[en] We report here the synthesis of 7-phenoxy-
substituted 3,4-dihydro-2H-1,2,4-benzothiadiazine 1,1-dioxides
and their evaluation as AMPA receptor positive allosteric
modulators (AMPApams). The impact of substitution on the
phenoxy ring and on the nitrogen atom at the 4-position was
examined. At GluA2(Q) expressed in HEK293 cells (calcium
flux experiment), the most potent compound was 11m (4-
cyclopropyl-7-(3-methoxyphenoxy)-3,4-dihydro-2H-1,2,4-ben-
zothiadiazine 1,1-dioxide, EC50 = 2.0 nM). The Hill coefficient
in the screening and the shape of the dimerization curve in
small-angle X-ray scattering (SAXS) experiments using isolated
GluA2 ligand-binding domain (GluA2-LBD) are consistent
with binding of one molecule of 11m per dimer interface, contrary to most benzothiadiazine dioxides developed to date. This observation was confirmed by the X-ray structure of 11m bound to GluA2-LBD and by NMR. This is the first benzothiadiazine dioxide AMPApam to reach the nanomolar range.
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Goffin, Eric ✱; Université de Liège - ULiège > GIGA-Research
Drapier, Thomas ✱; Université de Liège - ULiège > Département de pharmacie > Chimie pharmaceutique
Probst Larsen, Anja
Geubelle, Pierre ; Université de Liège - ULiège > Département de pharmacie > Chimie pharmaceutique
Ptak, Christopher P.
Laulumaa, Saara
Rovinskaja, Karoline
Gilissen, Julie
De Tullio, Pascal ; Université de Liège - ULiège > Département de pharmacie > Chimie pharmaceutique
Olsen, Lars
Frydenvang, Karla
Pirotte, Bernard ; Université de Liège - ULiège > Unités de recherche interfacultaires > Centre Interdisciplinaire de Recherche sur le Médicament (CIRM)
Hanson, Julien ; Université de Liège - ULiège > Département de pharmacie > Chimie pharmaceutique
Oswald, Robert E.
Sandholm Kastrup, Jette
Francotte, Pierre ; Université de Liège - ULiège > Département de pharmacie > Chimie pharmaceutique
Dingledine, R.; Borges, K.; Bowie, D.; Traynelis, S. F. The glutamate receptor ion channels Pharmacol. Rev. 1999, 51, 7-61
Traynelis, S. F.; Wollmuth, L. P.; McBain, C. J.; Menniti, F. S.; Vance, K. M.; Ogden, K. K.; Hansen, K. B.; Yuan, H.; Myers, S. J.; Dingledine, R. Glutamate receptor ion channels: structure, regulation, and function Pharmacol. Rev. 2010, 62, 405-496 10.1124/pr.109.002451
Huganir, R. L.; Nicoll, R. A. AMPARs and synaptic plasticity: the last 25 years Neuron 2013, 80, 704-717 10.1016/j.neuron.2013.10.025
Lynch, G.; Palmer, L. C.; Gall, C. M. The likelihood of cognitive enhancement Pharmacol., Biochem. Behav. 2011, 99, 116-129 10.1016/j.pbb.2010.12.024
Partin, K. M. AMPA receptor potentiators: from drug design to cognitive enhancement Curr. Opin. Pharmacol. 2015, 20, 46-53 10.1016/j.coph.2014.11.002
Jourdi, J.; Hsu, Y.-T.; Zhou, M.; Qin, Q.; Bi, X.; Baudry, M. Positive AMPA receptor modulation rapidly stimulates BDNF release and increases dendritic mRNA Transl J. Neurosci. 2009, 29, 8688-8697 10.1523/JNEUROSCI.6078-08.2009
Clarkson, A. N.; Overman, J. J.; Zhong, S.; Mueller, R.; Lynch, G.; Carmichael, S. T. AMPA receptor-induced local brain-derived neurotrophic factor signaling mediates motor recovery after stroke J. Neurosci. 2011, 31, 3766-3775 10.1523/JNEUROSCI.5780-10.2011
Gan, Q.; Salussolia, C. L.; Wollmuth, L. P. Assembly of AMPA receptors: mechanisms and regulation J. Physiol. 2015, 593, 39-48 10.1113/jphysiol.2014.273755
Sager, C.; Terhag, J.; Kott, S.; Hollmann, M. C-terminal domains of transmembrane α-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptor regulatory proteins not only facilitate trafficking but are major modulators of AMPA receptor Function J. Biol. Chem. 2009, 284, 32413-32424 10.1074/jbc.M109.039891
Arai, A. C.; Kessler, M.; Rodgers, G.; Lynch, G. Effects of the Potent Ampakine CX614 on hippocampal and recombinant AMPA receptors: interactions with cyclothiazide and GYKI 52466 Mol. Pharm. 2000, 58, 802-813
Ptak, C. P.; Hsieh, C. L.; Weiland, G. A.; Oswald, R. E. Role of stoichiometry in the dimer-stabilizing effect of ampa receptor allosteric modulators ACS Chem. Biol. 2014, 9, 128-133 10.1021/cb4007166
Ptak, C. P.; Ahmed, A. H.; Oswald, R. E. Probing the allosteric modulator binding site of GluR2 with thiazide derivatives Biochemistry 2009, 36, 8594-8602
Pirotte, B.; Francotte, P.; Goffin, E.; de Tullio, P. AMPA receptor positive allosteric modulators: a patent review Expert Opin. Ther. Pat. 2013, 23, 615-628 10.1517/13543776.2013.770840
Citti, C.; Battisti, U. M.; Cannazza, G.; Jozwiak, K.; Stasiak, N.; Puja, G.; Ravazzini, F.; Ciccarella, G.; Braghiroli, D.; Parenti, C.; Troisi, L.; Zoli, M. 7-Chloro-5-(furan-3-yl)-3-methyl-4H-benzo[e][1,2,4]thiadiazine 1,1-dioxide as positive allosteric modulator of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor. the end of the unsaturated-inactive paradigm? ACS Chem. Neurosci. 2016, 7, 149-160 10.1021/acschemneuro.5b00257
Battisti, U. M.; Citti, C.; Rastelli, G.; Pinzi, L.; Puja, G.; Ravazzini, F.; Ciccarella, G.; Braghiroli, D.; Cannazza, G. An unexpected reversal in the pharmacological stereoselectivity of benzothiadiazine AMPA positive allosteric modulators MedChemComm 2016, 7, 2410-2417 10.1039/C6MD00440G
Krintel, C.; Francotte, P.; Pickering, D. S.; Juknaitė, L.; Pøhlsgaard, J.; Olsen, L.; Frydenvang, K.; Goffin, E.; Pirotte, B.; Kastrup, J. S. Enthalpy-entropy compensation in the binding of modulators at ionotropic glutamate receptor GluA2 Biophys. J. 2016, 110, 2397-2406 10.1016/j.bpj.2016.04.032
Larsen, A. P.; Francotte, P.; Frydenvang, K.; Tapken, D.; Goffin, E.; Fraikin, P.; Caignard, D. H.; Lestage, P.; Danober, L.; Pirotte, B.; Kastrup, J. S. Synthesis and pharmacology of mono-, di-, and trialkyl-substituted 7-chloro-3,4-dihydro-2h-1,2,4-benzothiadiazine 1,1-dioxides combined with x-ray structure analysis to understand the unexpected structure-activity relationship at AMPA receptors ACS Chem. Neurosci. 2016, 7, 378-390 10.1021/acschemneuro.5b00318
Cordi, A.; Desos, P.; Lestage, P. N. Benzothia(dia)zine derivatives and their use as ampa modulators. U.S. Patent WO03053947 (A1), 2003.
Francotte, P.; de Tullio, P.; Goffin, E.; Dintilhac, G.; Graindorge, E.; Fraikin, P.; Lestage, P.; Danober, L.; Thomas, J.-Y.; Caignard, D.-H.; Pirotte, B. Design, synthesis, and pharmacology of novel 7-substituted 3,4-dihydro-2h-1,2,4-benzothiadiazine 1,1-dioxides as positive allosteric modulators of AMPA receptors J. Med. Chem. 2007, 50, 3153-3157 10.1021/jm070120i
Nørholm, A. B.; Francotte, P.; Goffin, E.; Botez, I.; Danober, L.; Lestage, P.; Pirotte, B.; Kastrup, J. S.; Olsen, L.; Oostenbrink, C. Thermodynamic characterization of new positive allosteric modulators binding to the glutamate receptor A2 ligand-binding domain: combining experimental and computational methods unravels differences in driving forces J. Chem. Inf. Model. 2014, 54, 3404-3416 10.1021/ci500559b
Francotte, P.; Nørholm, A. B.; Deva, T.; Olsen, L.; Frydenvang, K.; Goffin, E.; Fraikin, P.; de Tullio, P.; Challal, S.; Thomas, J. Y.; Iop, F.; Louis, C.; Botez-Pop, I.; Lestage, P.; Danober, L.; Kastrup, J. S.; Pirotte, B. positive allosteric modulators of 2-amino-3-(3-hydroxy-5-methylisoxazol-4-yl)propionic acid receptors belonging to 4-cyclopropyl-3,4-dihydro-2h-1,2,4-pyridothiadiazine dioxides and diversely chloro-substituted 4-cyclopropyl-3,4-dihydro-2H-1,2,4-benzothiadiazine 1,1-dioxides J. Med. Chem. 2014, 57, 9539-9553 10.1021/jm501268r
Weiss, J. N. The Hill equation revisited: uses and misuses FASEB J. 1997, 11, 835-841
Nørholm, A. B.; Francotte, P.; Olsen, L.; Krintel, C.; Frydenvang, K.; Goffin, E.; Challal, S.; Danober, L.; Botez-Pop, I.; Lestage, P.; Pirotte, B.; Kastrup, J. S. Synthesis, pharmacological and structural characterization, and thermodynamic aspects of GluA2-positive allosteric modulators with a 3,4-dihydro-2H-1,2,4-benzothiadiazine 1,1-dioxide scaffold J. Med. Chem. 2013, 56, 8736-8745 10.1021/jm4012092
Petoukhov, M. V.; Franke, D.; Shkumatov, A. V.; Tria, G.; Kikhney, A. G.; Gajda, M.; Gorba, C.; Mertens, H. D.; Konarev, P. V.; Svergun, D. I. New developments in the ATSAS program package for small-angle scattering data analysis J. Appl. Crystallogr. 2012, 45, 342-350 10.1107/S0021889812007662
Sun, Y.; Olson, R.; Horning, M.; Armstrong, N.; Mayer, M.; Gouaux, E. Mechanism of glutamate receptor desensitization Nature 2002, 417, 245-253 10.1038/417245a
Ahmed, A. H.; Ptak, C. P.; Oswald, R. E. Molecular mechanism of flop selectivity and subsite recognition for an AMPA receptor allosteric modulator: structures of GluA2 and GluA3 in complexes with PEPA Biochemistry 2010, 49, 2843-2850 10.1021/bi1000678
Dunitz, J. D.; Taylor, R. Organic Fluorine hardly ever accepts hydrogen bonds Chem.-Eur. J. 1997, 3, 89-98 10.1002/chem.19970030115
Francotte, P.; Goffin, E.; Fraikin, P.; Lestage, P.; Van Heugen, J.-C.; Gillotin, F.; Danober, L.; Thomas, J.-Y.; Chiap, P.; Caignard, D.-H.; Pirotte, B.; De Tullio, P. New fluorinated 1,2,4-benzothiadiazine 1,1-dioxides: discovery of an orally active cognitive enhancer acting through potentiation of the 2-amino-3-(3-hydroxy-5-methylisoxazol-4-yl)propionic acid receptors J. Med. Chem. 2010, 53, 1700-1711 10.1021/jm901495t
Clark, T.; Hennemann, M.; Murray, J. S.; Politzer, P. Halogen bonding: the σ-hole J. Mol. Model. 2007, 13, 291-296 10.1007/s00894-006-0130-2
Politzer, P.; Murray, J. S.; Clark, T. Halogen bonding: an electrostatically-driven highly directional noncovalent interaction Phys. Chem. Chem. Phys. 2010, 12, 7748-7757 10.1039/c004189k
Krintel, C.; Frydenvang, K.; Olsen, L.; Kristensen, M. T.; de Barrios, O.; Naur, P.; Francotte, P.; Pirotte, B.; Gajhede, M.; Kastrup, J. S. Thermodynamics and structural analysis of positive allosteric modulation of the ionotropic glutamate receptor GluA2 Biochem. J. 2012, 441, 173-178 10.1042/BJ20111221
Skou, S.; Gillilan, R. E.; Ando, N. Synchrotron-based small-angle X-ray scattering of proteins in solution Nat. Protoc. 2014, 9, 1727-1739 10.1038/nprot.2014.116
Acerbo, A. S.; Cook, M. J.; Gillilan, R. E. Upgrade of MacCHESS facility for X-ray scattering of biological macromolecules in solution J. Synchrotron Radiat. 2015, 22, 180-186 10.1107/S1600577514020360
Nielsen, S. S.; Møller, M.; Gillilan, R. E. High-throughput biological small-angle X-ray scattering with a robotically loaded capillary cell J. Appl. Crystallogr. 2012, 45, 213-223 10.1107/S0021889812000957
Hopkins, J. B.; Gillilan, R. E.; Skou, S. BioXTAS RAW: improvements to a free open-source program for small-angle X-ray scattering data reduction and analysis J. Appl. Crystallogr. 2017, 50, 1545-1553 10.1107/S1600576717011438
Ursby, T.; Unge, J.; Appio, R.; Logan, D. T.; Fredslund, F.; Svensson, C.; Larsson, K.; Labrador, A.; Thunnissen, M. M. G. M. The macromolecular crystallography beamline I911-3 at the MAX IV laboratory J. Synchrotron Radiat. 2013, 20, 648-653 10.1107/S0909049513011734
Evans, P. Scaling and assessment of data quality Acta Crystallogr., Sect. D: Biol. Crystallogr. 2006, 62, 72-82 10.1107/S0907444905036693
Winn, M. D.; Ballard, C. C.; Cowtan, K. D.; Dodson, E. J.; Emsley, P.; Evans, P. R.; Keegan, R. M.; Krissinel, E. B.; Leslie, A. G. W.; McCoy, A.; McNicholas, S. J.; Murshudov, G. N.; Pannu, N. S.; Potterton, E. A.; Powell, H. R.; Read, R. J.; Vagin, A.; Wilson, K. S. Overview of the CCP4 suite and current developments Acta Crystallogr., Sect. D: Biol. Crystallogr. 2011, 67, 235-242 10.1107/S0907444910045749
McCoy, A. J.; Grosse-Kunstleve, R. W.; Adams, P. D.; Winn, M. D.; Storoni, L. C.; Read, R. J. Phaser crystallographic software J. Appl. Crystallogr. 2007, 40, 658-674 10.1107/S0021889807021206
Terwilliger, T. C.; Grosse-Kunstleve, R. W.; Afonine, P. V.; Moriarty, N. W.; Zwart, P. H.; Hung, L. W.; Read, R. J.; Adams, P. D. Iterative model building, structure refinement and density modification with the PHENIX AutoBuild wizard Acta Crystallogr., Sect. D: Biol. Crystallogr. 2008, 64, 61-69 10.1107/S090744490705024X
Adams, P. D.; Afonine, P. V.; Bunkóczi, G.; Chen, V. B.; Davis, I. W.; Echols, N.; Headd, J. J.; Hung, L. W.; Kapral, G. J.; Grosse-Kunstleve, R. W.; McCoy, A. J.; Moriarty, N. W.; Oeffner, R.; Read, R. J.; Richardson, D. C.; Richardson, J. S.; Terwilliger, T. C.; Zwart, P. H. PHENIX: a comprehensive Python-based system for macromolecular structure solution Acta Crystallogr., Sect. D: Biol. Crystallogr. 2010, 66, 213-221 10.1107/S0907444909052925
Emsley, P.; Lohkamp, B.; Scott, W. G.; Cowtan, K. Features and development of Coot Acta Crystallogr., Sect. D: Biol. Crystallogr. 2010, 66, 486-501 10.1107/S0907444910007493
Moriarty, N. W.; Grosse-Kunstleve, R. W.; Adams, P. D. electronic Ligand Builder and Optimization Workbench (eLBOW): a tool for ligand coordinate and restraint generation Acta Crystallogr., Sect. D: Biol. Crystallogr. 2009, 65, 1074-1080 10.1107/S0907444909029436
Chen, V. B.; Arendall, W. B.; Headd, J. J.; Keedy, D. A.; Immormino, R. M.; Kapral, G. J.; Murray, L. W.; Richardson, J. S.; Richardson, D. C. MolProbity: all-atom structure validation for macromolecular crystallography Acta Crystallogr., Sect. D: Biol. Crystallogr. 2010, 66, 12-21 10.1107/S0907444909042073
Hayward, S.; Berendsen, H. J. C. Systematic analysis of domain motions in proteins from conformational change; new results on citrate synthase and T4 lysozyme Proteins: Struct., Funct., Genet. 1998, 30, 144-154 10.1002/(SICI)1097-0134(19980201)30:2<144::AID-PROT4>3.0.CO;2-N
Ptak, C. P.; Ahmed, A. H.; Oswald, R. E. NMR approaches to functional dynamics of genetically separated iGluR domains Neuromethods 2016, 106, 101-118 10.1007/978-1-4939-2812-5-8
Delaglio, F.; Grzesiek, S.; Vuister, G. W.; Zhu, G.; Pfeifer, J.; Bax, A. NMRPipe: a multidimensional spectral processing system based on UNIX pipes J. Biomol. NMR 1995, 6, 277-293 10.1007/BF00197809
Lee, W.; Tonelli, M.; Markley, J. L. NMRFAM-SPARKY: enhanced software for biomolecular NMR spectroscopy Bioinformatics 2015, 31, 1325-1327 10.1093/bioinformatics/btu830
Pettersen, E. F.; Goddard, T. D.; Huang, C. C.; Couch, G. S.; Greenblatt, D. M.; Meng, E. C.; Ferrin, T. E. UCSF Chimera-a visualization system for exploratory research and analysis J. Comput. Chem. 2004, 25, 1605-1612 10.1002/jcc.20084
