title: A novel, broad-spectrum inhibitor of enterovirus replication that targets host cell factor phosphatidylinositol 4-kinase IIIβ.
abstract: Despite their high clinical and socioeconomic impacts, there is currently no approved antiviral therapy for the prophylaxis or treatment of enterovirus infections. Here we report on a novel inhibitor of enterovirus replication, compound 1, 2-fluoro-4-(2-methyl-8-(3-(methylsulfonyl)benzylamino)imidazo[1,2-a]pyrazin-3-yl)phenol. This compound exhibited a broad spectrum of antiviral activity, as it inhibited all tested species of enteroviruses and rhinoviruses, with 50% effective concentrations ranging between 4 and 71 nM. After a lengthy resistance selection process, coxsackievirus mutants resistant to compound 1 were isolated that carried substitutions in their 3A protein. Remarkably, the same substitutions were recently shown to provide resistance to inhibitors of phosphatidylinositol 4-kinase IIIβ (PI4KIIIβ), a lipid kinase that is essential for enterovirus replication, suggesting that compound 1 may also target this host factor. Accordingly, compound 1 directly inhibited PI4KIIIβ in an in vitro kinase activity assay. Furthermore, the compound strongly reduced the PI 4-phosphate levels of the Golgi complex in cells. Rescue of coxsackievirus replication in the presence of compound 1 by a mutant PI4KIIIβ carrying a substitution in its ATP-binding pocket revealed that the compound directly binds the kinase at this site. Finally, we determined that an analogue of compound 1, 3-(3-fluoro-4-methoxyphenyl)-2-methyl-N-(pyridin-4-ylmethyl)imidazo[1,2-a]pyrazin-8-amine, is well tolerated in mice and has a dose-dependent protective activity in a coxsackievirus serotype B4-induced pancreatitis model.
authors.affiliation: Department of Infectious Diseases and Immunology, Virology Division, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Hilde M
authors.initials: HM
authors.lastname: van der Schaar
authors.forename: Pieter
authors.initials: P
authors.lastname: Leyssen
authors.forename: Hendrik J
authors.initials: HJ
authors.lastname: Thibaut
authors.forename: Armando
authors.initials: A
authors.lastname: de Palma
authors.forename: Lonneke
authors.initials: L
authors.lastname: van der Linden
authors.forename: Kjerstin H W
authors.initials: KH
authors.lastname: Lanke
authors.forename: Céline
authors.initials: C
authors.lastname: Lacroix
authors.forename: Erik
authors.initials: E
authors.lastname: Verbeken
authors.forename: Katja
authors.initials: K
authors.lastname: Conrath
authors.forename: Angus M
authors.initials: AM
authors.lastname: Macleod
authors.forename: Dale R
authors.initials: DR
authors.lastname: Mitchell
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Palmer
authors.forename: Hervé
authors.initials: H
authors.lastname: van de Poël
authors.forename: Martin
authors.initials: M
authors.lastname: Andrews
authors.forename: Johan
authors.initials: J
authors.lastname: Neyts
authors.forename: Frank J M
authors.initials: FJ
authors.lastname: van Kuppeveld
mesh_terms: D019870:1-Phosphatidylinositol 4-Kinase / Q000378:metabolism*; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology* / Q000627:therapeutic use*; D004770:Enterovirus / Q000187:drug effects* / Q000378:metabolism* / Q000472:pathogenicity; D004789:Enzyme Activation / Q000187:drug effects; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D008297:Male; D051379:Mice; D015394:Molecular Structure; D010195:Pancreatitis / Q000188:drug therapy / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D019870:1-Phosphatidylinositol 4-Kinase
references.citation: Biochem Pharmacol. 2012 Jan 15;83(2):185-92
references.pmid: 21889497
references.citation: Semin Pediatr Infect Dis. 2002 Jan;13(1):40-7
references.pmid: 12118843
references.citation: Cell. 2010 May 28;141(5):799-811
references.pmid: 20510927
references.citation: PLoS Pathog. 2008 Nov;4(11):e1000216
references.pmid: 19023417
references.citation: J Virol. 2012 Nov;86(21):11595-607
references.pmid: 22896614
references.citation: Virology. 2008 Dec 5;382(1):20-7
references.pmid: 18929380
references.citation: Emerg Infect Dis. 2011 Aug;17(8):1494-7
references.pmid: 21801632
references.citation: Pediatr Ann. 1998 Dec;27(12):790-5
references.pmid: 9866136
references.citation: Lancet. 1981 Jun 20;1(8234):1342-4
references.pmid: 6113314
references.citation: Antiviral Res. 2011 Mar;89(3):204-18
references.pmid: 21236302
references.citation: Antimicrob Agents Chemother. 1983 May;23(5):671-5
references.pmid: 6870216
references.citation: Antimicrob Agents Chemother. 2009 May;53(5):1850-7
references.pmid: 19237651
references.citation: J Virol. 2009 Oct;83(19):10058-74
references.pmid: 19605471
references.citation: Drug Des Devel Ther. 2013;7:105-15
references.pmid: 23440335
references.citation: J Biol Chem. 2011 May 27;286(21):18650-7
references.pmid: 21454700
references.citation: ACS Med Chem Lett. 2013 May 08;4(7):585-9
references.pmid: 24900715
references.citation: PLoS One. 2011;6(10):e26300
references.pmid: 22022594
references.citation: mBio. 2011 Oct 04;2(5):
references.pmid: 21972238
references.citation: Microbes Infect. 2004 Jun;6(7):702-13
references.pmid: 15158778
references.citation: Curr Med Chem. 2010;17(25):2804-21
references.pmid: 20586715
references.citation: Med Mal Infect. 2010 Apr;40(4):212-8
references.pmid: 19836177
references.citation: J Virol. 2011 Mar;85(5):2364-72
references.pmid: 21177810
references.citation: Cell Host Microbe. 2009 Mar 19;5(3):298-307
references.pmid: 19286138
references.citation: Biochem J. 2007 Dec 15;408(3):297-315
references.pmid: 17850214
references.citation: Antimicrob Agents Chemother. 2012 Oct;56(10):5149-56
references.pmid: 22825118
references.citation: J Biol Chem. 2006 Nov 24;281(47):36369-77
references.pmid: 17003043
references.citation: Curr Top Microbiol Immunol. 2010;343:43-89
references.pmid: 20397067
references.citation: Lancet. 2013 Mar 23;381(9871):1037-45
references.pmid: 23352749
references.citation: Antivir Chem Chemother. 2000 Jul;11(4):261-71
references.pmid: 10950388
references.citation: Cell Host Microbe. 2011 Jan 20;9(1):32-45
references.pmid: 21238945
references.citation: J Biol Chem. 2012 Mar 9;287(11):8457-67
references.pmid: 22253445
references.citation: Cell Res. 2012 Nov;22(11):1576-92
references.pmid: 22945356
references.citation: Cell. 2006 May 19;125(4):733-47
references.pmid: 16647110
references.citation: BMJ. 2011 Feb 03;342:d35
references.pmid: 21292721
references.citation: J Gen Virol. 2008 Oct;89(Pt 10):2518-2530
references.pmid: 18796721
references.citation: Biochemistry. 1996 Mar 19;35(11):3587-94
references.pmid: 8639510
references.citation: Mol Biol Cell. 2005 Mar;16(3):1282-95
references.pmid: 15635101
references.citation: Virology. 2009 Apr 25;387(1):5-10
references.pmid: 19304308
references.citation: Hepatology. 2009 Dec;50(6):1709-18
references.pmid: 19787809
references.citation: J Virol. 2012 Jan;86(1):302-12
references.pmid: 22072780
references.citation: Lancet Neurol. 2010 Nov;9(11):1097-105
references.pmid: 20965438
references.citation: Front Med. 2013 Mar;7(1):111-21
references.pmid: 23247645
references.citation: J Virol. 2005 Apr;79(8):5163-73
references.pmid: 15795300
references.citation: EMBO J. 2012 Feb 1;31(3):754-66
references.pmid: 22124328
references.citation: FASEB J. 2009 Nov;23(11):3780-9
references.pmid: 19608626
references.citation: Antimicrob Agents Chemother. 2013 Jul;57(7):3358-68
references.pmid: 23650168
references.citation: Dev Cell. 2006 Aug;11(2):191-201
references.pmid: 16890159
references.citation: Genes Dev. 2007 Jan 15;21(2):195-205
references.pmid: 17234885
references.citation: Biochemistry. 2008 Feb 12;47(6):1599-607
references.pmid: 18205404
references.citation: J Neurovirol. 2005 Oct;11(5):424-33
references.pmid: 16287683
references.citation: Nat Rev Microbiol. 2005 Oct;3(10):765-76
references.pmid: 16205710
references.citation: J Clin Virol. 2011 Dec;52(4):321-7
references.pmid: 21924672
references.citation: Proc Natl Acad Sci U S A. 2009 May 5;106(18):7577-82
references.pmid: 19376974
references.citation: PLoS One. 2010 Oct 27;5(10):e13687
references.pmid: 21060866
references.citation: Biochem Biophys Res Commun. 2007 Feb 16;353(3):628-32
references.pmid: 17194441
references.citation: J Virol. 2009 Nov;83(22):11940-9
references.pmid: 19740986
references.citation: Nat Cell Biol. 2004 May;6(5):393-404
references.pmid: 15107860
references.citation: Virology. 1987 Sep;160(1):220-6
references.pmid: 2820130