title: Ex vivo inhibition of Clostridium botulinum neurotoxin types B, C, E, and F by small molecular weight inhibitors.
abstract: Two small molecular weight inhibitors, compounds CB7969312 and CB7967495, that displayed inhibition of botulinum neurotoxin serotype A in a previous study, were evaluated for inhibition of botulinum neurotoxin serotypes B, C, E, and F. The small molecular weight inhibitors were assessed by molecular modeling, UPLC-based peptide cleavage assay; and an ex vivo assay, the mouse phrenic nerve - hemidiaphragm assay (MPNHDA). While both compounds were inhibitors of botulinum neurotoxin (BoNT) serotypes B, C, and F in the MPNHDA, compound CB7969312 was effective at lower molar concentrations than compound CB7967495. However, compound CB7967495 was significantly more effective at preventing BoNTE intoxication than compound CB7969312. In the UPLC-based peptide cleavage assay, CB7969312 was also more effective against LcC. Both compounds inhibited BoNTE, but not BoNTF, LcE, or LcF in the UPLC-based peptide cleavage assay. Molecular modeling studies predicted that both compounds would be effective inhibitors of BoNTs B, C, E, and F. But CB7967495 was predicted to be a more effective inhibitor of the four serotypes (B, C, E, and F) than CB7969312. This is the first report of a small molecular weight compound that inhibits serotypes B, C, E, and F in the ex vivo assay.
authors.affiliation: U.S. Army Medical Research Institute of Infectious Diseases, Division of Molecular and Translational Sciences, 1425 Porter St. Ft Detrick, Frederick, MD, USA. Electronic address: vicki.a.montgomery.civ@mail.mil.
authors.forename: Vicki A
authors.initials: VA
authors.lastname: Montgomery
authors.affiliation: U.S. Army Medical Research Institute of Infectious Diseases, Division of Molecular and Translational Sciences, 1425 Porter St. Ft Detrick, Frederick, MD, USA. Electronic address: syed.a.ahmed.civ@mail.mil.
authors.forename: S Ashraf
authors.initials: SA
authors.lastname: Ahmed
authors.affiliation: U.S. Army Medical Research Institute of Infectious Diseases, Division of Molecular and Translational Sciences, 1425 Porter St. Ft Detrick, Frederick, MD, USA. Electronic address: mark.a.olson1.civ@mail.mil.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Olson
authors.affiliation: U.S. Army Medical Research Institute of Infectious Diseases, Biosurety Division, 1430 Veterans Dr, Ft Detrick, Frederick, MD, USA(1). Electronic address: mdmizanur.rahman.civ@mail.mil.
authors.forename: Rahman M
authors.initials: RM
authors.lastname: Mizanur
authors.affiliation: U.S. Army Medical Research Institute of Infectious Diseases, Division of Molecular and Translational Sciences, 1425 Porter St. Ft Detrick, Frederick, MD, USA. Electronic address: robert.g.stafford2.civ@mail.mil.
authors.forename: Robert G
authors.initials: RG
authors.lastname: Stafford
authors.affiliation: U.S. Army Medical Research Institute of Infectious Diseases, Biosurety Division, 1430 Veterans Dr, Ft Detrick, Frederick, MD, USA(1). Electronic address: virginia.i.duncan.civ@mail.mil.
authors.forename: Virginia I
authors.initials: VI
authors.lastname: Roxas-Duncan
authors.affiliation: Medical Countermeasures Technology, U.S. Army Medical Research and Material Command, 1425 Porter St. Ft Detrick, Frederick, MD, USA. Electronic address: leonard.a.smith1.civ@mail.mil.
authors.forename: Leonard A
authors.initials: LA
authors.lastname: Smith
mesh_terms: D000631:Aminopyridines / Q000737:chemistry; D000818:Animals; D001905:Botulinum Toxins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D019274:Botulinum Toxins, Type A / Q000037:antagonists & inhibitors* / Q000737:chemistry; D002478:Cells, Cultured; D003014:Clostridium botulinum / Q000737:chemistry*; D006912:Hydroxyquinolines / Q000737:chemistry; D051379:Mice; D015394:Molecular Structure; D008970:Molecular Weight; D010791:Phrenic Nerve / Q000187:drug effects; D011487:Protein Conformation
chemical_list: D000631:Aminopyridines; C000604861:CB 7967495; C000604862:CB 7969312; D006912:Hydroxyquinolines; C096323:rimabotulinumtoxinB; D001905:Botulinum Toxins; D019274:Botulinum Toxins, Type A; C096324:botulinum toxin type C; C096321:botulinum toxin type F
keywords: C. botulinum neurotoxins; Inhibitors