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Background: Superficial fungal infections are extremely common in the tropical and subtropical regions all over the world. Although numerous antifungal drugs are available for the treatment, the pattern of susceptibility to the drugs being used for dermatophytes varies with time and place. Aims and Objectives: The objective of the study was to isolate, identify, and determine antifungal susceptibility pattern in dermatophytes isolated from patients attending dermatology OPD in a tertiary care hospital. Materials and Methods: This study was conducted on 150 dermatophytes isolated from clinically diagnosed patients with dermatomycosis of skin and nail attending the Department of Dermatology Outpatient Clinic of JJM Medical College over a period of 1 year. These cases were referred to the Department of Microbiology for fungal isolation, culture and sensitivity testing. Results: The most frequently affected age group was 21–30 years. The most common clinical manifestation was tinea corporis (52.56%) followed by tinea cruris (26%). The most common dermatophytes implicated were Trichophyton species in 88%. Irrespective of the species, about 96% of our isolates exhibited high minimum inhibitory concentration (MIC) (>1 ?g/ml) to fluconazole. The MIC of luliconazole was < 0.004 ?g/ml to all the dermatophytes isolates tested, compared to other antifungal agents that were ?0.25 ?g/ml. The MIC range was narrowest for luliconazole 0.002–0.128 ?g/ml and the widest for fluconazole, itraconazole, and ketoconazole (0.125–64, 0.0321–16, and 0.0321–16, respectively). Conclusion: Our study showed that luliconazole was the most active drug against all dermatophytes isolates, followed by itraconazole, ketoconazole, and fluconazole. The higher and wider range in MIC values for itraconazole, ketoconazole, and fluconazole found for some of our isolates raise the possibility of increasing resistance to these drugs.
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SUMMARY Aim: A new stability-indicating liquid chromatography method was developed and validated for the quantitative determination of luliconazole. Materials and methods: Preliminary forced degradation study demonstrated an additional peak of the degradation product at the same retention time to the drug, due to this, the method was developed optimizing the chromatographic conditions to provide sufficient peak resolution (R ≥ 2). The experimental design was evaluated to assess the robustness and the best chromatographic conditions to be used for the validation. Methodology: Luliconazole solutions were exposed to various stress conditions to evaluate the method indication stability, in which the degradation product (DP-1) formed was isolated, identified, and evaluated in silico to predict degradation pathway and toxicity. The procedure was validated by robustness, selectivity, linearity, precision, and accuracy. Liquid chromatography was performed in a Phenomenex® RP-18 column with a mixture of acetonitrile and 0.3% (v/v) triethylamine solution as a mobile phase in isocratic elution. Results and conclusions: The method demonstrated robustness, good recovery, precision, linear response over a range from 5.0 to 40.0 μg.mL-1- and to be stability indicating. The alkaline stress condition resulted in the formation of DP-1. HRMS studies identified this product as an hydroxyacetamide derivative, and in silico studies did not show toxic potential.
RESUMEN Objetivo: Un nuevo método indicativo de estabilidad por cromatografía líquida fue desarrollado y validado para la determinación cuantitativa de luliconazol. Materiales y métodos: Estudios preliminares de degradación forzada demostraron un pico adicional en el mismo tiempo de retención del fármaco. El método desarrollado para optimizar las condiciones cromatográicas proporcionó una adecuada resolución (R ≥ 2). El diseño experimental fue evaluado para verificar su robustez y la mejor condición cromatográica para validación. Metodología: Las soluciones de luliconazol fueron expuestas a diferentes condiciones de estrés para evaluar la indicación de estabilidad del método, el aislamiento del producto de degradación formado (DP-1), su identificación y análisis in silico para predecir su ruta de degradación y toxicidad. El procedimiento se validó por robustez, selectividad, linealidad, precisión y exactitud. Las condiciones cromatográficas incluyeron una columna Phenomenex® RP-18, como fase móvil una mezcla de acetonitrilo y solución 0,3% (v/v) de trietilamina en elución isocrática. Resultados y conclusiones: El método mostró ser robusto, con buena recuperación, precisión, respuesta lineal en el rango de 5,0 a 40,0 μg.mL-1 e indicativo de la estabilidad. La condición de estrés alcalina resultó en la formación de DP-1. Estudios por HRMS identificaron este producto como un derivado hidroxiacetamida y los estudios in silico no mostraron potencial de toxicidad.
RESUMO Objetivo: Um novo método indicativo de estabilidade por cromatograia líquida foi desenvolvido e validado para a determinação quantitativa de luliconazol. Materiais e métodos: Estudos preliminares de degradação forçada demonstraram um pico adicional no mesmo tempo de retenção do medicamento. O método desenvolvido para otimizar as condições cromatográficas proporcionou resolução adequada (R ≥ 2). O delineamento experimental foi avaliado para verificar sua robustez e a melhor condição cromatográica para validação. Metodologia: Soluções de luliconazol foram expostas a diferentes condições de estresse para avaliar a indicação da estabilidade do método, o isolamento do produto de degradação formado (DP-1), sua identificação e análise in silico para predizer sua rota de degradação e toxicidade. O procedimento foi validado quanto à robustez, seletividade, linearidade, precisão e exatidão. As condições cromatográficas incluíram uma coluna Phenomenex® RP-18, como fase móvel uma mistura de acetonitrila e solução de trietilamina 0,3% (v/v) em eluição isocrática. Resultados e conclusões: O método mostrou-se robusto, com boa recuperação, precisão, resposta linear na faixa de 5,0 a 40,0 μg.mL-1 e indicativo de estabilidade. A condição de estresse alcalino resultou na formação de DP-1. Os estudos da HRMS identificaram este produto como um derivado da hidroxiacetamida e os estudos in silico não mostraram nenhum potencial de toxicidade.
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Aims: Tinea corporis & cruris of skin respond well to topical antifungal therapy, but there is a need to apply cream 2- 3 times daily for up to four weeks will impair compliance & lead to treatment failure. Luliconazole is one of those drugs offering good efficacy & tolerability with a short duration of treatment. Terbinafine, an allylamine antifungal agent, acts by selective inhibition of fungal squalene epoxidase. Luliconazole, an imidazole antifungal agent is considered to be more effective in inhibition of ergosterol biosynthesis and its reservoir property in stratum corneum is greater than that of terbinafine. As there are lack of studies between terbinafine & luliconazole, the present study was undertaken to compare the clinical efficacy in tinea corporis/tinea cruris patients. Study Design: Prospective parallel study. Place and Duration of Study: Study was conducted on 60 patients presenting to the Dermatology out-patient department of RL Jalapa Hospital, Kolar, from 1st December 30th April 2012. Methodology: Patients alternatively assigned to either terbinafine or luliconazole & advised to apply test drugs topically for 14 days. Clinical symptoms & signs were assessed using 4-point (pruritus, erythema, scaling) scale & 10% KOH mount at base line, end of treatment visit (15th day) & later 30th day. The data was analysed based on age, gender distribution, duration of lesion, clinical score & KOH mount. Results: Of the 60 patients recruited, all came for 1st follow up (14th day) & 51 patients for 2nd follow-up (30th day). Mean age of the patients was 33.80± 9.58 years in terbinafine & 33.90 ± 9.58 years luliconazole group. Majority of patients were in 12- 40 years aged in both group. Sixty patients and 51 patients were negative for KOH mount preparation on 15th & 30th day respectively. At the end of first follow-up, the clinical score was reduced from 3 to zero (P=0.0001) in both the treatment groups. Mycological cure was 100% in both the drug groups. There was no relapse in 51 patients who came for 2nd follow-up. Four in terbinafine and 5 in luliconazole group were lost to follow up. Conclusion: Only mild forms of tinea infections were included as compared to other studies where moderate to severe (pustules, incrustations, vesiculation). Hence the onset of illness, treatment duration and severity of illness were favorable in this study for two weeks. In both the treatment arms, clinical & mycological cure was comparable, hence once a day application for two weeks of terbinafine & luliconazole were equally effective for treatment of tinea corporis/cruris infection.
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BACKGROUND: Luliconazole (LLCZ) is an imidazole antifungal agent widely used in Japan for treatment of tinea, cutaneous candidiasis, and pityriaisis versicolor, etc. Theoretically, its potent antifungal activity should make it a viable option for various conditions of dermatophytoses, but there has been no studies to support the notion thus far. OBJECTIVE: This prompted the authors to investigate the distribution of minimum inhibitory concentration (MIC) of luliconazole by carrying out in vitro antifungal susceptibility test on Trichophyton (T.) species clinically isolated from Korean subjects with tinea pedis. METHODS: In vitro antifungal susceptibility test, with broth macrodilution method of luliconazole and terbinafine hydrochloride (TBF) as reference drug on two clinically isolated Trichophton species (22 strains of T. rubrum, 5 of T. mentagrophytes), was carried out. RESULTS: The range of MIC for LLCZ on the total of 27 strains was 0.0110~0.0140 microg/mL (geometric mean: 0.0027 microg/mL), and the range for TBF was 0.0120~0.0250 microg/mL (geometric mean: 0.0042 microg/mL). Additionally, the geometric mean for MIC of LLCZ on T. mentagrophytes was 0.0125 microg/mL, and on T. rubrum, it was 0.0019 microg/mL. No strain showed resistance in vitro. CONCLUSIONS: The authors hence demonstrated that antifungal activity of LLCZ in vitro against Trichophyton species clinically isolated from Korean subjects is quite superior. These findings emphasize the usefulness of LLCZ, a promising new imidazole, for the topical management of dermatophytoses.