An on-flow assay for screening of β-secretase ligands by immobilised capillary reactor-mass spectrometry

Vilela, Adriana Ferreira Lopes; Cardoso, Carmen L.

doi:10.1039/c7ay00284j

Cited by 12 publications

(18 citation statements)

References 31 publications

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“…In addition, IMER can be coupled to different separation systems, such as high performance liquid chromatography (HPLC), which solve the possible problem of product and assayed compounds fluorescing at the same wavelength, since these analytes can be separated and analyzed individually (128). Active anti-cancer compounds (129), enzyme inhibitors (130)(131)(132)(133) and G protein-coupled receptors (GPCR) (134) binders have recently been identified using this approach.…”

Section: Target-based Screeningmentioning

confidence: 99%

Schistosomiasis Drug Discovery in the Era of Automation and Artificial Intelligence

et al. 2021

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Schistosomiasis is a parasitic disease caused by trematode worms of the genus Schistosoma and affects over 200 million people worldwide. The control and treatment of this neglected tropical disease is based on a single drug, praziquantel, which raises concerns about the development of drug resistance. This, and the lack of efficacy of praziquantel against juvenile worms, highlights the urgency for new antischistosomal therapies. In this review we focus on innovative approaches to the identification of antischistosomal drug candidates, including the use of automated assays, fragment-based screening, computer-aided and artificial intelligence-based computational methods. We highlight the current developments that may contribute to optimizing research outputs and lead to more effective drugs for this highly prevalent disease, in a more cost-effective drug discovery endeavor.

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Section: Target-based Screeningmentioning

confidence: 99%

Schistosomiasis Drug Discovery in the Era of Automation and Artificial Intelligence

et al. 2021

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“…Zonal elution has also been applied to monitor on-line enzyme activity in ligand screening assays, resulting in reliable and specific assays that allow the biocatalysis product to be directly quantified (da Silva et al, 2012; de Moraes et al, 2013; Calil et al, 2016; Lima et al, 2016; Magalhães et al, 2016; Ferreira Lopes Vilela and Cardoso, 2017; Cornelio et al, 2018; Vilela et al, 2018; Seidl et al, 2019). This approach prevents interferences in inhibitors screening and furnishes reliable data concerning the substrate- and inhibitor-enzyme binding.…”

Section: On-line Approachesmentioning

confidence: 99%

Solid-Supported Proteins in the Liquid Chromatography Domain to Probe Ligand-Target Interactions

2019

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Ligand-target interactions play a central role in drug discovery processes because these interactions are crucial in biological systems. Small molecules-proteins interactions can regulate and modulate protein function and activity through conformational changes. Therefore, bioanalytical tools to screen new ligands have focused mainly on probing ligand-target interactions. These interactions have been evaluated by using solid-supported proteins, which provide advantages like increased protein stability and easier protein extraction from the reaction medium, which enables protein reuse. In some specific approaches, precisely in the ligand fishing assay, the bioanalytical method allows the ligands to be directly isolated from complex mixtures, including combinatorial libraries and natural products extracts without prior purification or fractionation steps. Most of these screening assays are based on liquid chromatography separation, and the binding events can be monitored through on-line or off-line methods. In the on-line approaches, solid supports containing the immobilized biological target are used as chromatographic columns most of the time. Several terms have been used to refer to such approaches, such as weak affinity chromatography, high-performance affinity chromatography, on-flow activity assays, and high-performance liquid affinity chromatography. On the other hand, in the off-line approaches, the binding event occurs outside the liquid chromatography system and may encompass affinity and activity-based assays in which the biological target is immobilized on magnetic particles or monolithic silica, among others. After the incubation step, the supernatant or the eluate from the binding assay is analyzed by liquid chromatography coupled to various detectors. Regardless of the selected bioanalytical approach, the use of solid supported proteins has significantly contributed to the development of automated and reliable screening methods that enable ligands to be isolated and characterized in complex matrixes without purification, thereby reducing costs and avoiding time-laborious steps. This review provides a critical overview of recently developed assays.

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“…To overcome the frequently observed drawbacks of traditional FRET assays, the authors utilized an unlabeled substrate, thereby eliminating discrepancies. Similarly, Ferreira Lopes Vilela and Cardoso took advantage of an immobilized capillary enzyme reactor (ICER) that, when combined to MS, affords on‐flow screening of ligands.…”

Section: β‐Secretase Inhibitorsmentioning

confidence: 99%

Multidisciplinary approaches for targeting the secretase protein family as a therapeutic route for Alzheimer's disease

Schaduangrat

Prachayasittikul

Choomwattana

et al. 2019

Medicinal Research Reviews

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The continual increase of the aging population worldwide renders Alzheimer's disease (AD) a global prime concern. Several attempts have been focused on understanding the intricate complexity of the disease's development along with the on‐ andgoing search for novel therapeutic strategies. Incapability of existing AD drugs to effectively modulate the pathogenesis or to delay the progression of the disease leads to a shift in the paradigm of AD drug discovery. Efforts aimed at identifying AD drugs have mostly focused on the development of disease‐modifying agents in which effects are believed to be long lasting. Of particular note, the secretase enzymes, a group of proteases responsible for the metabolism of the β‐amyloid precursor protein (βAPP) and β‐amyloid (Aβ) peptides production, have been underlined for their promising therapeutic potential. This review article attempts to comprehensively cover aspects related to the identification and use of drugs targeting the secretase enzymes. Particularly, the roles of secretases in the pathogenesis of AD and their therapeutic modulation are provided herein. Moreover, an overview of the drug development process and the contribution of computational (in silico) approaches for facilitating successful drug discovery are also highlighted along with examples of relevant computational works. Promising chemical scaffolds, inhibitors, and modulators against each class of secretases are also summarized herein. Additionally, multitarget secretase modulators are also taken into consideration in light of the current growing interest in the polypharmacology of complex diseases. Finally, challenging issues and future outlook relevant to the discovery of drugs targeting secretases are also discussed.

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An on-flow assay for screening of β-secretase ligands by immobilised capillary reactor-mass spectrometry

Cited by 12 publications

References 31 publications

Schistosomiasis Drug Discovery in the Era of Automation and Artificial Intelligence

Schistosomiasis Drug Discovery in the Era of Automation and Artificial Intelligence

Solid-Supported Proteins in the Liquid Chromatography Domain to Probe Ligand-Target Interactions

Multidisciplinary approaches for targeting the secretase protein family as a therapeutic route for Alzheimer's disease

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