Evaluation of the In Vitro Antiparasitic Effect of the Essential Oil of Cymbopogon winterianus and Its Chemical Composition Analysis.

Cymbopogon winterianus, known as "citronella grass", is an important aromatic and medicinal tropical herbaceous plant. The essential oil of C. winterianus (EOCw) is popularly used to play an important role in improving human health due to its potential as a bioactive component. The present study aimed to identify the components of the essential oil of C. winterianus and verify its leishmanicidal and trypanocidal potential, as well as the cytotoxicity in mammalian cells, in vitro. The EOCw had geraniol (42.13%), citronellal (17.31%), and citronellol (16.91%) as major constituents. The essential oil only exhibited significant cytotoxicity in mammalian fibroblasts at concentrations greater than 250 µg/mL, while regarding antipromastigote and antiepimastigote activities, they presented values considered clinically relevant, since both had LC50 < 62.5 µg/mL. It can be concluded that this is a pioneer study on the potential of the essential oil of C. winterianus and its use against the parasites T. cruzi and L. brasiliensis, and its importance is also based on this fact. Additionally, according to the results, C. winterianus was effective in presenting values of clinical relevance and low toxicity and, therefore, an indicator of popular use.

Cytotoxicity of Essential Oil Cordia verbenaceae against Leishmania brasiliensis and Trypanosoma cruzi.

The species Cordia verbenacea DC (Boraginaceae), known as the whaling herb and camaradinha, is a perennial shrub species native to the Atlantic Forest. Its leaves are used in folk medicine as an anti-inflammatory, analgesic, antiulcerogenic and curative agent, in the form of teas or infusions for internal or topical use. The present study aimed to verify the cytotoxicity of the essential oil and the leishmanicidal and trypanocidal potential of C. verbenacea. The essential oil was characterized by GC-MS. The in vitro biological activity was determined by anti-Leishmania and anti-Trypanosoma assays. The cytotoxixity was determined using mammalian fibroblasts. The C. verbenacea species presented α-pinene (45.71%), ß-caryophyllene (18.77%), tricyclo[2,2,1-(2.6)]heptane (12.56%) as their main compounds. The essential oil exhibited strong cytotoxicity at concentrations below 250 µg/mL (LC50 138.1 µg/mL) in mammalian fibroblasts. The potent anti-trypanosome and anti-promastigote activities occurred from the concentration of 62.5 µg/mL and was considered clinically relevant. The results also demonstrate that at low concentrations (<62.5 µg/mL), the essential oil of C. verbenacea managed to be lethal for these activities. This can be considered an indication of the power used in daily human consumption. Therefore, it can be concluded that the essential oil of C. verbenacea contains a compound with remarkable antiparasitic activities and requires further research.

Ximenia americana L. enhances the antibiotic activity and inhibit the development of kinetoplastid parasites.

OBJECTIVE: The objective of this work was evaluate the cytotoxic, leishmanicidal and tripanocidal activity, as well as to evaluate its antimicrobial and modulatory activity in association with different antibiotics of the hydroethanolic extract of the Ximenia Americana stem bark (EHXA). METHOD: In vitro tests against Trypanosoma cruzi, Leishmania sp. and citotoxicity were performed. The evaluation of the antibacterial and bacterial resistance modulatory effect was given by the microdilution method. RESULTS: The chemical profile show different classes of compounds with significant presence of quercetrin and caffeic acid. The EHXA demonstrated activity only in the concentration of 1000 µg/mL against the L. infantum and L. brasiliensis promastigotes, causing mortality percentage of 40.66 and 27.62%, respectively. The extract presented a significant toxicity only in the concentration of 1000 µg/mL, causing a mortality of 55.42% of fibroblasts. The antibacterial activity of the EHXA demonstrated a MIC value ≥1024 µg/mL against all the tested bacteria. However, in the modulation assay with EHXA in association with different antibiotics the extract had a synergistic effect against S. aureus strains when associated with norfloxacin. CONCLUSION: The results of this investigation demonstrate for the first time the chemical composition of the hydroethanolic extract of the Ximenia Americana stem bark, your potential antiparasitic and modulatory effect. The low cytotoxic and biological potential against S. aureus open therapeutic perspectives against leishmaniosis and bacterial infections.

Antitrypanosomal, antileishmanial and cytotoxic activities of Brazilian red propolis and plant resin of Dalbergia ecastaphyllum (L) Taub.

The treatment for leishmaniasis and Chagas disease can be hard and painful, such that many patients give up on the treatment. In order to find an alternative path for the treatment of these diseases, researchers are using natural products to fight these parasites. The aim of this study was to evaluate the antiprotozoan and cytotoxic activities of red propolis samples collected from different Brazilian states and seasons whilst searching for possible activity differences. We also compared the red propolis results with the ones obtained for the plant resin extract collected from Dalbergia ecastaphyllum trees. The hydroethanolic red propolis extracts from Pernambuco and Alagoas, and the D. ecastaphyllum resin were evaluated regarding their antileishmanial, antitrypanosomal and cytotoxic activity. All extracts showed antiprotozoan and cytotoxic activity. RP-PER showed to be more cytotoxic against protozoan parasites and fibroblast cells. All propolis extracts showed a higher cytotoxic activity when compared to resin extracts. The propolis sample collected in Pernambuco during the rainy season killed the parasites with lower concentrations than the sample collected in the dry season. The IC50 observed against the parasites could be used without high fibroblast cell damage.

Antiparasitic effect of the Psidium guajava L. (guava) and Psidium brownianum MART. EX DC. (araçá-de-veado) extracts.

In the search for new therapeutic agents against neglected diseases, both aqueous and hydroethanolic extracts from Psidium guajava L. and P. brownianum Mart ex DC leaves were investigated regarding their antiparasitic effect and cytotoxic potential. The extracts were tested at three concentrations (250, 500 and 1000 µg/mL) against Trypanosoma cruzi epimastigote forms (Chagas, 1909), Leishmania braziliensis (Vianna, 1911) and L. infantum promastigotes forms (Nicolle, 1908), as well as against fibroblasts. P. guajava showed no activity against T. cruzi forms, while the hydroethanolic (PBHE), aqueous by decoction (PBAED) and aqueous by infusion (PBAEI) P. browninaum extracts were responsible, respectively, for inhibiting 100, 100 and 92.68% of T. cruzi epimastigote growth at the 1000 µg/mL concentration. The P. brownianum hydroethanolic extract (PBHE) at the highest concentration caused 58.46% death in L. braziliensis, thus demonstrating moderate activity, however when tested against L. infantum, the PBHE inhibited their growth by 37.16%, revealing its low activity. As for the cytotoxicity assays, the P. brownianum aqueous extract by decoction (PBAED) obtained the highest death percentage when compared to the others, causing 90.85% fibroblast mortality at the 1000 µg/mL concentration.

Multi-Anti-Parasitic Activity of Arylidene Ketones and Thiazolidene Hydrazines against Trypanosoma cruzi and Leishmania spp.

A series of fifty arylideneketones and thiazolidenehydrazines was evaluated against Leishmania infantum and Leishmania braziliensis. Furthermore, new simplified thiazolidenehydrazine derivatives were evaluated against Trypanosoma cruzi. The cytotoxicity of the active compounds on non-infected fibroblasts or macrophages was established in vitro to evaluate the selectivity of their anti-parasitic effects. Seven thiazolidenehydrazine derivatives and ten arylideneketones had good activity against the three parasites. The IC50 values for T. cruzi and Leishmania spp. ranged from 90 nM-25 µM. Eight compounds had multi-trypanocidal activity against T. cruzi and Leishmania spp. (the etiological agents of cutaneous and visceral forms). The selectivity of these active compounds was better than the three reference drugs: benznidazole, glucantime and miltefosine. They also had low toxicity when tested in vivo on zebrafish. Trying to understand the mechanism of action of these compounds, two possible molecular targets were investigated: triosephosphate isomerase and cruzipain. We also used a molecular stripping approach to elucidate the minimal structural requirements for their anti-T. cruzi activity.

The Paraguayan Rhinella toad venom: Implications in the traditional medicine and proliferation of breast cancer cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Toads belonging to genus Rhinella are used in Paraguayan traditional medicine to treat cancer and skin infections. AIM OF THE STUDY: The objective of the study was to determine the composition of venoms obtained from three different Paraguayan Rhinella species, to establish the constituents of a preparation sold in the capital city of Paraguay to treat cancer as containing the toad as ingredient, to establish the effect of the most active Rhinella schneideri venom on the cell cycle using human breast cancer cells and to assess the antiprotozoal activity of the venoms. METHODS: The venom obtained from the toads parotid glands was analyzed by HPLC-MS-MS. The preparation sold in the capital city of Paraguay to treat cancer that is advertised as made using the toad was analyzed by HPLC-MS-MS. The effect of the R. schneideri venom and the preparation was investigated on human breast cancer cells. The antiprotozoal activity was evaluated on Leishmania braziliensis, L. infantum and murine macrophages. RESULTS: From the venoms of R. ornata, R. schneideri and R. scitula, some 40 compounds were identified by spectroscopic and spectrometric means. Several minor constituents are reported for the first time. The preparation sold as made from the toad did not contained bufadienolides or compounds that can be associated with the toad but plant compounds, mainly phenolics and flavonoids. The venom showed activity on human breast cancer cells and modified the cell cycle proliferation. The antiprotozoal effect was higher for the R. schneideri venom and can be related to the composition and relative ratio of constituents compared with R. ornata and R. scitula. CONCLUSIONS: The preparation sold in the capital city of Paraguay as containing the toad venom, used popularly to treat cancer did not contain the toad venom constituents. Consistent with this, this preparation was inactive on proliferation of human breast cancer cells. In contrast, the toad venoms of Rhinella species altered the cell cycle progression, affecting the proliferation of malignant cells. The findings suggest that care should be taken with the providers of the preparation and that the crude drug present a strong activity towards human breast cancer cell lines. The antiprotozoal effect of the R. schneideri venom was moderate while the venom of R. ornata was devoid of activity and that of R. scitula was active at very high concentration.

A citotoxicidade e a avaliação da atividade antiprotozoária de Melissa officcinalis L (CIDRO-MELISA) / Actividad citotóxica y evaluación antiparasitaria de Melissa officcinalis L. (CIDRO-MELISA) / The cytotoxicity activity and evaluation of antiprotozoa Melissa officcinalis L (CIDRO-MELISA)

Introdução: as Leishmanioses correspondem a um complexo de doenças tropicais causada por mais de vinte diferentes espécies de protozoários intracelulares pertencentes ao gênero Leishmania. A Doença de Chagas é uma doença que representa um dos principais problemas de saúde pública em vários países, afetando sobretudo a população carente e rural. Melissa officinalis L., é uma erva perene, uma planta exótica, cuja família é Lamiaceae, que tem sido amplamente utilizada na medicina popular com finalidades terapêuticas. O seu óleo essencial tem sido usado como antioxidante, antimicótica, antifúngica, sedativa e antivirótica. Objetivo: avaliar a citotoxicidade, composição química e atividade antiparasitária do óleo essencial da espécie M. officinalis. Métodos: M. officcinalis foi coletada no município do Crato, CE, Brasil, e foi depositada no herbário da Universidade Regional do Cariri URCA. A análise da composição química do óleo essencial de M. officcinalis, foi realizada através de Cromatografia Gasosa acoplada à Espectrometria de Massas. No estudo da atividade leishmanicida in vitro, utilizou-se formas promastigotas de L. braziliensis e epimastigotas de Trypanosoma cruzi. Para a atividade citotóxica utilizou-se a linhagem de fibroblastos de mamífero NCTC clone 929. Resultados: a través da análise do óleo essencial de M. officcinalis feita por CG/EM, foi possível identificar 12 constituintes que totalizavam 99,83 por cento, dentre os quais alguns foram majoritários: Geranial, Z Citral, 1- Limonene e Carvone. Foi verificado baixa toxicidade contra os fibroblastos. M. officcinalis exibiu maior capacidade para impedir a replicação contra a forma promastigota do protozoário, nas concentrações testadas. Conclusão: a espécie pode ser uma fonte importante na busca de agentes novos e seletivos para o tratamento de doenças tropicais causadas por protozoários, cujas propriedades necessitam ser investigadas quanto aso seus mecanismos(AU)

Introducción: Leishmaniasis corresponde a un complejo de enfermedades tropicales causadas por más de veinte especies diferentes de protozoos intracelulares que pertenece al género Leishmania. La enfermedad de Chagas es una enfermedad que es uno de los principales problemas de salud pública en muchos países, que afectan especialmente a la población pobre y rural. Melissa officinalis L., es una hierba perenne, una planta exótica, cuya familia Lamiaceae, ha sido ampliamente utilizado en la medicina popular para fines terapéuticos. Su aceite esencial se ha usado como un antioxidante, antimicótico, antifúngica, antiviral y sedante. Objetivo: evaluar la citotoxicidad, la composición química y la actividad antiparasitaria del aceite esencial de la especie M. officinalis. Métodos: M. officinalis se recogió en el municipio de Crato, CE, Brasil, y fue depositado en el herbario de la Universidad Regional del Cariri URCA. El análisis de la composición química del aceite esencial de M. officcinalis se realizó por espectrometría de masa de cromatografía de gas. En el estudio de la actividad invitro leishmanicida se utilizó promastigotes L. braziliensis y Trypanosoma cruzi. Para la actividad citotóxica se utilizó la cepa de fibroblastos de mamífero NCTC clon 929. Resultados: a través de análisis de aceite esencial de M. officinalis hizo por GC/ EM se identificó un total de 12 constituyentes 99,83 por ciento, entre los cuales algunos eran mayoría: geranial, Z citral, limoneno y carvona 1. Se comprobó una baja toxicidad contra fibroblastos. M. officcinalis exhibió mayor capacidad para evitar la replicación contra la forma promastigote de parásito, a las concentraciones ensayadas. Conclusión: la especie puede ser una fuente importante en la búsqueda de nuevos agentes y selectivos para el tratamiento de enfermedades tropicales causadas por protozoos, cuyas propiedades deben ser investigados con respecto a sus mecanismos(AU)

Introduction: Leishmaniasis is a complex of tropical diseases caused by more than twenty different species of intracellular protozoa from the genus Leishmania. Chagas disease is one of the main public health problems in many countries. It mostly affects the poor, rural population. Melissa officinalis L. is a perennial herb, an exotic plant from the family Lamiaceae which has been widely used in folk medicine for therapeutic purposes. Its essential oil has been used as antioxidant, antifungal, antiviral and sedative. Objective: Evaluate the cytotoxicity, chemical composition and antiparasitic activity of essential oil of the species M. officinalis. Methods: M. officinalis specimens were collected from the municipality of Crato, CE, Brazil, and deposited in the herbarium at the Regional University of Cariri (URCA). Analysis of the chemical composition of essential oil of M. officinalis was performed by gas chromatography / mass spectrometry. L. braziliensis and Trypanosoma cruzi promastigotes were used for the study of in vitro leishmanicidal activity. Mammalian fibroblast strain NCTC clone 929 was used to evaluate cytotoxic activity. Results: Analysis of essential oil from M. officinalis by GC / MS led to identification of a total 12 constituents 99.83 percent, among which some were a majority: geranial, Z citral, limonene and 1-carvone. Toxicity against fibroblasts was found to be low. M. officinalis displayed greater capacity to avoid replication against the promastigote form of the parasite at the concentrations assayed. Conclusion: The species may be an important source of new agents and selective media for the treatment of tropical diseases caused by protozoa, whose properties should be studied with respect to their mechanisms(AU)

A citotoxicidade e a avaliação da atividade antiprotozoária de Melissa officcinalis L (CIDRO-MELISA) / The cytotoxicity activity and evaluation of antiprotozoa Melissa officcinalis L (CIDRO-MELISA)

Introdução: as Leishmanioses correspondem a um complexo de doenças tropicais causada por mais de vinte diferentes espécies de protozoários intracelulares pertencentes ao gênero Leishmania. A Doença de Chagas é uma doença que representa um dos principais problemas de saúde pública em vários países, afetando sobretudo a população carente e rural. Melissa officinalis L., é uma erva perene, uma planta exótica, cuja família é Lamiaceae, que tem sido amplamente utilizada na medicina popular com finalidades terapêuticas. O seu óleo essencial tem sido usado como antioxidante, antimicótica, antifúngica, sedativa e antivirótica. Objetivo: avaliar a citotoxicidade, composição química e atividade antiparasitária do óleo essencial da espécie M. officinalis. Métodos: M. officcinalis foi coletada no município do Crato, CE, Brasil, e foi depositada no herbário da Universidade Regional do Cariri URCA. A análise da composição química do óleo essencial de M. officcinalis, foi realizada através de Cromatografia Gasosa acoplada à Espectrometria de Massas. No estudo da atividade leishmanicida in vitro, utilizou-se formas promastigotas de L. braziliensis e epimastigotas de Trypanosoma cruzi. Para a atividade citotóxica utilizou-se a linhagem de fibroblastos de mamífero NCTC clone 929. Resultados: a través da análise do óleo essencial de M. officcinalis feita por CG/EM, foi possível identificar 12 constituintes que totalizavam 99,83 por cento, dentre os quais alguns foram majoritários: Geranial, Z Citral, 1- Limonene e Carvone. Foi verificado baixa toxicidade contra os fibroblastos. M. officcinalis exibiu maior capacidade para impedir a replicação contra a forma promastigota do protozoário, nas concentrações testadas. Conclusão: a espécie pode ser uma fonte importante na busca de agentes novos e seletivos para o tratamento de doenças tropicais causadas por protozoários, cujas propriedades necessitam ser investigadas quanto aso seus mecanismos(AU)

Introducción: Leishmaniasis corresponde a un complejo de enfermedades tropicales causadas por más de veinte especies diferentes de protozoos intracelulares que pertenece al género Leishmania. La enfermedad de Chagas es una enfermedad que es uno de los principales problemas de salud pública en muchos países, que afectan especialmente a la población pobre y rural. Melissa officinalis L., es una hierba perenne, una planta exótica, cuya familia Lamiaceae, ha sido ampliamente utilizado en la medicina popular para fines terapéuticos. Su aceite esencial se ha usado como un antioxidante, antimicótico, antifúngica, antiviral y sedante. Objetivo: evaluar la citotoxicidad, la composición química y la actividad antiparasitaria del aceite esencial de la especie M. officinalis. Métodos: M. officinalis se recogió en el municipio de Crato, CE, Brasil, y fue depositado en el herbario de la Universidad Regional del Cariri URCA. El análisis de la composición química del aceite esencial de M. officcinalis se realizó por espectrometría de masa de cromatografía de gas. En el estudio de la actividad invitro leishmanicida se utilizó promastigotes L. braziliensis y Trypanosoma cruzi. Para la actividad citotóxica se utilizó la cepa de fibroblastos de mamífero NCTC clon 929. Resultados: a través de análisis de aceite esencial de M. officinalis hizo por GC/ EM se identificó un total de 12 constituyentes 99,83 por ciento, entre los cuales algunos eran mayoría: geranial, Z citral, limoneno y carvona 1. Se comprobó una baja toxicidad contra fibroblastos. M. officcinalis exhibió mayor capacidad para evitar la replicación contra la forma promastigote de parásito, a las concentraciones ensayadas. Conclusión: la especie puede ser una fuente importante en la búsqueda de nuevos agentes y selectivos para el tratamiento de enfermedades tropicales causadas por protozoos, cuyas propiedades deben ser investigados con respecto a sus mecanismos(AU)

Introduction: Leishmaniasis is a complex of tropical diseases caused by more than twenty different species of intracellular protozoa from the genus Leishmania. Chagas disease is one of the main public health problems in many countries. It mostly affects the poor, rural population. Melissa officinalis L. is a perennial herb, an exotic plant from the family Lamiaceae which has been widely used in folk medicine for therapeutic purposes. Its essential oil has been used as antioxidant, antifungal, antiviral and sedative. Objective: Evaluate the cytotoxicity, chemical composition and antiparasitic activity of essential oil of the species M. officinalis. Methods: M. officinalis specimens were collected from the municipality of Crato, CE, Brazil, and deposited in the herbarium at the Regional University of Cariri (URCA). Analysis of the chemical composition of essential oil of M. officinalis was performed by gas chromatography / mass spectrometry. L. braziliensis and Trypanosoma cruzi promastigotes were used for the study of in vitro leishmanicidal activity. Mammalian fibroblast strain NCTC clone 929 was used to evaluate cytotoxic activity. Results: Analysis of essential oil from M. officinalis by GC / MS led to identification of a total 12 constituents 99.83 percent, among which some were a majority: geranial, Z citral, limonene and 1-carvone. Toxicity against fibroblasts was found to be low. M. officinalis displayed greater capacity to avoid replication against the promastigote form of the parasite at the concentrations assayed. Conclusion: The species may be an important source of new agents and selective media for the treatment of tropical diseases caused by protozoa, whose properties should be studied with respect to their mechanisms(AU)

Bond-based bilinear indices for computational discovery of novel trypanosomicidal drug-like compounds through virtual screening.

Two-dimensional bond-based bilinear indices and linear discriminant analysis are used in this report to perform a quantitative structure-activity relationship study to identify new trypanosomicidal compounds. A data set of 440 organic chemicals, 143 with antitrypanosomal activity and 297 having other clinical uses, is used to develop the theoretical models. Two discriminant models, computed using bond-based bilinear indices, are developed and both show accuracies higher than 86% for training and test sets. The stochastic model correctly indentifies nine out of eleven compounds of a set of organic chemicals obtained from our synthetic collaborators. The in vitro antitrypanosomal activity of this set against epimastigote forms of Trypanosoma cruzi is assayed. Both models show a good agreement between theoretical predictions and experimental results. Three compounds showed IC50 values for epimastigote elimination (AE) lower than 50 µM, while for the benznidazole the IC50 = 54.7 µM which was used as reference compound. The value of IC50 for cytotoxicity of these compounds is at least 5 times greater than their value of IC50 for AE. Finally, we can say that, the present algorithm constitutes a step forward in the search for efficient ways of discovering new antitrypanosomal compounds.

Phenol composition, cytotoxic and anti-kinetoplastidae activities of Lygodium venustum SW. (Lygodiaceae).

The search for new therapeutic agents has been a constant for the treatment of diseases such as leishmaniasis and Chagas disease. Most drugs used have side effects, justifying the need to evaluate the cytotoxicity of the tested products for candidates to new drugs. In this study, the bioactivity of Lygodium venustum, a cosmopolitan fern of Lygodiaceae, was assessed about their leishmanicidal and trypanocidal potential. The better activity was observed using methanol fraction, with inhibition percentage of 63% and 68% for promastigotes and epimastigotes, respectively, at a concentration of 500 µg/mL. The ethyl acetate and methanol fractions demonstrated a higher cytotoxic potential. This was the first report of leishmanicidal, trypanocidal and cytotoxic activities to L. venustum.

Atividade leishmanicida in vitro de Eugenia uniflora e Momordica charantia / Antileishmanial in vitro activity of Eugenia uniflora and Momordica charantia

A Leishmaniose Tegumentar Americana no Brasil é causada por uma variedade de espécies de Leishmania e uma grande diversidade destes parasitas pode ser encontrada na Região Amazônica. Revisões recentes na quimioterapia de leishmaniose enfatizam as deficiências dos agentes terapêuticos atualmente disponíveis e mostram a necessidade urgente de novos candidatos. Uma alternativa para substituir esses medicamentos são extratos naturais de Eugenia uniflora e Momordica charantia. Foram preparados extratos etanólicos das folhas de E. uniflora e M. charantia. Para os testes in vitro de Leishmania brasiliensis foram utilizadas formas promastigotas. O ensaio de citotoxicidade foi realizado com linhagens de fibroblastos. Nossos resultados indicam que E. uniflora foi eficaz contra a cepa de parasita testada, representando uma fonte alternativa de produtos naturais com atividade contra L. brasiliensis.

Cutaneous leishmaniasis is caused in Brazil by several species of the genus Leishmania and a wide variety of these protozoan parasites can be found in Brazil, mainly in the Amazon region. Recent reviews on the chemotherapy of leishmaniasis show the low effectiveness of the usual therapeutic agents, demonstrating the need for new drugs. An interesting possible alternative to the conventional drugs is offered by natural products extracted from Eugenia uniflora and Momordica charantia. Ethanol extracts were prepared from the leaves of Eugenia uniflora and Momordica charantia and assayed in vitro against Leishmania brasiliensis promastigotes and fibroblasts to assess their antileishmanial and cytotoxic activities, respectively. Our results indicate that E. uniflora was active against the parasitic forms of L. brasiliensis

Evaluation of the anti-Leishmania activity of ethanol extract and fractions of the leaves from Pityrogramma calomelanos (L.) link.

Leishmaniasis is caused by parasites of the genus Leishmania. Recent reports about leishmaniasis show a few number of drugs available, indicating the necessity of new drugs. In this study, the ethanol extract and fractions of Pityrogramma calomelanos (L.) link. (Pteridaceae) were assayed to verify the cytotoxicity and in vitro leishmanicidal activity against promastigote forms of Leishmania brasiliensis. The cytotoxic assay was performed using fibroblasts NCTC929. The studies indicated a leishmanicidal effect of the ethanol extract and the ethyl-acetate fraction. However, a high cytotoxic effect was observed. The hexane and methanol fractions did not show leishmanicidal activity, nor cytotoxic effect. The phytochemical screening detected the presence of alkaloids, a class of secondary metabolites with a known leishmanicidal activity. This is the first study reporting an anti-Leishmania and cytotoxic effect of P. calomelanos, being an interesting approach in the search for drugs against this disease.

Anti-Trypanosoma cruzi and cytotoxic activities of Eugenia uniflora L.

Chagas disease is caused by Trypanosoma cruzi, being considered a public health problem. An alternative to combat this pathogen is the use of natural products isolated from fruits such as Eugenia uniflora, a plant used by traditional communities as food and medicine due to its antimicrobial and biological activities. Ethanolic extract from E. uniflora was used to evaluate in vitro anti-epimastigote and cytotoxic activity. This is the first record of anti-Trypanosoma activity of E. uniflora, demonstrating that a concentration presenting 50% of activity (EC(50)) was 62.76 µg/mL. Minimum inhibitory concentration (MIC) was ≤ 1024 µg/mL. Our results indicate that E. uniflora could be a source of plant-derived natural products with anti-epimastigote activity with low toxicity.

Avaliação da atividade anti-Trypanosoma e anti-Leishmania de Mentha arvensis e Turnera ulmifolia / Evaluation of the anti-Trypanosoma and anti-Leishmania activity of Mentha arvensis and Turnera ulmifolia

Tripanosomiasis or “Chagas disease”, caused by Trypanosoma cruzi, affect 10 million people in Latin America. Today, the chemotherapy is the only specific treatment against this disease, being the most used drugs the nifurtimox and benznidazole. Leishmaniasis is a disease caused by parasites of the genus Leishmania, mainly founded in regions with forests, as the Amazonia. Recent reports about the Leishmaniasis indicate a deficit of therapeutical drugs available against this disease and reinforce the necessity of the discovering of new drugs. An interesting approach against these diseases is the use of natural products, as the extracts of plants as Mentha arvensis and Turnera ulmifolia. For the in vitro assays against T. cruzi and Leishmania, was used the clone CL-B5 and promastigote forms, respectively. The cytotoxic assay was performed using fibroblasts. Our results indicated that M. arvensis was active against all strains assayed, inhibiting 65 e 47 percent of the assayed strains (IC50 = 192.3 and 531.9 ug/mL respectively), representing an interesting and alternative source of natural products with anti-kinetoplastida activity.

Doença de Chagas, causada por Trypanosoma cruzi, afeta cerca de 10 milhões de pessoas nas Américas. Atualmente, a quimioterapia é o único tratamento específico disponível para esta doença, onde os medicamentos utilizados são nifurtimox e benzonidazol. Leishmaniose tegumentar Americana no Brasil é causada por uma variedade de espécies de Leishmania e uma grande diversidade destes parasitos pode ser encontrada na Região Amazônica. Revisões recentes na quimioterapia de leishmaniose enfatizam as deficiências dos agentes terapêuticos atualmente disponíveis e mostram a necessidade urgente de novos candidatos. Uma alternativa para substituir esses medicamentos são extratos naturais de Mentha arvensis e Turnera ulmifolia. Foram preparados extratos etanólicos das folhas de M. arvensis e T. ulmifolia. Para os testes in vitro de T. cruzi, foi utilizado o clone CL-B5 e para Leishmania brasiliensis foram utilizadas formas promastigotas. O ensaio de citotoxicidade foi realizado com linhagens de fibroblastos. Nossos resultados indicam que M. arvensis foi eficaz contra as cepas de parasitos testadas apresentando 65 e 47 por cento de inibição em uma concentração de 500 ug/mL (respectivamente, CE50 = 192.3 e 531.9 ug/mL), sendo considerada uma fonte alternativa de produtos naturais com atividade contra T. cruzi e L. brasiliensis.

Trypanocide, cytotoxic, and antifungal activities of Momordica charantia.

CONTEXT: Chagas disease, caused by Trypanosoma cruzi, is a public health problem. Currently, chemotherapy is the only available treatment for this disease, and the drugs used, nifurtimox and benzonidazol, present high toxicity levels. An alternative for replacing these drugs are natural extracts from Momordica charantia L. (Cucurbitaceae) used in traditional medicine because of their antimicrobial and biological activities. OBJECTIVE: In this study, we evaluated the extract of M. charantia for its antiepimastigote, antifungal, and cytotoxic activities. MATERIALS AND METHODS: An ethanol extract of leaves from M. charantia was prepared. To research in vitro antiepimastigote activity, T. cruzi CL-B5 clone was used. Epimastigotes were inoculated at a concentration of 1 × 10(5) cells/mL in 200 µl tryptose-liver infusion. For the cytotoxicity assay, J774 macrophages were used. The antifungal activity was evaluated by microdilution using strains of Candida albicans, Candida tropicalis, and Candida krusei. RESULTS: The effective concentration capable of killing 50% of parasites (IC(50)) was 46.06 µg/mL. The minimum inhibitory concentration (MIC) was ≤ 1024 µg/mL. Metronidazole showed a potentiation of its antifungal effect when combined with an extract of M. charantia. CONCLUSIONS: Our results indicate that M. charantia could be a source of plant-derived natural products with antiepimastigote and antifungal-modifying activity with moderate toxicity.

Cytotoxic, trypanocidal, and antifungal activities of Eugenia jambolana L.

Chagas' disease, caused by Trypanosoma cruzi, is considered a public health problem. Nowadays, chemotherapy is the only available treatment for this disease, and the drugs currently used, nifurtimox and benzonidazole, present high toxicity levels. Alternatives for replacing these drugs are natural extracts from Eugenia jambolana, a plant used in traditional medicine because of its antimicrobial and biological activities. An ethanol extract from E. jambolana was prepared. To research in vitro anti-epimastigote activity, T. cruzi CL-B5 clone was used. Epimastigotes were inoculated at a concentration of 1×10(5)/mL in 200 µL of tryptose-liver infusion. For the cytotoxicity assay J774 macrophages were used. To examine antifungal activity, Candida albicans, Candida tropicalis, and Candida krusei were used. This is the first record of trypanocide activity for E. jambolana. The effective concentration capable of killing 50% of the parasites was 56.42 µg/mL. The minimum inhibitory concentration was ≤1,024 µg/mL. Metronidazole showed a potentiation of its antifungal effect when combined with the ethanol extract of E. jambolana. Thus our results indicate that E. jambolana could be a source of plant-derived natural products with anti-epimastigote and antifungal modifying activity with moderate toxicity.

Zanthoxylum chiloperone leaves extract: first sustainable Chagas disease treatment.

ETHNOPHARMACOLOGICAL RELEVANCE: Zanthoxylum chiloperone var. angustifolium Engl. (Rutaceae) stem bark is used traditionally in Paraguay for its antiparasitic properties. Canthin-6-one is main compound isolated from Zanthoxylum chiloperone var angustifolium with broad spectrum antifungal, leishmanicidal and trypanocidal activities. AIM OF THE STUDY: The qualitative and quantitative characterization and the isolation of main alkaloidal components of different organs of Zanthoxylum chiloperone are investigated by HPLC-UV-MS. The in vitro biological activity of each extract against trypomastigote and amastigote forms of Trypanosoma cruzi parasites were evaluated, then comparison the in vivo efficacy of the ethanolic leaves extract of Zanthoxylum chiloperone with reference drug, benznidazole, in acute Trypanosoma cruzi infected mice when administered by oral route. We have also evaluated the mutagenic and cytotoxic activity of the main component of Zanthoxylum chiloperone, i.e. canthin-6-one, by mouse bone marrow micronucleus test. MATERIALS AND METHODS: The compositions of the ethanol extracts obtained after the maceration process were studied by HPLC-UV-MS methods. The quantitation analysis was performed by external standard method, using a calibration curve constructed utilizing solutions containing different concentrations of the reference samples. The anti-trypomastigote activity was evaluated by the lysis effect on mouse blood trypomastigotes (Y strain Trypanosoma cruzi). The anti-amastigote Trypanosoma cruzi activity was evaluated by a modified colorimetric method with chlorophenol red-ß-d-galactopyranoside (CPRG). The cytotoxicity of extracts and compounds was performed on NCTC 929 cells. The in vivo efficacy of the ethanolic leaves extract of Zanthoxylum chiloperone and benznidazole, in acute Trypanosoma cruzi (two different strains) was evaluated in Trypanosoma cruzi infected mice; the drugs were administered by oral route. The mortality rates were recorded and parasitaemias in control and treated mice were determined once weekly for 70 days. The mutagenic and cytotoxic activity of the main component of Zanthoxylum chiloperone, canthin-6-one, by mouse bone marrow micronucleus test. RESULTS: Canthin-6-one was the main compound of stem and root bark and 5-methoxy-canthin-6-one in leaves and fruits. The ethanolic leaves extract, canthin-6-one and benznidazole presented, approximately, the same level of in vitro activity against trypomastigote and amastigote forms of Trypanosoma cruzi. We have also evaluated the mutagenic and cytotoxic effects of canthin-6-one by micronucleus test in mice. This test showed any mutagenic and cytotoxic damages. The effects of oral or subcutaneous treatments at 10 mg/kg daily for 2 weeks with the ethanolic extract of leaves of Zanthoxylum chiloperone were examined in Balb/c mice infected acutely with Trypanosoma cruzi (CL or Y strain) and compared with benznidazole at 50 mg/kg for 2 weeks. In these experiments, 70 days after infection, parasitaemia and serological response were significantly reduced with the oral ethanolic extract treatment compared with reference drug. CONCLUSIONS: This study have shown the efficacy of the leaves extract of Zanthoxylum chiloperone in reducing Trypanosoma cruzi parasitaemia in vivo assays and could be welcomed by scientific and rural communities of Paraguay because it could help them towards the use of local resources to treat an endemic infection, Chagas disease, affecting 20% of the population of this country.