Prototipação de um novo método diagnóstico e avaliação in vitro de nova estratégia terapêutica para a leishmaniose

Abstract

Early and low-precision diagnosis decreases the chances of curing leishmaniasis, since the available treatments are limited, toxic and induce resistance. The objective of this work is to develop a new method of immunodiagnosis for tegumentary leishmaniasis, utilizing acustofluidic techniques, as well as to evaluate the leishmanicidal activity of 1,4- naphthoquinone derivatives in vitro. Thus, a technological prospection of new methods of immunodiagnosis for tegumentary leishmaniasis was conducted. Therefore, Leishmania amazonensis antigens were produced for application in performance tests in acustofluidic device, in order to evaluate the efficiency of the immunoassay with different biological samples. Furthermore, the cytotoxicity and leishmanicidal potential of 1,4-naphthoquinone derivatives and their leishmanicidal potential against promastigotes of different species of Leishmania spp. were determined. The technological prospection brought together 11 patent documents, most of which were deposited in Brazil. The evaluated patents presented different perspectives in the use of existing methods. As for the development of the new diagnostic method, the produced antigens were efficiently trapped in the acustofluidic device in a time interval of up to 10 minutes and using a low sample volume, 5 μL. The image pattern between negative and positive samples was very similar and reveals a limitation that the technique has in making this differentiation, since the device agglomerated the antigens in all samples. In the pharmacological studies, the results indicated that all 1,4-naphthoquinone derivatives showed potent leishmanicidal activity against L. chagasi and L. amazonensis, especially JN22, which showed an IC50 of 1.64 μg/mL and 5.07 μg/ mL, respectively, with selectivity superior or equivalent to amphotericin B, without causing a deleterious effect on macrophages. In this way, the present research brought innovative approaches for the development of new strategies for diagnosis and treatment of the disease, but there is a need for further studies to overcome the limitations observed in immunoagglutination tests, as well as to determine the antiamastigote activity and the mechanism of action of the compound JN22.