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 - 30 . (canceled) 
     
     
         31 . An albumin fusion protein comprising an atrial natriuretic polypeptide (ANP) fused to albumin, wherein the fusion protein has ANP activity, and wherein:
 (a) the ANP polypeptide is selected from a wild-type ANP, an ANP fragment, and an ANP variant, and   (b) the albumin is selected from a wild-type albumin, an albumin fragment, and an albumin variant, wherein the albumin increases the serum plasma half-life of the ANP polypeptide.   
     
     
         32 . The albumin fusion protein of  claim 31 , wherein the ANP polypeptide comprises the amino acid sequence selected from SEQ ID NO: 302 and SEQ ID NO: 560. 
     
     
         33 . The albumin fusion protein of  claim 31 , wherein the albumin is selected from:
 a) human albumin;   b) cow albumin;   c) sheep albumin;   d) pig albumin;   e) hen albumin;   f) salmon albumin;   j) an albumin fragment;   k) an albumin variant;   l) SEQ ID NO:1;   m) a fragment of albumin consisting of amino acids 1-194 of SEQ ID NO:1;   n) a fragment of albumin consisting of amino acids 195-387 of SEQ ID NO:1;   o) a fragment of albumin consisting of amino acids 388-585 of SEQ ID NO:1;   p) a fragment of albumin consisting of amino acids 1-387 of SEQ ID NO:1;   q) a fragment of albumin consisting of amino acids 195-585 of SEQ ID NO:1;   r) a fragment of albumin consisting of amino acids 1-105 of SEQ ID NO:1;   s) a fragment of albumin consisting of amino acids 120-194 of SEQ ID NO:1;   t) a fragment of albumin consisting of amino acids 195-291 of SEQ ID NO:1;   u) a fragment of albumin consisting of amino acids 316-387 of SEQ ID NO:1;   v) a fragment of albumin consisting of amino acids 388-491 of SEQ ID NO:1;   w) a fragment of albumin consisting of amino acids 512-585 of SEQ ID NO:1;   x) a fragment of albumin that is 10, 15, 20, 25, 30, 50, 100, or 150 amino acids in length;   y) a fragment of albumin consisting of one or more domains of albumin; and   z) a variant of SEQ ID NO:1 selected from
 i) L407A; 
 ii) L408V; 
 iii) V409A; 
 iv) R410A; 
 v) K413Q; and 
 vi) K414Q. 
   
     
     
         34 . The albumin fusion protein of  claim 31 , wherein the albumin fusion protein further comprises a leader sequence. 
     
     
         35 . The albumin fusion protein of  claim 34 , wherein the leader sequence is selected from:
 a) HSA;   b) kex2; and   c) a fusion of HSA and kex2.   
     
     
         36 . The albumin fusion protein of  claim 31 , further comprising one or more additional therapeutic polypeptides. 
     
     
         37 . The albumin fusion protein of  claim 31  comprising the amino acid sequence selected from SEQ ID NO: 227 and SEQ ID NO: 524. 
     
     
         38 . The albumin fusion protein of  claim 31  consisting of the amino acid sequence selected from SEQ ID NO: 227 and SEQ ID NO: 524. 
     
     
         39 . A nucleotide sequence encoding the albumin fusion protein of  claim 37 . 
     
     
         40 . The nucleotide sequence of  claim 39 , comprising a ANP nucleotide sequence selected from SEQ ID NO: 152 and SEQ ID NO: 488. 
     
     
         41 . A construct expressing the albumin fusion protein of  claim 31 . 
     
     
         42 . The construct of  claim 41  selected from construct ID 3484 and construct ID 4174. 
     
     
         43 . A host cell expressing the albumin fusion protein of  claim 31 . 
     
     
         44 . The host cell of  claim 43 , wherein the host cell is a mammalian cell, a yeast cell or a prokaryotic cell. 
     
     
         45 . A method for expressing an albumin fusion protein comprising culturing the host cell of  claim 43  under conditions suitable for the expression of the albumin fusion protein and recovering the albumin fusion protein. 
     
     
         46 . An albumin fusion protein expressed by the host cell of  claim 43 , wherein the albumin fusion protein is glycosylated, non-glycosylated or a glycosylation isomer. 
     
     
         47 . A composition comprising the albumin fusion protein of  claim 31  and a pharmaceutically acceptable carrier. 
     
     
         48 . A kit comprising the composition of  claim 47  and instructions for the use thereof. 
     
     
         49 . A method of treating, preventing, diagnosing or ameliorating a disease, disorder or condition in a subject in need thereof comprising administering an effective amount of an albumin fusion protein comprising a ANP polypeptide fused to albumin, wherein the fusion protein has ANP activity, and wherein:
 (a) the ANP polypeptide is selected from a wild-type ANP, an ANP fragment, and an ANP variant, and   (b) the albumin is selected from a wild-type albumin, an albumin fragment, and an albumin variant, wherein the albumin increases the serum plasma half-life of the ANP polypeptide.   
     
     
         50 . The method of  claim 49 , wherein the disease, disorder or condition is selected from hypertension; salt-sensitive hypertension; congestive heart failure; angina pectoris; peripheral artery disease; diabetic nephropathy; stroke; kidney failure; acute and/or chronic renal failure; acute tubular necrosis; acute renal failure; renal disease; renal glomerular disease; excess fluid in tissues; hypotension; cardiac volume overload; cardiac decompensation; left ventricular dysfunction; dyspnea; treatment for elevated aldosterone levels; vasoconstriction; impaired cardiac output and/or hypertension; cardiovascular disease; cardiac failure; myocardial reperfusion injury; left ventricular remodeling and post-myocardial infarction. 
     
     
         51 . An albumin fusion protein comprising a butyrylcholinesterase polypeptide (BChE) fused to albumin, wherein the fusion protein has BChE activity, and wherein:
 (a) the BChE polypeptide is selected from a wild-type BChE, a BChE fragment, and a BChE variant, and   (b) the albumin is selected from a wild-type albumin, an albumin fragment, and an albumin variant, wherein the albumin increases the serum plasma half-life of the BChE polypeptide.   
     
     
         52 . The albumin fusion protein of  claim 51 , wherein the BChE polypeptide comprises the amino acid sequence selected from SEQ ID NO: 708 and SEQ ID NO: 709. 
     
     
         53 . The albumin fusion protein of  claim 51 , wherein the albumin is selected from:
 a) human albumin;   b) cow albumin;   c) sheep albumin;   d) pig albumin;   e) hen albumin;   f) salmon albumin;   j) an albumin fragment;   k) an albumin variant;   l) SEQ ID NO:1;   m) a fragment of albumin consisting of amino acids 1-194 of SEQ ID NO:1;   n) a fragment of albumin consisting of amino acids 195-387 of SEQ ID NO:1;   o) a fragment of albumin consisting of amino acids 388-585 of SEQ ID NO:1;   p) a fragment of albumin consisting of amino acids 1-387 of SEQ ID NO:1;   q) a fragment of albumin consisting of amino acids 195-585 of SEQ ID NO:1;   r) a fragment of albumin consisting of amino acids 1-105 of SEQ ID NO:1;   s) a fragment of albumin consisting of amino acids 120-194 of SEQ ID NO:1;   t) a fragment of albumin consisting of amino acids 195-291 of SEQ ID NO:1;   u) a fragment of albumin consisting of amino acids 316-387 of SEQ ID NO:1;   v) a fragment of albumin consisting of amino acids 388-491 of SEQ ID NO:1;   w) a fragment of albumin consisting of amino acids 512-585 of SEQ ID NO:1;   x) a fragment of albumin that is 10, 15, 20, 25, 30, 50, 100, or 150 amino acids in length;   y) a fragment of albumin consisting of one or more domains of albumin; and   z) a variant of SEQ ID NO:1 selected from
 i) L407A; 
 ii) L408V; 
 iii) V409A; 
 iv) R410A; 
   v) K413Q; and   vi) K414Q.   
     
     
         54 . The albumin fusion protein of  claim 51 , wherein the albumin fusion protein further comprises a leader sequence. 
     
     
         55 . The albumin fusion protein of  claim 54 , wherein the leader sequence is selected from:
 a) HSA;   b) kex2;   c) a consensus signal sequence; and   d) a fusion of at least one of the leader sequences selected from HSA, kex2, and a consensus signal sequence.   
     
     
         56 . The albumin fusion protein of  claim 51 , further comprising one or more additional therapeutic polypeptides. 
     
     
         57 . The albumin fusion protein of  claim 51  comprising the amino acid sequence selected from SEQ ID NO: 669 and SEQ ID NO: 670. 
     
     
         58 . The albumin fusion protein of  claim 51  consisting of the amino acid sequence selected from SEQ ID NO: 669 and SEQ ID NO: 670. 
     
     
         59 . A nucleotide sequence encoding the albumin fusion protein of  claim 57 . 
     
     
         60 . The nucleotide sequence of  claim 59 , comprising a BChE nucleotide sequence selected from SEQ ID NO: 630 and SEQ ID NO: 631. 
     
     
         61 . A construct expressing the albumin fusion protein of  claim 51 . 
     
     
         62 . The construct of  claim 41  selected from construct ID 4258 and construct ID 4259. 
     
     
         63 . A host cell expressing the albumin fusion protein of  claim 51 . 
     
     
         64 . The host cell of  claim 63 , wherein the host cell is a mammalian cell, a yeast cell or a prokaryotic cell. 
     
     
         65 . A method for expressing an albumin fusion protein comprising culturing the host cell of  claim 63  under conditions suitable for the expression of the albumin fusion protein and recovering the albumin fusion protein. 
     
     
         66 . An albumin fusion protein expressed by the host cell of  claim 63 , wherein the albumin fusion protein is glycosylated, non-glycosylated or a glycosylation isomer. 
     
     
         67 . A composition comprising the albumin fusion protein of  claim 51  and a pharmaceutically acceptable carrier. 
     
     
         68 . A kit comprising the composition of  claim 67  and instructions for the use thereof. 
     
     
         69 . A method of treating, preventing, diagnosing or ameliorating a disease, disorder or condition in a subject in need thereof comprising administering an effective amount of an albumin fusion protein comprising a BChE polypeptide fused to albumin, wherein the fusion protein has BChE activity, and wherein:
 (a) the BChE polypeptide is selected from a wild-type BChE, a BChE fragment, and a BChE variant, and   (b) the albumin is selected from a wild-type albumin, an albumin fragment, and an albumin variant, wherein the albumin increases the serum plasma half-life of the BChE polypeptide.   
     
     
         70 . The method of  claim 69 , wherein the disease, disorder or condition is selected from detoxification for cocaine overdose, suxamethonium sensitivity and apnea. 
     
     
         71 . The method of  claim 70 , wherein the disease, disorder or condition is cocaine overdose. 
     
     
         72 . The method of  claim 70 , wherein the BChE polypeptide comprises the following amino acid substitutions: A227S, S315G, A356W and Y360G. 
     
     
         73 . The albumin fusion of  claim 52 , wherein the BChE polypeptide BChE polypeptide comprises the following amino acid substitutions: A227S, S315G, A356W and Y360G. 
     
     
         74 . A method of treating, preventing, diagnosing or ameliorating a disease, disorder or condition in a subject in need thereof comprising administering an effective amount of an albumin fusion protein, wherein the disease, disorder or condition comprises Alzheimer's disease.