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 . A pharmaceutical preparation, comprising a myostatin antagonist protein including a myostatin binding domain of a Cerberus/Dan/Gremlin polypeptide or variant thereof, which myostatin antagonist protein binds to and neutralizes one or more of nodal and/or myostatin, wherein said pharmaceutical preparation is substantially free of pyrogenic materials so as to be suitable for administration as a human or veterinarian therapeutic. 
     
     
         2 . The pharmaceutical preparation of  claim 1 , wherein the myostatin antagonist protein promotes growth of muscle tissue. 
     
     
         3 . The preparation of  claim 1 , wherein the myostatin binding domain is selected from the group consisting of: a Cerberus sequence or a variant sequence thereof, and a Coco sequence or a variant sequence thereof. 
     
     
         4 . The preparation of  claim 1 , wherein the myostatin antagonist protein has diminished potency, relative to a corresponding wild-type Cerberus/Dan/Gremlin polypeptide, for neutralizing BMP-4. 
     
     
         5 . The preparation of  claim 4 , wherein the myostatin binding domain is an N-terminally truncated derivative of wild-type Cerberus protein, said derivative comprising:
 (a) a sequence beginning at a position corresponding to any one of residues 106-119 of human Cerberus; and,   (b) a sequence ending at a position corresponding to any residue after residue 240 of human Cerberus,   wherein said derivative binds myostatin, GDF-11, and/or Nodal, but does not substantially bind BMP-4.   
     
     
         6 . The preparation of  claim 1 , wherein said myostatin binding domain is encoded by a polynucleotide that hybridizes under high stringency conditions to the coding sequence for human Cerberus or human Coco. 
     
     
         7 . The preparation of  claim 1 , wherein said myostatin antagonist protein binds myostatin with a K d  of 1 μM or less. 
     
     
         8 . The preparation of  claim 3 , wherein said said myostatin antagonist protein is a fusion protein including one additional polypeptide portion that enhance 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 preparation of  claim 8 , wherein said fusion protein includes an immunoglobulin Fc domain. 
     
     
         10 . The preparation of  claim 8 , wherein said fusion protein includes a purification subsequence selected from: an epitope tag, a FLAG tag, a polyhistidine sequence, and a GST fusion. 
     
     
         11 . The preparation of  claim 1 , wherein said myostatin antagonist protein includes one or more modified amino acid residues selected from: 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, and an amino acid conjugated to an organic derivatizing agent. 
     
     
         12 . The preparation of  claim 1 , wherein said myostatin antagonist protein does not substantially inhibit Activin A signaling in an A204 Reporter Gene Assay. 
     
     
         13 . The preparation of  claim 1 , wherein said myostatin antagonist protein is a fusion protein that further comprises a second myostatin inhibitor domain, which is a polypeptide affinity reagent that selectively binds to myostatin and competes with the binding of an ALK7 or ALK4 receptor. 
     
     
         14 . The preparation of  claim 13 , wherein said affinity reagent is an antibody agent. 
     
     
         15 . The preparation of  claim 14 , wherein said antibody agent is a recombinant antibody; a monoclonal antibody; a V H  domain; a V L  domain; an scFv; an Fab fragment; an Fab′ fragment; an F(ab') 2 ; an Fv; or a disulfide linked Fv. 
     
     
         16 . The preparation of  claim 15 , wherein said antibody agent is a fully human antibody or a humanized chimeric antibody, or an antigen binding fragment thereof. 
     
     
         17 . The preparation of  claim 13 , wherein said affinity reagent is a peptide or scaffolded peptide that selectively binds to myostatin and competes with the binding of an ALK7 or ALK4 receptor. 
     
     
         18 . The preparation of  claim 13 , wherein said affinity reagent is an myostatin binding domain of ALK7 or ALK4. 
     
     
         19 . The preparation of  claim 13 , wherein said affinity reagent is a small organic molecule that selectively binds to myostatin and competes with the binding of an ALK7 or ALK4 receptor. 
     
     
         20 - 52 . (canceled)