Method for suppressing spasmodic torticollis
The present invention relates to a modified polypeptide comprising a non-cytotoxic protease, a translocation domain, a destructive protease cleavage site and a Targeting Moiety that binds to a Binding Site on a nerve cell, wherein after cleavage of the destructive cleavage site the polypeptide has reduced potency. The destructive cleavage site is recognised and cleaved by a protease present at or in an off-site target cell, and, in one embodiment, the polypeptide is a modified clostridial neurotoxin. The present invention also relates to the use of said polypeptides for treating a range of conditions, and to nucleic acids encoding said polypeptides.
1. A method for suppressing spasmodic torticollis comprising administering to a patient in need thereof an effective amount of a polypeptide comprising:
a) a botulinum type A neurotoxin protease domain;
b) a botulinum type A neurotoxin translocation domain; and
c) a botulinum type A neurotoxin targeting moiety that binds to a binding site on a nerve cell of the neuromuscular junction, which binding site is capable of undergoing endocytosis to be incorporated into an endosome within the nerve cell;
wherein:
the polypeptide comprises an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 17, said sequence having protease, translocation, and targeting activity of a botulinum type A neurotoxin; and
the translocation domain is modified by modifying the amino acid sequence located at residues 478-483, 487-499, 511-547, 561-571, 580-584, 622-635, 647-654, 673-681, 755-771, 827-840, and/or 849-863 of SEQ ID NO: 17, or sequences corresponding to such amino acid sequences, to include a destructive cleavage site that is cleavable by a circulatory or tissue-associated protease and not by the botulinum type A neurotoxin protease.
2. The method of claim 1 , wherein the polypeptide comprises an amino acid sequence having at least 95% sequence identity with SEQ ID NO: 17.
3. The method of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 34.
4. The method of claim 1 , wherein:
the translocation domain is modified by modifying the sequence of SEQ ID NO: 64 or SEQ ID NO: 99 therein to include a destructive cleavage site cleavable by Factor Xa.