Albumin fusion proteins

Abstract

The present invention encompasses albumin fusion proteins. Nucleic acid molecules encoding the albumin fusion proteins of the invention are also encompassed by the invention, as are vectors containing these nucleic acids, host cells transformed with these nucleic acids vectors, and methods of making the albumin fusion proteins of the invention and using these nucleic acids, vectors, and/or host cells. Additionally the present invention encompasses pharmaceutical compositions comprising albumin fusion proteins and methods of treating, preventing, or ameliorating diseases, disorders or conditions using albumin fusion proteins of the invention.

1 .- 29 . (canceled) 
     
     
         30 . An albumin fusion protein comprising a follicle stimulating hormone (FSH) alpha subunit polypeptide, or a variant or fragment thereof, and a FSH beta subunit polypeptide, or a variant or fragment thereof, fused to an albumin polypeptide, wherein said variant or fragment of the FSH alpha subunit polypeptide and said variant or fragment of the FSH beta subunit polypeptide have FSH activity, and wherein said albumin polypeptide is selected from:
 (i) a full-length albumin;   (ii) a mature albumin;   (iii) a fragment of albumin;   (iv) a polypeptide consisting of an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO:18;   (v) amino acids 1 to 585 of SEQ ID NO:18; and   (vi) amino acids 1 to 387 of SEQ ID NO:18;   and further wherein said albumin polypeptide increases the serum plasma half-life of the FSH alpha and beta subunit polypeptides.   
     
     
         31 . The albumin fusion protein of  claim 30 , wherein said albumin fusion protein is sufficient to extend the shelf life of the FSH alpha subunit polypeptide compared to the shelf-life of the FSH alpha subunit polypeptide in an unfused state and the shelf life of the FSH beta subunit polypeptide compared to the shelf-life of the FSH beta subunit polypeptide in an unfused state. 
     
     
         32 . The albumin fusion protein of  claim 30 , wherein the albumin polypeptide is located at the N-terminus of the albumin fusion protein. 
     
     
         33 . The albumin fusion protein of  claim 32 , wherein the FSH alpha subunit polypeptide, or fragment or variant thereof, is located at the C-terminus of the albumin polypeptide and the FSH beta subunit polypeptide, or fragment or variant thereof, is located at the C-terminus of the FSH alpha subunit polypeptide, or fragment or variant thereof. 
     
     
         34 . The albumin fusion protein of  claim 32 , wherein the FSH beta subunit polypeptide, or fragment or variant thereof, is located at the C-terminus of the albumin polypeptide and the FSH alpha subunit polypeptide, or fragment or variant thereof, is located at the C-terminus of the FSH beta subunit polypeptide, or fragment or variant thereof. 
     
     
         35 . The albumin fusion protein of  claim 30 , wherein the albumin polypeptide is located at the C-terminus of the albumin fusion protein. 
     
     
         36 . The albumin fusion protein of  claim 35 , wherein the FSH alpha subunit polypeptide, or fragment or variant thereof, is located at the N-terminus of the albumin polypeptide and the FSH beta subunit polypeptide, or fragment or variant thereof, is located at the N-terminus of the FSH alpha subunit polypeptide, or fragment or variant thereof. 
     
     
         37 . The albumin fusion protein of  claim 35 , wherein the FSH beta subunit polypeptide, or fragment or variant thereof, is located at the N-terminus of the albumin polypeptide and the FSH alpha subunit polypeptide, or fragment or variant thereof, is located at the N-terminus of the FSH beta subunit polypeptide, or fragment or variant thereof. 
     
     
         38 . The albumin fusion protein of  claim 30 , wherein the FSH alpha subunit polypeptide, or fragment or variant thereof, is located at the N-terminus of the albumin polypeptide and the FSH beta subunit polypeptide, or fragment or variant thereof, is located at the C-terminus of the albumin polypeptide. 
     
     
         39 . The albumin fusion protein of  claim 30 , wherein the FSH alpha subunit polypeptide, or fragment or variant thereof, is located at the C-terminus of the albumin polypeptide and the FSH beta subunit polypeptide, or fragment or variant thereof, is located at the N-terminus of the albumin polypeptide. 
     
     
         40 . The albumin fusion protein of  claim 30 , wherein the albumin polypeptide is separated by a linker from the FSH alpha subunit polypeptide, or fragment or variant thereof; from the FSH beta subunit polypeptide, or fragment or variant thereof; or from both the FSH alpha subunit polypeptide, or fragment or variant thereof, and the FSH beta subunit polypeptide, or fragment or variant thereof. 
     
     
         41 . The albumin fusion protein of  claim 30 , wherein the albumin fusion protein has the following formula: R1-L-R2; R2-L-R1; or R1-L-R2-L-R1, wherein R1 comprises a FSH alpha subunit polypeptide, or fragment or variant thereof, a FSH beta subunit polypeptide, or fragment or variant thereof, or both a FSH alpha subunit polypeptide, or fragment or variant thereof, and a FSH beta subunit polypeptide, or fragment or variant thereof; L is a peptide linker; and R2 is an albumin polypeptide. 
     
     
         42 . The albumin fusion protein of  claim 30 , wherein the shelf-life of the albumin fusion protein is greater than the shelf-life of the FSH alpha subunit polypeptide in an unfused state or of the FSH beta subunit polypeptide in an unfused state. 
     
     
         43 . The albumin fusion protein of  claim 30 , wherein the in vitro biological activity of the albumin fusion protein is prolonged compared to the in vitro biological activity of the FSH alpha subunit polypeptide, or fragment or variant thereof, in an unfused state or of the FSH beta subunit polypeptide, or fragment or variant thereof, in an unfused state. 
     
     
         44 . The albumin fusion protein of  claim 30 , wherein the in vivo biological activity of the albumin fusion protein is prolonged compared to the in vivo biological activity of the FSH alpha subunit polypeptide, or fragment or variant thereof, in an unfused state or of the FSH beta subunit polypeptide, or fragment or variant thereof, in an unfused state. 
     
     
         45 . The albumin fusion protein of  claim 30 , wherein said fragment of the albumin polypeptide comprises:
 (a) amino acids 54 to 61 of SEQ ID NO: 18;   (b) amino acids 76 to 89 of SEQ ID NO: 18;   (c) amino acids 92 to 100 of SEQ ID NO: 18;   (d) amino acids 170 to 176 of SEQ ID NO: 18;   (e) amino acids 247 to 252 of SEQ ID NO: 18;   (f) amino acids 266 to 277 of SEQ ID NO: 18;   (g) amino acids 280 to 288 of SEQ ID NO: 18;   (h) amino acids 362 to 368 of SEQ ID NO: 18;   (i) amino acids 439 to 447 of SEQ ID NO: 18;   (j) amino acids 462 to 475 of SEQ ID NO: 18;   (k) amino acids 478 to 486 of SEQ ID NO: 18; or   (l) amino acids 560 to 566 of SEQ ID NO: 18.   
     
     
         46 . The albumin fusion protein of  claim 30 , which is non-glycosylated. 
     
     
         47 . The albumin fusion protein of  claim 30 , which is expressed from an isolated host cell. 
     
     
         48 . The albumin fusion protein of  claim 47 , wherein the isolated host cell is a yeast cell. 
     
     
         49 . The albumin fusion protein of  claim 48 , wherein the yeast cell is glycosylation deficient. 
     
     
         50 . The albumin fusion protein of  claim 48 , wherein the yeast cell is glycosylation and protease deficient. 
     
     
         51 . The albumin fusion protein of  claim 47 , wherein the isolated host cell is a mammalian cell. 
     
     
         52 . The albumin fusion protein of  claim 30 , wherein the albumin fusion protein further comprises a secretion leader sequence. 
     
     
         53 . A composition comprising the albumin fusion protein of  claim 30  and a pharmaceutically acceptable carrier. 
     
     
         54 . A kit comprising (a) the albumin fusion protein of  claim 30 , and (b) instructions for use. 
     
     
         55 . A method of treating a reproductive system disorder in a patient comprising administering to said patient a therapeutically effective amount of the composition of  claim 53 . 
     
     
         56 . A method of extending the shelf-life of a follicle stimulating hormone (FSH) alpha subunit polypeptide, or a fragment or variant thereof, and of a FSH beta subunit polypeptide, or a fragment or variant thereof, comprising the step of fusing the FSH alpha subunit polypeptide, or fragment or variant thereof, and the FSH beta subunit polypeptide, or fragment or variant thereof, to an albumin polypeptide, wherein said variant or fragment of the FSH alpha subunit polypeptide and said variant or fragment of the FSH beta subunit polypeptide have FSH activity, and wherein said albumin polypeptide is selected from:
 (i) a full-length albumin;   (ii) a mature albumin;   (iii) a fragment of albumin;   (iv) a polypeptide consisting of an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO:18;   (v) amino acids 1 to 585 of SEQ ID NO:18; and   (vi) amino acids 1 to 387 of SEQ ID NO:18;   and further wherein said albumin fusion protein is sufficient (1) to extend the shelf-life of the FSH alpha subunit polypeptide compared to the shelf-life of the FSH alpha subunit polypeptide in an unfused state and (2) to extend the shelf-life of the FSH beta subunit polypeptide compared to the shelf-life of the FSH beta subunit polypeptide in an unfused state.   
     
     
         57 . A nucleic acid molecule comprising a polynucleotide sequence encoding the albumin fusion protein of  claim 30 . 
     
     
         58 . A vector comprising the nucleic acid molecule of  claim 57 . 
     
     
         59 . An isolated host cell comprising the nucleic acid molecule of  claim 57 .