Long-acting coagulation factors and methods of producing same

Abstract

Polypeptides comprising at least one carboxy-terminal peptide (CTP) of chorionic gonadotropin attached to the carboxy terminus but not to the amino terminus of a coagulation factor and polynucleotides encoding the same are disclosed. Pharmaceutical compositions and pharmaceutical formulations comprising the polypeptides and polynucleotides of the disclosure and methods of using and producing same are also disclosed.

What is claimed is: 
     
         1 . Use of a pharmaceutical formulation for preventing or treating a blood clotting or coagulation disorder in a subject, the preventing or treating comprising the step of administering said pharmaceutical formulation to said subject, wherein said pharmaceutical formulation comprises a buffer, a tonicity agent, and a chorionic gonadotropin carboxy terminal peptide (CTP)-modified polypeptide consisting of a Factor VII coagulation factor or activated Factor VII coagulation factor and three chorionic gonadotropin carboxy terminal peptides (CTPs) attached to the carboxy terminus of said coagulation factor, wherein said polypeptide does not include a signal peptide. 
     
     
         2 . The use of  claim 1 , wherein said buffer comprises 20 mM citrate, said tonicity agent is 150 mM sodium chloride, or said formulation is at a pH of 6.4. 
     
     
         3 . The use of  claim 1 , wherein said formulation comprises 20 mM citrate, 13.3 mM glycine, 150 mM sodium chloride, and is at a pH of 6.4. 
     
     
         4 . The use of  claim 1 , wherein said formulation is a liquid formulation. 
     
     
         5 . The use of  claim 1 , wherein the amino acid sequence of at least one CTP is set forth in SEQ ID NO: 1 or SEQ ID NO: 2. 
     
     
         6 . The use of  claim 1 , wherein at least one of said CTPs is truncated or glycosylated. 
     
     
         7 . The use of  claim 1 , wherein at least one CTP is attached to said coagulation factor via a linker, for example wherein the linker is a peptide bond. 
     
     
         8 . The use of  claim 1 , wherein the sequence of said CTP-modified polypeptide is set forth in SEQ ID NO: 46. 
     
     
         9 . The use of  claim 1 , wherein said CIP-modified polypeptide comprises a light chain and a heavy chain linked by a disulfide bond. 
     
     
         10 . The use of  claim 9 , wherein separation of said light chain and said heavy chain on an SDS-PAGE gel occurs under denaturing conditions, and wherein said light chain migrates at 25 kDa molecular weight and said heavy chain migrates at 60 kDa molecular weight. 
     
     
         11 . The use of  claim 9 , wherein the disulfide bond occurs between cysteine residue 135 and cysteine residue 262 of SEQ ID NO: 46. 
     
     
         12 . The use of  claim 9 , wherein the light chain comprises amino acids 1-152 of SEQ ID: 46 and the heavy chain comprises amino acids 153-490 of SEQ ID NO: 46. 
     
     
         13 . The use of  claim 1 , wherein said disorder is congenital FVII deficiency, hemophilia, hemophilia A, hemophilia B, hemophilia A with inhibitors to FVIII, or hemophilia B with inhibitors to FIX. 
     
     
         14 . The use of  claim 1 , wherein said subject is a human adult or is a human child. 
     
     
         15 . The use of  claim 1 , wherein said administering is via a subcutaneous or intravenous route. 
     
     
         16 . The use of  claim 1 , wherein said polypeptide is administered daily, every other day, every three days, once weekly, twice weekly, or once every other week, or any combination thereof. 
     
     
         17 . The use of  claim 1 , wherein said pharmaceutical formulation is administered at about 50 μg/Kg to 400 μg/Kg of said CTP-modified polypeptide.