Drug target identification in protozoan parasites

Abstract

Introduction Despite the fact that diseases caused by protozoan parasites represent serious challenges for public health, animal production and welfare, only a limited panel of drugs has been marketed for clinical applications. Areas covered Herein, the authors investigate two strategies, namely whole organism screening and target-based drug design. The present pharmacopoeia has resulted from whole organism screening, and the mode of action and targets of selected drugs are discussed. However, the more recent extensive genome sequencing efforts and the development of dry and wet lab genomics and proteomics that allow high-throughput screening of interactions between micromolecules and recombinant proteins has resulted in target-based drug design as the predominant focus in anti-parasitic drug development. Selected examples of target-based drug design studies are presented, and calcium-dependent protein kinases, important drug targets in apicomplexan parasites, are discussed in more detail. Expert opinion Despite the enormous efforts in target-based drug development, this approach has not yet generated market-ready antiprotozoal drugs. However, whole-organism screening approaches, comprising of both in vitro and in vivo investigations, should not be disregarded. The repurposing of already approved and marketed drugs could be a suitable strategy to avoid fastidious approval procedures, especially in the case of neglected or veterinary parasitoses.

Comparative assessment of the effects of bumped kinase inhibitors on early zebrafish embryo development and pregnancy in mice

“…HFF were cultured in Dulbecco's modified Eagle (DMEM) with phenol red, supplemented with 10% fetal calf serum (FCS) and 50U of penicillin/ml and were maintained as previously described [32]. For measuring cytotoxicity, HFF monolayers were grown in 96 well plates and were exposed to medium containing serial dilutions of BKIs (0.0009-20 μM).…”

“…After culture at 37°C/5%CO 2 for 3 days, medium was discarded and cells were washed once with PBS. Cytotoxicity was measured by Alamar blue assay as described [32]. HepG2 (hepatocyte) or CRL-8155 (lymphocyte) cells were cultured as previously described [33] with minor changes.…”

Leishmaniasis y Enfermedad de Chagas: Herramientas para Nuevos Enfoques en su Tratamiento

“…Esta área, desarrollada mediante las herramientas biomoleculares (biología molecular, bioquímica, genética, etc. ), ha aumentado el número de blancos moleculares para pruebas en screening de alto rendimiento para identificación de agentes innovadores en las últimas décadas (7). Los blancos moleculares se definen como aquellas proteínas o ácidos nucleicos cuya actividad biológica es específica de un organismo y que pueden ser susceptibles a la acción de compuestos o moléculas con la capacidad de alterar sustancialmente su actividad.…”

Gene Cloning, Recombinant Expression, Characterization, and Molecular Modeling of the Glycolytic Enzyme Triosephosphate Isomerase from Fusarium oxysporum

“…The design and chemical synthesis of antifungal molecules have been used as a method to evaluate the inhibition of mycelial growth or to inhibit the activity of specific proteins, as evaluated in F. graminearum [12][13][14]. In recent years, the search for new molecules has been based on the analysis of specific targets such as proteins, of which intracellular signaling proteins, cytoskeleton, membrane, or enzymes that participate in the metabolism have been proposed for the treatment of pathogenic species [15].…”

“…The main function of the TPI enzyme is to catalyze the interconversion of dihydroxyacetone phosphate into (D)-glyceraldehyde-3-phosphate; this process allows the two three-carbon molecules to continue being processed in the glycolytic pathway, since, without this reaction, there would be no net ATP production generated by the pathway [19]. Due to the relevance of this enzyme, some researchers have proposed the TPI as a potential target for the search (or synthesis) for new drugs [15][16][17][18], and even propose it as targets for the development of vaccines, of parasites that affect humans [16,20,21]. According to the aforementioned, it was in our interest to study the TPI of the fungus identified as F. oxysporum, since, despite being a pathogenic fungus of plants and/or humans, the analysis of biochemical characterization of this or another enzyme of the glycolysis had not been performed previously.…”