Abstract

The prevalence of invasive fungal infections has increased steadily worldwide in the last few decades, particularly in immunocompromised patient. Many advances have been made in antifungal drug development in the past decade. However, the search for more specific drugs, in an effort to overcome the global problem of resistance to antifungal agents and serious side effects, is increasingly relevant and necessary. Genomics and proteomics have created new ways of examining genes and proteins, which open new strategies for drug development and control of human diseases. In silico analyses and manual mining selected initially 57 potential drug targets, 55 essential genes and 2 genes relevant for fungal survival within the host. Of those, 10 genes were present in eight pathogenic fungi analyzed and absent in the human genome. For initial validation were selected four candidates: trr1 that encodes for thioredoxin reductase, rim8 that encodes for a protein involved in the proteolytic activation of a transcriptional factor in response to alkaline pH, kre2 that encodes for α-1,2-mannosyltransferase and erg6 that encodes for Δ(24)-sterol C-methyltransferase. Two proteins, Kre2 and Trr1 showed a reasonable sequence identity to the templates in PDB, necessary to homology modeling and virtual screening in chemical libraries.The recombinants proteins Erg6 and Trr1 have been produced by heterologous expression in Escherichia coli system. For the Kre2 and Rim8 proteins the system selected was heterologous expression in Pichia pastoris. These proteins have been used for monoclonal antibody production, enzymatic assay, antifungal activity in vitro and crystallization. Our data show that the comparative genomics analysis of eight fungal pathogens enabled the identification of four new potential drug targets, conserved among fungi and absent in the human. The news perspectives were generated for technological development and innovation of antifungal agents.਀ Financial Support: CNPq, FAPDF.਀㰀⼀昀漀渀琀㸀㰀⼀瀀㸀㰀戀爀㸀㰀戀㸀䬀攀礀眀漀爀搀㨀 㰀⼀戀㸀☀渀戀猀瀀㬀搀爀甀最 琀愀爀最攀琀猀Ⰰ 愀渀琀椀昀甀渀最愀氀Ⰰ 挀漀洀瀀愀爀愀琀椀瘀攀 最攀渀漀洀椀挀猀㰀⼀琀搀㸀㰀⼀琀爀㸀㰀⼀琀愀戀氀攀㸀㰀⼀琀爀㸀㰀⼀琀搀㸀㰀⼀琀愀戀氀攀㸀㰀⼀戀漀搀礀㸀㰀⼀栀琀洀氀㸀