Process and system for obtaining botulinum neurotoxin

Abstract

Rapid, animal protein free, chromatographic processes and systems for obtaining high potency, high yield botulinum neurotoxin for research, therapeutic and cosmetic use.

1 - 12 . (canceled) 
     
     
         13 . A powder pharmaceutical composition comprising:
 a) a substantially animal protein free (APF) drug substance comprising about 1.5-1.9 ng of onabotulinumtoxinA;   b) human serum albumin (HSA); and   c) sodium chloride;   
       wherein:
 the onabotulinumtoxinA is obtained by a substantially animal product free (APF) culture, fermentation, and purification process utilizing a plurality of chromatography columns; 
 the composition is reconstituted with normal saline prior to administration to a human patient; and 
 the composition comprises an amount of onabotulinumtoxinA sufficient to afford 100 Units of onabotulinumtoxinA upon reconstitution with normal saline; wherein 1 Unit of onabotulinumtoxinA is an amount that kills 50% of a group (LD 50 ) of female Swiss Webster mice weighing about 18-20 grams each upon intraperitoneal injection. 
 
     
     
         14 . The composition of  claim 13 , wherein the drug substance is essentially APF. 
     
     
         15 . The composition of  claim 13 , wherein the drug substance is entirely APF. 
     
     
         16 . The composition of  claim 13 , wherein the substantially APF culture, fermentation, and purification process utilizes a Hall A strain of  Clostridium botulinum.    
     
     
         17 . The composition of  claim 13 , wherein the plurality of chromatography columns comprises an anion exchange chromatography (AEX) column followed by a cation exchange chromatography (CEX) column. 
     
     
         18 . The composition of  claim 17 , wherein the composition is free of benzamidine. 
     
     
         19 . The composition of  claim 17 , wherein the composition comprises about 0.5 mg of HSA. 
     
     
         20 . The composition of  claim 17 , wherein the composition comprises about 0.9 mg of sodium chloride. 
     
     
         21 . The composition of  claim 13 , wherein the composition is lyophilized or vacuum dried. 
     
     
         22 . A vacuum-dried or lyophilized pharmaceutical composition comprising:
 a) a substantially animal protein free (APF) drug substance comprising about 1.5-1.9 ng of onabotulinumtoxinA;   b) about 0.5 mg of human serum albumin (HSA); and   c) about 0.9 mg sodium chloride;   
       wherein:
 the onabotulinumtoxinA is obtained by a substantially animal product free (APF) culture, fermentation, and purification process utilizing a plurality of chromatography columns; 
 the substantially APF culture, fermentation, and purification process utilizes a Hall A strain of  Clostridium botulinum;    
 the substantially APF drug substance has a residual nucleic acid content below a limit of detection (LOD) as measured by qPCR; 
 the composition is reconstituted with normal saline prior to administration to a human patient; and 
 the composition comprises an amount of onabotulinumtoxinA sufficient to afford 100 Units of onabotulinumtoxinA upon reconstitution with normal saline; wherein 1 Unit of onabotulinumtoxinA is an amount that kills 50% of a group (LD 50 ) of female Swiss Webster mice weighing about 18-20 grams each upon intraperitoneal injection. 
 
     
     
         23 . The composition of  claim 13 , wherein the drug substance is essentially APF. 
     
     
         24 . The composition of  claim 13 , wherein the drug substance is entirely APF. 
     
     
         25 . The composition of  claim 13 , wherein the plurality of chromatography columns comprises an anion exchange chromatography (AEX) column followed by a cation exchange chromatography (CEX) column. 
     
     
         26 . The composition of  claim 25 , wherein the composition is free of benzamidine. 
     
     
         27 . A powder pharmaceutical composition comprising:
 a) a substantially animal protein free (APF) drug substance comprising about 1.5-1.9 ng of onabotulinumtoxinA;   b) human serum albumin (HSA); and   c) sodium chloride;   
       wherein:
 the onabotulinumtoxinA is obtained by a substantially animal product free (APF) culture, fermentation, and purification process utilizing a plurality of chromatography columns; 
 the substantially APF drug substance has a residual nucleic acid content of less than 1 ng per mg of onabotulinumtoxinA or the substantially APF drug substance has a residual nucleic acid content below a limit of detection (LOD) as measured by qPCR; 
 the composition is reconstituted with normal saline prior to administration to a human patient; and 
 the composition comprises an amount of onabotulinumtoxinA sufficient to afford 100 Units of onabotulinumtoxinA upon reconstitution with normal saline; wherein 1 Unit of onabotulinumtoxinA is an amount that kills 50% of a group (LD 50 ) of female Swiss Webster mice weighing about 18-20 grams each upon intraperitoneal injection. 
 
     
     
         28 . The composition of  claim 27 , wherein the drug substance is essentially APF. 
     
     
         29 . The composition of  claim 27 , wherein the drug substance is entirely APF. 
     
     
         30 . The composition of  claim 27 , wherein the substantially APF culture, fermentation, and purification process utilizes a Hall A strain of  Clostridium botulinum.    
     
     
         31 . The composition of  claim 27 , wherein the plurality of chromatography columns comprises an anion exchange chromatography (AEX) column followed by a cation exchange chromatography (CEX) column. 
     
     
         32 . The composition of  claim 31 , wherein the composition is free of benzamidine. 
     
     
         33 . The composition of  claim 31 , wherein the composition comprises about 0.5 mg of HSA. 
     
     
         34 . The composition of  claim 31 , wherein the composition comprises about 0.9 mg of sodium chloride. 
     
     
         35 . The composition of  claim 27 , wherein the composition is lyophilized or vacuum dried. 
     
     
         36 . A vacuum-dried or lyophilized pharmaceutical composition comprising:
 a) a substantially animal protein free (APF) drug substance comprising about 1.5-1.9 ng of onabotulinumtoxinA;   b) about 0.5 mg of human serum albumin (HSA); and   c) about 0.9 mg sodium chloride;   
       wherein:
 the onabotulinumtoxinA is obtained by a substantially animal product free (APF) culture, fermentation, and purification process utilizing a plurality of chromatography columns; 
 the substantially APF culture, fermentation, and purification process utilizes a Hall A strain of  Clostridium botulinum;    
 the substantially APF drug substance has a residual nucleic acid content of less than 1 ng per mg of onabotulinumtoxinA; 
 the composition is reconstituted with normal saline prior to administration to a human patient; and 
 the composition comprises an amount of onabotulinumtoxinA sufficient to afford 100 Units of onabotulinumtoxinA upon reconstitution with normal saline; wherein 1 Unit of onabotulinumtoxinA is an amount that kills 50% of a group (LD 50 ) of female Swiss Webster mice weighing about 18-20 grams each upon intraperitoneal injection. 
 
     
     
         37 . The composition of  claim 36 , wherein the drug substance is essentially APF. 
     
     
         38 . The composition of  claim 36 , wherein the drug substance is entirely APF. 
     
     
         39 . The composition of  claim 36 , wherein the plurality of chromatography columns comprises an anion exchange chromatography (AEX) column followed by a cation exchange chromatography (CEX) column. 
     
     
         40 . The composition of  claim 39 , wherein the composition is free of benzamidine.