Biomadison, Inc.

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Ward C Tucker

Compositions and methods for analyzing intracellular BoNT protease activity, and especially BoNT/B, BoNT/G, BoNT/D, and/or BoNT/F protease activity are provided. Most preferably, cells express one or more recombinant hybrid proteins that include one or more fluorescent proteins and at least one BoNT protease recognition and cleavage sequence, and analysis is performed using FRET analysis.

Resonance energy transfer assay with synaptobrevin substrate moiety

Compositions and methods for inhibiting BoNT protease activity are presented. In most preferred aspects, inhibitors comprise a thiol reactive group, a zinc binding group, a redox active group, an alkylating group, and/or an electrophilic Michael addition acceptor group. Particularly preferred inhibitors include an isothiazolone ring, a thiadiazolidine dione ring, a (hydro)quinone ring, an iminophenol group, and/or a hydrazonophenol group, and inhibit BoNT/A at &#x3bc;M and sub-&#x3bc;M concentrations.

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Del Mar, CA, United States of America

Füsûn N Zeytin