RESUMEN
The objective of the present study was to evaluate the action of the crude hydroalcoholic extract of Piper cubeba fruits and isolated lignans (cubebin, dihydrocubebin, ethylcubebin, hinokinin and methylcubebin) on head and neck cancer cells. We evaluated the influence of the Piper cubeba extract and isolated lignans (10, 50 e 100 µg/mL) for 4, 24, 48 and 72 h, in the larynx (Hep-2) and oral (SCC-25) squamous cell carcinoma cells and normal fibroblasts, on morphology, cell proliferation and migration, cytotoxicity, genotoxicity and gene and protein expression (PTGS2, PTGER3, PTGER4, MMP2, MMP9). The results showed that the P. cubeba extract and different lignans do not alter the cellular morphology, but decrease cell proliferation and migration, have low cytotoxic and genotoxic effects, probably due to the alteration of the expression of genes and proteins involved with inflammatory process. From these data, we can conclude that the lignans cubebin and methylcubebin had a greater effect on head and neck cancer cells in the antiproliferative, antimigratory and genotoxic action, and could be the target of the development of new therapies including possible new drugs as a therapeutic resource for the treatment of head and neck cancer due to its immense range of biological properties.
Asunto(s)
Carcinoma de Células Escamosas/tratamiento farmacológico , Carcinoma de Células Escamosas/patología , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Fibroblastos/fisiología , Neoplasias de Cabeza y Cuello/tratamiento farmacológico , Neoplasias de Cabeza y Cuello/patología , Lignanos/aislamiento & purificación , Lignanos/farmacología , Fitoterapia , Piper/química , Extractos Vegetales/química , Extractos Vegetales/farmacología , Carcinoma de Células Escamosas/genética , Línea Celular Tumoral , Células Cultivadas , Relación Dosis-Respuesta a Droga , Expresión Génica/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/genética , Neoplasias de Cabeza y Cuello/genética , Humanos , Lignanos/química , Lignanos/uso terapéutico , Terapia Molecular Dirigida , Extractos Vegetales/uso terapéutico , Factores de TiempoRESUMEN
This paper reports the synthesis of (±)-licarin A 1, a dihydrobenzofuran neolignan, resultant of an oxidative coupling reaction of isoeugenol and horseradish peroxidase (HRP) enzyme. Following, three semi-synthetic derivatives from this compound were obtained: benzylated (±)-licarin A 2, methylated (±)-licarin A 3 and acetylated (±)-licarin A 4. After structural elucidation and assignment by Nuclear Magnetic Resonance of 1H, 13C and DEPT, all compounds were evaluated in vitro against Trypomastigote forms of Trypanosoma cruzi (T. cruzi), the etiologic agent of Chagas disease, and Schistosoma mansoni (S. mansoni) worms, the etiologic agent of schistosomiasis. Compound (4) was the most active against S. mansoni adult worms, displaying worm viability reduction at 25 µM and mortality of all worms at 100 and 200 µM within 24 h. Compound 1 was the second most active, showing worm viability reduction at 50 µM and mortality of 25% and 100% of worms in 24h at concentrations of 100 and 200 µM, respectively. In addition, theoretical calculations aiming at finding molecular properties that showed the correlation for schistosomicidal and trypanocidal activities of (±)-licarin A and three of its semi-synthetic derivatives were also performed.
Asunto(s)
Lignanos/síntesis química , Schistosoma mansoni/efectos de los fármacos , Esquistosomicidas/síntesis química , Tripanocidas/síntesis química , Trypanosoma cruzi/efectos de los fármacos , Animales , Enfermedad de Chagas/tratamiento farmacológico , Simulación por Computador , Lignanos/farmacología , Lignanos/uso terapéutico , Esquistosomiasis/tratamiento farmacológico , Esquistosomicidas/farmacología , Tripanocidas/farmacologíaRESUMEN
Piper cubeba L. f. is a food seasoning, which contains secondary metabolites displaying several biological properties, such as cytotoxic, anti-inflammatory, and antiparasitic activities. The lignans (+)-dihydroclusin, (-)-clusin, (-)-cubebin, (-)-yatein, and (-)-haplomyrfolin were isolated, with (-)-haplomyrfolin reported for the first time in P. cubeba seeds. Chromatographic standards were used to develop a reliable reverse-phase high-performance liquid chromatography analytical method according to the Agência Nacional de Vigilância Sanitária and International Conference on Harmonization guidelines to quantitate these lignans in both P. cubeba seeds and their extracts. The extraction of the lignans was also optimized, with the best conditions being ultrasound-assisted extraction, with 84% aqueous ethanol for 38 min in a single extraction. This procedure allows for the extraction of more than 80% of the total lignans, which is better in comparison to other techniques, such as maceration and Soxhlet extraction.
Asunto(s)
Fraccionamiento Químico/métodos , Cromatografía Líquida de Alta Presión/métodos , Lignanos/análisis , Lignanos/aislamiento & purificación , Piper/química , Extractos Vegetales/análisis , Extractos Vegetales/aislamiento & purificación , Ultrasonido/métodos , Semillas/químicaRESUMEN
Encouraged by the anti-inflammatory activity of hinokinin inâ vivo, which is also observed for the analogues dinitrohinokinin and diidrocubebin, herein we used inâ vitro and in silico methods to assess their selectivity profiles and predict their binding modes with Cyclooxygenases (COX-1 and 2). The inâ vitro assays demonstrated dinitrohinokinin is about 13 times more selective for COX-2 than for COX-1, a similar profile observed for the drugs celecoxib (selective index ≈9) and meloxicam (selective index ≈11). Predictions of the binding modes suggested dinitrohinokinin interacts with COX-2 very similarly to rofecoxib, exploring residues at the hydrophilic pocket of the enzyme that accessible to ligands only in this isoform. This lignan also interacts with COX-1 in a similar mode to meloxicam, blocking the access of the substrate to the catalytic cleft. Therefore, dinitrohinokinin is a promising lead for the design of selective COX-2 inhibitors.
Asunto(s)
4-Butirolactona/análogos & derivados , Benzodioxoles/farmacología , Inhibidores de la Ciclooxigenasa/farmacología , Lignanos/farmacología , Simulación del Acoplamiento Molecular , 4-Butirolactona/química , 4-Butirolactona/farmacología , Benzodioxoles/química , Sitios de Unión , Ciclooxigenasa 1/metabolismo , Ciclooxigenasa 2/metabolismo , Inhibidores de la Ciclooxigenasa/química , Humanos , Lactonas/química , Lactonas/farmacología , Lignanos/química , Meloxicam/farmacología , Unión Proteica , Relación Estructura-Actividad Cuantitativa , Especificidad por Sustrato , Sulfonas/química , Sulfonas/farmacologíaRESUMEN
Reduction in the parasitemic levels of the Y strain of Trypanosoma cruzi in mice treated with oral or intraperitoneal ursolic (UA) and oleanolic (OA) acids was evaluated during the acute phase of Chagas' disease. Oral administration of UA and OA (50mg/kg/day) provided the most significant reduction in the parasitemic peak, while intraperitoneal administration of UA and OA did not significantly affect the biological activity of the Y strain of T. cruzi. Interleukin levels in mice treated by the intraperitoneal route were compared to untreated chagasic mice. Reduced γ-IFN levels and enhanced IL-10 concentrations potentially explain the exacerbated parasitemia. Our data suggests an immunosuppressive effect for UA and OA, which could interfere with host control of parasitemia. Optimal results were achieved with oral administration. This observation may be explained by the low intestinal absorption of UA and OA, could cause a reduced immune response and promote parasite control. Taken together, these data demonstrate that triterpenes could be interesting compounds to develop therapeutically for the treatment of Chagas' disease.
Asunto(s)
Antiinfecciosos/uso terapéutico , Enfermedad de Chagas/tratamiento farmacológico , Ácido Oleanólico/uso terapéutico , Triterpenos/uso terapéutico , Enfermedad Aguda , Administración Oral , Animales , Antiinfecciosos/administración & dosificación , Antiinfecciosos/química , Enfermedad de Chagas/inmunología , Modelos Animales de Enfermedad , Infusiones Parenterales , Interferón gamma/sangre , Interleucina-10/sangre , Masculino , Melastomataceae/química , Ratones , Ratones Endogámicos BALB C , Nitroimidazoles/administración & dosificación , Nitroimidazoles/uso terapéutico , Ácido Oleanólico/administración & dosificación , Ácido Oleanólico/química , Parasitemia/tratamiento farmacológico , Parasitemia/inmunología , Distribución Aleatoria , Triterpenos/administración & dosificación , Triterpenos/química , Tripanocidas/administración & dosificación , Tripanocidas/uso terapéutico , Ácido UrsólicoRESUMEN
Even though the Chagas' disease, caused by the protozoan Trypanosoma cruzi, was described 100years ago by Carlos Chagas, it still represents a major public health concern and is found in 18 developing countries in South and Central America. In Brazil, Benznidazole (Rochagan) is the only drug with trypanocidal activity available in the market, despite its several side effects and limited efficacy in the chronic phase of the infection. In view of the need for new substances displaying biological activity against T. cruzi, there has been growing interest in research toward the attainment of compounds capable of acting on the parasite while being devoid of serious side effects. In this context, this study aims to evaluate the in vivo therapeutic activity of dibenzylbutyrolactone lignans (-)-cubebin and (-)-hinokinin during the acute phase of infection by T. cruzi. As a study criterion, animals with acute parasitemia were investigated by tissue morphometric analysis. There was significant parasitemia reduction in the groups of animals treated with (-)-cubebin or (-)-hinokin oral administration, compared to the negative control. Values close to those of the uninfected control were found in the groups treated with (-)-cubebin and (-)-hinokinin via kariometry, showing that there was positive cellular response compared to the infected control.
Asunto(s)
4-Butirolactona/análogos & derivados , Enfermedad de Chagas/tratamiento farmacológico , Dioxoles/uso terapéutico , Lignanos/uso terapéutico , Tripanocidas/farmacología , Tripanocidas/uso terapéutico , Trypanosoma cruzi/efectos de los fármacos , 4-Butirolactona/química , 4-Butirolactona/farmacología , 4-Butirolactona/uso terapéutico , Animales , Benzodioxoles , Dioxoles/química , Dioxoles/farmacología , Lignanos/química , Lignanos/farmacología , Masculino , Ratones , Ratones Endogámicos BALB C , Nitroimidazoles/farmacología , Nitroimidazoles/uso terapéutico , Piper/química , Tripanocidas/químicaRESUMEN
The (-)-hinokinin display high activity against Trypanosoma cruzi in vitro and in vivo. (-)-Hinokinin-loaded poly(D,L-lactide-co-glycolide) microparticles were prepared and characterized in order to protect (-)-hinokinin of biological interactions and promote its sustained release for treatment of Chagas disease. The microparticles contain (-)-hinokinin were prepared by the classical method of the emulsion/solvent evaporation. The scanning electron microscopy, light-scattering analyzer were used to study the morphology and particle size, respectively. The encapsulation efficiency was determined, drug release studies were kinetically evaluated, and the trypanocidal effect was evaluated in vivo. (-)-Hinokinin-loaded microparticles obtained showed a mean diameter of 0.862 microm with smooth surface and spherical shape. The encapsulation efficiency was 72.46 +/- 2.92% and developed system maintained drug release with Higuchi kinetics. The preparation method showed to be suitable, since the morphological characteristics, encapsulation efficiency, and in vitro release profile were satisfactory. In vivo assays showed significant reduction of mice parasitaemia after administration of (-)-hinokinin-loaded microparticles. Thus, the developed microparticles seem to be a promising system for sustained release of (-)-hinokinin for treatment of Chagas disease.
Asunto(s)
4-Butirolactona/análogos & derivados , Antiprotozoarios/uso terapéutico , Enfermedad de Chagas/tratamiento farmacológico , Preparaciones de Acción Retardada/uso terapéutico , Dioxoles/uso terapéutico , Ácido Láctico/uso terapéutico , Lignanos/uso terapéutico , Microesferas , Ácido Poliglicólico/uso terapéutico , 4-Butirolactona/farmacocinética , 4-Butirolactona/uso terapéutico , Animales , Benzodioxoles , Preparaciones de Acción Retardada/farmacocinética , Dioxanos , Dioxoles/farmacocinética , Modelos Animales de Enfermedad , Humanos , Ácido Láctico/farmacocinética , Lignanos/farmacocinética , Ratones , Parasitemia/tratamiento farmacológico , Ácido Poliglicólico/farmacocinética , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Resultado del Tratamiento , Trypanosoma cruzi/efectos de los fármacosRESUMEN
The trypanocidal effect of six lignan lactones, (-)-cubebin (1), (-)-O-methyl cubebin (2), (-)-O-benzyl cubebin (3), (-)-6,6'-dinitrohinokinin (4), (-)-hinokinin (5) and dimethoxymorelensin (6), previously synthesized by our research group, was evaluated in vitro and in vivo. The compounds with higher anti-epimastigote activity were screened against intracellular amastigote of Trypanosoma cruzi. Among these, compound 5 was selected to be assayed in vivo. It was observed that compounds 5, 6 and 2 showed higher trypanocidal activity against epimastigote forms of T. cruzi, displaying inhibitory concentration (IC(50)) values of 0.67, 3.89 and 31.35 muM, respectively. These compounds were also evaluated against intracellular amastigote forms of T. cruzi, with five displaying similar activity to benznidazole. In vivo assays showed significant reduction of parasitaemia after administration of five in mice infected.