Conjugated Factor VIII Molecules

Abstract

The present invention relates to B-domain truncated Factor VIII molecules with a modified circulatory half life, said molecule being covalently conjugated with a hydrophilic polymer. The invention furthermore relates to methods for obtaining such molecules as well as use of such molecules.

1 . A B-domain truncated Factor VIII molecule with a modified circulatory half life, said molecule being covalently conjugated with a hydrophilic polymer via an O-linked oligosaccharide in the truncated B domain, wherein Factor VIII activation results in removal of the covalently conjugated hydrophilic polymer. 
     
     
         2 . A molecule according to  claim 1 , wherein the O-linked oligosaccharide is attached to an O-glycosylation site that is made by truncation of the B-domain. 
     
     
         3 . A molecule according to  claim 1 , wherein the hydrophilic polymer is PEG. 
     
     
         4 . A molecule according to  claim 3 , wherein the size of the PEG is from about 10,000 to about 160,000 Da. 
     
     
         5 . A molecule according to  claim 4 , wherein the size of the PEG is about 40,000 Da. 
     
     
         6 . A molecule according to  claim 5 , comprising the amino acid sequence as set forth in SEQ ID NO 2. 
     
     
         7 . A pharmaceutical composition comprising a molecule according to  claim 1 . 
     
     
         8 . A method of making a B-domain truncated Factor VIII molecule with a modified circulatory half life that is covalently conjugated with a hydrophilic polymer via an O-linked oligosaccharide in the truncated B domain, wherein activation of the truncated Factor VIII molecule results in removal of the covalently conjugated hydrophilic polymer said method comprising conjugating a B-domain truncated Factor VIII molecule with a hydrophilic polymer via an O3 linked oligosaccharide in the truncated B domain. 
     
     
         9 . (canceled) 
     
     
         10 . A method of treatment of a haemophilic disease comprising administering to a patient in need thereof a therapeutically effective amount of a molecule according to  claim 1 . 
     
     
         11 . (canceled) 
     
     
         12 . (canceled) 
     
     
         13 . A method of producing a B-domain truncated Factor VIII molecule with a modified circulatory half-life, comprising (i) preparing a B-domain-truncated Factor VIII molecule; and optionally subjecting the amino acid sequence of the truncated Factor VIII molecule to an analysis to identify potential O-linked glycosylation sites, (ii) producing the molecule in a suitable host cell, (iii) selecting molecules having O-linked glycans in the truncated B-domain, and (iv) covalently conjugating the truncated Factor VIII molecule with a hydrophilic polymer via an O-linked oligosaccharide in the truncated B domain. 
     
     
         14 . A molecule according to  claim 1  comprising the amino acid sequence as set forth in SEQ ID NO 2. 
     
     
         15 . A molecule according to  claim 2  comprising the amino acid sequence as set forth in SEQ ID NO 2. 
     
     
         16 . A molecule according to  claim 1  comprising the amino acid sequence as set forth in SEQ ID NO 2. 
     
     
         17 . A pharmaceutical composition comprising a molecule according to  claim 6 . 
     
     
         18 . A pharmaceutical composition comprising a molecule according to  claim 3 . 
     
     
         19 . A method of treatment of a haemophilic disease comprising administering to a patient in need thereof a therapeutically effective amount of a molecule according to  claim 3 . 
     
     
         20 . A method of treatment of a haemophilic disease comprising administering to a patient in need thereof a therapeutically effective amount of a molecule according to  claim 6 .