RESUMEN
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
Parasitic diseases continue to be a major worldwide health problem, and there is an urgent need for development of therapeutic drugs. This paper describes synthesis of dehydrodiferulic acid dilactone 1 and dehydrodisinapic acid dilactone 2 furofuran lignans by oxidative coupling of ferulic and sinapic acids, respectively. Their schistosomicidal, trypanocidal, and leishmanicidal activities were evaluated in vitro against Schistosoma mansoni adult worms, trypomastigote and amastigotes forms of Trypanosoma cruzi, and promastigote forms of Leishmania amazonensis. Compound 1 did not display significant schistosomicidal activity, but it presented potent trypanocidal activity, since it induced death of trypomastigotes and amastigotes with IC50/24h of 9.3µM and 7.3µM, respectively. Compound 2 had slight trypanocidal and schistosomicidal activities. None of the compounds were active against L. amazonensis. These results demonstrated that furofuran lignans are potentially useful for anti-parasitic drugs development and should be further investigated.
Asunto(s)
Furanos/síntesis química , Furanos/farmacología , Lignanos/síntesis química , Lignanos/farmacología , Esquistosomicidas/síntesis química , Esquistosomicidas/farmacología , Tripanocidas/síntesis química , Tripanocidas/farmacología , Animales , Compuestos Bicíclicos Heterocíclicos con Puentes/síntesis química , Compuestos Bicíclicos Heterocíclicos con Puentes/química , Furanos/química , Humanos , Concentración 50 Inhibidora , Lactonas/síntesis química , Lactonas/química , Leishmania/efectos de los fármacos , Leishmania mexicana/efectos de los fármacos , Lignanos/química , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Pruebas de Sensibilidad Parasitaria , Schistosoma mansoni/efectos de los fármacos , Esquistosomicidas/química , Tripanocidas/química , Trypanosoma cruzi/efectos de los fármacosRESUMEN
Protozoans of the trypanosomatid family cause the neglected tropical diseases leishmaniasis and trypanosomiasis, for which few drugs are available. In this context our group has recently reported that the essential oil obtained by steam distillation of the fruits of Piper cubeba is active against Schistosoma mansoni. Therefore, we have investigated the in vitro effects of the essential oil against the trypomastigote and amastigote forms of Trypanosoma cruzi isolated from an LLCMK2 cell line culture and the promastigote forms of Leishmania amazonensis. The in vitro activity of the essential oil against trypomastigotes of T. cruzi increased upon rising concentrations, giving IC50 values of 45.5 and 87.9 µgâ·âmL⻹ against trypomastigotes and amastigotes, respectively. The essential oil was not active against L. amazonensis, since it displayed lyses of only 24â% at 400 µgâ·âmL⻹, and an IC50 of 326.5 µgâ·âmL⻹. Therefore, the essential oil should be further investigated to determine the compounds responsible for the observed activities, as well as its mechanism of action.
Asunto(s)
Antiparasitarios/farmacología , Leishmania/efectos de los fármacos , Aceites Volátiles/farmacología , Piper/química , Extractos Vegetales/farmacología , Trypanosoma cruzi/efectos de los fármacos , Línea Celular , Frutas/química , Concentración 50 Inhibidora , Leishmaniasis/microbiología , Estadios del Ciclo de Vida , Macrófagos , Pruebas de Sensibilidad ParasitariaRESUMEN
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
Nifurtimox and benznidazole, medications currently used for the treatment of the Chagas disease, are not always successful. We determine whether (-)-cubebin and (-)-hinokinin could be used as alternative drugs for the treatment of parasitic infections by Trypanosoma cruzi. To this end, male BALB/c mice were treated with both drugs, and the nuclear parameters (largest diameter, smallest diameter, and perimeter) were determined from slides prepared from the spleen, liver, and heart. The cytotoxicity of the substances was determined after 24-h treatment. Results revealed increased cell nuclei in untreated infected animals as compared to uninfected mice. The values obtained for infected animals treated with (-)-cubebin and (-)-hinokinin were close to those observed for uninfected mice. For the spleen, perimeter values of 10.85 µm (p < 0.01) and 10.90 µm (p < 0.05) were obtained for mice treated with (-)-cubebin 50 mg/kg and (-)-hinokinin 20 mg/kg, respectively, whereas untreated infected animals furnished a perimeter of 11.76 µm. As for the liver, perimeter values of 19.06 µm (p < 0.01) and 18.61 µm (p < 0.001) were achieved for mice treated with (-)-cubebin 50 mg/kg and (-)-hinokinin 20 mg/kg, respectively, whereas a perimeter of 18.54 µm was obtained for untreated infected animals. The cytotoxicity assays demonstrated that (-)-cubebin and (-)-hinokinin does not display toxicity. Therefore, (-)-cubebin and (-)-hinokinin are promising therapeutic agents and could be used in future clinical studies concerning treatment of the Chagas disease. Even if the karyometry is not used frequently, it can complement other methods, such as PCR, and furthermore, it is a simple method which is easily possible to analyze the activity of substances in the tissues of treated infected animals compared to uninfected animals.
Asunto(s)
4-Butirolactona/análogos & derivados , Enfermedad de Chagas/tratamiento farmacológico , Enfermedad de Chagas/patología , Dioxoles/administración & dosificación , Lignanos/administración & dosificación , 4-Butirolactona/administración & dosificación , 4-Butirolactona/efectos adversos , Animales , Antiprotozoarios/administración & dosificación , Antiprotozoarios/efectos adversos , Benzodioxoles , Biometría , Línea Celular , Enfermedad de Chagas/parasitología , Dioxoles/efectos adversos , Modelos Animales de Enfermedad , Fibroblastos/efectos de los fármacos , Cariotipificación , Lignanos/efectos adversos , Hígado/patología , Macaca mulatta , Masculino , Ratones , Ratones Endogámicos BALB C , Bazo/patología , Trypanosoma cruzi/patogenicidadRESUMEN
The reduction of parasitism tissue upon treatment with two lignano lactones, namely (-)- cubebin (CUB) and (-)-hinokinin (HNK), was evaluated in the chronic phase of Chagas' disease by quantifying the enzyme beta-galactosidase expressed by the CL B5 clone strain of Trypanosoma cruzi. Tissue karyometry was also performed. Treatment with the assessed lignans led to a larger reduction in parasitism tissue in all evaluated organs, compared with benznidazole (BZN). Oral treatment with CUB or HNK was more effective. Karyometry results demonstrated that the infected control animals had increased nuclear area compared with uninfected controls, indicating cellular hypertrophy. Results also revealed that use of CUB or HNK was able to significantly prevent this increase, and a slight decrease in the nuclear area was observed, compared with mice treated with BZN. Taken together, these data demonstrate that CUB and HNK could be considered as potential compounds for the development of new drugs for treatment of Chagas' disease.
Asunto(s)
4-Butirolactona/análogos & derivados , Enfermedad de Chagas/tratamiento farmacológico , Dioxoles/uso terapéutico , Lignanos/uso terapéutico , Tripanocidas/uso terapéutico , Trypanosoma cruzi/efectos de los fármacos , 4-Butirolactona/química , 4-Butirolactona/farmacología , 4-Butirolactona/uso terapéutico , Animales , Benzodioxoles , Enfermedad de Chagas/parasitología , Enfermedad Crónica , Dioxoles/química , Dioxoles/farmacología , Corazón/efectos de los fármacos , Corazón/parasitología , Cariometría , Lactonas/química , Lactonas/farmacología , Lactonas/uso terapéutico , Lignanos/química , Lignanos/farmacología , Hígado/efectos de los fármacos , Hígado/parasitología , Ratones , Ratones Endogámicos BALB C , Bazo/efectos de los fármacos , Bazo/parasitología , Resultado del Tratamiento , Tripanocidas/química , Tripanocidas/farmacología , Trypanosoma cruzi/enzimología , Trypanosoma cruzi/aislamiento & purificación , beta-Galactosidasa/metabolismoRESUMEN
The present study evaluates the in vitro and in vivo trypanocidal activity of ursolic acid and oleanolic acid against the Bolivia strain of Trypanosoma cruzi. Their acute toxicity is also assessed on the basis of median lethal dose (DL50) determination and quantification of biochemical parameters. Ursolic acid is the most active compound in vitro, furnishing IC50 of 25.5 microM and displaying 77% of trypomastigote lysis at a concentration of 128 microM. In agreement with in vitro assays, the results obtained for the in vivo assay reveals that ursolic acid (at a dose of 20 mg/Kg/day) provides the most significant reduction in the number of parasites at the parasitemic peak. Results concerning the LD50 assay and the biochemical parameters evaluated in the present study demonstrate that these substances can be safely used on an experimental basis.
Asunto(s)
Enfermedad de Chagas/tratamiento farmacológico , Triterpenos/farmacología , Triterpenos/uso terapéutico , Tripanocidas/farmacología , Tripanocidas/uso terapéutico , Trypanosoma cruzi/efectos de los fármacos , Animales , Supervivencia Celular/efectos de los fármacos , Enfermedad de Chagas/parasitología , Humanos , Concentración 50 Inhibidora , Dosificación Letal Mediana , Masculino , Parasitemia , Ratas , Ratas Wistar , Triterpenos/administración & dosificación , Triterpenos/toxicidad , Tripanocidas/toxicidad , Ácido UrsólicoRESUMEN
We developed a new method for the quantification of parasites in tissue. Trypanosoma cruzi strain CL parasites were genetically engineered to express the Escherichia coli beta-galactosidase gene, lacZ and this enzyme is able to catalyze a colorimetric reaction with chlorophenol red beta-D: galactopyranoside (CPRG) as the substrate. The animals were infected with clone CL Brener strain B5 of T. cruzi and treated with benznidazole in order to verify the reduction in the number of parasites in tissue study by quantifying the enzyme beta-galactosidase. The assay demonstrates a reduction in the number of parasites in the groups treated. Thus, this test can be used to test other substances with the aim of verifying the effectiveness in the chronic phase of experimental Chagas' disease.
Asunto(s)
Enfermedad de Chagas/parasitología , Trypanosoma cruzi/aislamiento & purificación , Animales , Clorofenoles/metabolismo , Colorimetría/métodos , Escherichia coli/enzimología , Galactósidos/metabolismo , Genes Reporteros/genética , Ratones , Ratones Endogámicos BALB C , Coloración y Etiquetado/métodos , beta-Galactosidasa/genética , beta-Galactosidasa/metabolismoRESUMEN
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.