Liquid neurotoxin formulation stabilized with tryptophan or tyrosine

Abstract

The invention relates to stable liquid neurotoxin formulations which are free of animal proteins, comprising a surfactant, an amino acid selected from tryptophan and tyrosine, a buffer comprising sodium, chloride and phosphate ions, which have a pH between 5.5 and 8, and which are stable for 2 months. These compositions are suitable for use in therapy and in particular for administration to a patient to achieve a desired therapeutic or aesthetic effect. The invention also relates to the use of an amino acid selected from tryptophan and tyrosine to protect a proteinaceous neurotoxin from degradation in a liquid composition which is free of animal derived proteins.

1 . A liquid composition comprising: (i) a botulinum neurotoxin; (ii) a surfactant; (iii) an amino acid selected from tryptophan and tyrosine; and (iv) a buffer comprising sodium, chloride, and phosphate ions; wherein:
 the liquid composition has a pH between 5.5 and 8 and is free of animal derived proteins; and   the amino acid is present at a concentration of from about 0.5 mg/ml to about 8 mg/ml and protects the botulinum neurotoxin from degradation, thereby providing a liquid composition that is stable for at least 2 months at 2 to 8° C.   
     
     
         2 . The liquid composition of  claim 1 , wherein the surfactant is a non-ionic surfactant. 
     
     
         3 . The liquid composition of  claim 2 , wherein the non-ionic surfactant is a polysorbate, preferably Polysorbate 20, Polysorbate 60 or Polysorbate 80. 
     
     
         4 . The liquid composition of  claim 1 , wherein the amino acid is tryptophan. 
     
     
         5 . The liquid composition of  claim 1 , wherein the buffer further comprises potassium ions. 
     
     
         6 . The liquid composition of  claim 1 , wherein the composition has a pH between 6.0 and 7.5. 
     
     
         7 . The liquid composition of  claim 1 , wherein no more than a 30% loss in extracellular proteolytic activity occurs over 2, 3, 6, 12, 18, 24 or 36 months at 5° C. 
     
     
         8 . The liquid composition of  claim 1 , wherein the botulinum neurotoxin is a natural botulinum neurotoxin in complex form, a high purity natural botulinum neurotoxins and a recombinant botulinum neurotoxin. 
     
     
         9 . The liquid composition of  claim 8 , wherein the botulinum neurotoxin is a recombinant botulinum neurotoxin selected from a botulinum neurotoxin A, B, C, D, E, F or G, a modified botulinum neurotoxin, or a chimeric botulinum neurotoxin. 
     
     
         10 . The liquid composition of  claim 1 , comprising:
 to 10000 LD50 units of botulinum neurotoxin per mL;   0.001 to 15% v/v polysorbate;   0.1 to 5 mg/mL tryptophan;   10 to 500 mM NaCl;   1 to 50 mM KCl; and   1 to 100 mM sodium phosphate;   
       wherein the composition has a pH between 5.5 and 8 and is stable for 6 months at 5° C. 
     
     
         11 . The liquid composition of  claim 10 , comprising:
 10 to 2000 LD50 units of botulinum neurotoxin per mL;   0.05 to 0.2% v/v polysorbate 80;   0.1 to 5 mg/mL tryptophan;   25 to 300 mM NaCl;   1 to 10 mM KCl; and   2 to 50 mM sodium phosphate;   
       wherein the composition has a pH between 6.0 and 7.5 and is stable for 12 months at 5° C. 
     
     
         12 - 18 . (canceled) 
     
     
         19 . The liquid composition of  claim 1 , wherein the amino acid is L-tryptophan. 
     
     
         20 . The liquid composition of  claim 1 , wherein the amino acid is present at a concentration of greater than 0.5 mg/ml to about 8 mg/ml. 
     
     
         21 . The liquid composition of  claim 1 , wherein the amino acid is present at a concentration of about 0.5 mg/ml to about 5 mg/ml. 
     
     
         22 . The liquid composition of  claim 1 , wherein the amino acid is present at a concentration of about 0.5 mg/ml to about 3 mg/ml. 
     
     
         23 . The liquid composition of  claim 1 , wherein the amino acid is present at a concentration of about 0.74 mg/ml to about 5 mg/ml. 
     
     
         24 . The liquid composition of  claim 1 , wherein the amino acid is present at a concentration of about 0.74 mg/ml to about 3 mg/ml. 
     
     
         25 . A stabilized, botulinum neurotoxin ready-to-use (RTU) liquid composition comprising: (i) a botulinum neurotoxin; (ii) a surfactant; (iii) an amino acid selected from tryptophan and tyrosine; and (iv) a buffer comprising sodium, chloride, and phosphate ions; wherein:
 the liquid composition has a pH between 5.5 and 8 and is free of animal derived proteins;   the amino acid is present at a concentration of at least about 0.5 mg/ml to about 8 mg/ml and protects the botulinum neurotoxin from degradation, thereby providing a liquid composition that is stable for at least 2 months at 2 to 8° C.; and   no more than a 30% loss in extracellular proteolytic activity occurs over 2, 3, 6, 12, 18, 24 or 36 months at 5° C.   
     
     
         26 . A stabilized, botulinum neurotoxin ready-to-use (RTU) liquid composition comprising:
 4 to 10000 LD50 units of botulinum neurotoxin per mL;   0.001 to 15% v/v polysorbate;   0.5 to 8 mg/mL amino acid selected from tryptophan and tyrosine;   10 to 500 mM NaCl;   1 to 50 mM KCl; and   1 to 100 mM sodium phosphate;   
       wherein: the composition has a pH between 5.5 and 8; and the amino acid at a concentration of 0.5 to 8 mg/mL protects the botulinum neurotoxin from degradation, thereby providing a liquid composition that is stable for at least 2 months at 2 to 8° C. wherein no more than a 30% loss in extracellular proteolytic activity occurs over 2, 3, 6, 12, 18, 24 or 36 months at 5° C.