Studies towards the therapeutic potential of drugs and synthetic compounds free or loaded into liposomes: an in vitro and experimental approach against visceral leishmaniasis

Author(s)

Erika Gracielle Pinto

• Document type: Doctoral Thesis
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Instituto de Medicina Tropical de São Paulo (IMT)
• Defense date: 2016-02-24

Examining board members

André Gustavo Tempone Cardoso; Elizabeth Igne Ferreira; Daniela Gonçales Rando

• Advisor: André Gustavo Tempone Cardoso

Abstract

Included among the neglected tropical diseases (NTDs), leishmaniasis present itself as a complex of varying degrees of serious diseases, resulting from a skin lesion to a progressive fatal visceral infection. The development of new therapies is essential, due to the limited and complex therapeutic arsenal. Thus, this project conducted the in vitro and in vivo studies using infected hamsters with L. (L.) infantum for new therapies against visceral leishmaniasis, using the three following approaches: i) drug repurposing; ii) controlled and sustained systems for drug delivery, such as liposomes, and iii) synthetic analogs of naphthoquinones, as well as, analogs of synthetic iron chelators. The nitazoxanide drug (NTZ) has been studied extensively, showing anti-Leishmania activity against intracellular amastigotes and selectivity index (SI) of 2.5. When it was evaluated in an experimental model, was unable to reduce the parasite burden in the spleen and liver in free form. However, when was encapsulated in liposomes (NTZ-LP), it has conferred higher direction, resulting in an efficiency of 50 to 82% in the spleen and liver, respectively. The in vitro uptake of NTZ-LP was evaluated in healthy and infected macrophages demonstrating a broad distribution in the cytoplasm of both groups. The NTZ-LP physical-chemical parameters were evaluated and showed a unilamellar formulation, negative and with diameter lower than 200nm. Pharmacokinetics studies in the murine model with NTZ and NTZ-LP showed a half-life (T1/2) similar for both formulations (6.5 minutes to NTZ and 8.7 minutes to NTZ-LP). However, the exposure (AUC) and maximum plasma concentration (Cmax), resulted, for the liposomes values, four times higher, which justify the efficacy of liposomal formulation in preclinical study. Additionally, 10 antihistamines drugs were evaluated, but only cinnarizine showed activity in the intracellular amastigotes. The experimental study showed that the drug was unable to reduce the parasite burden in both, spleen and liver infected animals. Nevertheless, when it was encapsulated in liposomes, a 54% efficacy, only, in the liver was observed. Other two drugs were studied relating Leishmania activity: amiodarone (antiarrhythmic) and sertraline (antidepressant). Although amiodarone have shown potent activity against amastigotes with Effective Concentration 50% (EC50) of 0.5 ?M and a high SI of 56, no activity was seen in spleen and liver in the experimental model. On the other hand, sertraline was the most promising drug, showing an IC50 of 1.8 ?M and SI of 10, increasing the treatment levels of 58% in the spleen and 87% in the liver of the animals. Novel synthetic compounds were also studied. Among 36 synthetic analogues of naphthoquinones, 14 demonstrated activity against intracellular amastigotes, highlighting five compounds with SI>5. Moreover, synthetic analogues of iron chelators were not able to eliminate intracellular amastigotes. The project had as its primary objective the attainment of a new drug candidate, free or encapsulated into liposomes, and it could contribute with new perspectives for studies towards novel therapies against visceral leishmaniasis. (AU)

• FAPESP's process: 11/23703-1 - Study of the therapeutic potential of drugs and synthetic compounds, free or loaded into nanoliposomes: in vitro and experimental approach
• Grantee: Érika Gracielle Pinto
• Support Opportunities: Scholarships in Brazil - Doctorate