Partial filling affinity capillary electrophoresis as a useful tool for fragment-based drug discovery: A proof of concept on thrombin

Eléna Farcas, C. Bouckaert, Anne Catherine Servais, Julien Hanson, L. Pochet, Marianne Fillet

Résultats de recherche: Contribution à un journal/une revueArticle

Résumé

With the emergence of more challenging targets, a relatively new approach, fragment-based drug discovery (FBDD), proved its efficacy and gained increasing importance in the pharmaceutical industry. FBDD identifies low molecular-weight (MW) ligands (fragments) that bind to biologically important macromolecules, then a structure-guided fragment growing or merging approach is performed, contributing to the quality of the lead. However, to select the appropriate fragment to be evolved, sensitive analytical screening methods must be used to measure the affinity in the μM or even mM range. In this particular context, we developed a robust and selective partial filling affinity CE (ACE) method for the direct binding screening of a small fragment library in order to identify new thrombin inhibitors. To demonstrate the accuracy of our assay, the complex dissociation constants of three known thrombin inhibitors, namely benzamidine, p-aminobenzamidine and nafamostat were determined and found to be in good concordance with the previously reported values. Finally, the screening of a small library was performed and demonstrated the high discriminatory power of our method towards weak binders compared to classical spectrophotometric activity assay, proving the interest of our method in the context of FBDD.
langueAnglais
journalAnalytica chimica acta
Les DOIs
étatE-pub ahead of print - 2017

Empreinte digitale

Capillary electrophoresis
Capillary Electrophoresis
Drug Discovery
Thrombin
electrokinesis
Screening
drug
Assays
inhibitor
assay
Macromolecules
Merging
pharmaceutical industry
Binders
Drug Industry
Molecular weight
ligand
Libraries
Ligands
Molecular Weight

mots-clés

    Citer ceci

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    title = "Partial filling affinity capillary electrophoresis as a useful tool for fragment-based drug discovery: A proof of concept on thrombin",
    abstract = "With the emergence of more challenging targets, a relatively new approach, fragment-based drug discovery (FBDD), proved its efficacy and gained increasing importance in the pharmaceutical industry. FBDD identifies low molecular-weight (MW) ligands (fragments) that bind to biologically important macromolecules, then a structure-guided fragment growing or merging approach is performed, contributing to the quality of the lead. However, to select the appropriate fragment to be evolved, sensitive analytical screening methods must be used to measure the affinity in the μM or even mM range. In this particular context, we developed a robust and selective partial filling affinity CE (ACE) method for the direct binding screening of a small fragment library in order to identify new thrombin inhibitors. To demonstrate the accuracy of our assay, the complex dissociation constants of three known thrombin inhibitors, namely benzamidine, p-aminobenzamidine and nafamostat were determined and found to be in good concordance with the previously reported values. Finally, the screening of a small library was performed and demonstrated the high discriminatory power of our method towards weak binders compared to classical spectrophotometric activity assay, proving the interest of our method in the context of FBDD.",
    keywords = "Thrombin, Inhibitor, Fragment-based drug discovery, Affinity capillary electrophoresis",
    author = "El{\'e}na Farcas and C. Bouckaert and Servais, {Anne Catherine} and Julien Hanson and L. Pochet and Marianne Fillet",
    year = "2017",
    doi = "10.1016/j.aca.2017.06.035",
    language = "English",
    journal = "Analytica chimica acta",
    issn = "0003-2670",
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    Partial filling affinity capillary electrophoresis as a useful tool for fragment-based drug discovery: A proof of concept on thrombin. / Farcas, Eléna; Bouckaert, C.; Servais, Anne Catherine; Hanson, Julien; Pochet, L.; Fillet, Marianne.

    Dans: Analytica chimica acta, 2017.

    Résultats de recherche: Contribution à un journal/une revueArticle

    TY - JOUR

    T1 - Partial filling affinity capillary electrophoresis as a useful tool for fragment-based drug discovery: A proof of concept on thrombin

    AU - Farcas,Eléna

    AU - Bouckaert,C.

    AU - Servais,Anne Catherine

    AU - Hanson,Julien

    AU - Pochet,L.

    AU - Fillet,Marianne

    PY - 2017

    Y1 - 2017

    N2 - With the emergence of more challenging targets, a relatively new approach, fragment-based drug discovery (FBDD), proved its efficacy and gained increasing importance in the pharmaceutical industry. FBDD identifies low molecular-weight (MW) ligands (fragments) that bind to biologically important macromolecules, then a structure-guided fragment growing or merging approach is performed, contributing to the quality of the lead. However, to select the appropriate fragment to be evolved, sensitive analytical screening methods must be used to measure the affinity in the μM or even mM range. In this particular context, we developed a robust and selective partial filling affinity CE (ACE) method for the direct binding screening of a small fragment library in order to identify new thrombin inhibitors. To demonstrate the accuracy of our assay, the complex dissociation constants of three known thrombin inhibitors, namely benzamidine, p-aminobenzamidine and nafamostat were determined and found to be in good concordance with the previously reported values. Finally, the screening of a small library was performed and demonstrated the high discriminatory power of our method towards weak binders compared to classical spectrophotometric activity assay, proving the interest of our method in the context of FBDD.

    AB - With the emergence of more challenging targets, a relatively new approach, fragment-based drug discovery (FBDD), proved its efficacy and gained increasing importance in the pharmaceutical industry. FBDD identifies low molecular-weight (MW) ligands (fragments) that bind to biologically important macromolecules, then a structure-guided fragment growing or merging approach is performed, contributing to the quality of the lead. However, to select the appropriate fragment to be evolved, sensitive analytical screening methods must be used to measure the affinity in the μM or even mM range. In this particular context, we developed a robust and selective partial filling affinity CE (ACE) method for the direct binding screening of a small fragment library in order to identify new thrombin inhibitors. To demonstrate the accuracy of our assay, the complex dissociation constants of three known thrombin inhibitors, namely benzamidine, p-aminobenzamidine and nafamostat were determined and found to be in good concordance with the previously reported values. Finally, the screening of a small library was performed and demonstrated the high discriminatory power of our method towards weak binders compared to classical spectrophotometric activity assay, proving the interest of our method in the context of FBDD.

    KW - Thrombin

    KW - Inhibitor

    KW - Fragment-based drug discovery

    KW - Affinity capillary electrophoresis

    U2 - 10.1016/j.aca.2017.06.035

    DO - 10.1016/j.aca.2017.06.035

    M3 - Article

    JO - Analytica chimica acta

    T2 - Analytica chimica acta

    JF - Analytica chimica acta

    SN - 0003-2670

    ER -