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| Author(s): |
Natália Carvalho Pellison
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | Ribeirão Preto. |
| Institution: | Universidade de São Paulo (USP). Faculdade de Medicina de Ribeirão Preto (PCARP/BC) |
| Defense date: | 2018-05-28 |
| Examining board members: |
Dario Simões Zamboni;
Angela Kaysel Cruz;
Silvia Reni Bortolin Uliana
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| Advisor: | Dario Simões Zamboni |
| Abstract | |
Leishmaniasis is a neglected tropical disease caused by protozoan parasites of Leishmania genus. World Health Organization estimates that 700.000 new cases and almost 30.000 deaths occur every year. The most affected groups are those of lower socioeconomical status living in isolated areas, including conflict regions, which complicates access to treatment. Currently, the drugs available for treatment include pentavalent antimonials, amphotericin B, miltefosine and pentamidine. However, the long treatment course of the disease, numerous adverse effects of the drugs and extended follow-up period results in therapy interruption by the patients. These factors compounded with incorrect treatment plans facilitate the emergence of resistant strains. In view of these limitations and disadvantages of the available therapeutic options, the development of alternative treatments is essential in the treatment of Leishmania. Recognizing this global necessity, we screened a commercial drug library containing 2560 compounds, searching for hits with anti-leishmanial activity. For this, we used bone marrow-derived macrophages (BMDMs) from C57Bl/6 mice and Leishmania (L.) amazonensis expressing red fluorescent protein (RFP). Infected cells were treated with 10 µM of each small molecule present in the drug library. Using a high-content screening equipment, images of each well were acquired and analyzed after 24, 48 and 72 hours of infection. We accessed the number of cells and parasites using automatized analysis. BMDMs treated or not with 1% DMSO were used as negative control and Amphotericin B treatment was used as positive control. Both positive and negative controls were used to plate validation and to exclude unresponsive drugs. During the primary screening, 256 compounds showed significant statistical difference when compared with 1% DMSO control. Of them, 96 hits reduced more than 50% of parasites and were reanalyzed at 10 µM to validate the anti-leishmanial activity. Their cytotoxic concentration (CC50), effective concentration for intracellular amastigotes (EC50) and selectivity index (IS) were also determined. During this work, we identified 12 hits compound for anti-leishmanial actitivity, which had IS>20 and could represent potential leishmaniasis treatment. (AU) | |
| FAPESP's process: | 16/01794-9 - Investigation of the role of rapamicyn in restriction of Leishmania amazonensis replication in macrophages and in vivo. |
| Grantee: | Natália Carvalho Pellison |
| Support Opportunities: | Scholarships in Brazil - Master |