Subtilisin of Leishmania amazonensis as Potential Druggable Target: Subcellular Localization, In Vitro Leishmanicidal Activity and Molecular Docking of PF-429242, a Subtilisin Inhibitor

Autor: Pollyanna Stephanie Gomes, Monique Pacheco Duarte Carneiro, Patrícia de Almeida Machado, Valter Viana de Andrade-Neto, Alessandra Marcia da Fonseca-Martins, Amy Goundry, João Vitor Marques Pereira da Silva, Daniel Claudio Oliveira Gomes, Ana Paula Cabral de Araujo Lima, Vítor Ennes-Vidal, Ana Carolina Rennó Sodero, Salvatore Giovanni De-Simone, Herbert L. de Matos Guedes
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: Current Issues in Molecular Biology, Vol 44, Iss 5, Pp 2089-2106 (2022)
Druh dokumentu: article
ISSN: 1467-3045
1467-3037
DOI: 10.3390/cimb44050141
Popis: Subtilisin proteases, found in all organisms, are enzymes important in the post-translational steps of protein processing. In Leishmania major and L. donovani, this enzyme has been described as essential to their survival; however, few compounds that target subtilisin have been investigated for their potential as an antileishmanial drug. In this study, we first show, by electron microscopy and flow cytometry, that subtilisin has broad localization throughout the cytoplasm and membrane of the parasite in the promastigote form with foci in the flagellar pocket. Through in silico analysis, the similarity between subtilisin of different Leishmania species and that of humans were determined, and based on molecular docking, we evaluated the interaction capacity of a serine protease inhibitor against both life cycle forms of Leishmania. The selected inhibitor, known as PF-429242, has already been used against the dengue virus, arenaviruses, and the hepatitis C virus. Moreover, it proved to have antilipogenic activity in a mouse model and caused hypolipidemia in human cells in vitro. Here, PF-429242 significantly inhibited the growth of L. amazonensis promastigotes of four different strains (IC50 values = 3.07 ± 0.20; 0.83 ± 0.12; 2.02 ± 0.27 and 5.83 ± 1.2 µM against LTB0016, PH8, Josefa and LV78 strains) whilst having low toxicity in the host macrophages (CC50 = 170.30 µM). We detected by flow cytometry that there is a greater expression of subtilisin in the amastigote form; however, PF-429242 had a low effect against this intracellular form with an IC50 of >100 µM for intracellular amastigotes, as well as against axenic amastigotes (94.12 ± 2.8 µM for the LV78 strain). In conclusion, even though PF-429242 does not affect the intracellular forms, this drug will serve as a tool to explore pharmacological and potentially leishmanicidal targets.
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