title: Potent host-directed small-molecule inhibitors of myxovirus RNA-dependent RNA-polymerases.
abstract: Therapeutic targeting of host cell factors required for virus replication rather than of pathogen components opens new perspectives to counteract virus infections. Anticipated advantages of this approach include a heightened barrier against the development of viral resistance and a broadened pathogen target spectrum. Myxoviruses are predominantly associated with acute disease and thus are particularly attractive for this approach since treatment time can be kept limited. To identify inhibitor candidates, we have analyzed hit compounds that emerged from a large-scale high-throughput screen for their ability to block replication of members of both the orthomyxovirus and paramyxovirus families. This has returned a compound class with broad anti-viral activity including potent inhibition of different influenza virus and paramyxovirus strains. After hit-to-lead chemistry, inhibitory concentrations are in the nanomolar range in the context of immortalized cell lines and human PBMCs. The compound shows high metabolic stability when exposed to human S-9 hepatocyte subcellular fractions. Antiviral activity is host-cell species specific and most pronounced in cells of higher mammalian origin, supporting a host-cell target. While the compound induces a temporary cell cycle arrest, host mRNA and protein biosynthesis are largely unaffected and treated cells maintain full metabolic activity. Viral replication is blocked at a post-entry step and resembles the inhibition profile of a known inhibitor of viral RNA-dependent RNA-polymerase (RdRp) activity. Direct assessment of RdRp activity in the presence of the reagent reveals strong inhibition both in the context of viral infection and in reporter-based minireplicon assays. In toto, we have identified a compound class with broad viral target range that blocks host factors required for viral RdRp activity. Viral adaptation attempts did not induce resistance after prolonged exposure, in contrast to rapid adaptation to a pathogen-directed inhibitor of RdRp activity.
authors: Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, United States of America.
authors: Stefanie A
authors: SA
authors: Krumm
authors: J Maina
authors: JM
authors: Ndungu
authors: Jeong-Joong
authors: JJ
authors: Yoon
authors: Melanie
authors: M
authors: Dochow
authors: Aiming
authors: A
authors: Sun
authors: Michael
authors: M
authors: Natchus
authors: James P
authors: JP
authors: Snyder
authors: Richard K
authors: RK
authors: Plemper
references: Nat Biotechnol. 2007 Dec;25(12):1383-9
references: 18066032
references: J Biomol Screen. 2008 Aug;13(7):591-608
references: 18626114
references: Antimicrob Agents Chemother. 2010 Aug;54(8):3170-8
references: 20498326
references: Expert Rev Anti Infect Ther. 2006 Dec;4(6):953-6
references: 17181411
references: Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):8925-8
references: 7692440
references: J Virol. 2008 Feb;82(4):1946-58
references: 18057241
references: Clin Infect Dis. 2007 Jan 15;44(2):197-202
references: 17173216
references: J Virol. 2010 Feb;84(4):2180-3
references: 19939926
references: PLoS Negl Trop Dis. 2007 Nov 21;1(2):e86
references: 18060089
references: Mol Cell. 2008 Jan 18;29(1):1-7
references: 18206964
references: Antivir Chem Chemother. 2007;18(1):1-11
references: 17354647
references: Nature. 2010 Feb 11;463(7282):818-22
references: 20081832
references: Genes Cells. 2007 Feb;12(2):133-42
references: 17295834
references: Virology. 1995 Apr 20;208(2):800-7
references: 7747454
references: Sci Signal. 2008 Jul 22;1(29):re8
references: 18648074
references: J Virol. 2007 Apr;81(7):3058-67
references: 17229704
references: J Biotechnol. 2008 Jan 20;133(2):208-12
references: 17884219
references: EMBO J. 1995 Dec 1;14(23):5773-84
references: 8846771
references: J Virol. 2001 May;75(9):4399-401
references: 11287589
references: Org Biomol Chem. 2004 Sep 7;2(17):2530-7
references: 15326534
references: CMAJ. 2003 Jan 7;168(1):49-56
references: 12515786
references: J Infect Dis. 1997 Nov;176(5):1215-24
references: 9359721
references: Mol Cell Biol. 2008 Jul;28(14):4562-75
references: 18490444
references: Clin Infect Dis. 2006 Oct 15;43(8):1016-22
references: 16983614
references: J Virol. 1999 Jan;73(1):251-9
references: 9847328
references: Mol Carcinog. 2003 Apr;36(4):183-94
references: 12669310
references: Proc Natl Acad Sci U S A. 1985 Aug;82(16):5328-31
references: 2410919
references: Drug Discov Today. 2006 Nov;11(21-22):975-82
references: 17055406
references: J Med Chem. 2002 Apr 11;45(8):1712-22
references: 11931626
references: J Virol. 2008 Jan;82(1):105-14
references: 17959676
references: Science. 1994 Nov 4;266(5186):801-5
references: 7973634
references: Science. 2005 May 20;308(5725):1174-8
references: 15905402
references: J Virol. 2002 May;76(10):5051-61
references: 11967321
references: J Infect Dis. 2008 Jan 1;197(1):156-62
references: 18171299
references: J Gen Virol. 2008 Apr;89(Pt 4):984-993
references: 18343840
references: J Pediatr Health Care. 2004 Nov-Dec;18(6):297-301
references: 15523420
references: Proteomics. 2008 Feb;8(4):852-73
references: 18297655
references: J Inherit Metab Dis. 1991;14(4):421-30
references: 1749210
references: Nat Rev Genet. 2008 Apr;9(4):267-76
references: 18319742
references: Antimicrob Agents Chemother. 2007 Jul;51(7):2293-303
references: 17470652
references: J Gen Virol. 1999 Jul;80 ( Pt 7):1599-1608
references: 10423127
references: Antimicrob Agents Chemother. 2005 Nov;49(11):4721-32
references: 16251317
references: N Engl J Med. 1993 Nov 18;329(21):1524-30
references: 8413475
references: Nucleic Acids Res. 2009 Sep;37(17):e115
references: 19561200
references: Expert Opin Investig Drugs. 2008 Jan;17(1):43-60
references: 18095918
references: Comb Chem High Throughput Screen. 2008 Feb;11(2):135-45
references: 18336207
references: Pharmacogenomics J. 2008 Feb;8(1):61-70
references: 17505500
references: Nat Rev Genet. 2008 Feb;9(2):102-14
references: 18197166
references: Anal Bioanal Chem. 2008 Feb;390(4):989-1007
references: 17581746
references: J Med Chem. 2008 Jul 10;51(13):3731-41
references: 18529043
references: Nat Rev Drug Discov. 2006 Dec;5(12):1015-25
references: 17139286
references: Nature. 2010 Feb 11;463(7282):813-7
references: 20027183
references: Cardiovasc Clin. 1974;6(2):63-75
references: 4613464
references: J Infect Dis. 1990 Apr;161(4):640-6
references: 2319164
references: Antimicrob Agents Chemother. 2005 Sep;49(9):3755-61
references: 16127050
references: Proc Natl Acad Sci U S A. 1993 May 1;90(9):4171-5
references: 8387212
references: Proc Natl Acad Sci U S A. 2007 Jan 30;104(5):1604-9
references: 17242365
references: Gene. 2007 Dec 15;405(1-2):65-78
references: 17949927
references: Exp Cell Res. 1980 Apr;126(2):397-405
references: 6153987
references: J Virol. 2007 Aug;81(16):8821-6
references: 17553889
references: Comb Chem High Throughput Screen. 2007 Sep;10(8):706-18
references: 18045082
references: Methods Cell Biol. 1990;33:285-98
references: 1707487
references: Nat Protoc. 2007;2(7):1652-60
references: 17641630
references: Lancet. 2004 Aug 28-Sep 3;364(9436):759-65
references: 15337401
references: J Clin Invest. 2007 Dec;117(12):3633-41
references: 18060021
references: Immunity. 2007 Feb;26(2):133-7
references: 17307699
references: Proc Natl Acad Sci U S A. 2006 Aug 15;103(33):12481-6
references: 16885212
references: Cell Microbiol. 2009 Jun;11(6):863-71
references: 19290913
references: Antimicrob Agents Chemother. 2009 Sep;53(9):3860-70
references: 19528268
references: Proc Natl Acad Sci U S A. 2004 Apr 13;101(15):5628-33
references: 15056763
references: J Exp Med. 2008 Mar 17;205(3):585-94
references: 18299402
references: J Virol. 2008 Nov;82(21):10887-95
references: 18715928
references: BMC Immunol. 2005 Jan 18;6:2
references: 15655079
references: J Virol. 2005 Jul;79(14):9197-205
references: 15994814