RESUMO
The main form of control of leishmaniasis is the treatment, however various side effects and poor efficacy are associated with presently available drugs. The investigation of bioactive natural products for new antileishmanial drugs is a valid approach. The present study reports the in vitro efficacy of natural isoflavonoids and terpenes against Leishmania infantum and L. amazonensis and their cytotoxicity against HepG2 cells. L. infantum and L. amazonensis promastigotes were exposed to the terpenes kaurenoic acid, xylopic acid, and (-)-α-bisabolol and to the isoflavonoids (-)-duartin and (3R)-claussequinone for antileishmanial activity and to cytotoxicity to HepG2 cells. The most effective substance against both L. infantum and L. amazonensis species was (3R)-claussequinone (IC50 = 3.21 µg/mL and 2.47 µg/mL, respectively) that disclosed low cytotoxicity against HepG2 cells (CC50 = 387.79 µg/mL). The efficacy of (3R)-claussequinone against intracellular amastigotes of L. infantum and the externalization of phosphatidylserine in promastigotes of this isoflavanoid were investigated by infection of Raw 264.7 macrophages and marking with Annexin V-FITC and propidium Iodide for flow cytometry analysis. The results for amastigotes showed that (3R)-claussequinone was able to reduce the rate of infection with IC50 = 4.61 µg/mL and did not alter the externalization of phosphatidylserine. In conclusion it is presently reported, for the first time, the striking antileishmanial activity of (3R)-claussequinone against L. infantum and L. amazonensis associated to low cytotoxicity. Furthermore, these results suggest that (3R)-claussequinone is a new hit aiming to develop new therapeutic alternatives.
Assuntos
Antiprotozoários , Produtos Biológicos , Leishmania infantum , Camundongos , Animais , Terpenos/farmacologia , Fosfatidilserinas , Propídio , Camundongos Endogâmicos BALB C , Antiprotozoários/toxicidade , Antiprotozoários/uso terapêutico , Produtos Biológicos/farmacologiaRESUMO
A total of 33 extracts of eleven different plants species from Mata Atlântica biome, Brazil, and different fractions of the bioactive extracts were evaluated against chloroquine-resistant Plasmodium falciparum W2 strain by PfLDH method and cytotoxicity to HepG2 cells by the MTT assay, and chemically characterized by LC-DAD-ESI-MS/MS analysis. The results allowed the identification of Alchornea glandulosa, Miconia latecrenata, and Psychotria suterella as the most active plant species. Different flavonoids and tannins in Alchornea glandulosa and Miconia latecrenata besides alkaloids in Psychotria suterella were identified. Bioguided fractionation of A. glandulosa and M. latecrenata leaves extracts led to fractions exhibiting high parasite growth inhibition. Seven known alkaloids were identified in the P. suterella extract, and of these, only 5-carboxystrictosidine had been assayed for antiplasmodial activity what points to this species as the most promising among the eleven one assayed.
Assuntos
Antimaláricos/farmacologia , Bioprospecção , Ecossistema , Metaboloma , Plantas/química , Plasmodium falciparum/efeitos dos fármacos , Animais , Brasil , Fracionamento Químico , Cromatografia Líquida , Flavonoides/análise , Humanos , Extratos Vegetais/química , Folhas de Planta/química , Plantas/efeitos dos fármacos , Espectrometria de Massas em Tandem , Taninos/análiseRESUMO
Malaria is nowadays one of the most serious health concerns in a global scale and, although there is an evident increase in research studies in this area, pointed by the vast number of hits and leads, it still appears as a recurrent topic every year due to the drug resistance shown by the parasite exposing the urgent need to develop new antimalarial medications. In this work, 38 molecules were synthesized via copper(I)-catalyzed alkyne-azide cycloaddition (CuAAC) or "click" chemistry, following different routes to produce 2 different organic azides, obtained from a 4,7 dicholoquinoline, reacted with 19 different commercially available terminal alkynes. All those new compounds were evaluated for their in vitro activity against the chloroquine resistant malaria parasite Plasmodium falciparum (W2). The cytotoxicity evaluation was accomplished using Hep G2 cells and SI index was calculated for every molecule. Some of the quinoline derivatives have shown high antimalarial activity, with IC50 values in the range of 1.72-8.66 µM, low cytotoxicity, with CC50>1000 µM and selectivity index (SI) in the range of 20-100, with some compounds showing SI>800. Therefore, the quinolinotriazole hybrids could be considered a very important step on the development of new antimalarial drugs
Assuntos
Técnicas In Vitro/instrumentação , Cloroquina/administração & dosagem , Malária/tratamento farmacológico , Antimaláricos/análise , Plasmodium falciparum/metabolismo , Pesquisa/classificação , Resistência a Medicamentos/efeitos dos fármacos , Quimera/anormalidades , Concentração Inibidora 50 , Química ClickRESUMO
Aiming to investigate the antiplasmodial activity and the phytochemical composition of Xylopia sericea leaves, the essential oil and dichloromethane extract were analyzed by gas and liquid chromatography, respectively, both of them coupled to mass spectrometry, and were evaluated against the chloroquine-resistant Plasmodium falciparum strain (W2) and for cytotoxicity to HepG2 cells. Low growth inhibition of P. falciparum as well as low cytotoxicity to HepG2 cells were observed for the essential oil. The leaves dichloromethane extract showed moderate growth inhibition of P. falciparum and low cytotoxicity to HepG2 cells. Bioguided chromatographic fractionation of this extract led to fractions with increased antiplasmodial activity from which liriodenine (IC50 6.1 ± 0.1 µg/mL, CC50 > 1000.0 µg/mL, SI > 164), an aporphine alkaloid, and an acetogenin-rich fraction containing mainly isomers of annomontacin and 4-deoxy-annomontacin (IC50 22.7 ± 1.9 µg/mL, CC50 336.1 ± 15.5 µg/mL, SI = 15) might be highlighted for their antiplasmodial activity.
Assuntos
Antimaláricos/farmacologia , Plasmodium falciparum/efeitos dos fármacos , Xylopia/química , Animais , Antimaláricos/química , Antimaláricos/toxicidade , Aporfinas/química , Aporfinas/farmacologia , Cloroquina/farmacologia , Cromatografia Gasosa , Cromatografia Líquida , Avaliação Pré-Clínica de Medicamentos , Resistência Microbiana a Medicamentos , Furanos/farmacologia , Células Hep G2 , Humanos , Lactonas/farmacologia , Óleos Voláteis/química , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Folhas de Planta/químicaRESUMO
Leishmaniasis is one of the most important neglected diseases worldwide. It is a life-threatening disease and causes significant morbidity, long-term disability, and early death. Treatment involves disease control or use of intervention measures, although the currently used drugs require long-lasting therapy, and display toxicity and reduced efficacy. The use of natural products isolated from plants, such as lapachol, an abundant naphthoquinone naturally occurring in South American Handroanthus species (Tabebuia, Bignoniaceae), is a promising option for the treatment of leishmaniasis. In this study, we investigated the leishmanicidal activity of lapachol in vitro and in vivo against Leishmania infantum and L. amazonensis, causative agents of visceral and cutaneous leishmaniasis, respectively. Low cytotoxicity in HepG2 cells (3405.8⯱â¯261.33⯵M), good anti-Leishmania activity, and favorable selectivity indexes (SI) against promastigotes of both L. amazonensis (IC50â¯=â¯79.84⯱â¯9.10⯵M, SIâ¯=â¯42.65) and L. infantum (IC50â¯=â¯135.79⯱â¯33.04⯵M, SIâ¯=â¯25.08) were observed. Furthermore, anti-Leishmania activity assays performed on intracellular amastigotes showed good activity for lapachol (IC50â¯=â¯191.95⯵M for L. amazonensis and 171.26⯵M for L. infantum). Flow cytometric analysis demonstrated that the cytotoxic effect of lapachol in Leishmania promastigotes was caused by apoptosis-like death. Interestingly, the in vitro leishmanicidal effect of lapachol was confirmed in vivo in murine models of visceral and cutaneous leishmaniasis, as lapachol (25â¯mg/kg oral route for 24â¯h over 10 days) was able to significantly reduce the parasitic load in skin lesions, liver, and spleen, similar to amphotericin B, the reference drug. These results reinforce the therapeutic potential of lapachol, which warrants further investigations as an anti-leishmaniasis therapeutic.
Assuntos
Antiprotozoários/uso terapêutico , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Visceral/tratamento farmacológico , Naftoquinonas/uso terapêutico , Anfotericina B/farmacologia , Anfotericina B/uso terapêutico , Anfotericina B/toxicidade , Animais , Antiprotozoários/farmacologia , Antiprotozoários/toxicidade , Modelos Animais de Doenças , Feminino , Citometria de Fluxo , Células Hep G2/efeitos dos fármacos , Humanos , Concentração Inibidora 50 , Leishmania infantum/efeitos dos fármacos , Leishmania mexicana/efeitos dos fármacos , Fígado/parasitologia , Camundongos , Camundongos Endogâmicos BALB C , Naftoquinonas/farmacologia , Naftoquinonas/toxicidade , Carga Parasitária , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Extratos Vegetais/toxicidade , Células RAW 264.7/efeitos dos fármacos , Células RAW 264.7/parasitologia , Distribuição Aleatória , Pele/parasitologia , Baço/parasitologia , Tabebuia/químicaRESUMO
OBJECTIVES: To assess the antiplasmodial activity of the ethanol extract of Xylopia sericea leaves, Annonaceae, often associated with antimalarial use and to perform a bioguided isolation of active compounds. METHODS: Dereplication of ethanol extract by the UPLC-DAD-ESI-MS/MS technique allowed the identification of the major constituents, isolation and identification of alkaloids. The antiplasmodial and cytotoxic activity of the extract, fractions and isolated compounds was evaluated against the chloroquine-resistant W2 strain Plasmodium falciparum and HepG2 cells, respectively. KEY FINDINGS: Ethanol extract showed high reduction of parasitemia as well as moderate cytotoxicity (86.5 ± 3.0% growth inhibition at 50 µg/ml and CC50 72.1 ± 5.1 µg/ml, respectively). A total of eight flavonoids were identified, and two aporphine alkaloids, anonaine and O-methylmoschatoline, were isolated. Anonaine disclosed significant antiplasmodial effect and moderate cytotoxicity (IC50 23.2 ± 2.7 µg/ml, CC50 38.3 ± 2.3 µg/ml, SI 1.6) while O-methylmoschatoline was not active against P. falciparum and showed a low cytotoxicity (33.5 ± 1.9% growth inhibition at 50 µg/ml, CC50 274.4 ± 0.5 µg/ml). CONCLUSIONS: Characterization of Xylopia sericea leaves ethanol extract by UPLC-DAD-ESI-MS/MS as well as its antiplasmodial activity and the occurrence of anonaine and O-methylmoschatoline in this Xylopia species are reported by the first time.
Assuntos
Alcaloides/farmacologia , Antimaláricos/farmacologia , Extratos Vegetais/farmacologia , Xylopia/química , Alcaloides/isolamento & purificação , Alcaloides/toxicidade , Antimaláricos/isolamento & purificação , Antimaláricos/toxicidade , Aporfinas/isolamento & purificação , Aporfinas/farmacologia , Cromatografia Líquida de Alta Pressão/métodos , Dioxóis/isolamento & purificação , Dioxóis/farmacologia , Etanol/química , Células Hep G2 , Humanos , Concentração Inibidora 50 , Extratos Vegetais/administração & dosagem , Extratos Vegetais/toxicidade , Folhas de Planta , Plasmodium falciparum/efeitos dos fármacos , Espectrometria de Massas em Tandem/métodosRESUMO
Plant extracts exist as a complex matrix which serves as a source of numerous bioactive metabolites. The ultra performance liquid chromatography with diode-array detection-coupled electrospray ionization-mass spectrometry/mass spectrometry technique was used to characterize the aqueous extract from leaves of Alchornea glandulosa (EAG), a species popularly used to treat gastrointestinal problems as an antiulcer agent. Quantification of phenolic derivatives was determined using Folin-Ciocalteu and aluminum trichloride (AlCl3) methods. In addition, antioxidant (2,2-diphenyl-1-picrylhydrazyl [DPPHâ¢] radical scavenging, ß-carotene-linoleic acid, and lipid peroxidation), antibacterial (agar well diffusion method and minimum inhibitory concentration), antimutagenic (Ames test), and antigenotoxic (plasmid cleavage) assays were also performed on this plant extract. The ellagitannin tris-galloyl-hexahydroxydiphenic acid-glucose was identified as the predominant compound along with tannins as majority metabolites. EAG showed high antioxidant activity accompanied by moderate antibacterial activity against Staphylococcus aureus. The highest antimutagenic activity was observed for TA97 strain without metabolic activation (S9) and with metabolic activation, TA100 and TA102 were completely inhibited. In addition, EAG exhibited potential signs of antigenotoxic action. The high antioxidant and antimutagenic activity observed for EAG suggests important therapeutic uses that still need to be verified in future studies.
Assuntos
Antibacterianos/farmacologia , Antimutagênicos/farmacologia , Antioxidantes/farmacologia , Euphorbiaceae/química , Extratos Vegetais/farmacologia , Folhas de Planta/química , Staphylococcus aureus/efeitos dos fármacosRESUMO
Malaria is responsible for more deaths around the world than any other parasitic disease. Due to the emergence of strains that are resistant to the current chemotherapeutic antimalarial arsenal, the search for new antimalarial drugs remains urgent though hampered by a lack of knowledge regarding the molecular mechanisms of artemisinin resistance. Semisynthetic compounds derived from diterpenes from the medicinal plant Wedelia paludosa were tested in silico against the Plasmodium falciparum Ca2+-ATPase, PfATP6. This protein was constructed by comparative modelling using the three-dimensional structure of a homologous protein, 1IWO, as a scaffold. Compound 21 showed the best docking scores, indicating a better interaction with PfATP6 than that of thapsigargin, the natural inhibitor. Inhibition of PfATP6 by diterpene compounds could promote a change in calcium homeostasis, leading to parasite death. These data suggest PfATP6 as a potential target for the antimalarial ent-kaurane diterpenes.
Assuntos
ATPases Transportadoras de Cálcio/metabolismo , Diterpenos do Tipo Caurano/uso terapêutico , Desenho de Fármacos , Plasmodium falciparum/enzimologia , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/metabolismo , Wedelia/química , Antimaláricos/metabolismo , Artemisininas/metabolismo , Cálcio/metabolismo , Diterpenos do Tipo Caurano/síntese química , Diterpenos do Tipo Caurano/farmacologia , Interações Medicamentosas , Inibidores Enzimáticos/farmacologia , Simulação de Acoplamento Molecular , Estrutura Molecular , Plasmodium falciparum/efeitos dos fármacos , Tapsigargina/farmacologia , Wedelia/classificaçãoRESUMO
Malaria is responsible for more deaths around the world than any other parasitic disease. Due to the emergence of strains that are resistant to the current chemotherapeutic antimalarial arsenal, the search for new antimalarial drugs remains urgent though hampered by a lack of knowledge regarding the molecular mechanisms of artemisinin resistance. Semisynthetic compounds derived from diterpenes from the medicinal plant Wedelia paludosa were tested in silico against the Plasmodium falciparum Ca2+-ATPase, PfATP6. This protein was constructed by comparative modelling using the three-dimensional structure of a homologous protein, 1IWO, as a scaffold. Compound 21 showed the best docking scores, indicating a better interaction with PfATP6 than that of thapsigargin, the natural inhibitor. Inhibition of PfATP6 by diterpene compounds could promote a change in calcium homeostasis, leading to parasite death. These data suggest PfATP6 as a potential target for the antimalarial ent-kaurane diterpenes.
Assuntos
Idoso , Feminino , Humanos , Masculino , Neoplasias Gastrointestinais/fisiopatologia , Promoção da Saúde/organização & administração , Sobreviventes , República da CoreiaRESUMO
Ethanol extracts of eighteen Bignoniaceae species have been evaluated by the MTT assay for cytotoxicity in Vero cells and for antiviral activity against Human herpes virus type 1, Vaccinia virus and murine Encephalomyocarditis virus. Among such species, seven are reported to be of traditional medicinal use No cytotoxicity was observed for most of the extracts up to the concentration of 500 μg/mL. Fourteen (50 percent) of the 28 extracts assayed have disclosed antiviral activity with EC50 values in the range of 4.6+0.3 to 377.2+17.7 μg/mL. Only two species, Arrabidaea samydoides and Callichlamys latifolia, have shown activity against all the three viruses. The extracts were chemically characterized by their TLC and HPLC-DAD profiles. Mangiferin is the major constituent of A. samydoides but the isolated compound has been less active than the crude extract. This is the first report on the antiviral evaluation of the eighteen Bignoniaceae species assayed.
Extratos etanólicos de dezoito espécies vegetais pertencentes à família Bignoniaceae, das quais sete são descritas como de uso medicinal, foram avaliados, pelo ensaio colorimétrico do MTT, para atividades citotóxica, em células Vero, e antiviral, frente aos vírus herpes simplex-tipo 1, vaccinia e encefalomiocardite murina. A maior parte dos extratos não apresentou citotoxicidade até a concentração de 500 μg/mL. Dos 28 extratos testados quatorze (50 por cento) apresentaram atividade antiviral com valores de CE50 na faixa de 4,6+03 a 377,2+17,7 μg/mL. Somente duas espécies, Arrabidaea samydoides e Callichlamys latifolia, foram ativas frente aos três vírus. Os extratos foram caracterizados pelos seus perfís cromatográficos em CCD e CLAE-FR. Análises por CLAE-FR mostraram que a mangiferina é o constituinte majoritário em A. samydoides mas a substância isolada foi menos ativa do que o extrato bruto. Esta é a primeira vez que se relata a atividade antiviral de extratos das dezoito espécies avaliadas.