Evaluation of in vivo or in vitro of butenafine free or in nanocarrier on L. (L.) infantum e L. (L.) amazonensis

Author(s)

Adriana Bezerra de Souza

• Document type: Master's Dissertation
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Faculdade de Medicina (FM/SBD)
• Defense date: 2019-11-06

Examining board members

Luiz Felipe Domingues Passero; Thais Alves da Costa Silva

• Advisor: Luiz Felipe Domingues Passero

Abstract

Leishmaniasis are diseases caused by different species of the protozoan parasites belonging to the genus Leishmania. These diseases occur in 98 countries, so it is a global health problem. Although, the treatment is performed with two main drugs that are pentavalent antimonial and amphotericin B. These drugs have a number of side effects to patients, causing the abandonment of the therapy. Therefore, the search for new therapeutic targets for leishmaniasis is urgently needed. An interesting target drug is butenafine chloride, that presented leishmanicidal potential in vitro against dermatotropic species of Leishmania sp. Thus, the in vitro efficacy of free or nanoencapsulated butenafine against promastigote and amastigote forms of the viscerotropic species L. (L.) infantum was evaluated, and the in vivo efficacy for topical use was evaluated in male BALB/c mice infected with L. (L.) amazonensis. For this, nanoformulations were produced in a self-emulsifying nanoformulation system (SNEDD) and nanogel, from which a skin absorption study was also performed. BALB/c mice were infected with L. (L.) amazonensis at the base of the tail and after four weeks topical treatment with free or nanoencapsulated butenafine was administered once daily for a total of 15 days. As a positive control, infected animals received intralesionally meglumine antimoniate (Glucantime®). Lesion sizes of all groups were measured weekly during disease progression and two weeks after the end of treatment, the animals were euthanized, tissue parasitism was quantified at the cutaneous point of parasite inoculation and histological analyzes of the skin parasitism were performed. IFN-Gamma and IL-4 concentrations were also evaluated in the supernatant of lymph node mononuclear cells stimulated with L. (L.) amazonensis total antigen. The obtained data showed that F1, F3 and F4 nanoformulations were effective against L. (L.) infantum (in vitro). In the skin absorption test, it was found that nanoencapsulated butenafine presented better drug distribution and rapid diffusion capacity in the skin, compared to free butenafine. The groups of animals treated with the butenafine formulations presented smaller lesion size and decreased parasite load, characterized by moderate and diffuse inflammatory infiltrate when compared to the untreated infected control group. The nanogel showed similar activity to the animals treated with Glucantime. The cellular immune response of the animals showed that the groups of animals treated with free and nanogel-formulated animals showed increased IFN-Gamma production and cells of animals treated with nanogel also produced increased levels of IL-4. In the histological study of the skin of healthy animals treated with nanoencapsulated or free butenafine showed no changes in the epidermis or dermis, whereas animals treated with Glucantime showed an inflammatory process in the dermis consisting of mononuclear and polymorphonuclear cells. Taken together, these data suggest that repurposing butenafine in leishmaniasis treatment can be effective (AU)

• FAPESP's process: 17/09405-4 - Evaluation of the therapeutic effect of butenafine in the American Tegumentar Leishmaniasis
• Grantee: Adriana Bezerra de Souza
• Support Opportunities: Scholarships in Brazil - Master