Efficacy of a novel metalloprotease inhibitor on botulinum neurotoxin B activity.
The novel inhibitor 7-N-phenylcarbamoylamino-4-chloro-3-propyloxyisocoumarin (ICD 1578) was tested for its ability to antagonize the zinc metalloprotease activity of botulinum toxin B (BoNT/B). The efficacy of this compound was tested in a cell-free system using a 50-mer synaptobrevin peptide as substrate. The peptide, designated as [Pya88] S 39-88, had a fluorescent amino acid analog, L-pyrenylalanine (Pya), substituted for the normal Phe88 of synaptobrevin-2. Cleavage by BoNT light chain yielded fragments of 38 and 11 amino acids, respectively. The smaller fragment, containing the Pya fluorophore, was readily separated and quantified by fluorescence spectroscopy at 377 nm. In the presence of 7-200 microM ICD 1578, cleavage of [Pya88] S 39-88 was progressively reduced (IC50 = 27.6 microM), and 100 microM ICD 1578 produced >95% inhibition. For comparison, captopril, a well-known zinc metalloprotease inhibitor, generated less than 10% inhibition at a concentration of 5 mM. ICD 1578 is the most potent antagonist of BoNT/B light chain thus far described.
Adler
M
M
Neurotoxicology Branch, Pharmacology Division, U.S. Army Medical Research Institute of Chemical Defense, Aberdeen Proving Ground, MD 21010, USA.
Nicholson
J D
JD
Cornille
F
F
Hackley
B E
BE
D001905:Botulinum Toxins / Q000037:antagonists & inhibitors*; D019274:Botulinum Toxins, Type A; D002216:Captopril / Q000494:pharmacology; D003374:Coumarins / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D049934:Isocoumarins; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors*; D011480:Protease Inhibitors / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence; D015032:Zinc
C113253:7-N-phenylcarbamoylamino-4-chloro-3-propyloxyisocoumarin; D003374:Coumarins; D049934:Isocoumarins; D011480:Protease Inhibitors; C096323:rimabotulinumtoxinB; D002216:Captopril; D008666:Metalloendopeptidases; D001905:Botulinum Toxins; D019274:Botulinum Toxins, Type A; D015032:Zinc