Conjugates of a polypeptide and an oligosaccharide

Abstract

The present invention relates to conjugates of a polypeptide and an oligosaccharide, wherein the polypeptide is conjugated to at least one oligosaccharide-spacer residue, the oligosaccharide being a synthetic sulfated oligosaccharide comprising 4-18 monosaccharide units and per se having affinity to antithrombin III and the spacer being a bond or an essentially pharmacologically inactive flexible linking residue, or a pharmaceutically acceptable salt thereof. The conjugates of the invention have improved pharmacokinetic properties when compared to the original polypeptides (i.e. the corresponding non-conjugated polypeptides per se).

1 . A conjugate of a calcitonin, ganirelix, GLP-1, [D-Ala 8 ]-GLP-1(7-36), adrenomedullin, ADM(27-52), kisspeptin-10, octreotide or interleukin-2 polypeptide and an oligosaccharide, wherein the polypeptide is conjugated to at least one oligosaccharide-spacer residue having the structure (II) 
       
         
           
           
               
               
           
         
         wherein R is independently or (1-8C)alkoxy, or a pharmacologically inactive flexible linking residue, the charge of OSO 3   −  and COO −  being compensated by positively charged counter ions and wherein one pharmacologically inactive flexible linking residue is present or a pharmaceutically acceptable salt thereof. 
       
     
     
         2 - 3 . (canceled) 
     
     
         4 . The conjugate of  claim 1 , having a circulating plasma level of≦50 nM. 
     
     
         5 . The conjugate of  claim 1 , wherein the oligosaccharide per se has an anticoagulant activity which is of subtherapeutic level when compared to the pharmacological activity of the polypeptide per se. 
     
     
         6 - 7 . (canceled) 
     
     
         8 . The conjugate of  claim 1 , wherein the oligosaccharide residue has the structure (III) 
       
         
           
           
               
               
           
         
         wherein R is independently OSO 3   −  or (1-8C)alkoxy, the charge of OSO 3   −  and COO −  being compensated by positively charged counterions. 
       
     
     
         9 . The conjugate of  claim 8 , wherein the oligosaccharide residue has the structure (IV) 
       
         
           
           
               
               
           
         
       
       wherein R is independently OCH 3  or OSO 3   − . 
     
     
         10 . The conjugate of  claim 9 , wherein both R groups in (IV) are OSO 3   − . 
     
     
         11 - 21 . (canceled) 
     
     
         22 . The conjugate of  claim 1 , having the structure 
       
         
           
           
               
               
           
         
       
     
     
         23 . The conjugate of  claim 1 , having the structure 
       
         
           
           
               
               
           
         
       
     
     
         24 . A pharmaceutical composition comprising the conjugate of  claim 1  and pharmaceutically suitable auxiliaries. 
     
     
         25 . The conjugate of  claim 1  for use in therapy. 
     
     
         26 . (canceled) 
     
     
         27 . A process for the preparation of a conjugate of a calcitonin ganirelix, GLP-1, [D-Ala 8 ]-GLP-1(7-36), adrenomedullin, ADM(27-52), kisspeptin-10, octreotide or interleukin-2 polypeptide and an oligosaccharide, wherein the polypeptide is conjugated to at least one oligosaccharide-spacer residue having the structure (II) 
       
         
           
           
               
               
           
         
         wherein R is independently or (1-8C)alkoxy, or a pharmacologically inactive flexible linking residue, the charge of OSO 3   −  and COO −  being compensated by positively charged counter ions and wherein one pharmacologically inactive flexible linking residue is present, or a pharmaceutically acceptable salt thereof, comprising
 (a) an optional step wherein the polypeptide is adapted for conjugation, and 
 (b) a coupling step wherein the optionally adapted polypeptide is reacted with the oligosaccharide-spacer residue. 
 
       
     
     
         28 - 29 . (canceled) 
     
     
         30 . A process for the preparation of a therapeutically active conjugate comprising a calcitonin, ganirelix, GLP-1, [D-Ala 8 ]-GLP-1(7-36), adrenomedullin, ADM(27-52), kisspeptin-10, octreotide or interleukin-2 polypeptide the conjugate having negligible anticoagulant activity, wherein the conjugate has a longer plasma half-life than the original polypeptide while the biological activity is essentially retained, comprising a step wherein an oligosaccharide having the structure (II) 
       
         
           
           
               
               
           
         
       
       wherein R is independently or (1-8C)alkoxy, or a pharmacologically inactive flexible linking residue, the charge of OSO 3   −  and COO −  being compensated by positively charged counter ions and wherein one pharmacologically inactive flexible linking residue is present, is covalently attached to a polypeptide . 
     
     
         31 - 32 . (canceled)