title: Anthrax lethal factor inhibition.
abstract: The primary virulence factor of Bacillus anthracis is a secreted zinc-dependent metalloprotease toxin known as lethal factor (LF) that is lethal to the host through disruption of signaling pathways, cell destruction, and circulatory shock. Inhibition of this proteolytic-based LF toxemia could be expected to provide therapeutic value in combination with an antibiotic during and immediately after an active anthrax infection. Herein is shown the crystal structure of an intimate complex between a hydroxamate, (2R)-2-[(4-fluoro-3-methylphenyl)sulfonylamino]-N-hydroxy-2-(tetrahydro-2H-pyran-4-yl)acetamide, and LF at the LF-active site. Most importantly, this molecular interaction between the hydroxamate and the LF active site resulted in (i) inhibited LF protease activity in an enzyme assay and protected macrophages against recombinant LF and protective antigen in a cell-based assay, (ii) 100% protection in a lethal mouse toxemia model against recombinant LF and protective antigen, (iii) approximately 50% survival advantage to mice given a lethal challenge of B. anthracis Sterne vegetative cells and to rabbits given a lethal challenge of B. anthracis Ames spores and doubled the mean time to death in those that died in both species, and (iv) 100% protection against B. anthracis spore challenge when used in combination therapy with ciprofloxacin in a rabbit "point of no return" model for which ciprofloxacin alone provided 50% protection. These results indicate that a small molecule, hydroxamate LF inhibitor, as revealed herein, can ameliorate the toxemia characteristic of an active B. anthracis infection and could be a vital adjunct to our ability to combat anthrax.
authors.affiliation: Merck Research Laboratories, Rahway, NJ 07065, USA. weseley_shoop@merck.com
authors.forename: W L
authors.initials: WL
authors.lastname: Shoop
authors.forename: Y
authors.initials: Y
authors.lastname: Xiong
authors.forename: J
authors.initials: J
authors.lastname: Wiltsie
authors.forename: A
authors.initials: A
authors.lastname: Woods
authors.forename: J
authors.initials: J
authors.lastname: Guo
authors.forename: J V
authors.initials: JV
authors.lastname: Pivnichny
authors.forename: T
authors.initials: T
authors.lastname: Felcetto
authors.forename: B F
authors.initials: BF
authors.lastname: Michael
authors.forename: A
authors.initials: A
authors.lastname: Bansal
authors.forename: R T
authors.initials: RT
authors.lastname: Cummings
authors.forename: B R
authors.initials: BR
authors.lastname: Cunningham
authors.forename: A M
authors.initials: AM
authors.lastname: Friedlander
authors.forename: C M
authors.initials: CM
authors.lastname: Douglas
authors.forename: S B
authors.initials: SB
authors.lastname: Patel
authors.forename: D
authors.initials: D
authors.lastname: Wisniewski
authors.forename: G
authors.initials: G
authors.lastname: Scapin
authors.forename: S P
authors.initials: SP
authors.lastname: Salowe
authors.forename: D M
authors.initials: DM
authors.lastname: Zaller
authors.forename: K T
authors.initials: KT
authors.lastname: Chapman
authors.forename: E M
authors.initials: EM
authors.lastname: Scolnick
authors.forename: D M
authors.initials: DM
authors.lastname: Schmatz
authors.forename: K
authors.initials: K
authors.lastname: Bartizal
authors.forename: M
authors.initials: M
authors.lastname: MacCoss
authors.forename: J D
authors.initials: JD
authors.lastname: Hermes
mesh_terms: D000818:Animals; D000881:Anthrax / Q000188:drug therapy*; D000942:Antigens, Bacterial / Q000378:metabolism / Q000633:toxicity*; D001408:Bacillus anthracis / Q000378:metabolism / Q000472:pathogenicity*; D001427:Bacterial Toxins / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000633:toxicity*; D002939:Ciprofloxacin / Q000627:therapeutic use; D003461:Crystallography; D003601:Cytotoxicity Tests, Immunologic; D017931:DNA Primers; D004359:Drug Therapy, Combination; D006877:Hydroxamic Acids / Q000378:metabolism / Q000494:pharmacology* / Q000627:therapeutic use; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D011817:Rabbits
chemical_list: D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D017931:DNA Primers; D006877:Hydroxamic Acids; C030325:anthrax toxin; D002939:Ciprofloxacin