title: Identification of novel non-hydroxamate anthrax toxin lethal factor inhibitors by topomeric searching, docking and scoring, and in vitro screening.
abstract: Anthrax is an infectious disease caused by Bacillus anthracis, a Gram-positive, rod-shaped, anaerobic bacterium. The lethal factor (LF) enzyme is secreted by B. anthracis as part of a tripartite exotoxin and is chiefly responsible for anthrax-related cytotoxicity. As LF can remain in the system long after antibiotics have eradicated B. anthracis from the body, the preferred therapeutic modality would be the administration of antibiotics together with an effective LF inhibitor. Although LF has garnered a great deal of attention as an attractive target for rational drug design, relatively few published inhibitors have demonstrated activity in cell-based assays and, to date, no LF inhibitor is available as a therapeutic or preventive agent. Here we present a novel in silico high-throughput virtual screening protocol that successfully identified 5 non-hydroxamic acid small molecules as new, preliminary LF inhibitor scaffolds with low micromolar inhibition against that target, resulting in a 12.8% experimental hit rate. This protocol screened approximately 35 million nonredundant compounds for potential activity against LF and comprised topomeric searching, docking and scoring, and drug-like filtering. Among these 5 hit compounds, none of which has previously been identified as a LF inhibitor, three exhibited experimental IC(50) values less than 100 microM. These three preliminary hits may potentially serve as scaffolds for lead optimization as well as templates for probe compounds to be used in mechanistic studies. Notably, our docking simulations predicted that these novel hits are likely to engage in critical ligand-receptor interactions with nearby residues in at least two of the three (S1', S1-S2, and S2') subsites in the LF substrate binding area. Further experimental characterization of these compounds is in process. We found that micromolar-level LF inhibition can be attained by compounds with non-hydroxamate zinc-binding groups that exhibit monodentate zinc chelation as long as key hydrophobic interactions with at least two LF subsites are retained.
authors.affiliation: Department of Medicinal Chemistry, College of Pharmacy, Institute for Therapeutics Discovery and Development, and Minnesota Supercomputing Institute for Advanced Computational Research, University of Minnesota, Minneapolis, Minnesota 55414-2959, USA.
authors.forename: Ting-Lan
authors.initials: TL
authors.lastname: Chiu
authors.forename: Jonathan
authors.initials: J
authors.lastname: Solberg
authors.forename: Satish
authors.initials: S
authors.lastname: Patil
authors.forename: Todd W
authors.initials: TW
authors.lastname: Geders
authors.forename: Xia
authors.initials: X
authors.lastname: Zhang
authors.forename: Subhashree
authors.initials: S
authors.lastname: Rangarajan
authors.forename: Rawle
authors.initials: R
authors.lastname: Francis
authors.forename: Barry C
authors.initials: BC
authors.lastname: Finzel
authors.forename: Michael A
authors.initials: MA
authors.lastname: Walters
authors.forename: Derek J
authors.initials: DJ
authors.lastname: Hook
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Amin
mesh_terms: D000942:Antigens, Bacterial / Q000737:chemistry / Q000378:metabolism; D000992:Antitoxins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D001427:Bacterial Toxins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D006877:Hydroxamic Acids / Q000737:chemistry; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D017434:Protein Structure, Tertiary; D015203:Reproducibility of Results; D014584:User-Computer Interface
chemical_list: D000942:Antigens, Bacterial; D000992:Antitoxins; D001427:Bacterial Toxins; D004791:Enzyme Inhibitors; D006877:Hydroxamic Acids; D061965:Matrix Metalloproteinase Inhibitors; C030325:anthrax toxin; D020782:Matrix Metalloproteinases