title: Structure-based design of a heptavalent anthrax toxin inhibitor.
abstract: The design of polyvalent molecules, consisting of multiple copies of a biospecific ligand attached to a suitable scaffold, represents a promising approach to inhibit pathogens and oligomeric microbial toxins. Despite the increasing interest in structure-based drug design, few polyvalent inhibitors based on this approach have shown efficacy in vivo. Here we demonstrate the structure-based design of potent biospecific heptavalent inhibitors of anthrax lethal toxin. Specifically, we illustrate the ability to design potent polyvalent ligands by matching the pattern of binding sites on the biological target. We used a combination of experimental studies based on mutagenesis and computational docking studies to identify the binding site for an inhibitory peptide on the heptameric subunit of anthrax toxin. We developed an approach based on copper-catalyzed azide-alkyne cycloaddition (click-chemistry) to facilitate the attachment of seven copies of the inhibitory peptide to a β-cyclodextrin core via a polyethylene glycol linker of an appropriate length. The resulting heptavalent inhibitors neutralized anthrax lethal toxin both in vitro and in vivo and showed appreciable stability in serum. Given the inherent biocompatibility of cyclodextrin and polyethylene glycol, these potent well-defined heptavalent inhibitors show considerable promise as anthrax antitoxins.
authors.affiliation: The Howard P. Isermann Department of Chemical and Biological Engineering, Troy, New York 12180, USA.
authors.forename: Amit
authors.initials: A
authors.lastname: Joshi
authors.forename: Sandesh
authors.initials: S
authors.lastname: Kate
authors.forename: Vincent
authors.initials: V
authors.lastname: Poon
authors.forename: Dhananjoy
authors.initials: D
authors.lastname: Mondal
authors.forename: Mohan B
authors.initials: MB
authors.lastname: Boggara
authors.forename: Arundhati
authors.initials: A
authors.lastname: Saraph
authors.forename: Jacob T
authors.initials: JT
authors.lastname: Martin
authors.forename: Ryan
authors.initials: R
authors.lastname: McAlpine
authors.forename: Ryan
authors.initials: R
authors.lastname: Day
authors.forename: Angel E
authors.initials: AE
authors.lastname: Garcia
authors.forename: Jeremy
authors.initials: J
authors.lastname: Mogridge
authors.forename: Ravi S
authors.initials: RS
authors.lastname: Kane
mesh_terms: D000942:Antigens, Bacterial; D000992:Antitoxins / Q000737:chemistry*; D001427:Bacterial Toxins / Q000037:antagonists & inhibitors*; D001665:Binding Sites; D003505:Cyclodextrins / Q000737:chemistry*; D015195:Drug Design; D004355:Drug Stability; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D013329:Structure-Activity Relationship
chemical_list: D000942:Antigens, Bacterial; D000992:Antitoxins; D001427:Bacterial Toxins; D003505:Cyclodextrins; D010455:Peptides; C030325:anthrax toxin