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| Author(s): |
Marina Berardi
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | Ribeirão Preto. |
| Institution: | Universidade de São Paulo (USP). Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (PCARP/BC) |
| Defense date: | 2010-08-26 |
| Examining board members: |
Amando Siuiti Ito;
Antonio Alonso;
Katia Regina Perez Daghastanli
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| Advisor: | Amando Siuiti Ito |
| Abstract | |
This work is about the aggregation of the leishmanicidal drug miltefosine and a fluorescent analogue and their interaction with phospholipid vesicles. Leishmaniasis is a complex of tropical diseases caused by different species of the genus Leishmania which reaches almost the whole world, including Brazil, and, on the last decades, its visceral form reappeared with hundreds of million people at risk of infection, and the mortality rate increases every year. Several drugs have been used to treat the disease, and miltefosine, a synthetic phospholipid analogue, has been tested and it is already used in some countries. This drug has a confirmed antitumor and orally antileishmanial action on the cell membranes, which is the first local of interaction of a phospholipid analogue. The cytotoxic activity is not specific on concentrations above its critical micelle concentration (cmc), and the knowledge of the aggregation properties of the drug in aqueous medium becomes important, as well as its interaction with other aggregates presents in the environment. Fluorescent analogues of miltefosine allow the use of fluorescent techniques to characterize the antileishmanial activity of miltefosine. In this work we have investigated the aggregation properties of the drug miltefosine (MT), and the fluorescent analogue MT-BODIPY in aqueous medium, by using techniques of surface tension measurements and both, steady-state and time-resolved fluorescence spectroscopy. The values of miltefosine cmc at 25°C from different methods were about 60M in pure aqueous medium (Milli-Q water), 50M in phosphate buffer 10mM (pH 7,4), and 35M in phosphate buffer with the addition of NaCl 150mM. The fluorescent probe amino-hexadecyl-benzamide (Ahba) was used to study the interaction of miltefosine with phospholipid vesicles of DMPC and DPPC, from absorption and fluorescent emission spectra, steady-state anisotropy and fluorescence and anisotropy decays. The results have shown that the effects of miltefosine addition in the vesicles, with molar ratios between 1:100 and 5:100, are not monittored by the probe Ahba, which is located on the polar head groups region on the bilayer. On the other hand, analysing the intrinsic fluorescence of the analogue MT-BODIPY, we concluded that when the molecule is added, in the same molar ratio intervals, there is a disorder in the lipidic bilayer. The fluorescent group BODIPY is located on the aliphatic chain of MT, therefore, the more accentuated effects of miltefosine in the bilayers occur in the region of the apolar tails. (AU) | |