The Design and Synthesis of Orally Active Inhibitors of Botilinum Toxin metalloproteases

This proposal describes strategies to synthesize potent inhibitors of botulinum neurotoxin metalloproteases. These metalloproteases [BoTx protease] have been shown to be the molecular effectors of the paralytic effects of these toxins. Mechanism-based drug design will be applied to develop peptide-derived inhibitors of these proteases. At the same time, experiments to develop non-peptide templates for creating orally active or injectable inhibitors of these proteases will begin, based on use of combinatorial chemistry on known orally active inhibitors of metalloproteases. During Phase, I we will characterize inhibitor activity with a set of 10 15 lead compounds using a procedure based on incubation of various toxin proteolytic substrates with clostridial neurotoxins. These substrates are cleaved by the toxin metalloprotease generating fragments with an electrophoretic mobility different from those of the uncleaved substrates. These compounds are expected to function as pharmacological antagonists and to serve as important probes useful for neurobiological research as adjuncts for reversing effects of BoTx in treating achalasia and related disorders in humans, and as important lead compounds in antitoxin drug development. The product of our work from our biological screen will be provided to investigators at USAMRICD and USANRID for evaluation. These botulinum inhibitors have potential as antidotes under battle field conditions as well as to detoxify military equipment and environments that have been exposed to the toxin. They also would benefit consumer food safety as well as create new applications in medical therapeutics for the treatment of muscular disorders. Additionally there is potential to use these inhibitors for toxin detection and as novel cell biology research tools.