Polygonaceae), and Baccharis dracunculifolia DC. (Asteraceae). Hexane fractions from hydroalcoholic extracts of Piper regnellii and Baccharis dracunculifolia were the most active against the fungus, displaying minimum inhibitory concentration (MIC) values of 7.8 microg/mL and 7.8-30 mug/mL, respectively. Additionally, neither of the extracts exhibited any apparent cytotoxic effects on murine macrophages at 20 microg/mL. Analyses of these fractions using gas chromatography-mass spectrometry (GC-MS) showed that the major components of B. dracunculifolia were ethyl hydrocinnamate (14.35%) and spathulenol (16.02%), while the major components of the hexane fraction of Piper regnellii were 1-methoxy-4-(1-propenyl) benzene (21.94%) and apiol (21.29%). The activities of these fractions against P. brasiliensis without evidence of cytotoxicity to macrophages justify their investigation as a potential source of new chemical agents for the treatment of PCM.

Antifungal properties of plants used in Brazilian traditional medicine against clinically relevant fungal pathogens

Antifungal properties of extracts from eight Brazilian plants traditionally used in popular Brazilian medicine were tested against five clinically relevant Candida species, Cryptococcus neoformans, and Sporothrix schenckii. Results demonstrate that almost all extracts exhibited antifungal activity, at least against one of the microorganisms tested. The ethanolic extract from the leaves of Schinus terebinthifolius exhibited potential antifungal activity against C. glabrata and S. schenckii. Preliminary phytochemical analysis of extract from S. terebinthifolius showed the presence of biologically active compounds, namely saponins, flavonoids, triterpenes, steroids and tannins.

A propriedade antifúngica de extratos de oito plantas utilizadas na medicina tradicional brasileira foi testada contra cinco espécies de Candida, com relevância clínica, Cryptococcus neoformans e Sporothrix schenckii. Os resultados mostraram que todos os extratos exibiram atividade antifúngica contra pelo menos um dos microrganismos testados. O extrato etanólico das folhas de Schinus terebinthifolius apresentou potencial atividade antifúngica contra C. glabrata e S. schenckii. Na análise fitoquímica preliminar dos extratos de S. terebinthifolius observou-se a presença de compostos biologicamente ativos como, flavonóides, triterpenos, esteróides e taninos.

Antimicrobial activity of wax and hexane extracts from Citrus spp. peels.

Antibacterial and antifungal properties of wax and hexane extracts of Citrus spp. peels were tested using bioautographic and microdilution techniques against three plant pathogenic fungi (Penicillium digitatum, Curvularia sp., and Colletotrichum sp.), two human pathogens (Trichophyton mentagrophytes and Microsporum canis), and two opportunistic bacteria (Escherichia coli and Staphylococcus aureus). Two polymethoxylated flavonoids and a coumarin derivative, were isolated and identified from peel extracts, which presented antimicrobial activity especially against M. canis and T. mentagrophytes: 4',5,6,7,8-pentamethoxyflavone (tangeritin) and 3',4',5,6,7,8-hexamethoxyflavone (nobiletin) from C. reticulata; and 6,7-dimethoxycoumarin (also known as escoparone, scoparone or scoparin) from C. limon.

Antimicrobial activity of wax and hexane extracts from Citrus spp. peels

Antibacterial and antifungal properties of wax and hexane extracts of Citrus spp. peels were tested using bioautographic and microdilution techniques against three plant pathogenic fungi (Penicillium digitatum, Curvularia sp., and Colletotrichum sp.), two human pathogens (Trichophyton mentagrophytes and Microsporum canis), and two opportunistic bacteria (Escherichia coli and Staphylococcus aureus). Two polymethoxylated flavonoids and a coumarin derivative, were isolated and identified from peel extracts, which presented antimicrobial activity especially against M. canis and T. mentagrophytes: 4',5,6,7,8-pentamethoxyflavone (tangeritin) and 3',4',5,6,7,8-hexamethoxyflavone (nobiletin) from C. reticulata; and 6,7-dimethoxycoumarin (also known as escoparone, scoparone or scoparin) from C. limon.