Ion exchange chromatography of GLP-1, analogs and derivatives thereof

Abstract

The present invention relates to an ion exchange chromatography process for purifying GLP-1 or an analog or a derivative thereof from a mixture containing said GLP-1 and related impurities, and to an industrial method including such ion exchange chromatography process.

1 . A cation exchange chromatography process for purifying a peptide or GLP-1 peptide from a mixture comprising said peptide or GLP-1 peptide and related or unrelated impurities, comprising the step of: 
 separating said peptide or GLP-1 peptide and said related or unrelated impurities of said mixture by elution in a solution comprising an organic modifier, water, optionally a salt component, and optionally a buffer, with a linear or step gradient or isocratically in salt component and/or with a linear or step pH-gradient or at a constant pH-value, wherein the pH-gradient or pH-value should be in the range where said peptide or GLP-1 peptide has a positive local or overall net charge different from the local or overall positive net charge of said related or unrelated impurities so as to remove said related or unrelated impurities.    
     
     
         2 . A cation exchange chromatography process for purifying a peptide or GLP-1 peptide from a mixture comprising said peptide or GLP-1 peptide and related impurities, comprising the step of: 
 separating said peptide or GLP-1 peptide and said related impurities of said mixture by elution in a solution consisting essentially of an organic modifier, water, optionally a salt component, and optionally a buffer, with a linear or step gradient or isocratically in salt component and/or with a linear or step pH-gradient or at a constant pH-value, wherein the pH-gradient or pH-value should be in the range where said peptide or GLP-1 peptide has a positive local or overall net charge different from the local or overall positive net charge of said related impurities so as to remove said related impurities.    
     
     
         3 . An anion exchange chromatography process for purifying a peptide or GLP-1 peptide from a mixture comprising said peptide or GLP-1 peptide and related or unrelated impurities, comprising the step of: 
 separating said peptide or GLP-1 peptide and said related or unrelated impurities of said mixture by elution in a solution comprising an organic modifier, water, optionally a salt component, and optionally a buffer, with a linear or step gradient or isocratically in salt component and/or with a linear or step pH-gradient or at a constant pH-value, wherein the pH-gradient or pH-value should be in the range where said peptide or GLP-1 peptide has a negative local or overall net charge different from the local or overall negative net charge of said related or unrelated impurities so as to remove said related or unrelated impurities.    
     
     
         4 . An anion exchange chromatography process for purifying a peptide or GLP-1 peptide from a mixture comprising said peptide or GLP-1 peptide and related impurities, comprising the step of: 
 separating said peptide or GLP-1 peptide and said related impurities of said mixture by elution in a solution consisting essentially of an organic modifier, water, optionally a salt component, and optionally a buffer, with a linear or step gradient or isocratically in salt component and/or with a linear or step pH-gradient or at a constant pH-value, wherein the pH-gradient or pH-value should be in the range where said peptide or GLP-1 peptide has a negative local or overall net charge different from the local or overall negative net charge of said related impurities so as to remove said related impurities.    
     
     
         5 . An industrial method for producing a pure peptide or GLP-1 peptide, the method including a cation exchange chromatography process for purifying a peptide from a mixture comprising said peptide or GLP-1 peptide and related impurities, comprising the steps of: 
 separating said peptide or GLP-1 peptide and said related impurities of said mixture by elution in a solution comprising an organic modifier, water, optionally a salt component, and optionally a buffer, with a linear or step gradient or isocratically in salt component and/or with a linear or step pH-gradient or at a constant pH-value, wherein the pH-gradient or pH-value should be in the range where said peptide or GLP-1 peptide has a positive local or overall net charge different from the local or overall positive net charge of said related impurities so as to remove said related impurities.    
     
     
         6 . An industrial method for producing a pure peptide or GLP-1 peptide, the method including an anion exchange chromatography process for purifying a peptide from a mixture comprising said peptide or GLP-1 peptide and related impurities, comprising the steps of: 
 separating said peptide or GLP-1 peptide and said related impurities of said mixture by elution in a solution comprising an organic modifier, water, optionally a salt component, and optionally a buffer, with a linear or step gradient or isocratically in salt component and/or with a linear or step pH-gradient or at a constant pH-value, wherein the pH-gradient or pH-value should be in the range where said peptide or GLP-1 peptide has a negative local or overall net charge different from the local or overall negative net charge of said related impurities so as to remove said related impurities.    
     
     
         7 . A method for isolating a peptide or GLP-1 peptide, the method including purification of a peptide or GLP-1 peptide from a mixture comprising said peptide or GLP-1 peptide and related impurities via a cation exchange chromatography process, the cation exchange chromatography process comprising the steps of: 
 separating said peptide or GLP-1 peptide and said related impurities of said mixture by elution in a solution comprising an organic modifier, water, optionally a salt component, and optionally a buffer, with a linear or step gradient or isocratically in salt component and/or with a linear or step pH-gradient or at a constant pH-value, wherein the pH-gradient or pH-value should be in the range where said peptide or GLP-1 peptide has a positive local or overall net charge different from the local or overall positive net charge of said related impurities so as to remove said related impurities;    and subsequently, if necessary, subjecting to analytical tests and/or further purification, and isolating said peptide or GLP-1 peptide in a conventional manner.    
     
     
         8 . A method for isolating a peptide or GLP-1 peptide, the method including purification of a peptide or GLP-1 peptide from a mixture comprising said peptide or GLP-1 peptide and related impurities via an anion exchange chromatography process, the anion exchange chromatography process comprising the steps of: 
 separating said peptide or GLP-1 peptide and said related impurities of said mixture by elution in a solution comprising an organic modifier, water, optionally a salt component, and optionally a buffer, with a linear or step gradient or isocratically in salt component and/or with a linear or step pH-gradient or at a constant pH-value, wherein the pH-gradient or pH-value should be in the range where said peptide or GLP-1 peptide has a negative local or overall net charge different from the local or overall negative net charge of said related impurities so as to remove said related impurities;    and subsequently, if necessary, subjecting to analytical tests and/or further purification, and isolating said peptide or GLP-1 peptide in a conventional manner.    
     
     
         9 . The process or method according to any one of claims  1 - 8  wherein said GLP-1 peptide to be purified is selected from GLP-1 (7-37), GLP-1 (7-36) amide as well as analogues and derivatives thereof, such as Arg 34 GLP-1(7-37); Arg 26 GLP-1(7-37); Arg 34 Lys 26 (N ε -(γ-Glu-(N α -tetradecanoyl)))GLP-1 (7-37); Arg 34 Lys 26 (N ε -(γ-Glu-(N α -hexadecanoyl)))GLP-1 (7-37); Arg 26 Lys 34 (N ε -(γ-Glu-(N α -tetradecanoyl)))GLP-1 (7-37); Arg 26 Lys 34 (N ε -(γ-Glu-(N α -hexadecanoyl)))GLP-1 (7-37); Val 8 GLP-1(7-37); Thr 8 GLP-1(7-37); Met 8 GLP-1(7-37); Gly 8 GLP-1(7-37); Val 8 GLP-1(7-36) amide; Thr 8 GLP-1(7-36) amide; Met 8 GLP-1(7-36) amide; Gly 8 GLP-1(7-36) amide.  
     
     
         10 . The process or method according to any one of claims  1 - 8  wherein said peptide to be purified is selected from FactorVII, FactorVIIa, FactorVIIai, and FFR-FactorVIIa.  
     
     
         11 . The process or method according to any one of claims  1 - 8  wherein said peptide to be purified is selected from a GLP-2 peptide as well as analogues and derivatives thereof.  
     
     
         12 . The process or method according to any one of claims  1 - 11  wherein the ratio of organic modifier to water on a weight percent basis is from 1:99 to 99:1.  
     
     
         13 . The process or method according to any one of claims  1 - 12  wherein said organic modifier is selected from C 1-6 -alkanol, C 1-6 -alkenol or C 1-6 -alkynol, urea, guanidine, or C 1-6 -alkanoic acid, C 2-6 -glycol, or C 3-7 -polyalcohol including sugars.