Dados do Trabalho

Título

ANTILEISHMANIAL ACTIVE AZELASTINE DRUG DID NOT CHANGE TARGET GENE EXPRESSION IN Leishmania infantum PROMASTIGOTES

Introdução

The search for new drugs for Leishmaniasis is becoming increasingly necessary in view of the unfeasibility of current therapeutic regimens. The drug repositioning strategy has garnered attention for its promising outcomes in research. Azelastine is a second-generation antihistamine with anti-allergic and anti-inflammatory properties that acts by blocking H1 histamine receptors. In previous studies, its antileishmanial activity was investigated against Leishmania infantum, demonstrating lower active concentrations compared to the standard drug (miltefosine). However, its mechanism of action remains unclear. In previous studies, the antileishmanial activity of azelastine was identified in both forms of L.infantum. With this, the use of RT-qPCR to study gene expression profiles has become an important tool for identifying potential molecular targets related to the therapeutic response of investigational compounds, allowing for a more precise and rapid assessment of pharmacological potential.

Objetivo (s)

Characterize the gene expression profile of promastigote forms of L. infantum treated with azelastine and identify the relationship between the changes found and the biological activity of this drug in the parasite.

Material e Métodos

Total RNA was extracted from promastigotes treated with azelastine at their 50% effective concentration (EC50) for 1 hour and cDNA was generated for amplification, a panel of 20 genes involved in different cellular functions was used to evaluate relative expression. The reference genes were selected using five different algorithms. The relative expression of the genes was analyzed using the comparative 2-ΔΔCq method and normalized with the reference genes and in relation to the calibrator sample (untreated parasite).

Resultados e Conclusão

Azelastine presented an EC50 of 2.09 µM in promastigote forms after 1h of incubation. Among all assessed genes, the algorithms identified two reference genes, CBS and TRYP. Azelastine did not change relative gene expression levels of all investigated genes. As a second-generation antihistamine, azelastine exhibits enhanced chemical specificity in cellular interactions, reflecting in potential implications for its interaction with the parasite's gene expression profile. The possible mechanism of action of this drug may be involved with other lethal processes of the parasite.

Palavras Chave

Leishmania infantum; drug repositioning; azelastine; molecular targets; RTqPCR