RESUMEN
Chemotherapeutic agents used in cancer treatment associated to nanoparticles (LDE) that mimic the composition of low-density lipoprotein and buffer their toxicity can have strong anti-atherosclerosis action, as we showed in cholesterol-fed rabbits. Here, a novel preparation of docetaxel (DTX) carried in LDE was evaluated. Eighteen rabbits were fed 1% cholesterol during 8â¯weeks. After the first 4â¯weeks, 9 animals were treated for 4â¯weeks with intravenous LDE-DTX (1â¯mg/kg/week) and 9 with LDE only (controls) once a week for 4â¯weeks. Animals were then euthanized and the aortas were analyzed for morphometry, immunohistochemistry and Western blot. LDE-DTX treated group showed 80% reduction of atheroma area compared to controls. LDE-DTX treatment reduced in 60% the protein expression of macrophage marker CD68 and of MCP-1 in 80%. LDE-DTX pronouncedly lowered expression of pro-inflammatory markers NF-κB, TNF-α, IL-1ß, IL-6 and von Willebrand factor and elicited 40% reduction in cell proliferation marker PCNA. The presence of smooth muscle cells in the intima was 85% smaller than in controls. Pro-apoptotic caspase 3, caspase 9, Bax, and anti-apoptotic Bcl-2 all were reduced by LDE-DTX. Protein expression of MMP-2 and MMP-9, TGF-ß, and collagen 1 and 3 were also markedly lowered by the LDE-DTX treatment. Animals showed no hematological, hepatic or renal toxicity consequent to LDE-DTX treatment. In conclusion, LDE-DTX showed a wide array of strong effects on pro-inflammatory and proliferation-promoting factors that drive the lesion development. These findings and the lack of observable toxicity indicate that LDE-DTX can be a candidate for future clinical trials.
Asunto(s)
Antiinflamatorios/farmacología , Aorta/efectos de los fármacos , Aortitis/prevención & control , Aterosclerosis/prevención & control , Proliferación Celular/efectos de los fármacos , Docetaxel/farmacología , Lípidos/química , Nanopartículas , Placa Aterosclerótica , Animales , Antiinflamatorios/química , Aorta/metabolismo , Aorta/patología , Aortitis/metabolismo , Aortitis/patología , Proteínas Reguladoras de la Apoptosis/metabolismo , Aterosclerosis/metabolismo , Aterosclerosis/patología , Muerte Celular/efectos de los fármacos , Colesterol en la Dieta , Dieta Alta en Grasa , Modelos Animales de Enfermedad , Docetaxel/química , Composición de Medicamentos , Colágenos Fibrilares/metabolismo , Mediadores de Inflamación/metabolismo , Masculino , Antígeno Nuclear de Célula en Proliferación/metabolismo , Factor de von Willebrand/metabolismoRESUMEN
Fungal resistance is the major problem related to fluconazole treatments. This study aims to develop innovative lipid core nanocapsules and nanostructured lipid carriers containing fluconazole, to study in vitro antifungal activity and to assess the possibility of resistance reversion in Candida albicans, C. glabrata, C. krusei, and C. tropicalis isolates. The action mechanism of nanoparticles was investigated through efflux pumps and scanning electron microscopy studies. The lipid core nanocapsules and nanostructured lipid carriers were prepared by interfacial deposition of preformed polymer and high-pressure homogenization methods, respectively. Both nanostructures presented sizes below 250 nm, SPAN < 1.6, negative zeta potential, pH slightly acid, high drug content and controlled drug release. The nanostructured lipid carriers were unable to reverse the fungal resistance. Lipid core nanoparticles displayed advantages such as a reduction in the effective dose of fluconazole and resistance reversion in all isolates tested - with multiple mechanisms of resistance. The main role of the supramolecular structure and the composition of the nanoparticles on antifungal mechanisms of action were discussed. The results achieved through this study have an impact on clinical therapy, with a potential application in the treatment of fungal infections caused by resistant isolates of Candida spp.
Asunto(s)
Antifúngicos/farmacología , Candida/efectos de los fármacos , Preparaciones de Acción Retardada/química , Farmacorresistencia Fúngica/efectos de los fármacos , Fluconazol/farmacología , Proteínas Fúngicas/antagonistas & inhibidores , Nanopartículas/química , Transportadoras de Casetes de Unión a ATP/antagonistas & inhibidores , Transportadoras de Casetes de Unión a ATP/genética , Transportadoras de Casetes de Unión a ATP/metabolismo , Candida/genética , Candida/crecimiento & desarrollo , Candida/metabolismo , Candida albicans/efectos de los fármacos , Candida albicans/genética , Candida albicans/crecimiento & desarrollo , Candida albicans/metabolismo , Candida glabrata/efectos de los fármacos , Candida glabrata/genética , Candida glabrata/crecimiento & desarrollo , Candida glabrata/metabolismo , Candida tropicalis/efectos de los fármacos , Candida tropicalis/genética , Candida tropicalis/crecimiento & desarrollo , Candida tropicalis/metabolismo , Caprilatos/química , Composición de Medicamentos/métodos , Farmacorresistencia Fúngica/genética , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Expresión Génica , Genes MDR/efectos de los fármacos , Hexosas/química , Concentración de Iones de Hidrógeno , Pruebas de Sensibilidad Microbiana , Nanopartículas/ultraestructura , Palmitatos/química , Tamaño de la Partícula , Triglicéridos/química , Verapamilo/farmacologíaRESUMEN
ABSTRACT Halcinonide is a high-potency topical glucocorticoid used for skin inflammation treatments that presents toxic systemic effects. A simple and quick analytical method to quantify the amount of halcinonide encapsulated into lipid nanoparticles, such as polymeric lipid-core nanoparticles and solid lipid nanoparticles, was developed and validated regarding the drug's encapsulation efficiency and in vitro permeation. The development and validation of the analytical method were carried out using the high performance liquid chromatography with the UV detection at 239 nm. The validation parameters were specificity, linearity, precision and accuracy, limits of detection and quantitation, and robustness. The method presented an isocratic flow rate of 1.0 mL.min-1, a mobile phase methanol:water (85:15 v/v), and a retention time of 4.21 min. The method was validated according to international and national regulations. The halcinonide encapsulation efficiency in nanoparticles was greater than 99% and the in vitro drug permeation study showed that less than 9% of the drug permeated through the membrane, indicating a nanoparticle reservoir effect, which can reduce the halcinonide's toxic systemic effects. These studies demonstrated the applicability of the developed and validated analytical method to quantify halcinonide in lipid nanoparticles.