Compositions and Methods for Topical Application and Transdermal Delivery of Botulinum Toxins

Abstract

A composition for topical application of a botulinum toxin (including botulinum toxin derivatives) comprises a botulinum toxin and a carrier comprising a polymeric backbone comprising a long-chain polypeptide or nonpeptidyl polymer having attached positively charged branching or “efficiency” groups. The invention also relates to methods for reducing muscle paralysis and other conditions that may be treated with a botulinum toxin, particularly paralysis of subcutaneous, and most particularly, facial, muscles, by topically applying an effective amount of the botulinum toxin and carrier, in conjunction, to the subject's skin or epithelium. Kits for administration are also described.

What is claimed is: 
     
         1 . A method of preventing, treating, or reducing symptoms associated with a bladder disease or disorder, the method comprising:
 topically administering to the bladder of a subject in need thereof a pharmaceutically acceptable composition comprising an effective amount of  botulinum  toxin and an effective amount of a positively charged carrier comprising a positively charged polymeric backbone having one or more positively charged efficiency groups covalently attached thereto;   wherein the  botulinum  toxin is not covalently modified;   wherein the  botulinum  toxin and the positively charged polymeric backbone of the carrier directly contact to form a non-covalent complex; and   wherein the topically administered composition prevents, treats, or reduces the symptoms associated with the bladder disease or disorder.   
     
     
         2 . The method according to  claim 1 , wherein the carrier comprises a positively charged polymeric backbone having covalently attached positively charged efficiency groups selected from -(gly) n1 -(arg) n2  (SEQ ID NO. 1), HIV-TAT, or Antennapedia protein transduction domain (PTD) peptide sequences having activity for promoting translocation through a tissue or across a cell membrane, wherein the subscript n1 is an integer of from 0 to about 20, and the subscript n2 is independently an odd integer of from about 5 to about 25. 
     
     
         3 . The method according to  claim 2 , wherein the carrier comprises a positively charged polypeptide backbone having positively charged efficiency groups selected from -(gly) n1 -(arg) n2  (SEQ ID NO. 1), wherein the subscript n1 is an integer of from about 0 to about 20 and the subscript n2 is independently an odd integer of from about 5 to about 25. 
     
     
         4 . The method according to  claim 3 , wherein the subscript n1 is an integer of from 0 to about 8. 
     
     
         5 . The method according to  claim 3 , wherein the subscript n1 is an integer of from about 2 to about 5. 
     
     
         6 . The method according to  claim 3 , wherein the subscript n2 is an odd integer of from about 7 to about 17. 
     
     
         7 . The method according to  claim 3 , wherein the subscript n2 is an odd integer from about 7 to about 13. 
     
     
         8 . The method according to  claim 2 , wherein the carrier comprises a positively charged polypeptide backbone having one or more covalently attached positively charged efficiency groups which are HIV-TAT peptide sequences having activity for promoting translocation through a tissue or across a cell membrane. 
     
     
         9 . The method according to  claim 8 , wherein the one or more positively charged efficiency groups have the amino acid sequences (gly) p -RGRDDRRQRRR-(gly) q  (SEQ ID NO. 2), (gly) p -YGRKKRRQRRR-(gly) q  (SEQ ID NO. 3), or (gly) p -RKKRRQRRR-(gly) q  (SEQ ID NO. 4), wherein the subscripts p and q are each independently an integer of from 0 to 20. 
     
     
         10 . The method according to  claim 9 , wherein the positively charged efficiency groups comprise at least about 0.05% by weight of the total carrier weight. 
     
     
         11 . The method according to  claim 9 , wherein the positively charged efficiency groups comprise from about 0.5% to about 45% by weight of the total carrier weight. 
     
     
         12 . The method according to  claim 9 , wherein the positively charged efficiency groups comprise from about 0.1% to about 30% by weight of the total carrier weight. 
     
     
         13 . The method according to  claim 1 , wherein the positively charged polymeric backbone comprises a positively charged polylysine. 
     
     
         14 . The method according to  claim 9 , wherein the positively charged polypeptide backbone comprises a positively charged polylysine. 
     
     
         15 . The method according to  claim 14 , wherein the polylysine has a molecular weight of from about 10,000 to 1.5 million. 
     
     
         16 . The method according to  claim 15 , wherein the polylysine has a molecular weight of from about 25,000 to about 1,200,000. 
     
     
         17 . The method according to  claim 16 , wherein the polylysine has a molecular weight of from about 100,000 to about 1,000,000. 
     
     
         18 . The method according to  claim 1 , wherein the positively charged polymeric backbone comprises a positively charged nonpeptidyl carrier. 
     
     
         19 . The method according to  claim 18 , wherein the positively charged nonpeptidyl carrier is a polyalkyleneimine. 
     
     
         20 . The method according to  claim 19 , wherein the polyalkyleneimine is a polyethyleneimine. 
     
     
         21 . The method according to  claim 20 , wherein the polyethyleneimine has a molecular weight of from about 10,000 to about 2,500,000. 
     
     
         22 . The method according to  claim 21 , wherein the polyethyleneimine has a molecular weight of from about 100,000 to about 1,800,000. 
     
     
         23 . The method according to  claim 22 , wherein the polyethyleneimine has a molecular weight of from about 500,000 to about 1,400,000. 
     
     
         24 . The method according to  claim 18 , wherein the  botulinum  toxin comprises a recombinant  botulinum  toxin. 
     
     
         25 . The method according to  claim 1 , wherein the  botulinum  toxin comprises a recombinant protein or a fusion protein. 
     
     
         26 . The method according to  claim 1 , wherein the  botulinum  toxin is of serotype A, B, C, D, E, F, or G. 
     
     
         27 . The method according to  claim 14 , wherein the  botulinum  toxin is of serotype A. 
     
     
         28 . The method according to  claim 27 , wherein the serotype A  botulinum  toxin has a molecular weight of about 150,000. 
     
     
         29 . The method of  claim 27 , wherein the  botulinum  toxin is in the form of a  botulinum  toxin complex. 
     
     
         30 . The method according to  claim 28 , wherein the  botulinum  toxin is administered in a composition selected from the group consisting of a controlled release composition; a liquid composition; a gel composition; a cream composition; a lotion composition; and an ointment composition. 
     
     
         31 . The method according to  claim 30 , wherein the composition comprises saline. 
     
     
         32 . The method according to  claim 30 , wherein the composition further comprises a pH buffer system. 
     
     
         33 . The method according to  claim 32 , wherein the composition comprising a pH buffer system having a pH of from about 4.5 to about 6.3. 
     
     
         34 . The method according to  claim 30 , wherein the composition comprising the  botulinum  toxin is contained in a device for dispensing the composition, which device topically applies the composition to the bladder epithelium of the subject. 
     
     
         35 . The method according to  claim 13 , wherein the polylysine has a molecular weight less than 75,000. 
     
     
         36 . The method according to  claim 35 , wherein the polylysine has a molecular weight less than 30,000. 
     
     
         37 . The method according to  claim 36 , wherein the polylysine has a molecular weight less than 25,000. 
     
     
         38 . The method according to  claim 1 , wherein the positively charged polymeric backbone comprises polylysine. 
     
     
         39 . The method according to  claim 38 , wherein the polylysine has a molecular weight less than 75,000. 
     
     
         40 . The method according to  claim 39 , wherein the polylysine has a molecular weight less than 30,000. 
     
     
         41 . The method according to  claim 40 , wherein the polylysine has a molecular weight less than 25,000. 
     
     
         42 . The method according to  claim 1 , wherein the  botulinum  toxin is of serotype A; wherein the positively charged polymeric backbone comprises polylysine; and wherein at least one of the positively charged efficiency groups has the amino acid sequence (gly) p -RGRDDRRQRRR-(gly) q  (SEQ ID NO. 2) and the subscripts p and q are independently an integer in the range of 0 to 20. 
     
     
         43 . The method according to  claim 42 , wherein the polylysine has a molecular weight less than 75,000. 
     
     
         44 . The method according to  claim 43 , wherein the polylysine has a molecular weight less than 30,000. 
     
     
         45 . The method according to  claim 44 , wherein the polylysine has a molecular weight less than 25,000. 
     
     
         46 . The method according to  claim 42 , wherein the  botulinum  toxin has a molecular weight of about 150,000. 
     
     
         47 . The method according to  claim 42 , wherein the subscripts p and q are independently an integer in the range of 0 to 8. 
     
     
         48 . The method according to  claim 1 , wherein the  botulinum  toxin is of serotype A; wherein the positively charged polymeric backbone comprises polylysine; and wherein at least one of the positively charged efficiency groups has the amino acid sequence (gly) p -YGRKKRRQRRR-(gly) q  (SEQ ID NO. 3) and the subscripts p and q are independently an integer in the range of 0 to 20. 
     
     
         49 . The method according to  claim 48 , wherein the polylysine has a molecular weight less than 75,000. 
     
     
         50 . The method according to  claim 49 , wherein the polylysine has a molecular weight less than 30,000. 
     
     
         51 . The method according to  claim 50 , wherein the polylysine has a molecular weight less than 25,000. 
     
     
         52 . The method according to  claim 48 , wherein the  botulinum  toxin has a molecular weight of about 150,000. 
     
     
         53 . The method according to  claim 48 , wherein the subscripts p and q are independently an integer in the range of 0 to 8. 
     
     
         54 . The method according to  claim 1 , wherein the  botulinum  toxin is of serotype A; wherein the positively charged polymeric backbone comprises polylysine; and wherein at least one of the positively charged efficiency groups has the amino acid sequence (gly) p -RKKRRQRRR-(gly) q  (SEQ ID NO. 4) and the subscripts p and q are independently an integer in the range of 0 to 20. 
     
     
         55 . The method according to  claim 54 , wherein the polylysine has a molecular weight less than 75,000. 
     
     
         56 . The method according to  claim 55 , wherein the polylysine has a molecular weight less than 30,000. 
     
     
         57 . The method according to  claim 56 , wherein the polylysine has a molecular weight less than 25,000. 
     
     
         58 . The method according to  claim 54 , wherein the  botulinum  toxin has a molecular weight of about 150,000. 
     
     
         59 . The method according to  claim 54 , wherein the subscripts p and q are independently an integer in the range of 0 to 8. 
     
     
         60 . The method according to  claim 58 , wherein the composition comprising  botulinum  toxin is selected from the group consisting of a controlled release composition; a liquid composition; a gel composition; a cream composition, a lotion composition and an ointment composition. 
     
     
         61 . The method according to  claim 60 , wherein the composition further comprises saline. 
     
     
         62 . The method according to  claim 60 , wherein the composition further comprises a pH buffer system. 
     
     
         63 . The method according to  claim 62 , wherein the pH buffer system has a pH of from about 4.5 to about 6.3. 
     
     
         64 . The method according to  claim 1 , wherein the one or more positively charged efficiency groups covalently attached to the positively charged polymeric backbone are present in an amount sufficient to enhance transdermal delivery of  botulinum  toxin relative to transdermal delivery of  botulinum  toxin using the same positively charged polymeric backbone but without the covalently attached one or more efficiency groups. 
     
     
         65 . The method according to  claim 1 , wherein the composition comprising the  botulinum  toxin is contained in a device for dispensing the composition, which device topically applies the composition to the bladder epithelium of the subject.