Injectable Botulinum Toxin Formulations And Methods Of Use Thereof Having Long Duration Of Therapeutic Or Cosmetic Effect

Abstract

This invention provides novel injectable compositions comprising botulinum toxin that may be administered to a subject for various therapeutic, aesthetic and/or cosmetic purposes. The injectable compositions embraced by the invention exhibit one or more advantages over conventional botulinum toxin formulations, including reduced antigenicity, a reduced tendency to undergo unwanted localized diffusion following injection, increased duration of clinical efficacy or enhanced potency relative, faster onset of clinical efficacy, and/or improved stability. According to the invention, single treatment of the compositions by injection affords significant clinical responses and at least a 6-month duration of effect in a subject undergoing treatment, as provided by the described treatment methods.

1 . A method of administering botulinum toxin to achieve an extended duration therapeutic or cosmetic effect in an individual, the method comprising:
 administering by injection a first treatment dose of a sterile injectable composition into an area of the individual in need of treatment to achieve the therapeutic or cosmetic effect following a first treatment with the composition;   wherein the composition comprises a pharmaceutically acceptable diluent suitable for injection; and   a botulinum toxin component selected from the group consisting of a botulinum toxin, a botulinum toxin complex, or a reduced botulinum toxin complex; and   a positively charged carrier component comprising a positively charged polylysine backbone having covalently attached thereto one or more positively charged efficiency groups having an amino acid sequence of (gly) p -RGRDDRRQRRR-(gly) q , (gly) p -YGRKKRRQRRR-(gly) q  or (gly) p -RKKRRQRRR-(gly) q , wherein the subscripts p and q are each independently an integer of from 0 to 20;   wherein the botulinum toxin component is administered to the individual in a treatment dose of 20 U to 60 U;   wherein the positively charged carrier is non-covalently associated with the botulinum toxin component; and   wherein the first treatment dose of the composition administered by injection to the individual achieves the extended duration therapeutic or cosmetic effect having at least about a 6 month to about a 10 month duration of effect, optionally, before a second or subsequent treatment dose is administered.   
     
     
         2 . A method of reducing wrinkles, lines, or furrows in an individual in need thereof, the method comprising:
 administering to the individual as a single dose injection a composition comprising:   a pharmaceutically acceptable diluent for injection;   a botulinum toxin component selected from the group consisting of a botulinum toxin, a botulinum toxin complex, or a reduced botulinum toxin complex; and   a positively charged carrier component comprising a positively charged polylysine backbone having covalently attached thereto one or more positively charged efficiency groups having an amino acid sequence of (gly) p -RGRDDRRQRRR-(gly) q , (gly) p -YGRKKRRQRRR-(gly) q  or (gly) p -RKKRRQRRR-(gly) q , wherein the subscripts p and q are each independently an integer of from 0 to 20;   wherein the botulinum toxin is administered to the individual in an amount of 20 U to 60 U;   wherein the positively charged carrier is non-covalently associated with the botulinum component; and   wherein the single dose injection of the composition provides a single treatment having at least about a six month to about a 10 month duration of effect in reducing the wrinkles, lines, or furrows in the individual, thereby extending treatment interval duration for the individual.   
     
     
         3 . A pharmaceutical composition in a sterile injectable formulation for use in administering botulinum toxin to achieve an extended duration therapeutic or cosmetic effect in an individual,
 said composition comprising a pharmaceutically acceptable diluent suitable for injection;   a botulinum toxin component in a treatment dose of 20 U to 60 U, wherein said botulinum toxin component is selected from the group consisting of a botulinum toxin complex, a reduced botulinum toxin complex, or a botulinum toxin; and   a positively charged carrier component comprising a positively charged polylysine backbone having covalently attached thereto one or more positively charged efficiency groups having an amino acid sequence of (gly) p -RGRDDRRQRRR-(gly) q , (gly) p -YGRKKRRQRRR-(gly) q  or (gly) p -RKKRRQRRR-(gly) q , wherein the subscripts p and q are each independently an integer of from 0 to 20;   wherein the positively charged carrier is non-covalently associated with the botulinum toxin component; and   wherein said treatment dose of the composition achieves the extended duration therapeutic or cosmetic effect having at least about a 6 month to about a 10 month duration of effect in the individual administered said formulation by injection.   
     
     
         4 . A pharmaceutical composition in a sterile injectable formulation for use in reducing wrinkles, lines, or furrows in an individual in need thereof, said composition comprising:
 a botulinum toxin component in a dose of 20 U to 60 U, said botulinum toxin component selected from the group consisting of a botulinum toxin complex, a reduced botulinum toxin complex, or a botulinum toxin,   a positively charged carrier component comprising a positively charged polylysine backbone having covalently attached thereto one or more positively charged efficiency groups having an amino acid sequence of (gly) p -RGRDDRRQRRR-(gly) q , (gly) p -YGRKKRRQRRR-(gly) q  or (gly) p -RKKRRQRRR-(gly) q , wherein the subscripts p and q are each independently an integer of from 0 to 20; and   a pharmaceutically acceptable diluent for injection;   wherein the positively charged carrier is non-covalently associated with the botulinum toxin component; and   wherein said dose of the composition provides a single treatment having at least about a six month to about a 10 month duration of effect in reducing the wrinkles, lines, or furrows in the individual, thereby extending treatment interval duration for the individual.   
     
     
         5 . The method according to  claim 1  or  claim 2 , or the pharmaceutical composition for use according to  claim 3  or  claim 4 , wherein the composition reduces glabellar lines in the face of the individual. 
     
     
         6 . The method or pharmaceutical composition for use according to  claim 5 , wherein the composition comprises botulinum toxin of serotype A. 
     
     
         7 . The method or pharmaceutical composition for use according to  claim 6 , wherein the composition comprises botulinum toxin of serotype A having a molecular weight of 150 kDa. 
     
     
         8 . The method or pharmaceutical composition for use according to any one of  claims 1  to  7 , wherein the positively charged polylysine backbone has covalently attached thereto one or more positively charged efficiency groups having the amino acid sequence (gly) p -RGRDDRRQRRR-(gly) q , wherein the subscripts p and q are each independently an integer of from 0 to 20. 
     
     
         9 . The method or pharmaceutical composition for use according to any one of  claims 1  to  7 , wherein the positively charged polylysine backbone has covalently attached thereto one or more positively charged efficiency groups having the amino acid sequence (gly) p -YGRKKRRQRRR-(gly) q , wherein the subscripts p and q are each independently an integer of from 0 to 20. 
     
     
         10 . The method or pharmaceutical composition for use according to any one of  claims 1  to  7 , wherein the positively charged polylysine backbone has covalently attached thereto one or more positively charged efficiency groups having the amino acid sequence (gly) p -RKKRRQRRR-(gly) q , wherein the subscripts p and q are each independently an integer of from 0 to 20. 
     
     
         11 . The method or pharmaceutical composition for use according to any one of  claims 1  to  10 , wherein (i) the subscripts p and q are each independently an integer of from 0 to 8; or (ii) are each independently an integer of from 2 to 5. 
     
     
         12 . The method or pharmaceutical composition for use according to any one of  claims 1  to  11 , wherein the one or more positively charged efficiency groups are attached to either end, or both ends, of the positively charged polylysine backbone of the positively charged carrier. 
     
     
         13 . The method or pharmaceutical composition for use according to  claim 12 , wherein the positively charged carrier has the amino acid sequence RKKRRQRRRG-(K) 15 -GRKKRRQRRR. 
     
     
         14 . The method or pharmaceutical composition for use according to any one of  claims 1  to  13 , wherein the composition does not locally diffuse from the site of injection following injection. 
     
     
         15 . The method or pharmaceutical composition for use according to any one of  claims 1  to  14 , wherein the botulinum toxin is administered to the individual in an amount of 20 U. 
     
     
         16 . The method or pharmaceutical composition for use according to any one of  claims 1  to  14 , wherein the botulinum toxin is administered to the individual in an amount of 40 U. 
     
     
         17 . The method or pharmaceutical composition for use according to any one of  claims 1  to  14 , wherein the botulinum toxin is administered to the individual in an amount of 60 U. 
     
     
         18 . The method according to  claim 1  or the or pharmaceutical composition for use according to  claim 3 , wherein the therapeutic effect is reduction of a symptom associated with a disorder selected from the group consisting of hemifacial spasm, adult onset spasmodic torticollis, anal fissure, blepharospasm, cerebral palsy, headaches, strabismus, temporomandibular joint disorder, neurologic pain, overactive bladder, rhinitis, sinusitis, acne, dystonia, dystonic contractions, hyperhidrosis, and hypersecretion of a gland controlled by the cholinergic nervous system. 
     
     
         19 . The method according to  claim 1 , or pharmaceutical composition for use according to  claim 3 , wherein the therapeutic or cosmetic effect is treatment or reduction of wrinkles, lines, or furrows in the individual. 
     
     
         20 . The method or pharmaceutical composition for use according to any one of  claims 1  to  19 , wherein the duration of treatment effect comprises greater than 3 months. 
     
     
         21 . The method or pharmaceutical composition for use according to any one of  claims 1  to  19 , wherein the duration of treatment effect comprises greater than 4 months. 
     
     
         22 . The method or pharmaceutical composition for use according to any one of  claims 1  to  19 , wherein the duration of treatment effect comprises greater than 5 months. 
     
     
         23 . The method or pharmaceutical composition for use according to any one of  claims 1  to  19 , wherein the duration of treatment effect comprises greater than 6 months. 
     
     
         24 . The method or pharmaceutical composition for use according to any one of  claims 1  to  19 , wherein the duration of treatment effect comprises greater than 7 months. 
     
     
         25 . The method or pharmaceutical composition for use according to any one of  claims 1  to  19 , wherein the duration of treatment effect comprises greater than 8 months. 
     
     
         26 . The method or pharmaceutical composition for use according to any one of  claims 1  to  19 , wherein the duration of treatment effect comprises greater than 9 months. 
     
     
         27 . The method or pharmaceutical composition for use according to any one of  claims 1  to  19 , wherein the duration of treatment effect comprises at least 6 months through 10 months. 
     
     
         28 . A sterile injectable composition comprising:
 a botulinum toxin component selected from the group consisting of a botulinum toxin, a botulinum toxin complex, or a reduced botulinum toxin complex, in a dosage amount selected from 20 U, 40 U, or 60 U; and   a positively charged carrier component comprising a positively charged polylysine backbone having covalently attached thereto one or more positively charged efficiency groups having an amino acid sequence of (gly) p -RGRDDRRQRRR-(gly) q , (gly) p -YGRKKRRQRRR-(gly) q  or (gly) p -RKKRRQRRR-(gly) q , wherein the subscripts p and q are each independently an integer of from 0 to 20; and   a pharmaceutically acceptable diluent for injection;   wherein the positively charged carrier is non-covalently associated with the botulinum toxin component; and   wherein the composition provides a cosmetic or therapeutic effect which endures for at least 20 to 24 weeks following a single treatment of an individual with the injectable composition.   
     
     
         29 . The composition according to  claim 28 , wherein the positively charged carrier has the amino acid sequence RKKRRQRRRG-(K) 15 -GRKKRRQRRR. 
     
     
         30 . The composition according to  claim 28  or  claim 29 , wherein the composition comprises botulinum toxin of serotype A. 
     
     
         31 . The composition according to  claim 30 , wherein the composition comprises botulinum toxin of serotype A having a molecular weight of 150 kDa. 
     
     
         32 . The composition according to any one of  claims 28  to  31 , wherein the botulinum toxin is administered to the individual in a dosage amount of 20 U. 
     
     
         33 . The composition according to any one of  claims 28  to  31 , wherein the botulinum toxin is administered to the individual in a dosage amount of 40 U. 
     
     
         34 . The composition according to any one of  claims 28  to  31 , wherein the botulinum toxin is administered to the individual in a dosage amount of 60 U. 
     
     
         35 . A method of treating an individual in need of treatment with injectable botulinum toxin, wherein the method of treatment comprises a treatment course having multiple treatment intervals with prolonged duration of effect and duration time between each treatment interval, the treatment course comprising:
 administering by injection an initial treatment dose of a sterile injectable composition into an area of the individual in need of treatment to achieve a therapeutic or a cosmetic effect following the initial treatment with the composition;   wherein the composition comprises a pharmaceutically acceptable diluent suitable for injection;   a botulinum toxin component selected from the group consisting of a botulinum toxin, a botulinum toxin complex, or a reduced botulinum toxin complex; and   a positively charged carrier component comprising a positively charged polylysine backbone having covalently attached thereto one or more positively charged efficiency groups having an amino acid sequence of (gly) p -RGRDDRRQRRR-(gly) q , (gly) p -YGRKKRRQRRR-(gly) q  or (gly) p -RKKRRQRRR-(gly) q , wherein the subscripts p and q are each independently an integer of from 0 to 20; and   wherein the botulinum toxin component is administered to the individual in a treatment dose of 20 U to 60 U;   wherein the positively charged carrier is non-covalently associated with the botulinum toxin component;   wherein the initial treatment dose of the composition administered by injection to the individual provides a therapeutic or cosmetic duration of effect lasting through at least about 10 months; and   administering subsequent treatment doses of the composition by injection to the individual at treatment intervals comprising a duration of greater than or equal to 3 months to at least about 10 months following the initial treatment dose and between each subsequent treatment dose.   
     
     
         36 . The method according to  claim 35 , wherein the therapeutic or cosmetic effect is treatment or reduction of wrinkles, lines, or furrows. 
     
     
         37 . The method according to  claim 36 , wherein the therapeutic or cosmetic effect is reduction of glabellar lines in the face of the individual. 
     
     
         38 . The method according to any one of  claim 35  to  claim 37 , wherein the composition comprises botulinum toxin of serotype A. 
     
     
         39 . The method according to  claim 38 , wherein the composition comprises botulinum toxin of serotype A having a molecular weight of 150 kDa. 
     
     
         40 . The method according to any one of  claims 35  to  39 , wherein the positively charged polylysine backbone has covalently attached thereto one or more positively charged efficiency groups having the amino acid sequence (gly) p -RGRDDRRQRRR-(gly) q , wherein the subscripts p and q are each independently an integer of from 0 to 20. 
     
     
         41 . The method according to any one of  claims 35  to  39 , wherein the positively charged polylysine backbone has covalently attached thereto one or more positively charged efficiency groups having the amino acid sequence (gly) p -YGRKKRRQRRR-(gly) q , wherein the subscripts p and q are each independently an integer of from 0 to 20. 
     
     
         42 . The method according to any one of  claims 35  to  39 , wherein the positively charged polylysine backbone has covalently attached thereto one or more positively charged efficiency groups having the amino acid sequence (gly) p -RKKRRQRRR-(gly) q , wherein the subscripts p and q are each independently an integer of from 0 to 20. 
     
     
         43 . The method according to any one of  claims 35  to  42 , wherein (i) the subscripts p and q are each independently an integer of from 0 to 8; or (ii) are each independently an integer of from 2 to 5. 
     
     
         44 . The method according to any one of  claims 35  to  43 , wherein the one or more positively charged efficiency groups are attached to either end, or both ends, of the positively charged polylysine backbone of the positively charged carrier. 
     
     
         45 . The method according to any one of  claims 35  to  39 , wherein the positively charged carrier has the amino acid sequence RKKRRQRRRG-(K) 15 -GRKKRRQRRR. 
     
     
         46 . The method according to any one of  claims 35  to  45 , wherein the composition does not locally diffuse from the site of injection following injection. 
     
     
         47 . The method according to any one of  claims 35  to  46 , wherein the botulinum toxin is administered to the individual in an amount of 20 U. 
     
     
         48 . The method according to any one of  claims 35  to  46 , wherein the botulinum toxin is administered to the individual in an amount of 40 U. 
     
     
         49 . The method according to any one of  claims 35  to  46 , wherein the botulinum toxin is administered to the individual in an amount of 60 U. 
     
     
         50 . The method according to  claim 35 , wherein the therapeutic effect is reduction of a symptom associated with a disorder selected from the group consisting of hemifacial spasm, adult onset spasmodic torticollis, anal fissure, blepharospasm, cerebral palsy, headaches, strabismus, temporomandibular joint disorder, neurologic pain, overactive bladder, rhinitis, sinusitis, acne, dystonia, dystonic contractions, hyperhidrosis, and hypersecretion of a gland controlled by the cholinergic nervous system. 
     
     
         51 . The method according to any one of  claims 35  to  50 , wherein the duration of the treatment interval comprises greater than 3 months. 
     
     
         52 . The method according to any one of  claims 35  to  50 , wherein the duration of the treatment interval comprises greater than 4 months. 
     
     
         53 . The method according to any one of  claims 35  to  50 , wherein the duration of the treatment interval comprises greater than 5 months. 
     
     
         54 . The method according to any one of  claims 35  to  50 , wherein the duration of the treatment interval comprises greater than 6 months. 
     
     
         55 . The method according to any one of  claims 35  to  50 , wherein the duration of the treatment interval comprises greater than 7 months. 
     
     
         56 . The method according to any one of  claims 35  to  50 , wherein the duration of the treatment interval comprises greater than 8 months. 
     
     
         57 . The method according to any one of  claims 35  to  50 , wherein the duration of the treatment interval comprises greater than 9 months. 
     
     
         58 . The method according to any one of  claims 35  to  50 , wherein the duration of the treatment interval comprises at least 6 months through 10 months.