Author: "Hélène Munier" - Searchworks@Jio Institute Digital Library Search Results

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145 results on '"Hélène Munier"'

1. An equine iPSC-based phenotypic screening platform identifies pro- and anti-viral molecules against West Nile virus

Author: Marielle Cochet, François Piumi, Kamila Gorna, Noémie Berry, Gaëlle Gonzalez, Anne Danckaert, Nathalie Aulner, Odile Blanchet, Stéphan Zientara, Francesc Xavier Donadeu, Hélène Munier-Lehmann, Jennifer Richardson, Alexandra Benchoua, and Muriel Coulpier
Subjects: Equine, brain, neural progenitors, Flavivirus, antiviral, statin, Veterinary medicine, SF600-1100
Abstract: Abstract Outbreaks of West Nile virus (WNV) occur periodically, affecting both human and equine populations. There are no vaccines for humans, and those commercialised for horses do not have sufficient coverage. Specific antiviral treatments do not exist. Many drug discovery studies have been conducted, but since rodent or primate cell lines are normally used, results cannot always be transposed to horses. There is thus a need to develop relevant equine cellular models. Here, we used induced pluripotent stem cells to develop a new in vitro model of WNV-infected equine brain cells suitable for microplate assay, and assessed the cytotoxicity and antiviral activity of forty-one chemical compounds. We found that one nucleoside analog, 2′C-methylcytidine, blocked WNV infection in equine brain cells, whereas other compounds were either toxic or ineffective, despite some displaying anti-viral activity in human cell lines. We also revealed an unexpected proviral effect of statins in WNV-infected equine brain cells. Our results thus identify a potential lead for future drug development and underscore the importance of using a tissue- and species-relevant cellular model for assessing the activity of antiviral compounds.
Published: 2024
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2. A journey into the regulatory secrets of the de novo purine nucleotide biosynthesis

Author: Nour Ayoub, Antoine Gedeon, and Hélène Munier-Lehmann
Subjects: allostery, antibacterial agents, chemical compounds, enzyme regulation, IMP dehydrogenase, nucleotide biosynthesis, Therapeutics. Pharmacology, RM1-950
Abstract: De novo purine nucleotide biosynthesis (DNPNB) consists of sequential reactions that are majorly conserved in living organisms. Several regulation events take place to maintain physiological concentrations of adenylate and guanylate nucleotides in cells and to fine-tune the production of purine nucleotides in response to changing cellular demands. Recent years have seen a renewed interest in the DNPNB enzymes, with some being highlighted as promising targets for therapeutic molecules. Herein, a review of two newly revealed modes of regulation of the DNPNB pathway has been carried out: i) the unprecedent allosteric regulation of one of the limiting enzymes of the pathway named inosine 5′-monophosphate dehydrogenase (IMPDH), and ii) the supramolecular assembly of DNPNB enzymes. Moreover, recent advances that revealed the therapeutic potential of DNPNB enzymes in bacteria could open the road for the pharmacological development of novel antibiotics.
Published: 2024
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3. Discovery of Bis-Imidazoline Derivatives as New CXCR4 Ligands

Author: Zhicheng Zhou, Isabelle Staropoli, Anne Brelot, Peggy Suzanne, Aurélien Lesnard, Fanny Fontaine, Serge Perato, Sylvain Rault, Olivier Helynck, Fernando Arenzana-Seisdedos, Jana Sopkova-de Oliveira Santos, Bernard Lagane, Hélène Munier-Lehmann, and Philippe Colin
Subjects: CXCR4, CXCL12, bis-imidazoline, antagonist, therapeutic target, HIV-1 infection, Organic chemistry, QD241-441
Abstract: The chemokine receptor CXCR4 and its ligand CXCL12 regulate leukocyte trafficking, homeostasis and functions and are potential therapeutic targets in many diseases such as HIV-1 infection and cancers. Here, we identified new CXCR4 ligands in the CERMN chemical library using a FRET-based high-throughput screening assay. These are bis-imidazoline compounds comprising two imidazole rings linked by an alkyl chain. The molecules displace CXCL12 binding with submicromolar potencies, similarly to AMD3100, the only marketed CXCR4 ligand. They also inhibit anti-CXCR4 mAb 12G5 binding, CXCL12-mediated chemotaxis and HIV-1 infection. Further studies with newly synthesized derivatives pointed out to a role of alkyl chain length on the bis-imidazoline properties, with molecules with an even number of carbons equal to 8, 10 or 12 being the most potent. Interestingly, these differ in the functions of CXCR4 that they influence. Site-directed mutagenesis and molecular docking predict that the alkyl chain folds in such a way that the two imidazole groups become lodged in the transmembrane binding cavity of CXCR4. Results also suggest that the alkyl chain length influences how the imidazole rings positions in the cavity. These results may provide a basis for the design of new CXCR4 antagonists targeting specific functions of the receptor.
Published: 2023
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4. Screening of potential antiviral molecules against equid herpesvirus-1 using cellular impedance measurement: Dataset of 2,891 compounds.

Author: Côme Thieulent, Christine Fortier, Hélène Munier-Lehmann, Peggy Suzanne, Patrick Dallemagne, Stephan Zientara, Aymeric Hans, Romain Paillot, Pierre-Olivier Vidalain, Stéphane Pronost, and Erika Hue
Subjects: Real-time cell assay, Chemical library screening, Antiviral, Equid herpesvirus-1, Computer applications to medicine. Medical informatics, R858-859.7, Science (General), Q1-390
Abstract: Data presented in this article are associated with the research article “Identification of antiviral compounds against equid herpesvirus-1 using real-time cell assay screening: efficacy of decitabine and valganciclovir alone and in combination” [1]. These data correspond to the in vitro screening of 2,891 potential antiviral compounds against equid herpesvirus-1 (EHV-1) based on impedance measurements using the xCELLigence® RTCA MP System. This dataset includes compounds from three different libraries: i) 1,199 compounds from the Prestwick® Chemical Library, which contains mostly US Food and Drug Administration approved drugs (Prestwick® Chemical, Illkirch, France); ii) 1,651 compounds from the Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN, Caen, France); iii) 41 compounds (called herein in-house antiviral library) selected for their effects against different human viruses. Compounds effective against EHV-1 were selected using the area under normalised curves (AUCn) and the time required for the Cell Index to decrease by 50% after virus infection (CIT50). The full dataset from the screen is made publicly available for further analyses.
Published: 2020
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5. 1-(Piperidin-3-yl)thymine amides as inhibitors of M. tuberculosis thymidylate kinase

Author: Yanlin Jian, Martijn D. P. Risseeuw, Mathy Froeyen, Lijun Song, Davie Cappoen, Paul Cos, Hélène Munier-Lehmann, and Serge van Calenbergh
Subjects: thymidylate kinase, inhibitors, mycobacterium tuberculosis, modelling, Therapeutics. Pharmacology, RM1-950
Abstract: A series of readily accessible 1-(piperidin-3-yl)thymine amides was designed, synthesised and evaluated as Mycobacterium tuberculosis TMPK (MtbTMPK) inhibitors. In line with the modelling results, most inhibitors showed reasonable MtbTMPK inhibitory activity. Compounds 4b and 4i were slightly more potent than the parent compound 3. Moreover, contrary to the latter, amide analogue 4g was active against the avirulent M. tuberculosis H37Ra strain (MIC50=35 µM). This finding opens avenues for future modifications.
Published: 2019
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6. Identification of a small molecule that primes the type I interferon response to cytosolic DNA

Author: Samira Khiar, Marianne Lucas-Hourani, Sébastien Nisole, Nikaïa Smith, Olivier Helynck, Maryline Bourgine, Claude Ruffié, Jean-Philippe Herbeuval, Hélène Munier-Lehmann, Frédéric Tangy, and Pierre-Olivier Vidalain
Subjects: Medicine, Science
Abstract: Abstract The type I interferon response plays a pivotal role in host defense against infectious agents and tumors, and promising therapeutic approaches rely on small molecules designed to boost this system. To identify such compounds, we developed a high-throughput screening assay based on HEK-293 cells expressing luciferase under the control of Interferon-Stimulated Response Elements (ISRE). An original library of 10,000 synthetic compounds was screened, and we identified a series of 1H-benzimidazole-4-carboxamide compounds inducing the ISRE promoter sequence, specific cellular Interferon-Stimulated Genes (ISGs), and the phosphorylation of Interferon Regulatory Factor (IRF) 3. ISRE induction by ChX710, a prototypical member of this chemical series, was dependent on the adaptor MAVS and IRF1, but was IRF3 independent. Although it was unable to trigger type I IFN secretion per se, ChX710 efficiently primed cellular response to transfected plasmid DNA as assessed by potent synergistic effects on IFN-β secretion and ISG expression levels. This cellular response was dependent on STING, a key adaptor involved in the sensing of cytosolic DNA and immune activation by various pathogens, stress signals and tumorigenesis. Our results demonstrate that cellular response to cytosolic DNA can be boosted with a small molecule, and potential applications in antimicrobial and cancer therapies are discussed.
Published: 2017
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7. 1-(1-Arylethylpiperidin-4-yl)thymine Analogs as Antimycobacterial TMPK Inhibitors

Author: Yanlin Jian, Fabian Hulpia, Martijn D. P. Risseeuw, He Eun Forbes, Guy Caljon, Hélène Munier-Lehmann, Helena I. M. Boshoff, and Serge Van Calenbergh
Subjects: tuberculosis, Mycobacterium tuberculosis, MtbTMPK, 1-(1-arylethylpiperidin-4-yl)thymine, Organic chemistry, QD241-441
Abstract: A series of Mycobacterium tuberculosis TMPK (MtbTMPK) inhibitors based on a reported compound 3 were synthesized and evaluated for their capacity to inhibit MtbTMPK catalytic activity and the growth of a virulent M. tuberculosis strain (H37Rv). Modifications of the scaffold of 3 failed to afford substantial improvements in MtbTMPK inhibitory activity and antimycobacterial activity. Optimization of the substitution pattern of the D ring of 3 resulted in compound 21j with improved MtbTMPK inhibitory potency (three-fold) and H37Rv growth inhibitory activity (two-fold). Moving the 3-chloro substituent of 21j to the para-position afforded isomer 21h, which, despite a 10-fold increase in IC50-value, displayed promising whole cell activity (minimum inhibitory concentration (MIC) = 12.5 μM).
Published: 2020
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8. Identification of novel leishmanicidal molecules by virtual and biochemical screenings targeting Leishmania eukaryotic translation initiation factor 4A.

Author: Emna Harigua-Souiai, Yosser Zina Abdelkrim, Imen Bassoumi-Jamoussi, Ons Zakraoui, Guillaume Bouvier, Khadija Essafi-Benkhadir, Josette Banroques, Nathan Desdouits, Hélène Munier-Lehmann, Mourad Barhoumi, N Kyle Tanner, Michael Nilges, Arnaud Blondel, and Ikram Guizani
Subjects: Arctic medicine. Tropical medicine, RC955-962, Public aspects of medicine, RA1-1270
Abstract: Leishmaniases are neglected parasitic diseases in spite of the major burden they inflict on public health. The identification of novel drugs and targets constitutes a research priority. For that purpose we used Leishmania infantum initiation factor 4A (LieIF), an essential translation initiation factor that belongs to the DEAD-box proteins family, as a potential drug target. We modeled its structure and identified two potential binding sites. A virtual screening of a diverse chemical library was performed for both sites. The results were analyzed with an in-house version of the Self-Organizing Maps algorithm combined with multiple filters, which led to the selection of 305 molecules. Effects of these molecules on the ATPase activity of LieIF permitted the identification of a promising hit (208) having a half maximal inhibitory concentration (IC50) of 150 ± 15 μM for 1 μM of protein. Ten chemical analogues of compound 208 were identified and two additional inhibitors were selected (20 and 48). These compounds inhibited the mammalian eIF4I with IC50 values within the same range. All three hits affected the viability of the extra-cellular form of L. infantum parasites with IC50 values at low micromolar concentrations. These molecules showed non-significant toxicity toward THP-1 macrophages. Furthermore, their anti-leishmanial activity was validated with experimental assays on L. infantum intramacrophage amastigotes showing IC50 values lower than 4.2 μM. Selected compounds exhibited selectivity indexes between 19 to 38, which reflects their potential as promising anti-Leishmania molecules.
Published: 2018
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9. Two classes of bacterial IMPDHs according to their quaternary structures and catalytic properties.

Author: Thomas Alexandre, Bertrand Raynal, and Hélène Munier-Lehmann
Subjects: Medicine, Science
Abstract: Inosine-5'-monophosphate dehydrogenase (IMPDH) occupies a key position in purine nucleotide metabolism. In this study, we have performed the biochemical and physico-chemical characterization of eight bacterial IMPDHs, among which six were totally unexplored. This study led to a classification of bacterial IMPDHs according to the regulation of their catalytic properties and their quaternary structures. Class I IMPDHs are cooperative enzymes for IMP, which are activated by MgATP and are octameric in all tested conditions. On the other hand, class II IMPDHs behave as Michaelis-Menten enzymes for both substrates and are tetramers in their apo state or in the presence of IMP, which are shifted to octamers in the presence of NAD or MgATP. Our work provides new insights into the IMPDH functional regulation and a model for the quaternary structure modulation is proposed.
Published: 2015
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10. Correction: Two Classes of Bacterial IMPDHs according to Their Quaternary Structures and Catalytic Properties.

Author: Thomas Alexandre, Bertrand Raynal, and Hélène Munier-Lehmann
Subjects: Medicine, Science
Published: 2015
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11. Inhibition of pyrimidine biosynthesis pathway suppresses viral growth through innate immunity.

Author: Marianne Lucas-Hourani, Daniel Dauzonne, Pierre Jorda, Gaëlle Cousin, Alexandru Lupan, Olivier Helynck, Grégory Caignard, Geneviève Janvier, Gwénaëlle André-Leroux, Samira Khiar, Nicolas Escriou, Philippe Desprès, Yves Jacob, Hélène Munier-Lehmann, Frédéric Tangy, and Pierre-Olivier Vidalain
Subjects: Immunologic diseases. Allergy, RC581-607, Biology (General), QH301-705.5
Abstract: Searching for stimulators of the innate antiviral response is an appealing approach to develop novel therapeutics against viral infections. Here, we established a cell-based reporter assay to identify compounds stimulating expression of interferon-inducible antiviral genes. DD264 was selected out of 41,353 compounds for both its immuno-stimulatory and antiviral properties. While searching for its mode of action, we identified DD264 as an inhibitor of pyrimidine biosynthesis pathway. This metabolic pathway was recently identified as a prime target of broad-spectrum antiviral molecules, but our data unraveled a yet unsuspected link with innate immunity. Indeed, we showed that DD264 or brequinar, a well-known inhibitor of pyrimidine biosynthesis pathway, both enhanced the expression of antiviral genes in human cells. Furthermore, antiviral activity of DD264 or brequinar was found strictly dependent on cellular gene transcription, nuclear export machinery, and required IRF1 transcription factor. In conclusion, the antiviral property of pyrimidine biosynthesis inhibitors is not a direct consequence of pyrimidine deprivation on the virus machinery, but rather involves the induction of cellular immune response.
Published: 2013
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12. AutomiG, a biosensor to detect alterations in miRNA biogenesis and in small RNA silencing guided by perfect target complementarity.

Author: Clément Carré, Caroline Jacquier, Anne-Laure Bougé, Fabrice de Chaumont, Corinne Besnard-Guerin, Hélène Thomassin, Josette Pidoux, Bruno Da Silva, Eleftheria Chalatsi, Sarah Zahra, Jean-Christophe Olivo-Marin, Hélène Munier-Lehmann, and Christophe Antoniewski
Subjects: Medicine, Science
Abstract: Defects in miRNA biogenesis or activity are associated to development abnormalities and diseases. In Drosophila, miRNAs are predominantly loaded in Argonaute-1, which they guide for silencing of target RNAs. The miRNA pathway overlaps the RNAi pathway in this organism, as miRNAs may also associate with Argonaute-2, the mediator of RNAi. We set up a gene construct in which a single inducible promoter directs the expression of the GFP protein as well as two miRNAs perfectly matching the GFP sequences. We show that self-silencing of the resulting automiG gene requires Drosha, Pasha, Dicer-1, Dicer-2 and Argonaute-2 loaded with the anti-GFP miRNAs. In contrast, self-silencing of the automiG gene does not involve Argonaute-1. Thus, automiG reports in vivo for both miRNA biogenesis and Ago-2 mediated silencing, providing a powerful biosensor to identify situations where miRNA or siRNA pathways are impaired. As a proof of concept, we used automiG as a biosensor to screen a chemical library and identified 29 molecules that strongly inhibit miRNA silencing, out of which 5 also inhibit RNAi triggered by long double-stranded RNA. Finally, the automiG sensor is also self-silenced by the anti-GFP miRNAs in HeLa cells and might be easily used to identify factors involved in miRNA biogenesis and silencing guided by perfect target complementarity in mammals.
Published: 2013
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13. High content analysis of primary macrophages hosting proliferating Leishmania amastigotes: application to anti-leishmanial drug discovery.

Author: Nathalie Aulner, Anne Danckaert, Eline Rouault-Hardoin, Julie Desrivot, Olivier Helynck, Pierre-Henri Commere, Hélène Munier-Lehmann, Gerald F Späth, Spencer L Shorte, Geneviève Milon, and Eric Prina
Subjects: Arctic medicine. Tropical medicine, RC955-962, Public aspects of medicine, RA1-1270
Abstract: Background/objectivesHuman leishmaniases are parasitic diseases causing severe morbidity and mortality. No vaccine is available and numerous factors limit the use of current therapies. There is thus an urgent need for innovative initiatives to identify new chemotypes displaying selective activity against intracellular Leishmania amastigotes that develop and proliferate inside macrophages, thereby causing the pathology of leishmaniasis.Methodology/principal findingsWe have developed a biologically sound High Content Analysis assay, based on the use of homogeneous populations of primary mouse macrophages hosting Leishmania amazonensis amastigotes. In contrast to classical promastigote-based screens, our assay more closely mimics the environment where intracellular amastigotes are growing within acidic parasitophorous vacuoles of their host cells. This multi-parametric assay provides quantitative data that accurately monitors the parasitic load of amastigotes-hosting macrophage cultures for the discovery of leishmanicidal compounds, but also their potential toxic effect on host macrophages. We validated our approach by using a small set of compounds of leishmanicidal drugs and recently published chemical entities. Based on their intramacrophagic leishmanicidal activity and their toxicity against host cells, compounds were classified as irrelevant or relevant for entering the next step in the drug discovery pipeline.Conclusions/significanceOur assay represents a new screening platform that overcomes several limitations in anti-leishmanial drug discovery. First, the ability to detect toxicity on primary macrophages allows for discovery of compounds able to cross the membranes of macrophage, vacuole and amastigote, thereby accelerating the hit to lead development process for compounds selectively targeting intracellular parasites. Second, our assay allows discovery of anti-leishmanials that interfere with biological functions of the macrophage required for parasite development and growth, such as organelle trafficking/acidification or production of microbicidal effectors. These data thus validate a novel phenotypic screening assay using virulent Leishmania amastigotes growing inside primary macrophage to identify new chemical entities with bona fide drug potential.
Published: 2013
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14. Interaction network among de novo purine nucleotide biosynthesis enzymes in Escherichia coli

Author: Antoine Gedeon, Gouzel Karimova, Nour Ayoub, Julien Dairou, Quentin Giai Gianetto, Sophie Vichier‐Guerre, Pierre‐Olivier Vidalain, Daniel Ladant, Hélène Munier‐Lehmann, Chimie et Biocatalyse, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Université Paris Cité (UPCité), Biochimie des Interactions Macromoléculaires / Biochemistry of Macromolecular Interactions, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB, Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Plateforme de Protéomique / Proteomics platform, Université Paris Cité (UPCité)-Spectrométrie de Masse pour la Biologie – Mass Spectrometry for Biology (UTechS MSBio), Institut Pasteur [Paris] (IP)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité)-Institut Pasteur [Paris] (IP)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Spectrométrie de Masse pour la Biologie – Mass Spectrometry for Biology (UTechS MSBio), Institut Pasteur [Paris] (IP)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Chimie biologique épigénétique - Epigenetic Chemical Biology (EpiCBio), Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and This work was supported in part by the Centre National de la Recherche Scientifique (CNRS), the Institut National de la Santé Et de la Recherche Médicale (INSERM), and the Institut Pasteur.Antoine Gedeon and Nour Ayoub acknowledge PhD fellowships from the Médicament, Toxicologie, Chimieet Imageries PhD school (MTCI, ED 563), Université Paris Cité.
Subjects: bacterial two-hybrid system, [SDV]Life Sciences [q-bio], Escherichia coli, Cell Biology, de novo purine biosynthesis, Molecular Biology, Biochemistry, supramolecular assembly
Abstract: International audience; In human cells, de novo purine nucleotide biosynthesis is known to be regulated through the formation of a metabolon called purinosome. Here, we employed a bacterial two-hybrid approach to characterize the protein-protein interactions network among the corresponding enzymes of Escherichia coli. Our study revealed a dense network of binary interactions that connect most purine nucleotide biosynthesis enzymes. Notably, PurK, an exclusive prokaryotic enzyme, appears as one of the central hubs of this network. We further showed that modifications in PurK, which disrupted several interactions in the network, affected the purine nucleotide pools and altered the bacterial fitness. Our data suggest that the bacterial de novo purine nucleotide biosynthesis enzymes can assemble in a supramolecular complex and that proper interactions among the components of this complex can contribute to bacterial fitness.
Published: 2023
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15. Author response for 'Interaction network among de novo purine nucleotide biosynthesis enzymes in Escherichia coli'

Author: null Antoine Gedeon, null Gouzel Karimova, null Nour Ayoub, null Julien Dairou, null Quentin Giai Gianetto, null Sophie Vichier‐Guerre, null Pierre‐Olivier Vidalain, null Daniel Ladant, and null Hélène Munier‐Lehmann
Published: 2023
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16. SigH stress response mediates killing of Mycobacterium tuberculosis by activating nitronaphthofuran prodrugs via induction of Mrx2 expression

Author: Laura Cioetto-Mazzabò, Francesca Boldrin, Claire Beauvineau, Martin Speth, Alberto Marina, Amine Namouchi, Greta Segafreddo, Mena Cimino, Sandrine Favre-Rochex, Seetha Balasingham, Beatriz Trastoy, Hélène Munier-Lehmann, Gareth Griffiths, Brigitte Gicquel, Marcelo E Guerin, Riccardo Manganelli, Noelia Alonso-Rodríguez, Università degli Studi di Padova = University of Padua (Unipd), Chimie et modélisation pour la biologie du cancer (CMBC), Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Physiopathologie et traitement des maladies du foie, Hôpital Paul Brousse-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay, University of Oslo (UiO), Basque Research and Technology Alliance (BRTA), Génétique mycobactérienne - Mycobacterial genetics, Institut Pasteur [Paris] (IP), Oslo University Hospital [Oslo], Hospital Universitario Cruces = Cruces University Hospital, Département de Biologie structurale et Chimie - Department of Structural Biology and Chemistry, Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Chimie Biologique pour le Vivant / Chemistry for Life Sciences (CNRS - UMR3523 - Chem4Life), Institut Pasteur [Paris] (IP)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Shenzhen Nanshan Center for Chronic Disease Control [Shenzhen, China] (ShenZhenCDC), Ikerbasque - Basque Foundation for Science, European Seventh Framework Program Nanotherapeutics against Resistant Emerging Bacterial Pathogens [NAREB Project 604237], European Union's Horizon 2020 research and innovation program under the Marie Skłodowska-Curie [609020 to N.A.R., 844905 to B.T.], Norwegian Research Council [275873, 273319], MINECO/FEDER EU contracts [PID2019-105649RB-I00], Severo Ochoa Excellence Accreditation [SEV-2016-0644], Basque Government [KK-2019/00076], NIH [R01AI149297 to M.E.G.], Innovative Medicines Initiative 2 Joint Undertaking (JU) [853989 to R.M.]. Funding for open access charge: University of Oslo., European Project: 604237,EC:FP7:NMP,FP7-NMP-2013-LARGE-7,NAREB(2014), and European Project: 609020,EC:FP7:PEOPLE,FP7-PEOPLE-2013-COFUND,SCIENTIA-FELLOWS(2014)
Subjects: [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Genetics, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology
Abstract: The emergence of drug-resistant Mycobacterium tuberculosis strains highlights the need to discover anti-tuberculosis drugs with novel mechanisms of action. Here we discovered a mycobactericidal strategy based on the prodrug activation of selected chemical derivatives classified as nitronaphthofurans (nNFs) mediated by the coordinated action of the sigH and mrx2 genes. The transcription factor SigH is a key regulator of an extensive transcriptional network that responds to oxidative, nitrosative, and heat stresses in M. tuberculosis. The nNF action induced the SigH stress response which in turn induced the mrx2 overexpression. The nitroreductase Mrx2 was found to activate nNF prodrugs, killing replicating, non-replicating and intracellular forms of M. tuberculosis. Analysis of SigH DNA sequences obtained from spontaneous nNF-resistant M. tuberculosis mutants suggests disruption of SigH binding to the mrx2 promoter site and/or RNA polymerase core, likely promoting the observed loss of transcriptional control over Mrx2. Mutations found in mrx2 lead to structural defects in the thioredoxin fold of the Mrx2 protein, significantly impairing the activity of the Mrx2 enzyme against nNFs. Altogether, our work brings out the SigH/Mrx2 stress response pathway as a promising target for future drug discovery programs.
Published: 2023
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17. Steroid hormone catabolites activate the pyrin inflammasome through a non-canonical mechanism

Author: Flora Magnotti, Daria Chirita, Sarah Dalmon, Amandine Martin, Pauline Bronnec, Jeremy Sousa, Olivier Helynck, Wonyong Lee, Daniel L. Kastner, Jae Jin Chae, Michael F. McDermott, Alexandre Belot, Michel Popoff, Pascal Sève, Sophie Georgin-Lavialle, Hélène Munier-Lehmann, Tu Anh Tran, Ellen De Langhe, Carine Wouters, Yvan Jamilloux, Thomas Henry, Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Chimie et Biocatalyse, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Université Paris Cité (UPCité), National Institutes of Health [Bethesda] (NIH), University of Leeds, Lyon Immunopathology Federation (LIFe), Hospices Civils de Lyon (HCL), Toxines bactériennes - Bacterial Toxins, Institut Pasteur [Paris] (IP), Centre de Recherche Saint-Antoine (CRSA), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Hôpital Universitaire Carémeau [Nîmes] (CHU Nîmes), Centre Hospitalier Universitaire de Nîmes (CHU Nîmes), University Hospitals Leuven [Leuven], Catholic University of Leuven - Katholieke Universiteit Leuven (KU Leuven), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon, This work is supported by an ANR grant (FMFgeneToDiag). This project has received funding from the European Union's Horizon 2020 research and innovation program under grant agreement no. 779295. T.H.’s team is supported by The Fondation pour la Recherche Médicale (FRM EQU202103012640). D.C. is supported by a fellowship from The Fondation pour la Recherche Médicale (FDT202106012874). E.D.L. and C.W. are members of the European Reference Network for Rare Immunodeficiency, Autoinflammatory and Autoimmune Diseases—Project ID no. 739543., ANR-17-CE17-0021,FMFgeneToDiag,Fièvre Méditerranéenne Familiale (FMF) et maladies apparentées : des bases moléculaires et cellulaires à la mise au point de tests diagnostiques(2017), Centre International de Recherche en Infectiologie - UMR (CIRI), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Henry, Thomas, and Fièvre Méditerranéenne Familiale (FMF) et maladies apparentées : des bases moléculaires et cellulaires à la mise au point de tests diagnostiques - - FMFgeneToDiag2017 - ANR-17-CE17-0021 - AAPG2017 - VALID
Subjects: Molecular biology [CP], MESH: Mutation, [SDV.IMM] Life Sciences [q-bio]/Immunology, Inflammasomes, MESH: Testosterone, PAAND, progesterone, General Biochemistry, Genetics and Molecular Biology, catabolite, B30.2, autoinflammatory disease, familial Mediterranean fever, inflammasome, pyrin, MESH: Progesterone, Etiocholanolone, Humans, Testosterone, MESH: Humans, steroid, CP: Immunology, MESH: Pregnanolone, MESH: Etiocholanolone, Mutation, Immunology [CP], [SDV.IMM]Life Sciences [q-bio]/Immunology, CP: Molecular biology, pregnanolone
Abstract: The pyrin inflammasome acts as a guard of RhoA GTPases and is central to immune defenses against RhoA-manipulating pathogens. Pyrin activation proceeds in two steps. Yet, the second step is still poorly understood. Using cells constitutively activated for the pyrin step 1, a chemical screen identifies etiocholanolone and pregnanolone, two catabolites of testosterone and progesterone, acting at low concentrations as specific step 2 activators. High concentrations of these metabolites fully and rapidly activate pyrin, in a human specific, B30.2 domain-dependent manner and without inhibiting RhoA. Mutations in MEFV, encoding pyrin, cause two distinct autoinflammatory diseases pyrin-associated autoinflammation with neutrophilic dermatosis (PAAND) and familial Mediterranean fever (FMF). Monocytes from PAAND patients, and to a lower extent from FMF patients, display increased responses to these metabolites. This study identifies an unconventional pyrin activation mechanism, indicates that endogenous steroid catabolites can drive autoinflammation, through the pyrin inflammasome, and explains the "steroid fever" described in the late 1950s upon steroid injection in humans. ispartof: CELL REPORTS vol:41 issue:2 ispartof: location:United States status: published
Published: 2022
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18. Structural basis for the allosteric inhibition of UMP kinase from Gram‐positive bacteria, a promising antibacterial target

Author: Patrick Walter, Ariel Mechaly, Julien Bous, Ahmed Haouz, Patrick England, Joséphine Lai‐Kee‐Him, Aurélie Ancelin, Sylviane Hoos, Bruno Baron, Stefano Trapani, Patrick Bron, Gilles Labesse, Hélène Munier‐Lehmann, Chimie et Biocatalyse, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Cristallographie (Plateforme) - Crystallography (Platform), Centre de Biologie Structurale [Montpellier] (CBS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Biophysique Moléculaire (Plate-forme), Biophysique Moléculaire (plateforme) - Molecular Biophysics (platform), This work was supported by the European Union’s Framework Programme for Research and Innovation Horizon 2020 (2014-2020) under the Marie Skłodowska-Curie Grant Agreement No. 675555, Accelerated Early staGe drug discovery (AEGIS) project (to HML). The project was also supported in part by the Centre National de la Recherche Scientifique (CNRS), the Institut National de la Santé Et de la Recherche Médicale (INSERM), the Institut Pasteur and the French Infrastructure for Integrated Structural Biology (FRISBI) ANR-10-INBS-05. PW was the recipient of a PhD fellowship from the EU H2020 (2014-2020) under the Marie Skłodowska-Curie Grant Agreement No. 675555, Accelerated Early staGe drug discovery (AEGIS)., ANR-10-INBS-0005,FRISBI,Infrastructure Française pour la Biologie Structurale Intégrée(2010), and European Project: 675555,H2020,H2020-MSCA-ITN-2015,AEGIS(2016)
Subjects: crystal structure, allostery, [SDV]Life Sciences [q-bio], Cryoelectron Microscopy, Uridine Triphosphate, Cell Biology, nucleotide metabolism, Gram-Positive Bacteria, Biochemistry, Anti-Bacterial Agents, Adenosine Triphosphate, effector, Allosteric Regulation, cryo-EM single particle analysis, Humans, Amino Acid Sequence, Guanosine Triphosphate, enzyme regulation, Nucleoside-Phosphate Kinase, Uridine Monophosphate, Molecular Biology, UMP kinase
Abstract: International audience; Tuberculosis claims significantly more than one million lives each year. A feasible way to face the issue of drug resistance is the development of new antibiotics. Bacterial uridine 5’-monophosphate (UMP) kinase is a promis- ing target for novel antibiotic discovery as it is essential for bacterial sur- vival and has no counterpart in human cells. The UMP kinase from M. tuberculosis is also a model of particular interest for allosteric regula- tion with two effectors, GTP (positive) and UTP (negative). In this study, using X-ray crystallography and cryo-electron microscopy, we report for the first time a detailed description of the negative effector UTP-binding site of a typical Gram-positive behaving UMP kinase. Comparison between this snapshot of low affinity for Mg-ATP with our previous 3D-structure of the GTP-bound complex of high affinity for Mg-ATP led to a better understanding of the cooperative mechanism and the allosteric regulation of UMP kinase. Thermal shift assay and circular dichroism experiments corroborate our model of an inhibition by UTP linked to higher flexibility of the Mg-ATP-binding domain. These new structural insights provide valuable knowledge for future drug discovery strategies targeting bacterial UMP kinases.
Published: 2022
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19. 2-((3,5-Dinitrobenzyl)thio)quinazolinones: Potent Antimycobacterial Agents Activated by Deazaflavin (F420)-Dependent Nitroreductase (Ddn)

Author: Serge Van Calenbergh, Fabian Hulpia, Helena I. Boshoff, Martijn D. P. Risseeuw, He Eun Forbes, Guy Caljon, Yanlin Jian, and Hélène Munier-Lehmann
Subjects: biology, Stereochemistry, medicine.drug_class, Pharmacology. Therapy, Thio, Antimycobacterial, Thymidylate kinase, Cofactor, Ames test, Nitroreductase, chemistry.chemical_compound, chemistry, Drug Discovery, medicine, biology.protein, Molecular Medicine, Structure–activity relationship, Quinazolinone
Abstract: Swapping the substituents in positions 2 and 4 of the previously synthesized but yet undisclosed 5-cyano-4-(methylthio)-2-arylpyrimidin-6-ones 4, ring closure, and further optimization led to the identification of the potent antitubercular 2-thio-substituted quinazolinone 26. Structure-activity relationship (SAR) studies indicated a crucial role for both meta-nitro substituents for antitubercular activity, while the introduction of polar substituents on the quinazolinone core allowed reduction of bovine serum albumin (BSA) binding (63c, 63d). While most of the tested quinazolinones exhibited no cytotoxicity against MRC-5, the most potent compound 26 was found to be mutagenic via the Ames test. This analogue exhibited moderate inhibitory potency against Mycobacterium tuberculosis thymidylate kinase, the target of the 3-cyanopyridones that lies at the basis of the current analogues, indicating that the whole-cell antimycobacterial activity of the present S-substituted thioquinazolinones is likely due to modulation of alternative or additional targets. Diminished antimycobacterial activity was observed against mutants affected in cofactor F-420 biosynthesis (fbiC), cofactor reduction (fgd), or deazaflavin-dependent nitroreductase activity rv3547 , indicating that reductive activation of the 3,5-dinitrobenzyl analogues is key to antimycobacterial activity.
Published: 2020
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20. Replication of Equine arteritis virus is efficiently suppressed by purine and pyrimidine biosynthesis inhibitors

Author: Patrick Dallemagne, Pierre-Olivier Vidalain, Aymeric Hans, Anthony Madeline, D. Gaudaire, José Carlos Valle-Casuso, Lydie Martin-Faivre, Stéphane Pronost, Hélène Munier-Lehmann, Stéphan Zientara, Physiopathologie et épidémiologie des maladies équines (PhEED), Laboratoire de santé animale, sites de Maisons-Alfort et de Normandie, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), LABÉO, Pôle d’analyses et de recherche de Normandie (LABÉO), Biologie, génétique et thérapies ostéoarticulaires et respiratoires (BIOTARGEN), Chimie et Biocatalyse, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Virologie UMR1161 (VIRO), École nationale vétérinaire - Alfort (ENVA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Fonds Eperon, AnsesFonds EperonRegion Normandie, IFCE (Institut Francais du Cheval et de l'Equitation), Région Normandie, ANSES, Laboratoire de Santé Animale - site de Dozulé, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Université Paris-Est (UPE), Laboratoire de santé animale, sites de Maisons-Alfort et de Dozulé, Centre International de Recherche en Infectiologie - UMR (CIRI), Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), This work was supported by Anses, IFCE (Institut Français du Cheval et de l’Equitation), Fonds Eperon and Région Normandie., AnsesIFCE (Institut Francais du Cheval et de l'Equitation)Fonds EperonRegion Normandie, Bodescot, Myriam, École nationale vétérinaire - Alfort (ENVA)-Laboratoire de santé animale, sites de Maisons-Alfort et de Normandie, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0301 basic medicine, Dihydroorotate Dehydrogenase, lcsh:Medicine, Virus Replication, chemistry.chemical_compound, Cytopathogenic Effect, Viral, Coronaviridae, lcsh:Science, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, 0303 health sciences, education.field_of_study, Multidisciplinary, Arterivirus Infections, Drug discovery, 3. Good health, [SDV.SP.PHARMA] Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Pyrimidine metabolism, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Oxidoreductases Acting on CH-CH Group Donors, 030106 microbiology, Population, Biology, Antiviral Agents, Microbiology, Article, Cell Line, 03 medical and health sciences, Equartevirus, Ribavirin, Animals, Horses, education, 030304 developmental biology, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, 030306 microbiology, lcsh:R, [SDV.BA.MVSA] Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, RNA, RNA virus, biology.organism_classification, Virology, 030104 developmental biology, Pyrimidines, chemistry, Viral replication, Purines, Dihydroorotate dehydrogenase, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Horse Diseases, lcsh:Q
Abstract: RNA viruses are responsible for a large variety of animal infections. Equine Arteritis Virus (EAV) is a positive single-stranded RNA virus member of the family Arteriviridae from the order Nidovirales like the Coronaviridae. EAV causes respiratory and reproductive diseases in equids. Although two vaccines are available, the vaccination coverage of the equine population is largely insufficient to prevent new EAV outbreaks around the world. In this study, we present a high-throughput in vitro assay suitable for testing candidate antiviral molecules on equine dermal cells infected by EAV. Using this assay, we identified three molecules that impair EAV infection in equine cells: the broad-spectrum antiviral and nucleoside analog ribavirin, and two compounds previously described as inhibitors of dihydroorotate dehydrogenase (DHODH), the fourth enzyme of the pyrimidine biosynthesis pathway. These molecules effectively suppressed cytopathic effects associated to EAV infection, and strongly inhibited viral replication and production of infectious particles. Since ribavirin is already approved in human and small animal, and that several DHODH inhibitors are in advanced clinical trials, our results open new perspectives for the management of EAV outbreaks.
Published: 2020
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21. Identification of antiviral molecules against West-Nile virus by an approach combining cell imaging and equine neural cells derived from induced-pluripotent stem cells

Author: Marielle Cochet-Bernoin, François Piumi, Kamila Gorna, Gaelle Gonzalez, Anne Danckaert, Nathalie Aulner, Hélène Munier-Lehmann, Stephan Zientara, Xavier Donadeu, F., Alexandra Benchoua, Muriel Coulpier, Virologie UMR1161 (VIRO), École nationale vétérinaire d'Alfort (ENVA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-École nationale vétérinaire d'Alfort (ENVA), BioImagerie Photonique – Photonic BioImaging (UTechS PBI), Institut Pasteur [Paris], The Roslin Institute, CECS - I-STEM, Association française contre les myopathies (AFM-Téléthon), École nationale vétérinaire - Alfort (ENVA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), École nationale vétérinaire - Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Institut Pasteur [Paris] (IP), Biotechnology and Biological Sciences Research Council (BBSRC), Institut des cellules souches pour le traitement et l'étude des maladies monogéniques (I-STEM), and Université d'Évry-Val-d'Essonne (UEVE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay-Généthon
Subjects: [SDV.MHEP.ME]Life Sciences [q-bio]/Human health and pathology/Emerging diseases, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology, ComputingMilieux_MISCELLANEOUS
Abstract: International audience
Published: 2021

22. 2-((3,5-Dinitrobenzyl)thio)quinazolinones: Potent Antimycobacterial Agents Activated by Deazaflavin (F

Author: Yanlin, Jian, He Eun, Forbes, Fabian, Hulpia, Martijn D P, Risseeuw, Guy, Caljon, Hélène, Munier-Lehmann, Helena I M, Boshoff, and Serge, Van Calenbergh
Subjects: Structure-Activity Relationship, Mutagenicity Tests, Riboflavin, Antitubercular Agents, Microbial Sensitivity Tests, Mycobacterium tuberculosis, Nitroreductases, High-Throughput Screening Assays, Quinazolinones
Abstract: Swapping the substituents in positions 2 and 4 of the previously synthesized but yet undisclosed 5-cyano-4-(methylthio)-2-arylpyrimidin-6-ones
Published: 2020

23. Screening of potential antiviral molecules against equid herpesvirus-1 using cellular impedance measurement: dataset of 2,891 compounds

Author: Christine Fortier, Erika Hue, Aymeric Hans, Pierre-Olivier Vidalain, Patrick Dallemagne, Romain Paillot, Peggy Suzanne, Stéphane Pronost, Côme Thieulent, Hélène Munier-Lehmann, Stéphan Zientara, LABÉO, Pôle d’analyses et de recherche de Normandie (LABÉO), Biologie, génétique et thérapies ostéoarticulaires et respiratoires (BIOTARGEN), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), ImpedanCELL, Interactions Cellules Organismes Environnement (ICORE), CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-CHU Caen, Normandie Université (NU), Chimie et Biocatalyse, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN), Virologie UMR1161 (VIRO), École nationale vétérinaire d'Alfort (ENVA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire de pathologie équine de Dozulé, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Biologie cellulaire des infections virales – Cell biology of viral infection, Centre International de Recherche en Infectiologie - UMR (CIRI), Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), École nationale vétérinaire - Alfort (ENVA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire de santé animale, sites de Maisons-Alfort et de Normandie, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), HUE, Erika, École nationale vétérinaire - Alfort (ENVA)-Laboratoire de santé animale, sites de Maisons-Alfort et de Normandie, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Equid herpesvirus-1, Chemical library screening, Decitabine, lcsh:Computer applications to medicine. Medical informatics, Virus, Chemical library, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, medicine, Molecule, Research article, Antiviral, lcsh:Science (General), Data Article, 030304 developmental biology, Real-time cell assay, 0303 health sciences, Multidisciplinary, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, Focused Impedance Measurement, Chemistry, screening, [SDV.BA.MVSA] Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, Valganciclovir, Virology, In vitro, 3. Good health, lcsh:R858-859.7, 030217 neurology & neurosurgery, lcsh:Q1-390, medicine.drug
Abstract: International audience; Data presented in this article are associated with the research article “Identification of antiviral compounds against equid herpesvirus-1 using real-time cell assay screening: efficacy of decitabine and valganciclovir alone and in combination” [1]. These data correspond to the in vitro screening of 2,891 potential antiviral compounds against equid herpesvirus-1 (EHV-1) based on impedance measurements using the xCELLigence® RTCA MP System. This dataset includes compounds from three different libraries: i) 1,199 compounds from the Prestwick® Chemical Library, which contains mostly US Food and Drug Administration approved drugs (Prestwick® Chemical, Illkirch, France); ii) 1,651 compounds from the Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN, Caen, France); iii) 41 compounds (called herein in-house antiviral library) selected for their effects against different human viruses. Compounds effective against EHV-1 were selected using the area under normalised curves (AUCn) and the time required for the Cell Index to decrease by 50% after virus infection (CIT50). The full dataset from the screen is made publicly available for further analyses.
Published: 2020
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24. Un nouvel arsenal thérapeutique contre la rhinopneumonie équine

Author: Côme Thieulent, Erika HUE, Christine Fortier, Patrick Dallemagne, Stephan Zientara, Aymeric Hans, Hélène Munier-Lehmann, Pierre-Olivier Vidalain, Stéphane Pronost, LABÉO, Pôle d’analyses et de recherche de Normandie (LABÉO), Biologie, génétique et thérapies ostéoarticulaires et respiratoires (BIOTARGEN), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), ImpedanCELL, Interactions Cellules Organismes Environnement (ICORE), CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-CHU Caen, Normandie Université (NU), Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN), Virologie UMR1161 (VIRO), École nationale vétérinaire d'Alfort (ENVA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire de pathologie équine de Dozulé, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Chimie et Biocatalyse, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Centre International de Recherche en Infectiologie - UMR (CIRI), Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), École nationale vétérinaire - Alfort (ENVA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire de santé animale, sites de Maisons-Alfort et de Normandie, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), HUE, Erika, École nationale vétérinaire - Alfort (ENVA)-Laboratoire de santé animale, sites de Maisons-Alfort et de Normandie, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Épidémiologie, Herpèsvirus Équin Hve-1 Et Hve-4, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, [SDV.BA.MVSA] Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, Pharmacologie, Rhinopneumonie
Abstract: National audience; L’herpèsvirus équin1 (HVE-1)est l’un des deux agents responsables de la rhinopneumonie équine. Ce virus peut provoquer des troubles respiratoires, des avortements, une mort néonatale du jeune poulain et des troubles nerveux appelés myéloencéphalopathie équine. Les vaccins existant à l’heure actuelle permettent de réduire les problèmes respiratoires et donc la dissémination du virus. Cependant, ils ne préviennent pas suffisamment les formes graves de la maladie. Par ailleurs, aucune molécule antivirale ne possède d’autorisation de mise sur le marché en santé équine en France. Nous avons développé un système permettant de tester à haut débit un grand nombre de molécules contre l’HVE-1 en conditions de laboratoire. Parmi les 3000 molécules testées, 23 ont été identifiées comme potentiellement efficaces contre ce virus. Le ganciclovir et sa pro-drogue le valganciclovir se sont montrés parmi les molécules les plus efficaces contre l’HVE-1. Ces résultats encourageants, couplés aux données de pharmacocinétique déjà existantes de ces molécules sur le modèle équin nous ont conduits à envisager une étude expérimentale sur des poneys.
Published: 2020

25. Identification of antiviral compounds against equid herpesvirus-1 using real-time cell assay screening: Efficacy of decitabine and valganciclovir alone or in combination

Author: Pierre-Olivier Vidalain, Patrick Dallemagne, Aymeric Hans, Côme Thieulent, Christine Fortier, Erika Hue, Stéphan Zientara, Stéphane Pronost, Romain Paillot, Hélène Munier-Lehmann, Gabrielle Sutton, LABÉO, Pôle d’analyses et de recherche de Normandie (LABÉO), Biologie, génétique et thérapies ostéoarticulaires et respiratoires (BIOTARGEN), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), ImpedanCELL, Interactions Cellules Organismes Environnement (ICORE), CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-CHU Caen, Normandie Université (NU), Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN), Virologie UMR1161 (VIRO), École nationale vétérinaire d'Alfort (ENVA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Chimie et Biocatalyse, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Laboratoire de pathologie équine de Dozulé, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), ANSES, Laboratoire de Santé Animale - site de Dozulé, Centre International de Recherche en Infectiologie - UMR (CIRI), Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), This work was supported by LABÉO, IFCE (Institut Français du Cheval et de l'Equitation, project AMIE), Fonds Eperon (project N87–2014, N07–2015, N07–2016, N13–2017 and N62-2017), Région Normandie (CPER R25 P3) and CENTAURE European project co-funded by Normandy County Council, European Union in the framework of the ERDF-ESF operationnal programme 2014–2020., Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), École nationale vétérinaire - Alfort (ENVA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Physiopathologie et épidémiologie des maladies équines (PhEED), Laboratoire de santé animale, sites de Maisons-Alfort et de Normandie, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), HUE, Erika, École nationale vétérinaire - Alfort (ENVA)-Laboratoire de santé animale, sites de Maisons-Alfort et de Normandie, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0301 basic medicine, Aphidicolin, Ganciclovir, Equid herpesvirus-1, 030106 microbiology, Chemical library screening, Decitabine, Antiviral Agents, Cell Line, 03 medical and health sciences, chemistry.chemical_compound, Virology, Drug Discovery, Medicine, Animals, Valganciclovir, Synergism, Horses, Antiviral, Pharmacology, Real-time cell assay, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, business.industry, Idoxuridine, [SDV.BA.MVSA] Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, Drug Synergism, Deoxycytidine kinase, Herpesviridae Infections, 3. Good health, High-Throughput Screening Assays, Drug Combinations, 030104 developmental biology, chemistry, [SDV.SP.PHARMA] Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Deoxycytidine, Rabbits, business, Nucleoside, medicine.drug, Herpesvirus 1, Equid
Abstract: International audience; Equid herpesvirus-1 infections cause respiratory, neurological and reproductive syndromes. Despite preventive treatments with vaccines, resurgence of EHV-1 infection still constitutes a major threat to equine industry. However, no antiviral compound is available to treat infected horses. In this study, 2891 compounds were screened against EHV-1 using impedance measurement. 22 compounds have been found to be effective in vitro against EHV-1. Valganciclovir, ganciclovir, decitabine, aphidicolin, idoxuridine and pritelivir (BAY 57-1293) are the most effective compounds identified, and their antiviral potency was further assessed on E. Derm, RK13 and EEK cells and against 3 different field strains of EHV-1 (ORF30 2254 A/G/C). We also provide evidences of synergistic interactions between valganciclovir and decitabine in our in vitro antiviral assay as determined by MacSynergy II, isobologramm and Chou-Talalay methods. Finally, we showed that deoxycytidine reverts the antiviral effect of decitabine, thus supporting some competition at the level of nucleoside phosphorylation by deoxycytidine kinase and/or DNA synthesis. Deoxycitidine analogues, like decitabine, is a family of compounds identified for the first time with promising antiviral efficacy against herpesviruses.
Published: 2020
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26. Screening and evaluation of antiviral compounds against Equid alpha-herpesviruses using an impedance-based cellular assay

Author: Pierre-Hugues Pitel, Aymeric Hans, Côme Thieulent, Christine Fortier, Erika Hue, Guillaume Fortier, Stéphan Zientara, Patrick Dallemagne, Stéphane Pronost, Pierre-Olivier Vidalain, Hélène Munier-Lehmann, LABÉO, Pôle d’analyses et de recherche de Normandie (LABÉO), Biologie, génétique et thérapies ostéoarticulaires et respiratoires (BIOTARGEN), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), ImpedanCELL, Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN), Virologie UMR1161 (VIRO), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de la Recherche Agronomique (INRA)-École nationale vétérinaire d'Alfort (ENVA), Chimie et Biocatalyse, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Unité de virologie et parasitologie équine, Laboratoire de pathologie équine de Dozulé, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Equipe Chimie et Biologie, Modélisation et Immunologie pour la Thérapie (CBMIT), Université Paris Descartes - Paris 5 (UPD5)-Université Paris Descartes - Paris 5 (UPD5), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS), This work was supported by LABÉO, IFCE (Institut Français du Cheval et de l′Equitation, project AMIE), Fonds Eperon (project N87-2014, N07-2015, N07-2016 and N13-2017) and Région Normandie (CPER R25 P3)., Interactions Cellules Organismes Environnement (ICORE), CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-CHU Caen, Normandie Université (NU), Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-École nationale vétérinaire d'Alfort (ENVA), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), École nationale vétérinaire - Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de santé animale, sites de Maisons-Alfort et de Normandie, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Laboratoire de santé animale, sites de Maisons-Alfort et de Normandie, École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), and Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)
Subjects: Ganciclovir, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Population, Alpha (ethology), Cell analysis, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Spironolactone, Biology, Antiviral Agents, Equid herpesviruses, NERVOUS DISORDERS, Cell Line, Real-time cell analysis, 03 medical and health sciences, Cytopathogenic Effect, Viral, xCELLigence, Virology, Electric Impedance, medicine, Lack of efficacy, Animals, Horses, Aciclovir, Antiviral, education, 030304 developmental biology, 0303 health sciences, education.field_of_study, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, Cellular Assay, 030302 biochemistry & molecular biology, Impedance, Herpesviridae Infections, High-Throughput Screening Assays, 3. Good health, Screening, Herpesvirus 3, Equid, Herpesvirus 4, Equid, Herpesvirus 1, Equid, medicine.drug
Abstract: International audience; Equid alpha-herpesviruses (EHV) are responsible for different diseases in equine population. EHV-1 causes respiratory diseases, abortions and nervous disorders, EHV-4 causes respiratory diseases and sporadic abortion, while EHV-3 is responsible of equine coital exanthema. In view of the lack of efficacy of vaccines against EHV-1 and EHV-4 and in the absence of vaccines against EHV-3, the use of antiviral treatment is of great interest. In this study, we documented the interest of the Real-Time Cell Analysis (RTCA) technology to monitor the cytopathic effects induced by these viruses on equine dermal cells, and established the efficacy of this method to evaluate the antiviral effect of aciclovir (ACV) and ganciclovir (GCV). In addition, the RTCA technology has also been found appropriate for the high-throughput screening of small molecules against EHV, allowing the identification of spironolactone as a novel antiviral against EHV.
Published: 2019
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27. Structure-aided optimization of non-nucleoside M. tuberculosis thymidylate kinase inhibitors

Author: Savvas N. Savvides, Serge Van Calenbergh, Martijn D. P. Risseeuw, José A. Aínsa, Ainhoa Lucía, Begoña Gracia, Toon Verstraelen, Yanlin Jian, Hélène Munier-Lehmann, Romain Merceron, Paul Cos, Helena I. Boshoff, Lijun Song, and Fabian Hulpia
Subjects: Antitubercular Agents, Druggability, Drug design, Microbial Sensitivity Tests, Thymidylate kinase, Mycobacterium tuberculosis, Structure-Activity Relationship, Piperidines, Drug Discovery, Nucleotide, Enzyme Inhibitors, Pharmacology, chemistry.chemical_classification, Dose-Response Relationship, Drug, Molecular Structure, biology, Pharmacology. Therapy, Organic Chemistry, General Medicine, biology.organism_classification, Chemical space, chemistry, Biochemistry, Human medicine, Efflux, Nucleoside-Phosphate Kinase, Nucleoside, Thymine
Abstract: Mycobacterium tuberculosis thymidylate kinase (MtTMPK) has emerged as an attractive target for rational drug design. We recently investigated new families of non-nucleoside MtTMPK inhibitors in an effort to diversify MtTMPK inhibitor chemical space. We here report a new series of MtTMPK inhibitors by combining the Topliss scheme with rational drug design approaches, fueled by two co-crystal structures of MtTMPK in complex with developed inhibitors. These efforts furnished the most potent MtTMPK inhibitors in our assay, with two analogues displaying low micromolar MIC values against H37Rv Mtb. Prepared inhibitors address new sub-sites in the MtTMPK nucleotide binding pocket, thereby offering new insights into its druggability. We studied the role of efflux pumps as well as the impact of cell wall permeabilizers for selected compounds to potentially provide an explanation for the lack of correlation between potent enzyme inhibition and whole-cell activity.
Published: 2021
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28. Synthesis of sulfamide analogues of deoxthymidine monophosphate as potential inhibitors of mycobacterial cell wall biosynthesis

Author: Wanting Jiao, Antony J. Fairbanks, Hélène Munier-Lehmann, Kajitha Suthagar, University of Canterbury [Christchurch], Victoria University of Wellington, Chimie et Biocatalyse, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), The authors thank the University of Canterbury (PhD Scholarship to KS) and the Biomolecular Interaction Centre for financial support. Hélène Munier-Lehmann was supported by INSERM, CNRS and the Institut Pasteur., and Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris]
Subjects: Isostere, Stereochemistry, Antitubercular Agents, Protein Data Bank (RCSB PDB), Carbohydrates, 01 natural sciences, Biochemistry, Analytical Chemistry, Structure-Activity Relationship, chemistry.chemical_compound, Non-competitive inhibition, Thymidine monophosphate kinase, Biosynthesis, Cell Wall, Tuberculosis, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Sulfamide, chemistry.chemical_classification, biology, 010405 organic chemistry, Chemistry, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Organic Chemistry, Active site, Mycobacteria, Mycobacterium tuberculosis, General Medicine, 3. Good health, 0104 chemical sciences, 010404 medicinal & biomolecular chemistry, Enzyme, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Docking (molecular), Deoxythymidine, biology.protein, Sulfamides, Thymidine
Abstract: International audience; The recently discovered enzyme Mycobacterium tuberculosis thymidine monophosphate kinase (TMPKmt), which catalyses the phosphorylation of deoxythymidine monophosphate (dTMP) to give deoxythymidine diphosphate (dTDP), is indispensable for the growth and survival of M. tuberculosis as it plays an essential role in DNA synthesis. Inhibition of TMPKmt is an attractive avenue for the development of novel anti-tuberculosis agents. Based on the premise that sulfamide may be a suitable isostere of phosphate, deoxythymidine analogues comprising various substituted sulfamides at C5′ were modelled in silico into the active site of TMPKmt (PDB accession code: 1N5K) using induced-fit docking methods. A selection of modelled compounds was synthesized, and their activity as inhibitors of TMPKmt was evaluated. Three compounds showed competitive inhibition of TMPKmt in the micromolar range (10–50 μM). Compounds were tested in vitro for anti-mycobacterial activity against M. smegmatis: three compounds showed weak anti-mycobacterial activity (MIC 250 μg/mL).
Published: 2018
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29. Nitazoxanide Analogs Require Nitroreduction for Antimicrobial Activity in Mycobacterium smegmatis

Author: Gerald Larrouy-Maumus, Sandrine Favre-Rochex, Alessandro Cascioferro, Hélène Munier-Lehmann, Sonia Rebollo-Ramirez, Claire Beauvineau, Delphine Naud-Martin, Mena Cimino, Brigitte Gicquel, Maria Virginia Buchieri, Olivier Helynck, Génétique mycobactérienne - Mycobacterial genetics, Institut Pasteur [Paris], Imperial College London, Chimie, Modélisation et Imagerie pour la Biologie [Orsay], Université Paris-Sud - Paris 11 (UP11)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Pathogénomique mycobactérienne intégrée, Chimie et Biocatalyse, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), This work is part of the European Seventh Framework Program Nanotherapeutics against Resistant Emerging Bacterial Pathogens (NAREB Project 604237). This work was also supported by the 'Programme de Recherche sur les Nouvelles Molécules Intervenant dans la Lutte Contre la Tuberculose' of the Gabonese Republic, the Department of Life Sciences and the Faculty of Natural Sciences kick-start funding award from Imperial College London, U.K., and Agilent Technologies, U.K., concerning all LC–MS experiments performed on the Agilent 6545 QToF., We thank Yves Janin for fruitful discussions and technical support., European Project: 604237,EC:FP7:NMP,FP7-NMP-2013-LARGE-7,NAREB(2014), Institut Pasteur [Paris] (IP), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0301 basic medicine, Mycobacterium smegmatis, 030106 microbiology, Down-Regulation, Mycobacterium Infections, Nontuberculous, MESH: Thiazoles, MESH: Down-Regulation, Mycobacterium aurum, Mycobacterium, Microbiology, 03 medical and health sciences, Nitroreductase, chemistry.chemical_compound, Hydroxylamine, MESH: Anti-Bacterial Agents, Drug Resistance, Bacterial, MESH: Drug Resistance, Bacterial, Drug Discovery, MESH: Mycobacterium, medicine, Humans, [CHIM]Chemical Sciences, MESH: Mycobacterium Infections, Gene, MESH: Mycobacterium smegmatis, Mycobacterium Infections, MESH: Humans, biology, Nitazoxanide, Nitroreductases, Nitro Compounds, MESH: Mycobacterium Infections, Nontuberculous, Antimicrobial, biology.organism_classification, Anti-Bacterial Agents, 3. Good health, Thiazoles, MESH: Nitroreductases, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, 030104 developmental biology, chemistry, Biochemistry, Molecular Medicine, Antibacterial activity, medicine.drug
Abstract: International audience; In this study, we aimed to decipher the natural resistance mechanisms of mycobacteria against novel compounds isolated by whole-cell-based high-throughput screening (HTS). We identified active compounds using Mycobacterium aurum. Further analyses were performed to determine the resistance mechanism of M. smegmatis against one hit, 3-bromo-N-(5-nitrothiazol-2-yl)-4-propoxybenzamide (3), which turned out to be an analog of the drug nitazoxanide (1). We found that the repression of the gene nfnB coding for the nitroreductase NfnB was responsible for the natural resistance of M. smegmatis against 3. The overexpression of nfnB resulted in sensitivity of M. smegmatis to 3. This compound must be metabolized into hydroxylamine intermediate for exhibiting antibacterial activity. Thus, we describe, for the first time, the activity of a mycobacterial nitroreductase against 1 analogs, highlighting the differences in the metabolism of nitro compounds among mycobacterial species and emphasizing the potential of nitro drugs as antibacterials in various bacterial species.
Published: 2017
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30. A real-time cell-binding assay reveals dynamic features of STxB-Gb3 cointernalization and STxB-mediated cargo delivery into cancer cells

Author: Grzegorz M Popowicz, Jos Buijs, Valeria Napolitano, Giulia Opassi, U. Helena Danielson, Hanna Björkelund, Grzegorz Dubin, João Crispim Encarnação, Hélène Munier-Lehmann, Karl Andersson, Ridgeview Instruments AB [Uppsala, Sueden], Department of Immunology, Genetics and Pathology [Uppsala, Sueden] (IGP), Uppsala University, Uniwersytet Jagielloński w Krakowie = Jagiellonian University (UJ), Department of Chemistry [Uppsala, Sueden], Biomedical Center = Biomedicinskt centrum [Uppsala, Sueden] (BMC), Uppsala University-Uppsala University, Malopolska Centre of Biotechnology [Krakow] (MCB), Helmholtz-Zentrum München (HZM), Center for Integrated Protein Science (CIPSM), Technische Universität Munchen - Université Technique de Munich [Munich, Allemagne] (TUM)-Helmholtz-Zentrum München (HZM)-Ludwig Maximilian University of Munich [Germany] (LMU München), Chimie et Biocatalyse, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], This project has received funding from the European Union's Framework Programme for Research and Innovation Horizon 2020 (2014–2020) under the Marie Sklodowska‐Curie Grant Agreement No. 675555, Accelerated Early staGe drug discovery (AEGIS)., Microscopic imaging was performed with equipment maintained by the BioVis Platform at Uppsala University. We thank to Dr Arie Geerlof and Dr André Mourão (Institute of Structural Biology, Helmholtz Zentrum München) for providing help on protein expression and purification of the STxB constructs., European Project: 675555,H2020,H2020-MSCA-ITN-2015,AEGIS(2016), Helmholtz Zentrum München = German Research Center for Environmental Health, Technische Universität Munchen - Université Technique de Munich [Munich, Allemagne] (TUM)-Ludwig-Maximilians-Universität München (LMU)-Helmholtz Zentrum München = German Research Center for Environmental Health, and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
Subjects: media_common.quotation_subject, [SDV]Life Sciences [q-bio], Cell, education, Biophysics, Globotriaosylceramide, cell surface receptor, Biological Transport, Active, Shiga Toxins, Biochemistry, real-time cell-binding assays, 03 medical and health sciences, chemistry.chemical_compound, [SPI]Engineering Sciences [physics], Structural Biology, Cell surface receptor, receptor internalization, Neoplasms, Genetics, medicine, binding kinetics, cancer, Humans, Internalization, Molecular Biology, Binding Kinetics, Cancer, Cell Surface Receptor, Real-time Cell-binding Assays, Receptor Internalization, Shiga Toxin, real-time cell- binding assays, 030304 developmental biology, media_common, 0303 health sciences, Drug Carriers, Chemistry, Trihexosylceramides, 030302 biochemistry & molecular biology, Biochemistry and Molecular Biology, Cell Biology, Shiga toxin, Small molecule, Receptor–ligand kinetics, 3. Good health, Cell biology, ddc, medicine.anatomical_structure, Cancer cell, Biological Assay, K562 Cells, HT29 Cells, Intracellular, Biokemi och molekylärbiologi
Abstract: International audience; The interaction between the Shiga toxin B-subunit (STxB) and its globotriaosylceramide receptor (Gb3) has a high potential for being exploited for targeted cancer therapy. The primary goal of this study was to evaluate the capacity of STxB to carry small molecules and proteins as cargo into cells. For this purpose, an assay was designed to provide real-time information about the StxB-Gb3 interaction as well as the dynamics and mechanism of the internalization process. The assay revealed the ability to distinguish the process of binding to the cell surface from internalization and presented the importance of receptor and STxB clustering for internalization. The overall setup demonstrated that the binding mechanism is complex, and the concept of affinity is difficult to apply. Hence, time-resolved methods, providing detailed information about the interaction of STxB with cells, are critical for the optimization of intracellular delivery.
Published: 2020
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31. 1-(1-Arylethylpiperidin-4-yl)thymine analogs as antimycobacterial TMPK inhibitors

Author: Martijn D. P. Risseeuw, Guy Caljon, Fabian Hulpia, Serge Van Calenbergh, He Eun Forbes, Helena I. Boshoff, Yanlin Jian, and Hélène Munier-Lehmann
Subjects: Models, Molecular, 1-(1-arylethylpiperidin-4-yl)thymine, Antitubercular Agents, POTENT INHIBITORS, Pharmaceutical Science, Antimycobacterial, 01 natural sciences, Analytical Chemistry, chemistry.chemical_compound, MtbTMPK, BACTERIAL RNA-POLYMERASE, Drug Discovery, Medicine and Health Sciences, Enzyme Inhibitors, biology, Molecular Structure, TUBERCULOSIS THYMIDYLATE KINASE, Chemistry, tuberculosis, Chemistry (miscellaneous), Molecular Medicine, MYCOBACTERIUM-TUBERCULOSIS, Stereochemistry, medicine.drug_class, THYMIDINE, Substituent, Virulence, Inhibitory postsynaptic potential, Article, lcsh:QD241-441, Mycobacterium tuberculosis, Minimum inhibitory concentration, Structure-Activity Relationship, lcsh:Organic chemistry, Bacterial Proteins, medicine, Humans, Physical and Theoretical Chemistry, Biology, 010405 organic chemistry, Organic Chemistry, biology.organism_classification, 0104 chemical sciences, Thymine, 010404 medicinal & biomolecular chemistry, chemistry, DISCOVERY, CHIMERA, Thymidine, Nucleoside-Phosphate Kinase, RESISTANCE
Abstract: A series of Mycobacterium tuberculosis TMPK (MtbTMPK) inhibitors based on a reported compound 3 were synthesized and evaluated for their capacity to inhibit MtbTMPK catalytic activity and the growth of a virulent M. tuberculosis strain (H37Rv). Modifications of the scaffold of 3 failed to afford substantial improvements in MtbTMPK inhibitory activity and antimycobacterial activity. Optimization of the substitution pattern of the D ring of 3 resulted in compound 21j with improved MtbTMPK inhibitory potency (three-fold) and H37Rv growth inhibitory activity (two-fold). Moving the 3-chloro substituent of 21j to the para-position afforded isomer 21h, which, despite a 10-fold increase in IC50-value, displayed promising whole cell activity (minimum inhibitory concentration (MIC) = 12.5 &mu, M).
Published: 2020

32. First-in-class allosteric inhibitors of bacterial IMPDHs

Author: Gilles Labesse, Olivier Helynck, Sophie Vichier-Guerre, Thomas Alexandre, Muriel Gelin, Hélène Munier-Lehmann, Alexandru Lupan, Laurence Dugué, Ahmed Haouz, Université Sorbonne Paris Cité (USPC), Chimie et Biocatalyse, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Centre de Biochimie Structurale [Montpellier] (CBS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Cristallographie (Plateforme) - Crystallography (Platform), This work was supported in part by the Centre National de la Recherche Scientifique (CNRS), the Institut National de la Santé Et de la Recherche Médicale (INSERM), the Institut Pasteur, the Conseil Régional d'Ile-de-France (Chemical Library Project, grants n° I 06-222/R and I 09-1739/R, which included a postdoctoral fellowship for Alexandru Lupan) and the French Infrastructure for Integrated Structural Biology (FRISBI) ANR-10-INBS-05.Thomas Alexandre was a recipient of a PhD fellowship from the Conseil Régional d'Ile-de-France and the 'D.I.M. maladies infectieuses, parasitaires et nosocomiales émergentes' 2011., We thank Emilie Dufour for participation in the chemical library screening, Zakaria Jemouai for participation in the kinetic characterization of the allosteric inhibitors and Aurélie Bourderioux for performing the fluorescence experiments. We thank the Molecular Biophysics facility (PFBMI, Institut Pasteur, Paris) for their technical assistance and helpful suggestions. The authors are grateful to the staff of the Crystallographic platform from the Institut Pasteur for robot-driven crystallization screening. We thank Alexandre Chenal for fruitful discussion and technical assistance for fluorescence experiments, and Thomas J. Dougherty for careful reading of this manuscript. We acknowledge the Synchrotron SOLEIL (St Aubin, France) staff for assistance and advice during data collection on PROXIMA-1 (X-ray diffraction data) and SWING (SAXS data) beamlines., ANR-10-INBS-0005,FRISBI,Infrastructure Française pour la Biologie Structurale Intégrée(2010), Munier-Lehmann, Hélène, Infrastructure Française pour la Biologie Structurale Intégrée - - FRISBI2010 - ANR-10-INBS-0005 - INBS - VALID, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], and Cristallographie (Plate-forme)
Subjects: MESH: IMP Dehydrogenase, Drug target, Dehydrogenase, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, 01 natural sciences, Nucleotide metabolism, MESH: Allosteric Regulation, IMPDH, MESH: Apoproteins, Low affinity, Adenosine Triphosphate, IMP Dehydrogenase, Drug Discovery, MESH: Adenosine Triphosphate, Enzyme Inhibitors, MESH: Bacterial Proteins, ComputingMilieux_MISCELLANEOUS, chemistry.chemical_classification, 0303 health sciences, Effector, [CHIM.ORGA]Chemical Sciences/Organic chemistry, General Medicine, Bateman domain, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Biochemistry, MESH: Enzyme Inhibitors, MESH: Protein Domains, Allosteric inhibitor, [CHIM.THER] Chemical Sciences/Medicinal Chemistry, Allosteric regulation, Chemical library screening, Small Molecule Libraries, 03 medical and health sciences, Allosteric Regulation, Bacterial Proteins, Protein Domains, Oxidoreductase, MESH: Small Molecule Libraries, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Quaternary structure, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, 030304 developmental biology, Pharmacology, 010405 organic chemistry, Organic Chemistry, [CHIM.ORGA] Chemical Sciences/Organic chemistry, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, 0104 chemical sciences, Enzyme, chemistry, Protein quaternary structure, [SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Apoproteins
Abstract: International audience; Inosine-5'-monophosphate dehydrogenase (IMPDH) is an essential enzyme in many bacterial pathogens and is considered as a potential drug target for the development of new antibacterial agents. Our recent work has revealed the crucial role of one of the two structural domains (i.e. Bateman domain) in the regulation of the quaternary structure and enzymatic activity of bacterial IMPDHs. Thus, we have screened chemical libraries to search for compounds targeting the Bateman domain and identified first in-class allosteric inhibitors of a bacterial IMPDH. These inhibitors were shown to counteract the activation by the natural positive effector, MgATP, and to block the enzyme in its apo conformation (low affinity for IMP). Our structural studies demonstrate the versatility of the Bateman domain to accommodate totally unrelated chemical scaffolds and pave the way for the development of allosteric inhibitors, an avenue little explored until now.
Published: 2019
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33. Criblage et évaluation de l’effet antiviral de molécules contre les herpèsvirus équins 1 et 4 par la mesure d’impédance cellulaire : effet de la spironolactone

Author: Côme Thieulent, Erika HUE, Christine Fortier, Peggy Suzanne, Stephan Zientara, Aymeric Hans, Hélène Munier-Lehmann, Pierre-Olivier Vidalain, Stéphane Pronost, LABÉO, Pôle d’analyses et de recherche de Normandie (LABÉO), Biologie, génétique et thérapies ostéoarticulaires et respiratoires (BIOTARGEN), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), ImpedanCELL, Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN), Virologie UMR1161 (VIRO), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Laboratoire de pathologie équine de Dozulé, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Chimie et Biocatalyse, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Génomique Virale et Vaccination, Interactions Cellules Organismes Environnement (ICORE), CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-CHU Caen, Normandie Université (NU), Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-École nationale vétérinaire d'Alfort (ENVA), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), École nationale vétérinaire - Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Laboratoire de santé animale, sites de Maisons-Alfort et de Normandie, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
Subjects: [SDV]Life Sciences [q-bio], [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, [SDV.BC]Life Sciences [q-bio]/Cellular Biology
Abstract: International audience; Neuf herpèsvirus équins (HVE-1 à 9) sont actuellement décrits chez les équidés. Parmi les cinq herpèsvirus classiquement décrits chez les chevaux nous retrouvons trois alphaherpèsvirus (HVE-1, HVE-3 et HVE-4) et deux gammaherpèsvirus (HVE-2 et HVE-5). Les HVE-1 et HVE-4 sont responsables de maladies respiratoires mais également pour l’HVE-1 de formes secondaires graves comme l’avortement et l’encéphalomyélite, conduisant à de lourdes pertes économiques. Les vaccins disponibles actuellement contre ces deux virus atténuent les problèmes respiratoires et la dissémination du virus mais ne préviennent pas suffisamment les avortements et les formes nerveuses. Par ailleurs aucune molécule ne possède d’autorisation de mise sur le marché en santé équine malgré des besoins importants. Le but de notre étude est de développer de nouvelles molécules, ou de repositionner des molécules déjà existantes, capables de lutter contre l’HVE-1 et l’HVE-4. Pour cela nous avons développé un modèle cellulaire d’étude par la technologie RTCA xCELLigence® (ACEA Biosciences, Inc). Cette technique est basée sur la mesure d’impédance cellulaire en temps réel et permet le criblage moyen/haut débit de molécules (6 x 96 puits simultanément). Notre analyse est basée sur deux principaux paramètres que sont : le calcul d’aire sous la courbe (AUC) et le temps requis pour diminuer de 50% la valeur maximale de Cell Index (CIT50). Suite au criblage d’une première série de 1 200 molécules issues de la banque Prestwick®, 14 hits ont été identifiés comme potentiels antiviraux contre l’HVE-1. Parmi ces molécules, nous avons retrouvé l’aciclovir et le ganciclovir, dont les efficacités avaient déjà été démontrées contre l’HVE-1 et l’HVE-4, confirmant ainsi la pertinence de notre modèle. Nous avons également identifié la spironolactone parmi les 12 autres hits, comme molécule antivirale contre l’HVE-1. Une EC50 de 26,98 ± 1,22 µM mesurée avec le système RTCA xCELLigence et une EC50 de 9,75 ± 3,79 µM mesurée par quantification des charges virales d’HVE-1 (qPCR) ont été obtenues contre l’HVE-1 et des résultats similaires ont été obtenus contre l’HVE-4. Par ailleurs l’efficacité de la spironolactone a récemment été démontrée in vitro contre le virus d’Epstein-Barr [1] et sur les virus de l’immunodéficience humaine-1 et 2 [2]. Ces résultats laissent penser que la spironolactone a un effet antiviral à large spectre et pourrait être utilisée pour lutter contre un grand nombre de virus. Les autres hits sont actuellement étudiés par qPCR pour confirmer les effets obtenus par la technologie RTCA xCELLigence®.Références :1. Lacombe, B., Morel, M., Margottin-Goguet, F., Ramirez, B.C., 2016. Specific Inhibition of HIV Infection by the Action of Spironolactone in T Cells. Journal of Virology 90, 10972–10980. https://doi.org/10.1128/JVI.01722-162. Verma, D., Thompson, J., Swaminathan, S., 2016. Spironolactone blocks Epstein–Barr virus production by inhibiting EBV SM protein function. Proceedings of the National Academy of Sciences 113, 3609–3614.
Published: 2019

34. Endeavors towards transformation of M. tuberculosis thymidylate kinase (MtbTMPK) inhibitors into potential antimycobacterial agents

Author: Fabian Hulpia, Serge Van Calenbergh, He Eun Forbes, Romain Merceron, Steven De Munck, Martijn D. P. Risseeuw, Yanlin Jian, Hélène Munier-Lehmann, Savvas N. Savvides, Kristof Van Hecke, Helena I. Boshoff, Laboratorium voor Medicinale Chemie = Laboratory of Medicinal Chemistry [Ghent], Faculteit Farmaceutische Wetenschappen = Faculty of Pharmaceutical Sciences [Ghent] (FFW), Universiteit Gent = Ghent University (UGENT)-Universiteit Gent = Ghent University (UGENT), VIB-UGent Center for Inflammation Research [Gand, Belgique] (IRC), VIB [Belgium], Universiteit Gent = Ghent University (UGENT), Laboratory of Clinical Immunology and Microbiology [Bethesda, MA, USA] (LCIM), National Institute of Allergy and Infectious Diseases [Bethesda] (NIAID-NIH), National Institutes of Health [Bethesda] (NIH)-National Institutes of Health [Bethesda] (NIH), Chimie et Biocatalyse, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), This work was supported by the China Scholarship Council [grant number 201607060021] and in part by the Intramural Research Program of the National Institute of Allergy and Infectious Diseases (NIAID, NIH, US). KVH thanks the Hercules Foundation (project AUGE/11/029 '3D-SPACE: 3D Structural Platform Aiming for Chemical Excellence', Belgium) and the Special Research Fund (BOF) – UGent (project 01N03217, Belgium) for funding. S.D.M. was supported by a pre-doctoral fellowship from the Flanders Agency for Innovation and Entrepreneurship (VLAIO-Flanders, Belgium). S.N.S. acknowledges research support from the Hercules Foundation (no. AUGE- 11–029, Belgium), Ghent University (BOF17-GOA-028, Belgium) and the VIB., We thank the staff of Proxima 1 at synchrotron SOLEIL (Gif-sur-Yvette, France) for beam time allocation and excellent technical support, Universiteit Gent = Ghent University [Belgium] (UGENT)-Universiteit Gent = Ghent University [Belgium] (UGENT), Universiteit Gent = Ghent University [Belgium] (UGENT), and Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)
Subjects: SIDEROPHORES, [SDV]Life Sciences [q-bio], Antitubercular Agents, Uptake, Antimycobacterial, 0302 clinical medicine, THYMIDINE-5'-O-MONOPHOSPHATE ANALOGS, Siderophore, Drug Discovery, Enzyme Inhibitors, Structural motif, Cytotoxicity, chemistry.chemical_classification, 0303 health sciences, biology, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Chemistry, Drug discovery, Mycobacteria tuberculosis, General Medicine, 3. Good health, Biochemistry, 030220 oncology & carcinogenesis, MYCOBACTERIUM-TUBERCULOSIS, ANTIBIOTICS, NUCLEOSIDES, Thymidine monophosphate kinase (TMPK), medicine.drug_class, Microbial Sensitivity Tests, Thymidylate kinase, Cell Line, Mycobacterium tuberculosis, Structure-Activity Relationship, 03 medical and health sciences, medicine, Humans, 030304 developmental biology, Pharmacology, IDENTIFICATION, THYMIDINE ANALOGS, Organic Chemistry, IN-VITRO, biology.organism_classification, In vitro, Enzyme, PHYSICOCHEMICAL PROPERTIES, DISCOVERY, Drug Design, Nucleoside-Phosphate Kinase
Abstract: International audience; As the last enzyme in nucleotide synthesis as precursors for DNA replication, thymidylate kinase of M. tuberculosis (MtbTMPK) attracts significant interest as a target in the discovery of new anti-tuberculosis agents. Earlier, we discovered potent MtbTMPK inhibitors, but these generally suffered from poor antimycobacterial activity, which we hypothesize is due to poor bacterial uptake. To address this, we herein describe our efforts to equip previously reported MtbTMPK inhibitors with targeting moieties to increase the whole cell activity of the hybrid analogues. Introduction of a simplified Fe-chelating siderophore motif gave rise to analogue 17 that combined favorable enzyme inhibitory activity with significant activity against M. tuberculosis (MIC of 12.5 μM). Conjugation of MtbTMPK inhibitors with an imidazo[1,2-a]pyridine or 3,5-dinitrobenzamide scaffold afforded analogues 26, 27 and 28, with moderate MtbTMPK enzyme inhibitory potency, but sub-micromolar activity against mycobacteria without significant cytotoxicity. These results indicate that conjugation with structural motifs known to favor mycobacterial uptake may be a valid approach for discovering new antimycobacterial agents.
Published: 2020
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35. Synthesis and structure activity relationships of cyanopyridone based anti-tuberculosis agents

Author: He Eun Forbes, Martijn D. P. Risseeuw, Serge Van Calenbergh, Guy Caljon, Helena I. Boshoff, Fabian Hulpia, Yanlin Jian, Hélène Munier-Lehmann, Laboratorium voor Medicinale Chemie = Laboratory of Medicinal Chemistry [Ghent], Faculteit Farmaceutische Wetenschappen = Faculty of Pharmaceutical Sciences [Ghent] (FFW), Universiteit Gent = Ghent University [Belgium] (UGENT)-Universiteit Gent = Ghent University [Belgium] (UGENT), Laboratory of Clinical Immunology and Microbiology [Bethesda, MA, USA] (LCIM), National Institute of Allergy and Infectious Diseases [Bethesda] (NIAID-NIH), National Institutes of Health [Bethesda] (NIH)-National Institutes of Health [Bethesda] (NIH), Chimie et Biocatalyse, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Laboratory of Microbiology, Parasitology and Hygiene [Antwerpen] (LMPH), University of Antwerp (UA), National Institutes of Health [Bethesda] (NIH), This work was supported by the China Scholarship Council (grant number 201607060021), China and in part by the Intramural Research Program of the National Institute of Allergy and Infectious Diseases (NIAID), NIH, USA., Universiteit Gent = Ghent University (UGENT)-Universiteit Gent = Ghent University (UGENT), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
Subjects: [SDV]Life Sciences [q-bio], Antitubercular Agents, Thymidylate kinase, Pharmacology, Antimycobacterial, 01 natural sciences, chemistry.chemical_compound, Catalytic Domain, Drug Discovery, Enzyme Inhibitors, Cytotoxicity, Pathogen, chemistry.chemical_classification, 0303 health sciences, Molecular Structure, biology, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Chemistry, Pharmacology. Therapy, General Medicine, 3. Good health, Molecular Docking Simulation, Protein Binding, Pyridones, medicine.drug_class, Microbial Sensitivity Tests, Article, Mycobacterium tuberculosis, Structure-Activity Relationship, 03 medical and health sciences, Bacterial Proteins, Nitriles, medicine, Tuberculosis, [CHIM]Chemical Sciences, Biology, 030304 developmental biology, DNA synthesis, Inhibitors, 010405 organic chemistry, Organic Chemistry, biology.organism_classification, 0104 chemical sciences, Enzyme, Drug Design, Human medicine, Nucleoside-Phosphate Kinase, Thymidine
Abstract: International audience; Mycobacterium tuberculosis, the causative agent of tuberculosis, relies on thymidylate kinase (MtbTMPK) for the synthesis of thymidine triphosphates and thus also DNA synthesis. Therefore, this enzyme constitutes a potential Achilles heel of the pathogen. Based on a previously reported MtbTMPK 6-aryl-substituted pyridone inhibitor and guided by two co-crystal structures of MtbTMPK with pyridone- and thymine-based inhibitors, we report the synthesis of a series of aryl-shifted cyanopyridone analogues. These compounds generally lacked significant MtbTMPK inhibitory potency, but some analogues did exhibit promising antitubercular activity. Analogue 11i demonstrated a 10-fold increased antitubercular activity (MIC H37Rv, 1.2 μM) compared to literature compound 5. Many analogues with whole-cell antimycobacterial activity were devoid of significant cytotoxicity.
Published: 2020
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36. 1-(Piperidin-3-yl)thymine amides as inhibitors of M. tuberculosis thymidylate kinase

Author: Mathy Froeyen, Davie Cappoen, Lijun Song, Yanlin Jian, Hélène Munier-Lehmann, Martijn D. P. Risseeuw, Serge Van Calenbergh, and Paul Cos
Subjects: Tuberculosis, Stereochemistry, Antitubercular Agents, Thymidylate kinase, Microbial Sensitivity Tests, 01 natural sciences, modelling, Mycobacterium tuberculosis, Structure-Activity Relationship, chemistry.chemical_compound, inhibitors, Drug Discovery, medicine, Enzyme Inhibitors, mycobacterium tuberculosis, Pharmacology, Dose-Response Relationship, Drug, Molecular Structure, biology, thymidylate kinase, 010405 organic chemistry, Pharmacology. Therapy, lcsh:RM1-950, General Medicine, medicine.disease, biology.organism_classification, Amides, 0104 chemical sciences, Thymine, 010404 medicinal & biomolecular chemistry, lcsh:Therapeutics. Pharmacology, chemistry, Nucleoside-Phosphate Kinase
Abstract: A series of readily accessible 1-(piperidin-3-yl)thymine amides was designed, synthesised and evaluated as Mycobacterium tuberculosis TMPK (MtbTMPK) inhibitors. In line with the modelling results, most inhibitors showed reasonable MtbTMPK inhibitory activity. Compounds 4b and 4i were slightly more potent than the parent compound 3. Moreover, contrary to the latter, amide analogue 4g was active against the avirulent M. tuberculosis H37Ra strain (MIC50=35 µM). This finding opens avenues for future modifications. ispartof: JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY vol:34 issue:1 pages:1730-1739 ispartof: location:England status: published
Published: 2019
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37. Effets de l’aciclovir et du ganciclovir contre les herpèsvirus équins : suivi en temps réel par impédancemétrie

Author: Côme Thieulent, Erika Hue, Christine Fortier, Peggy Suzanne, Stephan Zientara, Hélène Munier-Lehmann, aymeric, Pierre-Hugues Pitel, Pierre-Olivier Vidalain, Stéphane Pronost, LABÉO, Pôle d’analyses et de recherche de Normandie (LABÉO), Biologie, génétique et thérapies ostéoarticulaires et respiratoires (BIOTARGEN), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN), Virologie UMR1161 (VIRO), Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-École nationale vétérinaire d'Alfort (ENVA), Chimie et Biocatalyse, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Laboratoire de pathologie équine de Dozulé, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Génomique Virale et Vaccination, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), École nationale vétérinaire - Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de santé animale, sites de Maisons-Alfort et de Normandie, and Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)
Subjects: [SDV]Life Sciences [q-bio], [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, [SDV.BC]Life Sciences [q-bio]/Cellular Biology
Abstract: International audience; Introduction : Trois alphaherpèsvirus (HVE-1, -3, et -4) sont décrits comme infectant les chevaux. L’herpèsvirus équin 1 (HVE-1), nommé virus abortif, est le plus pathogène d’entre eux. Trois formes de la maladie sont classiquement décrites : respiratoire, abortive et nerveuse. L’HVE-4, nommé virus de la rhinopneumonie, provoque principalement des symptômes respiratoires et occasionnellement des avortements. L’HVE-3 est quant à lui responsable de l'exanthème coïtal équin. L’infection des chevaux par ces virus provoque de lourdes pertes économiques pour la filière équine (exemple de l’épizootie d’ampleur observée depuis mars 2018) ce qui justifie une surveillance et une prophylaxie. Actuellement, aucune molécule antivirale ne dispose d'une autorisation de mise sur le marché pour les équidés, malgré des besoins critiques justifiant le développement ou l'adaptation de médicaments efficaces contre les virus équins.Objectifs : 1) Etudier l'efficacité du système xCELLigence pour suivre l’infection cellulaire par l’HVE-1, l’HVE-4 et l’HVE-3 ; 2) Evaluer l'efficacité de l'aciclovir (ACV) et du ganciclovir (GCV) contre les HVEs ; 3) Etudier la capacité du système xCELLigence® pour le criblage d’une chimiothèque de 1200 composés afin d'identifier de nouveaux antiviraux. Matériels et méthodes : L’infection des cellules dermiques équines par l’HVE-1, l’HVE-4 et l’HVE-3 est suivi par impédancemétrie (xCELLigence®, ACEA), par observation microscopique en temps réel et par quantification des charges virales en PCR quantitative.Résultats : L’évolution des valeurs d’impédance (Cell Index, CI) est corrélée à l’apparition et l’évolution de l’effet cytopathogène induit par les HVEs et à l’augmentation des charges virales mesurées. L’ACV et le GCV empêchent la diminution d'impédance induite par l'HVE-1 d'une manière dose-dépendante et les concentrations efficaces médiane (EC50) mesurées sont de 9,88 ± 2,14 μg/ml et de 0,62 ± 0,49 μg/ml, respectivement. Les EC50 de l'ACV et du GCV contre l’HVE-4 sont de 17,38 ± 5,95 μg/ml et de 2,79 ± 0,25 μg/ml, respectivement. Pour l’HVE-3, les EC50 de l'ACV et du GCV sont respectivement de 15,23 ± 2,85 μg/ml et 1,86 ± 0,46 μg/ml. Le criblage d’une première banque de 1200 composés a permis d'identifier plusieurs molécules ayant une activité antivirale contre l’HVE-1.Discussion-Conclusion : Cette étude confirme l'efficacité du système xCELLigence® pour cribler des molécules antivirales contre les alphaherpèsvirus équins. Le GCV s'est avéré être plus efficace que l’ACV contre les trois herpèsvirus étudiés. Comité d’éthique : Non applicableSources de financement : LABÉO, IFCE (Institut Français du Cheval et de l’Equitation, projet AMIE), Fonds Eperon (projets N87-2014, N07-2015, N07-2016, N13-2017 et N62-2017) et Région Normandie (CPER R25 P3).Conflit d’intérêt : Les auteurs déclarent aucun conflit d‘intérêt.
Published: 2018

38. Real-Time monitoring of Equid alphaherpesviruses infectivity in equine dermal cell based on impedance measurements: effects of aciclovir and ganciclovir treatments

Author: Côme Thieulent, Erika Hue, Christine Fortier, Peggy Suzanne, Stephan Zientara, aymeric, Hélène Munier-Lehmann, Guillaume Fortier, Pierre-Hugues Pitel, Pierre-Olivier Vidalain, Stéphane Pronost, LABÉO, Pôle d’analyses et de recherche de Normandie (LABÉO), Biologie, génétique et thérapies ostéoarticulaires et respiratoires (BIOTARGEN), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), ImpedanCELL, Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN), Virologie UMR1161 (VIRO), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Laboratoire de pathologie équine de Dozulé, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Chimie et Biocatalyse, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Interactions Cellules Organismes Environnement (ICORE), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), École nationale vétérinaire - Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Laboratoire de santé animale, sites de Maisons-Alfort et de Normandie, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-CHU Caen, Normandie Université (NU), Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-École nationale vétérinaire d'Alfort (ENVA), and Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris]
Subjects: [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, ComputingMilieux_MISCELLANEOUS
Abstract: International audience
Published: 2018

39. Real-Time monitoring of Equid alphaherpesviruses infectivity in equine dermal cell based on impedance measurements: effects of aciclovir and ganciclovir treatments (IBCA)

Author: Côme Thieulent, Erika Hue, Christine Fortier, Peggy Suzanne, Stephan Zientara, Hélène Munier-Lehmann, aymeric, Pierre-Hugues Pitel, Pierre-Olivier Vidalain, Stéphane Pronost, LABÉO, Pôle d’analyses et de recherche de Normandie (LABÉO), Biologie, génétique et thérapies ostéoarticulaires et respiratoires (BIOTARGEN), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), ImpedanCELL, Interactions Cellules Organismes Environnement (ICORE), CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-CHU Caen, Normandie Université (NU), Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN), Virologie UMR1161 (VIRO), Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-École nationale vétérinaire d'Alfort (ENVA), Chimie et Biocatalyse, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Laboratoire de pathologie équine de Dozulé, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Génomique Virale et Vaccination, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), École nationale vétérinaire - Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de santé animale, sites de Maisons-Alfort et de Normandie, and Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)
Subjects: viruses, [SDV]Life Sciences [q-bio], [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, [SDV.BC]Life Sciences [q-bio]/Cellular Biology
Abstract: International audience; Five herpesvirus have been reported to infect horses and 3 belong to the Alphaherpesvirinae subfamily (equine herpesvirus-1, EHV-4 and EHV-3). Among them, EHV-1 is the most pathogenic and causes respiratory disease in young horses, abortion in mares, neonatal death and neurologic damages, also call equine herpesvirus myeloencephalopathy. EHV-4, also named the rhinopneumonia virus, is closely related to EHV-1 but mostly causes respiratory disease and only sporadic abortion and neonatal infection in horses. EHV-3 is distinct from previous viruses and is responsible for equine coital exanthema, characterized by formation of papules, pustules, ulcers and vesicles on the external genitalia of horses. The equine industry economical loss linked to EHV infection is significant, which warrants surveillance and prophylaxis. Currently, no antiviral molecule has a marketing authorization for equine species despite critically needs, warranting the development or adaptation of drugs that are effective against equine viruses.The aims of this work were 1) to investigate the efficacy of Real-Time Cell Analysis (RTCA) system to monitor EHV infections in equine dermal cells 2) to assess the effectiveness of acyclovir (ACV) and ganciclovir (GCV) against EHV-1, EHV-4 and EHV-3 by RTCA system and 3) to investigate the capacity of RTCA system for the screening of compounds library in order to identified new inhibitors of these viruses. To confirm results achieved, monitoring of cell morphology by microscopy and quantification of viral loads were perform in parallel.Antiviral effect of ACV or GCV against the EHV-1 KyD strain was evaluated by RTCA. Results showed that ACV and GCV prevented the impedance decrease induced by EHV-1 infection in a dose-dependent manner and EC50 measured were 9.88 ± 2.14 µg/ml and 0.62 ± 0.49 µg/ml, respectively. The ACV and GCV EC50 against EHV-4 were 17.38 ± 5.95 µg/ml and 2.79 ± 0.25 µg/ml, respectively. Against EHV-3, the ACV and GCV EC50 were 15.23 ± 2.85 µg/ml and 1.86 ± 0.46 µg/ml, respectively. All the data were confirm by qPCR and microscopy observation. The screening of compounds library allowed identifying several molecules with an antiviral activity needing further investigation.This study confirms the efficiency of RTCA to monitor cytopathic effect formation induced by equine alphaherpesviruses on E. Derm cells in real-time. In this model, GCV was shown to be the most effective against all viruses used to infect E. Derm cells. This technology aims to complement and support conventional methods used in the field of virology.
Published: 2018

40. A Novel Polyaminocarboxylate Compound To Treat Murine Pulmonary Aspergillosis by Interfering with Zinc Metabolism

Author: Oumaïma Ibrahim-Granet, Daniel Mansuy, Olivier Helynck, Laurence du Merle, Rocío Vicentefranqueira, Grégory Jouvion, Jean-Paul Latgé, Frédéric Buron, Jean-Marc Cavaillon, Hélène Munier-Lehmann, Paris Laskaris, Franck Suzenet, José Antonio Calera, Rodolphe Alves de Sousa, Cytokines et Inflammation, Institut Pasteur [Paris], Universidad de Salamanca, Chimie et Biocatalyse, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Histopathologie humaine et Modèles animaux, Biologie des Bactéries pathogènes à Gram-positif, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique et Analytique (ICOA), Université d'Orléans (UO)-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut de Chimie du CNRS (INC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Aspergillus, O.I.-G., H.M.-L., and J.-P.L. were supported by the PTR468 funding program. P.L. received funding from PTR468 and CARNOT MS for a postdoctoral fellowship. J.A.C. was supported by the Spanish Ministry of Economy and Competitiveness through grant SAF2013-48382-R., We thank Yves Janin for the synthesis of the pyrazolones Pyr05 and Pyr11, Constance Bochot and Pierrette Battioni of UMR8601 for the supply of Por06 and Por07 compounds, and the National Library of National Chemistry for providing access to the complete database for screening. We thank Hervé Waxin from the Institut Pasteur education department and Marie-Anne Nicolas from the Institut Pasteur Photonic BioImaging (UTechS PBI) for their assistance in live imaging., Institut Pasteur [Paris] (IP), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université d'Orléans (UO)-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS), Institut Carnot Pasteur MS, and Ministerio de Economía y Competitividad (España)
Subjects: 0301 basic medicine, Fungal infection, [SDV]Life Sciences [q-bio], Aspergillosis, [SDV.MHEP.PSR]Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract, Aspergillus fumigatus, HeLa, chemistry.chemical_compound, Mice, Pharmacology (medical), aspergillosis, Pyrazolones, Antifungal susceptibility testing, zinc metabolism, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, immunosuppression, biology, Chemistry, antifungal susceptibility testing, bioluminescence, animal models, 3. Good health, Animal models, Zinc, Infectious Diseases, Drug screening, Toxicity, antifungal agents, Bioluminescence, Combination therapy, 030106 microbiology, Microbial Sensitivity Tests, Microbiology, 03 medical and health sciences, In vivo, medicine, Animals, Experimental Therapeutics, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, drug screening, Antifungal agents, Pharmacology, fungal infection, Zinc metabolism, Resazurin, biology.organism_classification, medicine.disease, In vitro, Disease Models, Animal, 030104 developmental biology, Luminescent Measurements, Pulmonary Aspergillosis, Immunosuppression
Abstract: Aspergillus fumigatus can cause pulmonary aspergillosis in immunocompromised patients and is associated with a high mortality rate due to a lack of reliable treatment options. This opportunistic pathogen requires zinc in order to grow and cause disease. Novel compounds that interfere with fungal zinc metabolism may therefore be of therapeutic interest. We screened chemical libraries containing 59,223 small molecules using a resazurin assay that compared their effects on an A. fumigatus wild-type strain grown under zinc-limiting conditions and on a zinc transporter knockout strain grown under zinc-replete conditions to identify compounds affecting zinc metabolism. After a first screen, 116 molecules were selected whose inhibitory effects on fungal growth were further tested by using luminescence assays and hyphal length measurements to confirm their activity, as well as by toxicity assays on HeLa cells and mice. Six compounds were selected following a rescreening, of which two were pyrazolones, two were porphyrins, and two were polyaminocarboxylates. All three groups showed good in vitro activity, but only one of the polyaminocarboxylates was able to significantly improve the survival of immunosuppressed mice suffering from pulmonary aspergillosis. This two-tier screening approach led us to the identification of a novel small molecule with in vivo fungicidal effects and low murine toxicity that may lead to the development of new treatment options for fungal infections by administration of this compound either as a monotherapy or as part of a combination therapy., O.I.-G., H.M.-L., and J.-P.L. were supported by the PTR468 funding program. P.L. received funding from PTR468 and CARNOT MS for a postdoctoral fellowship. J.A.C. was supported by the Spanish Ministry of Economy and Competitiveness through grant SAF2013-48382-R.
Published: 2018
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41. Structure Guided Lead Generation toward Nonchiral M. tuberculosis Thymidylate Kinase Inhibitors

Author: Paul Cos, Begoña Gracia, José A. Aínsa, Serge Van Calenbergh, Romain Merceron, Fabian Hulpia, Martijn D. P. Risseeuw, Ainhoa Lucía Quintana, Hélène Munier-Lehmann, Savvas N. Savvides, Lijun Song, Universiteit Gent = Ghent University (UGENT), VIB Inflammation Research Center, University of Zaragoza - Universidad de Zaragoza [Zaragoza], Centro de Investigación Biomédica en Red Enfermedades Respiratorias (CIBERES), University of Antwerp (UA), Chimie et Biocatalyse, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), L.S. thanks the financial support of the Chinese scholarship council (CSC) for a Ph.D. scholarship. F.H. is indebted to the FWO Flanders for a Ph.D. scholarship. The authors would also like to thank Aaron Vasseur for his help in the synthesis part of this project. R.M. and S.N.S. gratefully acknowledge infrastructural access and support at crystallographic beamlines PX1 at SOLEIL synchrotron (Gif-sur-Yvette, France) and P14 at PETRA3 synchrotron (Hamburg, Germany), as well as grant support from the Hercules Foundation (AUGE-11-029) and the Flanders Institute for Biotechnology (VIB)., Universiteit Gent = Ghent University [Belgium] (UGENT), and Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)
Subjects: Models, Molecular, Tuberculosis, Cell Survival, Antitubercular Agents, Molecular Conformation, Microbial Sensitivity Tests, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Pharmacology, 01 natural sciences, Thymidylate kinase, Mycobacterium tuberculosis, Gene Knockout Techniques, Structure-Activity Relationship, chemistry.chemical_compound, Biosynthesis, Drug Discovery, medicine, Humans, Transferase, Enzyme Inhibitors, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], chemistry.chemical_classification, Ligand efficiency, biology, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], 010405 organic chemistry, Pharmacology. Therapy, biology.organism_classification, medicine.disease, 0104 chemical sciences, 3. Good health, 010404 medicinal & biomolecular chemistry, Enzyme, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, chemistry, Drug Design, Molecular Medicine, Efflux, Nucleoside-Phosphate Kinase
Abstract: International audience; In recent years, thymidylate kinase (TMPK), an enzyme indispensable for bacterial DNA biosynthesis, has been pursued for the development of new antibacterial agents including against Mycobacterium tuberculosis, the causative agent for the widespread infectious disease tuberculosis (TB). In response to a growing need for more effective anti-TB drugs, we have built upon our previous efforts toward the exploration of novel and potent Mycobacterium tuberculosis TMPK ( MtTMPK) inhibitors, and reported here the design of a novel series of non-nucleoside inhibitors of MtTMPK. The inhibitors display hitherto unexplored interactions in the active site of MtTMPK, offering new insights into structure-activity relationships. To investigate the discrepancy between enzyme inhibitory activity and the whole-cell activity, experiments with efflux pump inhibitors and efflux pump knockout mutants were performed. The minimum inhibitory concentrations of particular inhibitors increased significantly when determined for the efflux pump mmr knockout mutant, which partly explains the observed dissonance
Published: 2018
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42. Identification of novel leishmanicidal molecules by virtual and biochemical screenings targeting Leishmania eukaryotic translation initiation factor 4A

Author: Imen Bassoumi-Jamoussi, Guillaume Bouvier, Michael Nilges, Mourad Barhoumi, N. Kyle Tanner, Josette Banroques, Arnaud Blondel, Ikram Guizani, Nathan Desdouits, Khadija Essafi-Benkhadir, Hélène Munier-Lehmann, Emna Harigua-Souiai, Yosser Zina Abdelkrim, Ons Zakraoui, Laboratoire d'Epidémiologie Moléculaire et de Pathologie Expérimentale Appliquée aux Maladies Infectieuses (LR11IPT04), Université Tunis El Manar (UTM)-Institut Pasteur de Tunis, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Bioinformatique structurale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Expression Génétique Microbienne (EGM (UMR_8261 / FRE_3630)), Institut de biologie physico-chimique (IBPC), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Faculté des Sciences de Bizerte, Université de Carthage - University of Carthage, Chimie et Biocatalyse, This work received financial support from the Institut Pasteur PTR program (grant PTR426), and partially from the Ministry of Higher Education and Research in Tunisia (LR00IPT04 & LR11IPT04, IG), MN received financial support from the Investissement d’Avenir bioinformatics programme (Grant Bip:Bip ANR-BINF-03-01) and BayCellS FP7-IDEAS-ERC 294809, NKT and JB are supported by a Centre National de la Recherche Scientifique (CNRS), HeliDEAD grant [ANR-13- BSV8-0009-01] from the Agence Nationale de la Recherche and the Initiative d’Excellence program from the French State [DYNAMO, ANR-11-LABX-0011-01]. EHS received support from the UNESCO-L’Oréal 'For Women in Science' international fellowship and the Institut Pasteur International Network (Calmette and Yersin programme). The non commercial compounds were obtained from Dr. Laïla El Kihel from Université de Caen de Basse-Normandie, Centre d’Études et de Recherche sur le Médicament de Normandie (CERMN), UFR des Sciences Pharmaceutiques, for the minimal costs of shipping fees. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript, ANR-10-BINF-03-01/10-BINF-0003,Bip:Bip,Bip:Bip(2010), ANR-13-BSV8-0009,HeliDEAD,Les ARN hélicases à boite DEAD: quels sont leurs rôles, leurs partenaires, leurs substrats ARN et comment elles fonctionnent.(2013), ANR-11-LABX-0011/11-LABX-0011,DYNAMO,Dynamique des membranes transductrices d'énergie : biogénèse et organisation supramoléculaire.(2011), European Project: 294809,EC:FP7:ERC,ERC-2011-ADG_20110310,BAYCELLS(2012), Université de Tunis El Manar (UTM)-Institut Pasteur de Tunis, Bioinformatique structurale - Structural Bioinformatics, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut de biologie physico-chimique (IBPC (FR_550)), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Faculté des Sciences de Bizerte [Université de Carthage], ANR-10-BINF-0003,Bip:Bip,Paradigme d'inference bayesienne pour la Biologie structurale in silico(2010), ANR-11-LABX-0011,DYNAMO,Dynamique des membranes transductrices d'énergie : biogénèse et organisation supramoléculaire.(2011), and Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris]
Subjects: Models, Molecular, 0301 basic medicine, Life Cycles, [SDV]Life Sciences [q-bio], Drug Evaluation, Preclinical, Protozoology, MESH: Parasitic Sensitivity Tests, Biochemistry, Chemical library, MESH: Eukaryotic Initiation Factor-4A, Database and Informatics Methods, White Blood Cells, chemistry.chemical_compound, Parasitic Sensitivity Tests, Animal Cells, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Medicine and Health Sciences, Leishmania infantum, MESH: Inhibitory Concentration 50, Protozoans, Leishmania, Adenosine Triphosphatases, biology, Chemistry, lcsh:Public aspects of medicine, Eukaryota, Adenosine triphosphatase, Parasitic diseases, MESH: Leishmania infantum, Enzymes, 3. Good health, Molecular Docking Simulation, Infectious Diseases, Helicases, MESH: Drug Evaluation, Preclinical, Protozoan Life Cycles, Cellular Types, Sequence Analysis, MESH: Models, Molecular, Research Article, Amastigotes, lcsh:Arctic medicine. Tropical medicine, Bioinformatics, lcsh:RC955-962, Immune Cells, Immunology, Antiprotozoal Agents, Research and Analysis Methods, Microbiology, Inhibitory Concentration 50, 03 medical and health sciences, Eukaryotic translation, MESH: Adenosine Triphosphatases, MESH: Molecular Docking Simulation, Initiation factor, Binding site, MESH: Antiprotozoal Agents, Amastigote, Virtual screening, Blood Cells, Binding Sites, Promastigotes, Macrophages, Phosphatases, Organisms, Public Health, Environmental and Occupational Health, Biology and Life Sciences, Proteins, lcsh:RA1-1270, Cell Biology, biology.organism_classification, RNA helicases, Parasitic Protozoans, 030104 developmental biology, MESH: Binding Sites, Sequence motif analysis, Eukaryotic Initiation Factor-4A, Enzymology, [INFO.INFO-BI]Computer Science [cs]/Bioinformatics [q-bio.QM], Developmental Biology
Abstract: Leishmaniases are neglected parasitic diseases in spite of the major burden they inflict on public health. The identification of novel drugs and targets constitutes a research priority. For that purpose we used Leishmania infantum initiation factor 4A (LieIF), an essential translation initiation factor that belongs to the DEAD-box proteins family, as a potential drug target. We modeled its structure and identified two potential binding sites. A virtual screening of a diverse chemical library was performed for both sites. The results were analyzed with an in-house version of the Self-Organizing Maps algorithm combined with multiple filters, which led to the selection of 305 molecules. Effects of these molecules on the ATPase activity of LieIF permitted the identification of a promising hit (208) having a half maximal inhibitory concentration (IC50) of 150 ± 15 μM for 1 μM of protein. Ten chemical analogues of compound 208 were identified and two additional inhibitors were selected (20 and 48). These compounds inhibited the mammalian eIF4I with IC50 values within the same range. All three hits affected the viability of the extra-cellular form of L. infantum parasites with IC50 values at low micromolar concentrations. These molecules showed non-significant toxicity toward THP-1 macrophages. Furthermore, their anti-leishmanial activity was validated with experimental assays on L. infantum intramacrophage amastigotes showing IC50 values lower than 4.2 μM. Selected compounds exhibited selectivity indexes between 19 to 38, which reflects their potential as promising anti-Leishmania molecules., Author summary Leishmaniases constitute a group of neglected parasitic diseases that inflict major burden on public health. Novel drugs and targets need to be identified since current therapies have adverse side effects. Herein, we focused on Leishmania infantum translation initiation factor 4A (LieIF), as a potential drug target. LieIF, a pivotal enzyme in the translation machinery, is also implicated in host-pathogen interactions. We modeled its 3D structure and identified two pockets, which were used in virtual screenings of a chemical compound library. Therefore, we selected and purchased 305 compounds. We established a reliable ATPase screening assay to test the molecules against the enzymatic activity of LieIF and its mammalian homologue. A promising hit was retained and further characterized. It inhibited both proteins but showed different kinetic properties. It was used as a basis to identify similar analogues and two additional inhibitors were identified. All three hits reduced the viability of the extracellular promastigote form of the parasite, but they had no significant cytotoxic effects on host cells. They also affected the viability of the intracellular amastigote form and reduced the macrophage infection. This selectivity is very promising and indicates that these inhibitors would constitute an avenue to develop strategies to fight leishmaniases.
Published: 2018
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43. Original 2-(3-Alkoxy-1H-pyrazol-1-yl)pyrimidine Derivatives as Inhibitors of Human Dihydroorotate Dehydrogenase (DHODH)

Author: Marianne Lucas-Hourani, Gigliola Zanghi, Sandrine Guillou, Anne Noel, Pierre-Olivier Vidalain, Yves L. Janin, Frédéric Tangy, Olivier Helynck, Hélène Munier-Lehmann, Chimie et Biocatalyse, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Génomique Virale et Vaccination, Medicen Initiative (Chemical Library Project, Région Ile de France Grants I 06-222/R and I 09-1739/R) The Institut Carnot-Pasteur Maladies Infectieuses (Programme STING) Agence Nationale pour la Recherche (ANR-RPIB, Programme STING 2.0) The 2013 Leonardo da Vinci program, ANR-11-RPIB-0006,STING-2.0,Molécules immunostimulatrices ayant une activité antivirale à large spectre: une preuve de concept dans un modèle de primate non-humain(2011), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], and Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)
Subjects: Oxidoreductases Acting on CH-CH Group Donors, Pyrimidine, Stereochemistry, Dihydroorotate Dehydrogenase, Antiviral Agents, Cofactor, law.invention, Measles virus, Structure-Activity Relationship, chemistry.chemical_compound, law, Drug Discovery, Humans, Enzyme Inhibitors, IC50, chemistry.chemical_classification, Coenzyme Q10, biology, [CHIM.ORGA]Chemical Sciences/Organic chemistry, biology.organism_classification, 3. Good health, Pyrimidines, Enzyme, chemistry, Biochemistry, Recombinant DNA, biology.protein, Dihydroorotate dehydrogenase, Molecular Medicine
Abstract: International audience; From a research program aimed at the design of new chemical entities followed by extensive screening on various models of infectious diseases, an original series of 2-(3-alkoxy-1H-pyrazol-1-yl)pyrimidines endowed with notable antiviral properties were found. Using a whole cell measles virus replication assay, we describe here some aspects of the iterative process that, from 2-(4-benzyl-3-ethoxy-5-methyl-1H-pyrazol-1-yl)pyrimidine, led to 2-(4-(2,6-difluorophenoxy)-3-isopropoxy-5-methyl-1H-pyrazol-1-yl)-5-ethylpyrimidine and a 4000-fold improvement of antiviral activity with a subnanomolar level of inhibition. Moreover, recent precedents in the literature describing antiviral derivatives acting at the level of the de novo pyrimidine biosynthetic pathway led us to determine that the mode of action of this series is based on the inhibition of the cellular dihydroorotate dehydrogenase (DHODH), the fourth enzyme of this pathway. Biochemical studies with recombinant human DHODH led us to measure IC50 as low as 13 nM for the best example of this original series when using 2,3-dimethoxy-5-methyl-6-(3-methyl-2-butenyl)-1,4-benzoquinone (coenzyme Q1) as a surrogate for coenzyme Q10, the cofactor of this enzyme.
Published: 2015
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44. MgATP Regulates Allostery and Fiber Formation in IMPDHs

Author: Thomas Alexandre, Hélène Munier-Lehmann, Isabelle Salard-Arnaud, Gilles Labesse, Laurène Vaupré, Joséphine Lai Kee Him, Bertrand Raynal, Patrick Bron, Centre de Biochimie Structurale [Montpellier] (CBS), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM), Chimie et Biocatalyse, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Université Paris Diderot - Paris 7 (UPD7), Biophysique des macromolécules et leurs interactions, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Models, Molecular, Protein Conformation, [SDV]Life Sciences [q-bio], Allosteric regulation, Mutant, Dehydrogenase, Crystallography, X-Ray, 03 medical and health sciences, Adenosine Triphosphate, Biopolymers, IMP Dehydrogenase, Allosteric Regulation, Structural Biology, Oxidoreductase, Molecular Biology, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, Binding Sites, Chemistry, 030302 biochemistry & molecular biology, Recombinant Proteins, Macromolecular assembly, Microscopy, Electron, Enzyme, Biochemistry, Nucleotide Biosynthesis, Pseudomonas aeruginosa
Abstract: This work was supported in part by the Centre National de la Recherche Scientifique (CNRS), the Institut National de la Santé Et de la Recherche Médicale (INSERM), theConseil Régional d'Ile-de-France (Chemical Library Project, grant nos. I 06- 222/R and I 09-1739/ R, which included a postdoctoral fellowship for I.S.-A.) and the French Infrastructure for Integrated Structural Biology (FRISBI) ANR- 10-INSB-05-01. T.A. was a recipient of a PhD fellowship from the Conseil Régional d'Ile-de-France and the ''D.I.M. maladies infectieuses, parasitaires et nosocomiales émergentes'' 2011; International audience; Inosine-5'-monophosphate dehydrogenase (IMPDH) is a rate-limiting enzyme in nucleotide biosynthesis studied as an important therapeutic target and its complex functioning in vivo is still puzzling and debated. Here, we highlight the structural basis for the regulation of IMPDHs by MgATP. Our results demonstrate the essential role of the CBS tandem, conserved among almost all IMPDHs. We found that Pseudomonas aeruginosa IMPDH is an octameric enzyme allosterically regulated by MgATP and showed that this octameric organization is widely conserved in the crystal structures of other IMPDHs. We also demonstrated that human IMPDH1 adopts two types of complementary octamers that can pile up into isolated fibers in the presence of MgATP. The aggregation of such fibers in the autosomal dominant mutant, D226N, could explain the onset of the retinopathy adRP10. Thus, the regulatory CBS modules in IMPDHs are functional and they can either modulate catalysis or macromolecular assembly.
Published: 2013
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45. Original Chemical Series of Pyrimidine Biosynthesis Inhibitors That Boost the Antiviral Interferon Response

Author: Marianne Lucas-Hourani, Philippe V. Afonso, Julien Dairou, Frédéric Tangy, Olivier Helynck, Sébastien Nisole, Daniel Dauzonne, Hélène Munier-Lehmann, Samira Khiar, Pierre-Olivier Vidalain, Génomique virale et vaccination, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Institut Curie, Chimie et Biocatalyse, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut de Recherche en Infectiologie de Montpellier (IRIM), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS), Epidémiologie et Physiopathologie des Virus Oncogènes (EPVO (UMR_3569 / U-Pasteur_3)), Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7)-Institut Pasteur [Paris], This work was supported by the Agence National de la Recherche (ChemInnate program to P.-O.V., S.N., and F.T.), the Institut Pasteur (https://www.pasteur.fr/fr), the Centre National de la Recherche Scientifique (CNRS [www.cnrs.fr]), the Institut National de la Santé Et de la Recherche Médicale (INSERM [www.inserm.fr]), the Institut Carnot–Pasteur Maladies Infectieuses (STING program to P.-O.V. and H.M.-L.), and the Fondation BNP-Paribas (to M.L.-H., S.K., and F.T.)., We thank Nicolas Vanthyne for the separation of chiral enantiomers (Laboratoire de Synthèse et Physico-Chimie de Molécules d'Intérêt Biologique, Plateforme de Chromatographie Chirale, Marseille, France). We thank Gaëlle Cousin, Pierre Jorda, and Chantal Combredet for their technical assistance and Stéphanie Dupuy from the flow cytometry core facility at Université Paris Descartes. We thank Pierre Talbot and Marc Desforges (INRS-Institut Armand-Frappier, Canada) for providing antibodies against the spike protein of HCoV-229E. We thank Philippe Desprès (Université de La Réunion, France) for providing the CHIKV/Ren recombinant virus. We thank Anastassia Komarova for the discussions on ssRNA structures., Institut Curie [Paris], Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris]-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0301 basic medicine, Dicumarol, Oxidoreductases Acting on CH-CH Group Donors, Indoles, Pyrimidine, [SDV]Life Sciences [q-bio], Dihydroorotate Dehydrogenase, Antiviral Agents, Cell Line, Interferon Lambda, 03 medical and health sciences, chemistry.chemical_compound, Structure-Activity Relationship, Interferon, medicine, [CHIM]Chemical Sciences, Humans, Pharmacology (medical), Enzyme Inhibitors, Gene, innate immunity, Pharmacology, Dicoumarol, Immunity, Innate, pyrimidine metabolism, 3. Good health, interferons, Metabolic pathway, 030104 developmental biology, Infectious Diseases, HEK293 Cells, Pyrimidines, chemistry, Biochemistry, Chromones, Measles virus, Pyrimidine metabolism, Interferon Type I, Dihydroorotate dehydrogenase, Chikungunya virus, medicine.drug
Abstract: De novo pyrimidine biosynthesis is a key metabolic pathway involved in multiple biosynthetic processes. Here, we identified an original series of 3-(1 H -indol-3-yl)-2,3-dihydro-4 H -furo[3,2- c ]chromen-4-one derivatives as a new class of pyrimidine biosynthesis inhibitors formed by two edge-fused polycyclic moieties. We show that identified compounds exhibit broad-spectrum antiviral activity and immunostimulatory properties, in line with recent reports linking de novo pyrimidine biosynthesis with innate defense mechanisms against viruses. Most importantly, we establish that pyrimidine deprivation can amplify the production of both type I and type III interferons by cells stimulated with retinoic acid-inducible gene 1 (RIG-I) ligands. Altogether, our results further expand the current panel of pyrimidine biosynthesis inhibitors and illustrate how the production of antiviral interferons is tightly coupled to this metabolic pathway. Functional and structural similarities between this new chemical series and dicoumarol, which was reported before to inhibit pyrimidine biosynthesis at the dihydroorotate dehydrogenase (DHODH) step, are discussed.
Published: 2017
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46. Administration of Zinc Chelators Improves Survival of Mice Infected with Aspergillus fumigatus both in Monotherapy and in Combination with Caspofungin

Author: Oumaïma Ibrahim-Granet, Paris Laskaris, Jean-Paul Latgé, Christophe d'Enfert, Ahmad Atrouni, José Antonio Calera, Hélène Munier-Lehmann, Jean-Marc Cavaillon, Cytokines et Inflammation, Institut Pasteur [Paris], Universidad de Salamanca, Biologie et Pathogénicité fongiques, Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris], Chimie et Biocatalyse, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Aspergillus, Institut Pasteur, Institut Pasteur [Paris]-Institut National de la Recherche Agronomique (INRA), Institut Pasteur [PTR 468], Institut Pasteur [Paris] (IP), Biologie et Pathogénicité fongiques (BPF), Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris] (IP), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0301 basic medicine, Male, Antifungal Agents, Pyridines, 030106 microbiology, Virulence, Aspergillosis, Aspergillus fumigatus, Microbiology, 03 medical and health sciences, chemistry.chemical_compound, Echinocandins, Immunocompromised Host, Lipopeptides, Pharmacotherapy, Caspofungin, medicine, Ethylamines, Animals, Pharmacology (medical), skin and connective tissue diseases, Pathogen, Mechanisms of Action: Physiological Effects, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, Chelating Agents, Pharmacology, Mice, Inbred BALB C, biology, Clioquinol, Immune Sera, Pneumonia, biology.organism_classification, medicine.disease, bacterial infections and mycoses, 3. Good health, Zinc, 030104 developmental biology, Infectious Diseases, Treatment Outcome, chemistry, Nasal administration, Drug Therapy, Combination, medicine.drug, Phenanthrolines
Abstract: Aspergillus fumigatus can infect immunocompromised patients, leading to high mortality rates due to the lack of reliable treatment options. This pathogen requires uptake of zinc from host tissues in order to successfully grow and cause virulence. Reducing the availability of that micronutrient could help treat A. fumigatus infections. In this study, we examined the in vitro effects of seven chelators using a bioluminescent strain of A. fumigatus. 1,10-Phenanthroline and N,N,N′,N′-tetrakis(2-pyridylmethyl) ethane-1,2-diamine (TPEN) proved to be the chelators most effective at inhibiting fungal growth. Intraperitoneal administration of either phenanthroline or TPEN resulted in a significant improvement in survival and decrease of weight loss and fungal burden for immunosuppressed mice intranasally infected with A. fumigatus. In vitro both chelators had an indifferent effect when employed in combination with caspofungin. The use of TPEN in combination with caspofungin also significantly increased survival compared to that when using these drugs individually. Our results suggest that zinc chelation may be a valid strategy for dealing with A. fumigatus infections and that both phenanthroline and TPEN could potentially be used either independently or in combination with caspofungin, indicating that their use in combination with other antifungal treatments might also be applicable., P.L., O.I.-G., H.M.-L., and J.-P.L. were supported by Institut Pasteur PTR 468 funding.
Published: 2016
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47. Real-time monitoring of Equid herpesviruses infectivity in equine dermal cell based on impedance measurements

Author: Stéphan Zientara, Aymeric Hans, Christine Fortier, Erika Hue, Hélène Munier-Lehmann, Peggy Suzanne, Stéphane Pronost, Pierre-Hugues Pitel, Guillaume Fortier, Pierre-Olivier Vidalain, LABÉO, Pôle d’analyses et de recherche de Normandie (LABÉO), Unité de Recherche Risques Microbiens (U2RM), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Fondation Hippolia, Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN), Virologie UMR1161 (VIRO), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Laboratoire de pathologie équine de Dozulé, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Chimie et Biocatalyse, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Institut National de la Recherche Agronomique (INRA)-École nationale vétérinaire d'Alfort (ENVA), Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)
Subjects: Infectivity, Programmed cell death, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, 040301 veterinary sciences, Cell growth, Chemistry, Equine, viruses, 0402 animal and dairy science, 04 agricultural and veterinary sciences, 040201 dairy & animal science, Virology, Virus, 3. Good health, 0403 veterinary science, Cell culture, Cell adhesion, Viral load, Cytopathic effect
Abstract: International audience; In virology, different conventional methods are commonly used for virus titration and infectivity studies. Most of them are based on direct examination of viral cytopathic effect, which is time consuming, tedious and requires an endpoint reading. The RTCA xCELLigence system (ACEA Biosciences) has been developed to follow cellular events and their dynamics in real time using a micro-electrical biosensor technology. The aim of this study was to evaluate the feasibility of using this biosensor technology to measure the effect of different equid herpesviruses on equine dermal cell cultures, with or without antiviral acyclovir treatment. Cells were infected with EHV-1 (VR700), EHV-4 (VR2230), EHV-2 (VR701) or EHV-3 (VR352). The viral effects were monitored by 3 different methods: impedance measurements (RTCA), cytopathic effects recording (microscopy) and viral load quantification (qPCR). The RTCA technology showed a dose dependent drop of the cellular index induced by specific impedance variations measured during growth of each virus tested. These results correlated with the cytopathic effect induced by the different herpesvirus species, as well as viral loads measured by qPCR. For example, after 3 days of incubation with EHV-1, RTCA results showed an increase of the cellular index from 0% (infection without antiviral) to 45% and 100% in presence of Acyclovir (10μl/mL and 100 μg/mL, respectively). These results are correlated with the decreased of viral load measured in the presence of Acyclovir: 1010 genome copies/mL with EHV-1 only, 106 copies/mL with EHV-1 + Acyclovir 10 μg/ mL and 105 copies/mL with EHV-1 + Acyclovir 100 μg/ mL. In conclusion, the cellular impedance monitoring system is a new real-time method to monitor cell growth and viral proliferation in cell cultures. This technology is well adapted for high-throughput screening of antiviral molecules. In fact, the xCELLigence system enables continuous impedance measurement and quantification of cell adhesion, proliferation, cell death and detachment during viral infection.
Published: 2016
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48. Anti-mycobacterial activity of thymine derivatives bearing boron clusters

Author: Edyta Paradowska, Anna Ruszczyńska, Agnieszka Jabłońska, Ewa Bulska, Anna Adamska, Agnieszka B. Olejniczak, Zbigniew J. Leśnikowski, Mirosława Studzińska, Anna Rumijowska-Galewicz, Jarosław Dziadek, Hélène Munier-Lehmann, Polish Academy of Sciences (PAN), University of Warsaw (UW), Warsaw University of Technology [Warsaw], Chimie et Biocatalyse, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], This work was supported by the following grants: the European Regional Development Fund under the Operational Programme Innovative Economy, grant POIG.01.01.02-10-107/09, and the Statutory Fund of IMB PAS to A. Adamska, A. Rumijowska-Galewicz, M. Studzińska, A. Jabłońska, E. Paradowska, J. Dziadek, Z. J. Leśnikowski, and A. B. Olejniczak. Hélene Munier-Lehmann was supported by CNRS, INSERM and the Institute Pasteur. The ICP MS study was carried out at the Biological and Chemical Research Centre, University of Warsaw, established within the project co-financed by European Union from the European Regional Development Found under Operational Programme Innovative Economy, 2007–2013., The authors thank Patrycja Król, Mateusz Pawlaczyk, and Dominika Staniszewska for assistance with the lipophilicity measurements., and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Boron Compounds, MESH: Thymine, Stereochemistry, [SDV]Life Sciences [q-bio], MESH: Boron Compounds, 010402 general chemistry, 01 natural sciences, Thymidylate kinase, Nucleobase, Mycobacterium, Mycobacterium tuberculosis, chemistry.chemical_compound, Structure-Activity Relationship, MESH: Structure-Activity Relationship, Bacterial Proteins, Anti-mycobacterial activity, MESH: Anti-Bacterial Agents, Drug Discovery, Borates, MESH: Mycobacterium, Structure–activity relationship, [CHIM]Chemical Sciences, Nucleoside, MESH: Bacterial Proteins, MESH: Borates, Pharmacology, biology, 010405 organic chemistry, Mycobacterium smegmatis, Organic Chemistry, MESH: Hydrophobic and Hydrophilic Interactions, General Medicine, biology.organism_classification, 3. Good health, 0104 chemical sciences, Thymine, Anti-Bacterial Agents, Boron cluster, chemistry, Nucleoside-Phosphate Kinase, MESH: Nucleoside-Phosphate Kinase, Hydrophobic and Hydrophilic Interactions
Abstract: International audience; A series of novel thymine derivatives bearing lipophilic, electron-neutral 1,2-dicarba-closo-dodecaborane, 1,12-dicarba-closo-dodecaborane or hydrophilic 7,8-dicarba-nido-undecaborate anions were synthesized. Synthesis was performed via copper(I)-catalysed Huisgen-Meldal-Sharpless 1,3-dipolar cycloaddition of N(1)-propargylthymine or N(1),N(3)-bispropargylthymine to 1-(3-azidopropyl)-1,2-dicarba-closo-dodecaborane. The obtained compounds were tested in vitro against Mycobacterium tuberculosis thymidylate kinase (TMPKmt) and as inhibitors of mycobacteria growth in culture using both saprophytic Mycobacterium smegmatis (M. smegmatis) and pathogenic Mycobacterium tuberculosis (M. tuberculosis) strains. The most potent TMPKmt inhibitor in the series contained two negatively charged 7,8-dicarba-nido-undecaborate modifications at positions 1 and 3 of thymine (9) and exhibited a Ki value of 1.5 μM. The most potent inhibitors of mycobacteria growth was compound 5 with one electron-neutral 1,2-dicarba-closo-dodecaborane modification at position 1 of thymine, and compound 8 with two modifications, at position 1 and 3. Both compounds completely inhibited M. smegmatis proliferation at a concentration of 100 μg/mL (0.25 mM and 0.15 mM, respectively).
Published: 2016
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49. The Toxicity of a Novel Antifungal Compound Is Modulated by Endoplasmic Reticulum-Associated Protein Degradation (ERAD) Components

Author: Aureslien Lesnard, Peggy Suzanne, Jean-Paul Latgé, David S. Askew, Ute Zeidler, Olivier Helynck, Sylvain Rault, Cosmin Saveanu, Karthik Krishnan, Christophe d'Enfert, Shriya Raj, Hélène Munier-Lehmann, Aspergillus, Institut Pasteur [Paris], University of Cincinnati (UC), Chimie et Biocatalyse, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Biologie et Pathogénicité fongiques, Institut Pasteur [Paris]-Institut National de la Recherche Agronomique (INRA), Génétique des Interactions macromoléculaires, Aviesan BAP109, Institut Carnot Pasteur Maladies Infectieuses FUNGI, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris], University of Cincinnati, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Centre d'Etudes et de Recherche sur le Médicament de Normandie ( CERMN ), Université de Caen Normandie ( UNICAEN ), Normandie Université ( NU ) -Normandie Université ( NU ), Institut National de la Recherche Agronomique ( INRA ) -Institut Pasteur [Paris], Latgé, Jean-Paul, Munier-Lehmann, Hélène, Saveanu, Cosmin, Institut Pasteur [Paris] (IP), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Biologie et Pathogénicité fongiques (BPF), Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris] (IP), and Génétique des Interactions macromoléculaires / Genetics of Macromolecular Interactions
Subjects: 0301 basic medicine, Antifungal Agents, Saccharomyces cerevisiae, Antifungal drug, Drug Evaluation, Preclinical, Mice, Inbred Strains, [ SDV.BBM.BM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Microbial Sensitivity Tests, UPR, Endoplasmic-reticulum-associated protein degradation, Biology, Protein degradation, Endoplasmic Reticulum, Aspergillus fumigatus, Microbiology, Small Molecule Libraries, 03 medical and health sciences, [ SDV.BBM.BC ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Candida albicans, Animals, Humans, Pharmacology (medical), [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], ER membrane protein complex, Mechanisms of Action: Physiological Effects, Pharmacology, Endoplasmic reticulum, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Endoplasmic Reticulum-Associated Degradation, ERAD, biology.organism_classification, Yeast, 3. Good health, Cell biology, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], 030104 developmental biology, Infectious Diseases, [ SDV.BBM.GTP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Mutation, Unfolded protein response, Cryptococcus neoformans, Unfolded Protein Response, Chemogenomics, HeLa Cells
Abstract: In a search for new antifungal compounds, we screened a library of 4,454 chemicals for toxicity against the human fungal pathogen Aspergillus fumigatus . We identified sr7575, a molecule that inhibits growth of the evolutionary distant fungi A. fumigatus , Cryptococcus neoformans , Candida albicans , and Saccharomyces cerevisiae but lacks acute toxicity for mammalian cells. To gain insight into the mode of inhibition, sr7575 was screened against 4,885 S. cerevisiae mutants from the systematic collection of haploid deletion strains and 977 barcoded haploid DAmP (decreased abundance by mRNA perturbation) strains in which the function of essential genes was perturbed by the introduction of a drug resistance cassette downstream of the coding sequence region. Comparisons with previously published chemogenomic screens revealed that the set of mutants conferring sensitivity to sr7575 was strikingly narrow, affecting components of the endoplasmic reticulum-associated protein degradation (ERAD) stress response and the ER membrane protein complex (EMC). ERAD-deficient mutants were hypersensitive to sr7575 in both S. cerevisiae and A. fumigatus , indicating a conserved mechanism of growth inhibition between yeast and filamentous fungi. Although the unfolded protein response (UPR) is linked to ERAD regulation, sr7575 did not trigger the UPR in A. fumigatus and UPR mutants showed no enhanced sensitivity to the compound. The data from this chemogenomic analysis demonstrate that sr7575 exerts its antifungal activity by disrupting ER protein quality control in a manner that requires ERAD intervention but bypasses the need for the canonical UPR. ER protein quality control is thus a specific vulnerability of fungal organisms that might be exploited for antifungal drug development.
Published: 2016
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50. The Composition of the Culture Medium Influences the β-1,3-Glucan Metabolism of Aspergillus fumigatus and the Antifungal Activity of Inhibitors of β-1,3-Glucan Synthesis

Author: Anne Beauvais, Jean-Paul Latgé, Cécile Clavaud, Lise Barbin, Hélène Munier-Lehmann, Aspergillus, Institut Pasteur [Paris] (IP), Centre hospitalier universitaire de Nantes (CHU Nantes), Chimie et Biocatalyse - Chemistry and Biocatalysis (group), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), This work was partly supported by an Institut Carnot Pasteur Maladies Infectieuses grant (FUNGI), an education grant from Merck (1007005/00), and the Era-Pathogenomic grant Antifun, Institut Pasteur [Paris], Chimie et Biocatalyse, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] - Centre National de la Recherche Scientifique (CNRS), and This work was partly supported by an Institut Carnot Pasteur Maladies Infectieuses grant (FUNGI), an education grant from Merck (1007005/00), and the Era-Pathogenomic grant Antifun.
Subjects: Antifungal Agents, beta-Glucans, Miconazole, [SDV]Life Sciences [q-bio], MESH: Glycosides, Aspergillus fumigatus, Terpene, Echinocandins, MESH: Lipopeptides, Caspofungin, Pharmacology (medical), Glycosides, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, chemistry.chemical_classification, Analytical Procedures, MESH: Microbial Sensitivity Tests, 0303 health sciences, MESH: beta-Glucans, biology, Chitin synthase, 3. Good health, Infectious Diseases, Biochemistry, MESH: Aspergillus fumigatus, medicine.drug, Microbial Sensitivity Tests, macromolecular substances, Microbiology, Lipopeptides, 03 medical and health sciences, medicine, [SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology, 030304 developmental biology, Pharmacology, MESH: Miconazole, 030306 microbiology, MESH: Echinocandins, MESH: Triterpenes, Glycoside, Metabolism, MESH: Antifungal Agents, biology.organism_classification, Triterpenes, In vitro, Culture Media, carbohydrates (lipids), chemistry, MESH: Culture Media, biology.protein
Abstract: In vitro testing of Aspergillus fumigatus susceptibility to echinocandins has always been a challenge. Using a simple and quick colorimetric method to analyze the activity of inhibitors of β-1,3-glucan synthesis, we found that the composition of the culture medium significantly influences glucan synthesis and consequently the antifungal properties of inhibitors of β-1,3-glucan synthesis when they are tested alone or in combination with chitin synthase inhibitors.
Published: 2012
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