Toward the identification of viral cap-methyltransferase inhibitors by fluorescence screening assay
Identifieur interne : 000D48 ( Main/Exploration ); précédent : 000D47; suivant : 000D49Toward the identification of viral cap-methyltransferase inhibitors by fluorescence screening assay
Auteurs : Wahiba Aouadi [France] ; Cécilia Eydoux [France] ; Bruno Coutard [France] ; Baptiste Martin [France] ; Françoise Debart [France] ; Jean Jacques Vasseur [France] ; Jean Marie Contreras [France] ; Christophe Morice [France] ; Gilles Quérat [France] ; Marie-Louise Jung [France] ; Bruno Canard [France] ; Jean-Claude Guillemot [France] ; Etienne Decroly [France]Source :
- Antiviral Research [ 0166-3542 ] ; 2017.
Descripteurs français
- KwdFr :
- Antiviraux (isolement et purification), Concentration inhibitrice 50, Exoribonucleases (antagonistes et inhibiteurs), Fluorimétrie, Humains, Methyltransferases (antagonistes et inhibiteurs), Protéines virales non structurales (antagonistes et inhibiteurs), Tests de sensibilité microbienne, Virus du SRAS (enzymologie), Évaluation préclinique de médicament ().
- MESH :
- antagonistes et inhibiteurs : Exoribonucleases, Methyltransferases, Protéines virales non structurales.
- enzymologie : Virus du SRAS.
- isolement et purification : Antiviraux.
- Concentration inhibitrice 50, Fluorimétrie, Humains, Tests de sensibilité microbienne, Évaluation préclinique de médicament.
English descriptors
- KwdEn :
- Antiviral Agents (isolation & purification), Drug Evaluation, Preclinical (methods), Exoribonucleases (antagonists & inhibitors), Fluorometry, Humans, Inhibitory Concentration 50, Methyltransferases (antagonists & inhibitors), Microbial Sensitivity Tests, SARS Virus (enzymology), Viral Nonstructural Proteins (antagonists & inhibitors).
- MESH :
- chemical , antagonists & inhibitors : Exoribonucleases, Methyltransferases, Viral Nonstructural Proteins.
- chemical , isolation & purification : Antiviral Agents.
- enzymology : SARS Virus.
- methods : Drug Evaluation, Preclinical.
- Fluorometry, Humans, Inhibitory Concentration 50, Microbial Sensitivity Tests.
- mix :
Abstract
Two highly pathogenic human coronaviruses associated with severe respiratory syndromes emerged since the beginning of the century. The severe acute respiratory syndrome SARS-coronavirus (CoV) spread first in southern China in 2003 with about 8000 infected cases in few months. Then in 2012, the Middle East respiratory syndrome (MERS-CoV) emerged from the Arabian Peninsula giving a still on-going epidemic associated to a high fatality rate. CoVs are thus considered a major health threat. This is especially true as no vaccine nor specific therapeutic are available against either SARS- or MERS-CoV. Therefore, new drugs need to be identified in order to develop antiviral treatments limiting CoV replication. In this study, we focus on the nsp14 protein, which plays a key role in virus replication as it methylates the RNA cap structure at the N7 position of the guanine. We developed a high-throughput N7-MTase assay based on Homogenous Time Resolved Fluorescence (HTRF®) and screened chemical libraries (2000 compounds) on the SARS-CoV nsp14. 20 compounds inhibiting the SARS-CoV nsp14 were further evaluated by IC50 determination and their specificity was assessed toward flavivirus- and human cap N7-MTases. Our results reveal three classes of compounds: 1) molecules inhibiting several MTases as well as the dengue virus polymerase activity unspecifically, 2) pan MTases inhibitors targeting both viral and cellular MTases, and 3) inhibitors targeting one viral MTase more specifically showing however activity against the human cap N7-MTase. These compounds provide a first basis towards the development of more specific inhibitors of viral methyltransferases.
Url:
- http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7113892
- https://hal-amu.archives-ouvertes.fr/hal-02349530
DOI: 10.1016/j.antiviral.2017.06.021
PubMed: 28676301
PubMed Central: 7113892
Affiliations:
- France
- Alsace (région administrative), Grand Est, Languedoc-Roussillon, Occitanie (région administrative), Provence-Alpes-Côte d'Azur
- Marseille, Montpellier, Strasbourg
- Université d'Aix-Marseille
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Le document en format XML
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first="Cécilia" last="Eydoux">Cécilia Eydoux</name><affiliation wicri:level="4"><nlm:aff id="aff1">Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille, France</nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille</wicri:regionArea><placeName><region type="region">Provence-Alpes-Côte d'Azur</region><region type="old region">Provence-Alpes-Côte d'Azur</region><settlement type="city">Marseille</settlement></placeName><orgName type="university">Université d'Aix-Marseille</orgName></affiliation></author><author><name sortKey="Coutard, Bruno" sort="Coutard, Bruno" uniqKey="Coutard B" first="Bruno" last="Coutard">Bruno Coutard</name><affiliation wicri:level="4"><nlm:aff id="aff1">Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille, France</nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille</wicri:regionArea><placeName><region type="region">Provence-Alpes-Côte d'Azur</region><region type="old region">Provence-Alpes-Côte d'Azur</region><settlement type="city">Marseille</settlement></placeName><orgName type="university">Université d'Aix-Marseille</orgName></affiliation></author><author><name sortKey="Martin, Baptiste" sort="Martin, Baptiste" uniqKey="Martin B" first="Baptiste" last="Martin">Baptiste Martin</name><affiliation wicri:level="4"><nlm:aff id="aff1">Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille, France</nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille</wicri:regionArea><placeName><region type="region">Provence-Alpes-Côte d'Azur</region><region type="old region">Provence-Alpes-Côte d'Azur</region><settlement type="city">Marseille</settlement></placeName><orgName type="university">Université d'Aix-Marseille</orgName></affiliation></author><author><name sortKey="Debart, Francoise" sort="Debart, Francoise" uniqKey="Debart F" first="Françoise" last="Debart">Françoise Debart</name><affiliation wicri:level="3"><nlm:aff id="aff2">IBMM, CNRS, Université Montpellier, ENSCM, Campus Triolet, Place E. Bataillon, 34095, Montpellier Cedex 05, France</nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>IBMM, CNRS, Université Montpellier, ENSCM, Campus Triolet, Place E. Bataillon, 34095, Montpellier Cedex 05</wicri:regionArea><placeName><region type="region" nuts="2">Occitanie (région administrative)</region><region type="old region" nuts="2">Languedoc-Roussillon</region><settlement type="city">Montpellier</settlement></placeName></affiliation></author><author><name sortKey="Vasseur, Jean Jacques" sort="Vasseur, Jean Jacques" uniqKey="Vasseur J" first="Jean Jacques" last="Vasseur">Jean Jacques Vasseur</name><affiliation wicri:level="3"><nlm:aff id="aff2">IBMM, CNRS, Université Montpellier, ENSCM, Campus Triolet, Place E. Bataillon, 34095, Montpellier Cedex 05, France</nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>IBMM, CNRS, Université Montpellier, ENSCM, Campus Triolet, Place E. Bataillon, 34095, Montpellier Cedex 05</wicri:regionArea><placeName><region type="region" nuts="2">Occitanie (région administrative)</region><region type="old region" nuts="2">Languedoc-Roussillon</region><settlement type="city">Montpellier</settlement></placeName></affiliation></author><author><name sortKey="Contreras, Jean Marie" sort="Contreras, Jean Marie" uniqKey="Contreras J" first="Jean Marie" last="Contreras">Jean Marie Contreras</name><affiliation wicri:level="3"><nlm:aff id="aff3">Prestwick Chemical, 67400, Illkirch, Strasbourg, France</nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Prestwick Chemical, 67400, Illkirch, Strasbourg</wicri:regionArea><placeName><region type="region">Grand Est</region><region type="old region">Alsace (région administrative)</region><settlement type="city">Strasbourg</settlement></placeName></affiliation></author><author><name sortKey="Morice, Christophe" sort="Morice, Christophe" uniqKey="Morice C" first="Christophe" last="Morice">Christophe Morice</name><affiliation wicri:level="3"><nlm:aff id="aff3">Prestwick Chemical, 67400, Illkirch, Strasbourg, France</nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Prestwick Chemical, 67400, Illkirch, Strasbourg</wicri:regionArea><placeName><region type="region">Grand Est</region><region type="old region">Alsace (région administrative)</region><settlement type="city">Strasbourg</settlement></placeName></affiliation></author><author><name sortKey="Querat, Gilles" sort="Querat, Gilles" uniqKey="Querat G" first="Gilles" last="Quérat">Gilles Quérat</name><affiliation wicri:level="3"><nlm:aff id="aff4">UMR “Emergence des Pathologies Virales” (EPV: Aix-Marseille Université - IRD 190 - Inserm 1207 - EHESP), Marseille, France</nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>UMR “Emergence des Pathologies Virales” (EPV: Aix-Marseille Université - IRD 190 - Inserm 1207 - EHESP), Marseille</wicri:regionArea><placeName><region type="region">Provence-Alpes-Côte d'Azur</region><region type="old region">Provence-Alpes-Côte d'Azur</region><settlement type="city">Marseille</settlement></placeName></affiliation></author><author><name sortKey="Jung, Marie Louise" sort="Jung, Marie Louise" uniqKey="Jung M" first="Marie-Louise" last="Jung">Marie-Louise Jung</name><affiliation wicri:level="3"><nlm:aff id="aff3">Prestwick Chemical, 67400, Illkirch, Strasbourg, France</nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Prestwick Chemical, 67400, Illkirch, Strasbourg</wicri:regionArea><placeName><region type="region">Grand Est</region><region type="old region">Alsace (région administrative)</region><settlement type="city">Strasbourg</settlement></placeName></affiliation></author><author><name sortKey="Canard, Bruno" sort="Canard, Bruno" uniqKey="Canard B" first="Bruno" last="Canard">Bruno Canard</name><affiliation wicri:level="4"><nlm:aff id="aff1">Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille, France</nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille</wicri:regionArea><placeName><region type="region">Provence-Alpes-Côte d'Azur</region><region type="old region">Provence-Alpes-Côte d'Azur</region><settlement type="city">Marseille</settlement></placeName><orgName type="university">Université d'Aix-Marseille</orgName></affiliation></author><author><name sortKey="Guillemot, Jean Claude" sort="Guillemot, Jean Claude" uniqKey="Guillemot J" first="Jean-Claude" last="Guillemot">Jean-Claude Guillemot</name><affiliation wicri:level="4"><nlm:aff id="aff1">Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille, France</nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille</wicri:regionArea><placeName><region type="region">Provence-Alpes-Côte d'Azur</region><region type="old region">Provence-Alpes-Côte d'Azur</region><settlement type="city">Marseille</settlement></placeName><orgName type="university">Université d'Aix-Marseille</orgName></affiliation></author><author><name sortKey="Decroly, Etienne" sort="Decroly, Etienne" uniqKey="Decroly E" first="Etienne" last="Decroly">Etienne Decroly</name><affiliation wicri:level="4"><nlm:aff id="aff1">Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille, France</nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille</wicri:regionArea><placeName><region type="region">Provence-Alpes-Côte d'Azur</region><region type="old region">Provence-Alpes-Côte d'Azur</region><settlement type="city">Marseille</settlement></placeName><orgName type="university">Université d'Aix-Marseille</orgName></affiliation></author></analytic><series><title level="j">Antiviral Research</title><idno type="ISSN">0166-3542</idno><idno type="eISSN">1872-9096</idno><imprint><date when="2017">2017</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antiviral Agents (isolation & purification)</term><term>Drug Evaluation, Preclinical (methods)</term><term>Exoribonucleases (antagonists & inhibitors)</term><term>Fluorometry</term><term>Humans</term><term>Inhibitory Concentration 50</term><term>Methyltransferases (antagonists & inhibitors)</term><term>Microbial Sensitivity Tests</term><term>SARS Virus (enzymology)</term><term>Viral Nonstructural Proteins (antagonists & inhibitors)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Antiviraux (isolement et purification)</term><term>Concentration inhibitrice 50</term><term>Exoribonucleases (antagonistes et inhibiteurs)</term><term>Fluorimétrie</term><term>Humains</term><term>Methyltransferases (antagonistes et inhibiteurs)</term><term>Protéines virales non structurales (antagonistes et inhibiteurs)</term><term>Tests de sensibilité microbienne</term><term>Virus du SRAS (enzymologie)</term><term>Évaluation préclinique de médicament ()</term></keywords><keywords scheme="MESH" type="chemical" qualifier="antagonists & inhibitors" xml:lang="en"><term>Exoribonucleases</term><term>Methyltransferases</term><term>Viral Nonstructural Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Antiviral Agents</term></keywords><keywords scheme="MESH" qualifier="antagonistes et inhibiteurs" xml:lang="fr"><term>Exoribonucleases</term><term>Methyltransferases</term><term>Protéines virales non structurales</term></keywords><keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Antiviraux</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Drug Evaluation, Preclinical</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Fluorometry</term><term>Humans</term><term>Inhibitory Concentration 50</term><term>Microbial Sensitivity Tests</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Concentration inhibitrice 50</term><term>Fluorimétrie</term><term>Humains</term><term>Tests de sensibilité microbienne</term><term>Évaluation préclinique de médicament</term></keywords><keywords scheme="mix" xml:lang="en"><term>Antiviral</term><term>Coronavirus</term><term>Flavivirus</term><term>HTRF</term><term>Inhibitor</term><term>Methyltransferase</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Two highly pathogenic human coronaviruses associated with severe respiratory syndromes emerged since the beginning of the century. The severe acute respiratory syndrome SARS-coronavirus (CoV) spread first in southern China in 2003 with about 8000 infected cases in few months. Then in 2012, the Middle East respiratory syndrome (MERS-CoV) emerged from the Arabian Peninsula giving a still on-going epidemic associated to a high fatality rate. CoVs are thus considered a major health threat. This is especially true as no vaccine nor specific therapeutic are available against either SARS- or MERS-CoV. Therefore, new drugs need to be identified in order to develop antiviral treatments limiting CoV replication. In this study, we focus on the nsp14 protein, which plays a key role in virus replication as it methylates the RNA cap structure at the N7 position of the guanine. We developed a high-throughput N7-MTase assay based on Homogenous Time Resolved Fluorescence (HTRF<sup>®</sup>) and screened chemical libraries (2000 compounds) on the SARS-CoV nsp14. 20 compounds inhibiting the SARS-CoV nsp14 were further evaluated by IC<sub>50</sub> determination and their specificity was assessed toward flavivirus- and human cap N7-MTases. Our results reveal three classes of compounds: 1) molecules inhibiting several MTases as well as the dengue virus polymerase activity unspecifically, 2) pan MTases inhibitors targeting both viral and cellular MTases, and 3) inhibitors targeting one viral MTase more specifically showing however activity against the human cap N7-MTase. These compounds provide a first basis towards the development of more specific inhibitors of viral methyltransferases.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Aouadi, W" uniqKey="Aouadi W">W. 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Fink</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>France</li></country><region><li>Alsace (région administrative)</li><li>Grand Est</li><li>Languedoc-Roussillon</li><li>Occitanie (région administrative)</li><li>Provence-Alpes-Côte d'Azur</li></region><settlement><li>Marseille</li><li>Montpellier</li><li>Strasbourg</li></settlement><orgName><li>Université d'Aix-Marseille</li></orgName></list><tree><country name="France"><region name="Provence-Alpes-Côte d'Azur"><name sortKey="Aouadi, Wahiba" sort="Aouadi, Wahiba" uniqKey="Aouadi W" first="Wahiba" last="Aouadi">Wahiba Aouadi</name></region><name sortKey="Canard, Bruno" sort="Canard, Bruno" uniqKey="Canard B" first="Bruno" last="Canard">Bruno Canard</name><name sortKey="Contreras, Jean Marie" sort="Contreras, Jean Marie" uniqKey="Contreras J" first="Jean Marie" last="Contreras">Jean Marie Contreras</name><name sortKey="Coutard, Bruno" sort="Coutard, Bruno" uniqKey="Coutard B" first="Bruno" last="Coutard">Bruno Coutard</name><name sortKey="Debart, Francoise" sort="Debart, Francoise" uniqKey="Debart F" first="Françoise" last="Debart">Françoise Debart</name><name sortKey="Decroly, Etienne" sort="Decroly, Etienne" uniqKey="Decroly E" first="Etienne" last="Decroly">Etienne Decroly</name><name sortKey="Eydoux, Cecilia" sort="Eydoux, Cecilia" uniqKey="Eydoux C" first="Cécilia" last="Eydoux">Cécilia Eydoux</name><name sortKey="Guillemot, Jean Claude" sort="Guillemot, Jean Claude" uniqKey="Guillemot J" first="Jean-Claude" last="Guillemot">Jean-Claude Guillemot</name><name sortKey="Jung, Marie Louise" sort="Jung, Marie Louise" uniqKey="Jung M" first="Marie-Louise" last="Jung">Marie-Louise Jung</name><name sortKey="Martin, Baptiste" sort="Martin, Baptiste" uniqKey="Martin B" first="Baptiste" last="Martin">Baptiste Martin</name><name sortKey="Morice, Christophe" sort="Morice, Christophe" uniqKey="Morice C" first="Christophe" last="Morice">Christophe Morice</name><name sortKey="Querat, Gilles" sort="Querat, Gilles" uniqKey="Querat G" first="Gilles" last="Quérat">Gilles Quérat</name><name sortKey="Vasseur, Jean Jacques" sort="Vasseur, Jean Jacques" uniqKey="Vasseur J" first="Jean Jacques" last="Vasseur">Jean Jacques Vasseur</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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