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A preparative solution-phase route was published by Ryakhovsky and co-workers in the Beilstein Journal of Organic Chemistry in 2008, assembling the hexapeptide by a [(2+2)+1+1] scheme, closing the lactam by carbodiimide-mediated lactamization between the epsilon-amino group of lysine and the side-chain carboxyl of aspartic acid, and finishing by appending N-acetylnorleucine

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64,65 Botulinum toxin type A (BoNT-A) produces sustained and prolonged relaxation of muscles by inhibiting release of Ach at the motor endplate and is itself an analgesic inhibiting central sensitization