Visualisation of penetration of topical antifungal drug substances through mycosis-infected nails by matrix-assisted laser desorption ionisation mass spectrometry imaging.

BACKGROUND: Matrix-assisted laser desorption ionisation mass spectrometry imaging (MALDI-MSI) is a mass spectrometry-based technique, which can be applied for compound-specific imaging of pharmaceuticals in tissues samples. MALDI-MSI technology is widely used to visualise penetration and distribution profile through different tissues but has never been used with nail tissue. OBJECTIVES: This study used MALDI-MSI technology to visualise distribution profile and penetration into ex vivo human mycosis-infected toenails of three antifungal active ingredients amorolfine, ciclopirox and naftifine contained in topical onychomycosis nail treatment preparations, marketed as Loceryl® , Ciclopoli® and Exoderil® . METHODS: Three mycosis-infected toenails were used for each treatment condition. Six and twenty-four hours after one single topical application of antifungal drugs, excess of formulation was removed, nails were cryo-sectioned at a thickness of 20 µm, and MALDI matrix was deposited on each nail slice. Penetration and distribution profile of amorolfine, ciclopirox and naftifine in the nails were analysed by MALDI-MSI. RESULTS: All antifungal actives have been visualised in the nail by MALDI-MSI. Ciclopirox and naftifine molecules showed a highly localised distribution in the uppermost layer of the nail plate. In comparison, amorolfine diffuses through the nail plate to the deep layers already 6 hours after application and keeps diffusing towards the lowest nail layers within 24 hours. CONCLUSIONS: This study shows for the first-time distribution and penetration of certain antifungal actives into human nails using MALDI-MSI analysis. The results showed a more homogeneous distribution of amorolfine to nail and a better penetration through the infected nails than ciclopirox and naftifine.

Gastroprotective effect of Lagenocarpus rigidus (Kunth) Ness (tirica-do-nativo) leaf extract against indomethacin-induced gastric injury / Efecto gastroprotector de Lagenocarpus rigidus (Kunth) Ness (tirica-do-Nativo) extracto de hoja contra las lesiones gástricas inducidas por indometacina

Introduction: species of the family Cyperaceae are commonly used by the population to treat gastric disorders.However, there are a few ethnopharmacological studies about this family Lagenocarpus rigidus (Kunth) Ness, Cyperaceae, is one of the most widespread swamp species. Objective: evaluate the gastric activity of L. rigidus and its chemical characterization. Methods: ethanolic extract of L. rigidus (ELR) leaves prepared by percolation was subjected to total polyphenol and flavonoid quantification, as well as HPLC quantification of some flavonoids. Angiotensin converting enzyme (ACE) inhibition was determined by colorimetric assays.The gastric effects of ELR were tested in male Wistar rats (n = 6 each group) treated with different doses (600, 60 and 6 mg/kg i.p.) ELR.Gastric lesions were induced by administration of indomethacin (30 mg/kg s.c.).The number of ulcers and the index of mucosal damage (IMD) were determined taking into account the color, edema and bleeding of gastric lesions, the number of petechiae, and the number and size of the ulcers. Statistical analysis of data was performed with one-way ANOVA followed by Tukey's test; significance was p < 0.05. Results: ELR inhibited the ACE (68.5±18.1%) at a concentration of 100 mg/mL.Oral administration of ELR (6, 60 and 600 mg/kg) showed protective activity against indomethacin-induced gastric injury. Total polyphenols in ELR were 157.7 ± 5.8 mg pirogalol/mg equivalent flavonoids and 66.9 ± 3.1 µg equivalent quercetin/mg. Conclusion: L. rigidus protects against acute gastric damage induced by indomethacin in an independent dose manner.

Introducción: las especies de la familia Cyperaceae popularmente se utilizan para tratar trastornos gástricos. Sin embargo, hay pocos estudios etnofarmacológicos sobre esta familia. Lagenocarpus rigidus (Kunth) Ness, Cyperaceae, es una de las especies más grandes de población en pantano. Objetivo: evaluar la actividad gástrica de L. rigidus, junto a su caracterización química. Métodos: el extracto etanólico de hojas de L. rigidus (ELR), preparado por percolación fue objeto de cuantificación de polifenoles y flavonoides totales, cuantificación por HPLC de algunos flavonoides. La inhibición de la enzima convertidora de angiotensina (ECA) se realizó por ensayos colorimétricos. Los efectos gástricos de ELR se llevó a cabo en ratas Wistar macho (n = 6, cada grupo), el tratamiento con diferentes dosis (600, 60 y 6 mg/kg ip) de ELR. Las lesiones gástricas se indujeron mediante la administración de indometacina (30 mg/kg sc). El número de úlceras y signos de puntuación del índice de lesión de las mucosas (IMD) se evaluó teniendo en cuenta el color, edema y hemorragia de las lesiones gástricas, el número de petequias, y el número y tamaño de las úlceras. El análisis estadístico de los datos se realizó mediante ANOVA 1 vía, seguido por el test de Tukey y significación fue de p < 0,05. Resultados: ELR inhiben la ACE (68,5 ± 18,1 %) a una concentración de 100 mg/mL. La administración oral de ELR (6, 60 y 600 mg/kg) mostró un efecto protector gástrico contra inducido por indometacina. Los polifenoles totales de ELR fue 157,7 ± 5,8 mg equivalente de pirogalol/mg de flavonoides y 66,9 ± 3,1 μg equivalentes de quercetina/mg. Conclusiones: L. rigidus protege contra el daño gástrico agudo inducido por la indometacina en una organización independiente de la dosis.