Cerberus/coco derivatives and uses thereof

Abstract

The invention relates to Cerberus/Dan/Gremlin polypeptides or variants thereof for use in treating a variety of disorders associated with myostatin, nodal and GDF-11. Preferred polypeptides are Coco or Cerberus derivatives.

1 . An isolated myostatin antagonist protein, the myostatin antagonist protein comprising the myostatin binding domain of a Coco polypeptide or variant thereof, which myostatin antagonist protein binds to and neutralizes one or both of nodal and myostatin. 
     
     
         2 . The isolated myostatin antagonist protein of  claim 1 , wherein the myostatin antagonist protein has diminished potency for binding BMP-4 relative to a corresponding wild-type Coco polypeptide. 
     
     
         3 . The isolated myostatin antagonist protein of  claim 1 , wherein the myostatin antagonist protein comprises an N-terminally truncated derivative of wild-type Coco protein. 
     
     
         4 . The isolated myostatin antagonist protein of  claim 1 , wherein the myostatin binding domain of Coco comprises the amino acid sequence of SEQ ID NO: 15. 
     
     
         5 . The isolated myostatin antagonist protein of  claim 1 , wherein the myostatin antagonist protein binds myostatin with a K D  of 1 μM of less. 
     
     
         6 . The isolated myostatin antagonist protein of  claim 1 , further comprising, in addition to the myostatin binding domain, a heterologous portion. 
     
     
         7 . The isolated myostatin antagonist protein of  claim 6 , wherein the heterologous portion is an immunoglobulin Fc domain. 
     
     
         8 . The isolated myostatin antagonist protein of  claim 6 , wherein the heterologous portion is a polypeptide that enhances one or more of in vivo stability, in vivo half life, uptake/administration, tissue localization or distribution, formation of protein complexes, and/or purification. 
     
     
         9 . The isolated myostatin antagonist protein of  claim 8 , wherein the heterologous portion is a polypeptide portion is a purification subsequence. 
     
     
         10 . The isolated myostatin antagonist protein of  claim 9 , wherein the purification subsequence is an epitope tag, a FLAG tag, a polyhistidine sequence, or as a GST fusion. 
     
     
         11 . The isolated myostatin antagonist protein of  claim 1 , wherein the myostatin antagonist protein comprises one or more modified amino acid residues, such as a glycosylated amino acid, a PEGylated amino acid, a farnesylated amino acid, an acetylated amino acid, a biotinylated amino acid, an amino acid conjugated to a lipid moiety, or an amino acid conjugated to an organic derivatizing agent. 
     
     
         12 . The isolated myostatin antagonist protein of  claim 1 , wherein the myostatin antagonist protein does not substantially inhibit Activin A signaling in an A204 Reporter Gene Assay. 
     
     
         13 . A method for inhibiting myostatin signal transduction in an animal, comprising: administering to an animal in need thereof a pharmaceutical preparation comprising an effective amount of an antagonist myostatin protein comprising the myostatin binding domain of Coco, wherein said pharmaceutical preparation is substantially free of pyrogenic materials so as to be suitable for injection as a human or veterinary therapeutic. 
     
     
         14 . The method of  claim 13 , wherein the animal has a pathologic condition, which is characterized, at least in part, by an abnormal amount, development or metabolic activity of muscle or adipose tissue in an animal. 
     
     
         15 . The method of  claim 13 , wherein the animal has a wasting disorder. 
     
     
         16 . The method of  claim 15 , wherein the wasting disorder is selected from cachexia, anorexia, Duchenne muscular dystrophy (DMD) syndrome, Becker's muscular dystrophy (BMD) syndrome, acquired immune deficiency syndrome (AIDS) wasting syndrome, age-related wasting, muscular dystrophies, and neuromuscular diseases. 
     
     
         17 . The method of  claim 13 , wherein the animal has a metabolic disorder. 
     
     
         18 . The method of  claim 17 , wherein the metabolic disorder is obesity or type II diabetes. 
     
     
         19 . The method of  claim 13 , wherein the myostatin antagonist protein comprises an N-terminally truncated derivative of wild-type Coco protein. 
     
     
         20 . The method of  claim 13 , wherein the myostatin binding domain of Coco comprises the amino acid sequence of SEQ ID NO: 15. 
     
     
         21 . The method of  claim 13 , wherein the myostatin antagonist protein is a fusion protein comprising, in addition to the myostatin binding domain of Coco, a heterologous portion. 
     
     
         22 . The method of  claim 21 , wherein the heterologous portion is a polypeptide that enhances one or more of in vivo stability, in vivo half life, uptake/administration, tissue localization or distribution, formation of protein complexes, and/or purification. 
     
     
         23 . The method of  claim 21 , wherein the heterologous portion is an immunoglobulin Fc domain. 
     
     
         24 . The method of  claim 21 , wherein the fusion protein has diminished potency for binding BMP-4 relative to a corresponding wild-type Coco polypeptide. 
     
     
         25 . The method of  claim 21 , wherein the fusion protein binds to and neutralizes one or both of nodal and myostatin. 
     
     
         26 . The method of  claim 21 , wherein the fusion protein binds myostatin with a K D  of 1 μM of less. 
     
     
         27 . The method of  claim 23 , wherein the fusion protein comprises an additional polypeptide portion that enhances one or more of in vivo stability, in vivo half life, uptake/administration, tissue localization or distribution, formation of protein complexes, and/or purification. 
     
     
         28 . The method of  claim 27 , wherein the additional polypeptide portion is a purification subsequence. 
     
     
         29 . The method of  claim 28 , wherein the purification subsequence is an epitope tag, a FLAG tag, a polyhistidine sequence, or as a GST fusion. 
     
     
         30 . The method of  claim 21 , wherein the fusion protein comprises one or more modified amino acid residues, such as a glycosylated amino acid, a PEGylated amino acid, a farnesylated amino acid, an acetylated amino acid, a biotinylated amino acid, an amino acid conjugated to a lipid moiety, or an amino acid conjugated to an organic derivatizing agent. 
     
     
         31 . The method of  claim 21 , wherein the fusion protein does not substantially inhibit Activin A signaling in an A204 Reporter Gene Assay.