RESUMEN
Morita-Baylis-Hillman adducts (MBHA) are synthetic molecules with several biological actions already described in the literature. It has been previously described that adduct 2-(3-hydroxy-2-oxoindolin-3-yl)acrylonitrile (ISACN) has anticancer potential in leukemic cells. Inflammation is often associated with the development and progression of cancer. Therefore, to better understand the effect of ISACN, this study aimed to evaluate the anti-inflammatory potential of ISACN both in vitro and in vivo. Results demonstrated that ISACN negatively modulated the production of inflammatory cytokines IL-1ß, TNF-α, and IL-6 by cultured macrophages. In vivo, ISACN 6 and 24 mg/kg treatment promoted reduced leukocyte migration, especially neutrophils, to the peritoneal cavity of zymosan-challenged animals. ISACN displays no anti-edematogenic activity, but it was able to promote a significant reduction in the production of inflammatory cytokines in the peritoneal cavity. These data show, for the first time, that MBHA ISACN negatively modulates several aspects of the inflammatory response, such as cell migration and cytokine production in vivo and in vitro, thus having an anti-inflammatory potential.
Asunto(s)
Acrilonitrilo/farmacología , Antiinflamatorios/farmacología , Citocinas/metabolismo , Mediadores de Inflamación/metabolismo , Macrófagos Peritoneales/efectos de los fármacos , Peritonitis/prevención & control , Acrilonitrilo/análogos & derivados , Animales , Movimiento Celular/efectos de los fármacos , Células Cultivadas , Modelos Animales de Enfermedad , Femenino , Macrófagos Peritoneales/inmunología , Macrófagos Peritoneales/metabolismo , Ratones , Infiltración Neutrófila/efectos de los fármacos , Neutrófilos/efectos de los fármacos , Neutrófilos/inmunología , Neutrófilos/metabolismo , Peritonitis/inducido químicamente , Peritonitis/inmunología , Peritonitis/metabolismo , ZimosanRESUMEN
BACKGROUND: The search for new active compounds from the Brazilian flora has intensified in recent years, especially for new drugs with antibiotic potential. Accordingly, the aim of this study was to determine whether riachin has antibiotic activity in itself or is able to modulate the activity of conventional antibiotics. METHODS: A non-cyanogenic cyanoglycoside known as riachin was isolated from Bauhinia pentandra, and was tested alone and in combination with three antibiotics (clindamycin, amikacin, and gentamicin) against multiresistant bacterial strains (Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus). RESULTS: Riachin did not show significant antibiotic activity when tested alone against any strain (P>0.05). However, when combined with conventional antibiotics, it showed drug-modifying activity against strains of S. aureus exposed to clindamycin (P<0.001) as well as against P. aeruginosa exposed to amikacin (P<0.001). Although riachin did not show direct antibiotic activity, it had synergistic activity when combined with amikacin or clindamycin. The mechanism of action of this synergism is under investigation. CONCLUSION: The results of this work demonstrate that some substances of natural origin can enhance the effectiveness of certain antibiotics, which means a substantial reduction in the drug dose required and possibly in consequent adverse events for patients.
Asunto(s)
Acrilonitrilo/análogos & derivados , Antibacterianos/farmacología , Bauhinia/química , Glucósidos/farmacología , Extractos Vegetales/farmacología , Acrilonitrilo/farmacología , Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Sinergismo Farmacológico , Escherichia coli/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Pseudomonas aeruginosa/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacosRESUMEN
Acrylonitrile (ACN) is a widely used chemical in the production of plastics, resins, nitriles, acrylic fibers, and synthetic rubber. Previous epidemiological investigations and animal studies have confirmed that ACN affects the lymphocytes and spleen. However, the immune toxicity mechanism is unknown. Lipid rafts are cell membrane structures that are rich in cholesterol and involved in cell signal transduction. The B cell lymophoma-10 (Bcl10) protein is a joint protein that is important in lymphocyte development and signal pathways. This study was conducted to examine the in vitro effects of ACN. We separated lipid rafts, and analyzed Bcl10 protein and caveolin. Western blotting was used to detect mitogen-activated protein kinase (MAPK) and phosphorylated MAPK levels. The results indicated that with increasing ACN concentration, the total amount of Bcl10 remained stable, but was concentrated mainly in part 4 to part 11 in electrophoretic band district which is high density in gradient centrifugation. Caveolin-1 was evaluated as a lipid raft marker protein; caveolin-1 content and position were relatively unchanged. Western blotting showed that in a certain range, MAPK protein was secreted at a higher level. At some ACN exposure levels, MAPK protein secretion was significantly decreased compared to the control group (P < 0.05). These results indicate that ACN can cause immune toxicity by damaging lipid raft structures, causing Bcl10 protein and lipid raft separation and restraining Ras-Raf-MAPK-extracellular signal-regulated kinase signaling pathways.
Asunto(s)
Acrilonitrilo/farmacología , Linfocitos/efectos de los fármacos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Microdominios de Membrana/efectos de los fármacos , Quinasas raf/metabolismo , Proteínas ras/metabolismo , Western Blotting , Humanos , Células Jurkat , Linfocitos/metabolismoRESUMEN
This study aimed to evaluate the effects of acrylonitrile (ACN) on neuronal morphology and apoptosis in rats. An ACN solution was administered to Wistar rats by gavage at doses of 0, 5, 10, or 20 mg/kg, 5 days a week for 13 weeks. The morphology of neurons and the presence of apoptosis was examined by light and electron microscope, DNA electrophoresis, immunohistochemistry, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling. Significant vacuolation and the widening of the interspaces around blood vessels were observed in the groups that received the highest dose. Disordered myelin sheaths, malformed neuronal nuclei, and chromatin condensation at the periphery of the nucleus that formed crescents were also observed in the treated rats. The number of apoptotic neurons was significantly decreased (P < 0.05) in the treated groups (5 mg/kg group: 1.5 ± 1.22 apoptotic neurons/slide; 10 mg/kg group: 2.5 ± 1.05 apoptotic neurons/slide; 20 mg/kg group: 2.34 ± 1.21 apoptotic neurons/slide) compared to the control group (4.5 ± 1.52 apoptotic neurons/slide). The number of Bcl-2-positive neurons and the levels of staining were increased in the treated rats compared to those of the control group. These results suggested that ACN may induce serious morphological changes in rat neurons and inhibit neuronal apoptosis in rats.
Asunto(s)
Acrilonitrilo/farmacología , Apoptosis/efectos de los fármacos , Neuronas/efectos de los fármacos , Neuronas/patología , Animales , Carcinógenos/farmacología , Femenino , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Hipocampo/patología , Masculino , Neuronas/metabolismo , Neuronas/ultraestructura , Antígeno Nuclear de Célula en Proliferación/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Células Piramidales/efectos de los fármacos , Células Piramidales/metabolismo , Células Piramidales/patología , Células Piramidales/ultraestructura , Ratas , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
Chagas disease, caused by the protozoan Trypanosoma cruzi, remains a serious health concern due to the lack of effective vaccines or satisfactory treatment. In the search for new compounds against this neglected disease, we have previously demonstrated that the compound 3-Hydroxy-2-methylene-3-(4-nitrophenylpropanenitrile) (MBHA3), derived from the Morita-Baylis-Hillman reaction, effectively caused a loss of viability in both the epimastigote and trypomastigote forms. However, the mechanisms of parasite death elicited by MBHA3 remain unknown. The aim of this study was to better understand the morphophysiological changes and the mechanism of cell death induced by MBHA3 treatment on T. cruzi. To perform this analysis, we used confocal microscopy and flow cytometry to monitor the fluorescent probes such as annexin-V/propidium iodide (AV/PI), calcein-AM/ethidium homodimer (CA/EH), acridine orange (AO) and rhodamine 123 (Rho 123). Lower concentrations of MBHA3 led to alterations in the mitochondrial membrane potential and AO labeling, but did not decrease the viability of the epimastiogote forms, as determined by the CA/EH and AV/PI assays. Conversely, treatment with higher concentrations of MBHA3 led to extensive plasma membrane damage, loss of mitochondrion membrane potential, DNA fragmentation and acidification of the cytoplasm. Our findings suggest that at higher concentrations, MBHA3 induces T. cruzi epimastigote death by necrosis in a mitochondrion-dependent manner.
Asunto(s)
Acrilonitrilo/análogos & derivados , Alcoholes Bencílicos/farmacología , Muerte Celular/efectos de los fármacos , Enfermedad de Chagas/tratamiento farmacológico , Trypanosoma cruzi/efectos de los fármacos , Acrilonitrilo/farmacología , Acrilonitrilo/uso terapéutico , Alcoholes Bencílicos/uso terapéutico , Membrana Celular/efectos de los fármacos , Enfermedad de Chagas/parasitología , Citoplasma/efectos de los fármacos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Microscopía Confocal , NitrilosRESUMEN
A doença de Chagas, causada pelo protozoário hemoflagelado Trypanosoma cruzi, consiste em um grave problema de saúde pública na América Latina, com cerca de 7 a 8 milhões de pessoas infectadas. O benznidazol é atualmente o único fármaco disponível para o tratamento, apresentando boa atividade na fase aguda, mas com eficácia questionada na fase crônica, além de apresentar severos efeitos colaterais e longo período de tratamento. Tendo em vista a sua fácil síntese e baixo custo, adutos aromáticos oriundos da reação Morita-Baylis-Hillman têm sido considerados promissores como quimioterápicos. Neste sentido, foi avaliada a atividade tripanocida e citotóxica de seis adutos Morita-Baylis-Hillman. Para analisar possíveis efeitos dos adutos sobre estruturas específicas do protozoário foi utilizado a microscopia confocal a laser, através da marcação com Rodamina 123, MitoSOXT, Laranja de Acridina e Kit Live/Dead(R). As alterações morfológicas em parasitas submetidos ao tratamento foram acompanhadas por microscopia eletrônica de transmissão. Nossos resultados mostraram que todos os compostos foram capazes de inibir o crescimento de formas epimastigotas e causaram uma diminuição da viabilidade em formas tripomastigotas, apresentando uma moderada citotoxicidade para células de mamíferos. Os adutos MBH1, MBH2, MBH5 e MBH7 também foram efetivos em inibir a infecção de macrófagos e diminuir a viabilidade das formas amastigotas. Os compostos não foram capazes de alterar os níveis de produção do óxido nítrico em macrófagos. Análises pela microscopia confocal e microscopia eletrônica de transmissão (MET) apontam a mitocôndria como principal alvo intracelular destes compostos. Alterações compatíveis com a perda da viabilidade e morte celular por necrose e autofagia foram observadas por MET. Os resultados obtidos neste trabalho colocam os adutos Morita -Baylis-Hillman em evidência como agentes promissores para o tratamento da tripanossomíase americana.
Asunto(s)
Enfermedad de Chagas/tratamiento farmacológico , Tripanocidas/farmacología , Tripanocidas/síntesis química , Trypanosoma cruzi , Trypanosoma cruzi/ultraestructura , Acrilonitrilo/análogos & derivados , Acrilonitrilo/farmacología , Acrilonitrilo/síntesis química , Trypanosoma cruzi/crecimiento & desarrolloRESUMEN
A doença de Chagas, causada pelo protozoário Trypanosoma cruzi, é um importante problema de saúde pública na América Latina, com cerca de 7,6 milhões de pessoas infectadas. O benznidazol e o nifurtimox são os únicos fármacos disponíveis para o tratamento, e além de apresentarem sérios efeitos colaterais, a sua eficácia na fase crônica da doença ainda é controversa. Neste sentido, nós sintetizamos o aduto Morita-Baylis-Hillman 3-hidroxi-2-metileno-3-(4-nitrofenilpropanonitrilo) (MBHA 3) e avaliamos sua atividade biológica sobre o T. cruzi. O MBHA 3 inibiu fortemente o crescimento das formas epimastigotas, com IC(50) / 72h de 28,5 µM e causou intensa lise de tripomastigotas, com IC(50)/ 24h de 25,5 µM. A análise ultraestrutural mostrou alterações morfológicas significantes, como arredondamento do corpo celular e desorganização intracelular. Alterações indicativas de apoptose, autofagia ou necrose também foram observadas nas células mais afetadas. A fim de melhor compreender o mecanismo envolvido na morte celular induzida pelo composto, nós utilizamos a microscopia confocal e a citometria de fluxo aliadas a sondas fluorescentes, como anexina-V (AV)/ iodeto de propídio (IP); Calceína-AM (CA)/ homodímero de etídio (HE); laranja de acridina (LA) e rodamina 123. O tratamento com 6 e 12 µg/ mL revelou uma alta porcentagem de células viáveis pela CA/ HE, mas também induziu alterações mitocondriais e aumento da marcação com a LA, sugerindo que um processo de morte celular programada (MCP) por autofagia poderia estar ocorrendo. Por outro lado, o tratamento com 24 µg/ mL levou à perda de viabilidade celular com danos excessivos sobre a membrana plasmática e mitocondrial e fragmentação inespecífica do DNA. Em conclusão, nossos achados sugerem que o MBHA 3, em alta concentração, induz MCP por necrose em T. cruzi.
Asunto(s)
Acrilonitrilo/análogos & derivados , Alcohol Bencilo/farmacología , Alcohol Bencilo/síntesis química , Enfermedad de Chagas/tratamiento farmacológico , Tripanocidas/farmacología , Tripanocidas/síntesis química , Trypanosoma cruzi , Acrilonitrilo/farmacología , Acrilonitrilo/síntesis química , Modelos Moleculares , Trypanosoma cruzi/crecimiento & desarrolloRESUMEN
We have synthesized the Morita-Baylis-Hillman adduct (MBHA) 3-hydroxy-2-methylene-3-(4-nitrophenyl)-propanenitrile (3) in quantitative yield and evaluated on Trypanosoma cruzi epimastigote and bloodstream trypomastigote forms. Compound 3 strongly inhibited epimastigote growth, with IC(50)/72hof 28.5 microM and also caused intense trypomastigotes lysis, with an IC(50)/24h of 25.5 microM. Ultrastructural analysis showed significant morphological changes on both parasite forms treated with 3, including increase of cell volume and rounding of cell body as well as intense intracellular disorganization. Morphological changes indicative of apoptosis, autophagy or necrosis were observed in most affected cells. Docking calculations of 1, 2 and 3 pointed out the possibility of T. cruzi Farnesyl Pyrophosphate Synthase (TcFPPS) enzyme inhibition in 3 mechanism of action.
Asunto(s)
Acrilonitrilo/análogos & derivados , Alcoholes Bencílicos/síntesis química , Alcoholes Bencílicos/farmacología , Enfermedad de Chagas/tratamiento farmacológico , Tripanocidas/síntesis química , Tripanocidas/farmacología , Trypanosoma cruzi/efectos de los fármacos , Acrilonitrilo/síntesis química , Acrilonitrilo/farmacología , Humanos , Concentración 50 Inhibidora , Modelos Moleculares , Nitrilos , Trypanosoma cruzi/crecimiento & desarrolloRESUMEN
A bio-guided screening against influenza A virus (FLUAV) was carried out with seven Euphorbiaceae species. The results showed that chromatographic fractions from Phyllantus niruri, Euphorbia pulcherrima and Codiaeum variegatum had relevant anti-FLUAV activity, although only chromatographical subfractions from C. variegatum kept the activity. From this plant, the active compound against FLUAV was isolated. Its structure was assigned as 2-(3,4,5)-trihydroxy-6-hydroxymethyltetrahydropyran-2-yloxymethyl)acrylonitrile (1) on the basis of NMR, mass spectrometry and X-ray diffraction analysis. The compound displayed virucidal activity without impairment of haemagglutination properties of the used virus strain. This is the first report indicating antiviral activity of a cyanoglucoside.
Asunto(s)
Acrilonitrilo/análogos & derivados , Antivirales/farmacología , Euphorbiaceae/química , Glucósidos/farmacología , Hexosas/farmacología , Virus de la Influenza A/efectos de los fármacos , Acrilonitrilo/química , Acrilonitrilo/aislamiento & purificación , Acrilonitrilo/farmacología , Animales , Antivirales/química , Antivirales/aislamiento & purificación , Línea Celular , Perros , Glucósidos/química , Glucósidos/aislamiento & purificación , Hexosas/química , Hexosas/aislamiento & purificación , Pruebas de Sensibilidad Microbiana , Resonancia Magnética Nuclear Biomolecular , Difracción de Rayos XRESUMEN
A series of 3-aryl-2-(2-thienyl)acrylonitriles 7 and 3-aryl-2-(3-thienyl)acrylonitriles 8 were synthesized by the reaction of aromatic aldehydes 6 with 2- and 3-thienylacetonitriles 4 and 5, and evaluated for antifungal and cytotoxic activities against a panel of opportunistic and pathogenic fungi and three different cancer cell lines, respectively.