Rational Design of Therapeutic and Diagnostic Against Botulinum Neurotoxin

Abstract : In September 2006, several cases of botulinum poisoning were reported in United States and Canada due to consumption of commercial organic carrot juice. This incident led to the hospitalization of several individuals who received intensive ventilator support. In spite of botulinum neurotoxin being the most poisonous material, little is known about its mechanism of binding, effective drugs are lacking, and correct diagnosis of botulinum poisoning is slow. Fast and accurate diagnosis of botulinum poisoning (through protein fingerprinting), and new rational drug designs are needed to supplement the current protocol for treatment of botulinum poisoning: administration of antitoxin, adequate mechanical ventilation, and meticulous and intensive care. This technical memorandum reviews past and present research & development efforts on botulinum neurotoxins and future pre-clinical drug discovery directions at DRDC Suffield. A comprehensive drug discovery process is described, including high throughput screening FRET assay, rapid and efficient CE-LIF enzymatic assay, equilibrium binding -affinity MS assay, cell-based assay, and in vivo mouse bioassay. Mechanistically-novel additions to current therapeutics could be in the form of a combination of antitoxin and drugs against cell binding and/or the proteolytic activity of botulinum