Estudo comparativo da atividade biológica de loções de uso externo de clobetasol em veículo convencional e lipossomal em um modelo de lesões psoriasiformes em camundongos
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Data
2021-09-29
Tipo de documento
Tese
Título da Revista
ISSN da Revista
Título de Volume
Área do conhecimento
Ciências da Saúde
Modalidade de acesso
Acesso aberto
Editora
Autores
Hilel, Alexandre Santana
Orientador
Bobinski, Franciane
Coorientador
Resumo
Introdução: Psoríase é uma doença multifatorial manifestada por hiperproliferação de queratinócitos na epiderme e inflamação local persistente associada a secreção de citocinas pró-inflamatórias.
Objetivo: Avaliar a eficácia, segurança e potência farmacológica do tratamento com clobetasol em veículo lipossomal (VL) em camundongos com lesões psoriasiformes, além de caracterizar o mecanismo de ação anti-inflamatório.
Métodos: Estudo pré-clínico experimental com análises in vivo e ex vivo. Lesões psoriasiformes foram induzidas em camundongos Swiss fêmeas (n = 12/grupo) com imiquimode (62,5 mg) aplicado em 2 cm2 de pele do dorso (ou veículo nos grupos Sham) por 5 dias. Duas horas após a aplicação os animais foram tratados por via tópica com clobetasol 0,5 mg/mL em veículo convencional (VC), comparando-se às doses de 0,1; 0,25 e 0,5 mg/mL em VL. Foram avaliados parâmetros de eficácia e segurança e quantificadas as concentrações de citocinas pelo método ELISA e proteínas cinase por Western Blotting em amostras de pele.
Resultados: Lesões induzidas foram reduzidas macroscopicamente e microscopicamente sem alterar a temperatura da pele entre os grupos tratados em VL ou VC. Clobetasol nos dois veículos, em diferentes concentrações, promoveu a limpeza completa da pele. No entanto, o tempo necessário ao desaparecimento das lesões psoriasiformes foi menor no grupo tratado com clobetasol a 0,5 mg/mL em VL. Clobetasol em VL a 0,1 e 0,5 mg/mL atenuou os níveis de IL17-A e IFN-γ, efeito similar ao observado com clobetasol em VC (0,5 mg/mL). Os níveis de IL-10, reduzidos nos animais com lesões psoriasiformes, aumentaram no grupo tratado com clobetasol (0,25 mg/mL) em VL. A fosforilação da proteína Akt foi maior nos animais com lesões psoriasiformes e menor após tratamento com clobetasol 0,5 mg/mL em VC ou VL, sem diferença de eficácia entre eles. Os tratamentos se mostraram seguros, não alterando a maioria dos parâmetros hematológicos, peso corporal e de órgãos ou locomoção.
Conclusão: VL em doses usuais de clobetasol (0,5 mg/mL) ou menores (0,1 e 0,25 mg/mL) demonstraram eficácia farmacológica e segurança, abrindo possibilidades para ensaios clínicos no sentido de aumentar a potência farmacológica e diminuir as doses utilizadas no tratamento tópico da psoríase através da utilização dos VL, em especial os nanolipossomas de fosfatidilcolina.
Introduction: A psoriasis is a multifactorial disease manifested by hyperproliferation of keratinocytes in the epidermis with persistent local inflammation associated with secretion of pro-inflammatory cytokines. Objective: To assess the efficacy, safety and pharmacological potency elicited by treatment with clobetasol in a liposomal vehicle (LV) in mice with psoriasiform lesions and to characterize the mechanism of anti-inflammatory action. Methods: Pre-clinical experimental study with in vivo and ex vivo analyzes. Lesions were induced in female Swiss mice (n=12/group) with imiquimode (62.5 mg) applied in 2 cm2 of back skin (or vehicle in Sham groups) for 5 days. Two hours after applying the imiquimode mice were treated by topical route with clobetasol 0.5 mg/mL in conventional vehicle (CV), compared with doses of 0.1; 0.25 and 0.5 mg/mL in LV. Efficacy and safety parameters were evaluated, cytokines and kinase proteins quantified by the ELISA method Western Blotting, respectively, in dorsal skin samples. Results: The induced-lesions macroscopically and microscopically reduced without changing the skin temperature between groups treated in LV or CV. Clobetasol promoted complete cleansing of the skin in both vehicles at different concentrations. The clearance time was shorter in the group treated with 0.5 mg/mL clobetasol in LV. Clobetasol in LV at 0.1 and 0.5 mg/mL attenuated the levels of IL17-A and IFN-γ, similar to the effects observed in CV (0.5 mg/mL). IL-10 levels, which were reduced in animals with psoriasiform lesions, were increased in the group treated with 0.25 mg/mL clobetasol in LV. Akt protein phosphorylation was higher in animals with psoriasiform lesions and lower following clobetasol 0.5 mg/mL in CV and LV, with no difference in efficacy between them. All treatments were shown to be safe, not altering hematological parameters, body and organ weight or locomotion. Conclusion: LV in usual doses of clobetasol (0.5 mg/mL) or lower doses (0.25 mg/mL and 0.1 mg/mL) demonstrated pharmacological efficacy and safety, suggesting the possibility for clinical trials to increase the pharmacological potency and decrease the doses of drugs used in the topical treatment of psoriasis through the use of LV, in particular, the phosphatidylcholine nanoliposomes.
Introduction: A psoriasis is a multifactorial disease manifested by hyperproliferation of keratinocytes in the epidermis with persistent local inflammation associated with secretion of pro-inflammatory cytokines. Objective: To assess the efficacy, safety and pharmacological potency elicited by treatment with clobetasol in a liposomal vehicle (LV) in mice with psoriasiform lesions and to characterize the mechanism of anti-inflammatory action. Methods: Pre-clinical experimental study with in vivo and ex vivo analyzes. Lesions were induced in female Swiss mice (n=12/group) with imiquimode (62.5 mg) applied in 2 cm2 of back skin (or vehicle in Sham groups) for 5 days. Two hours after applying the imiquimode mice were treated by topical route with clobetasol 0.5 mg/mL in conventional vehicle (CV), compared with doses of 0.1; 0.25 and 0.5 mg/mL in LV. Efficacy and safety parameters were evaluated, cytokines and kinase proteins quantified by the ELISA method Western Blotting, respectively, in dorsal skin samples. Results: The induced-lesions macroscopically and microscopically reduced without changing the skin temperature between groups treated in LV or CV. Clobetasol promoted complete cleansing of the skin in both vehicles at different concentrations. The clearance time was shorter in the group treated with 0.5 mg/mL clobetasol in LV. Clobetasol in LV at 0.1 and 0.5 mg/mL attenuated the levels of IL17-A and IFN-γ, similar to the effects observed in CV (0.5 mg/mL). IL-10 levels, which were reduced in animals with psoriasiform lesions, were increased in the group treated with 0.25 mg/mL clobetasol in LV. Akt protein phosphorylation was higher in animals with psoriasiform lesions and lower following clobetasol 0.5 mg/mL in CV and LV, with no difference in efficacy between them. All treatments were shown to be safe, not altering hematological parameters, body and organ weight or locomotion. Conclusion: LV in usual doses of clobetasol (0.5 mg/mL) or lower doses (0.25 mg/mL and 0.1 mg/mL) demonstrated pharmacological efficacy and safety, suggesting the possibility for clinical trials to increase the pharmacological potency and decrease the doses of drugs used in the topical treatment of psoriasis through the use of LV, in particular, the phosphatidylcholine nanoliposomes.
Palavras-chave
Psoríase, Lipossomas, Clobetasol, Nanotecnologia