Growth hormone polypeptides and methods of making and using same

Abstract

The present invention relates to compositions comprising growth hormone linked to extended recombinant polypeptide (XTEN), isolated nucleic acids encoding the compositions and vectors and host cells containing the same, and methods of making and using such compositions in treatment of growth hormone-related diseases, disorders, and conditions.

1 - 27 . (canceled) 
     
     
         28 . An isolated fusion protein comprising a growth hormone, wherein the growth hormone is linked to an extended recombinant polypeptide (“XTEN”),
 wherein at least about 80% of the XTEN sequence consists of non-overlapping sequence motifs, 
 wherein the sequence motifs are selected from one or more sequences of Table 2, and 
 wherein no three contiguous amino acids of the XTEN sequence are identical unless the amino acid is serine, in which case no more than three contiguous amino acids are serine residues. 
 
     
     
         29 . A fusion protein comprising:
 (i) an XTEN polypeptide with an unstructured conformation, the XTEN polypeptide having a non-repetitive sequence comprising 4 to 6 types of amino acids selected from glycine (G), alanine (A), serine (S), threonine (T), glutamate (E), and proline (P), wherein the sum of glycine (G), alanine (A), serine (S), threonine (T), glutamate (E), and proline (P) residues constitutes more than 90% of the total amino acid residues of the XTEN sequence; and   (ii) a growth hormone linked to the XTEN polypeptide.   
     
     
         30 . The fusion protein of  claim 28 , wherein the XTEN sequence is from about 400 to about 1000 amino acid residues in length. 
     
     
         31 . The fusion protein of  claim 28 , wherein the XTEN sequence is from about 100 to about 200 amino acid residues in length. 
     
     
         32 . The fusion protein of  claim 28 , wherein the XTEN sequence has at least about 90% random coil, as determined by Garnier-Osguthorpe-Robson algorithm. 
     
     
         33 . The fusion protein of  claim 29 , wherein the XTEN sequence is from about 100 to about 3000 amino acid residues, wherein at least about 80% of the XTEN sequence is formed from non-overlapping sequence motifs of 9 to 14 amino acid residues wherein the motifs consist of 4 to 6 types of amino acids selected from glycine (G), alanine (A), serine (S), threonine (T), glutamate (E), and proline (P), and wherein the sequence of any two contiguous amino acid residues in any one motif is not repeated more than twice in the sequence motif. 
     
     
         34 . The fusion protein of  claim 33 , wherein at least about 95% of the XTEN sequence consists of multiple units of two or more non-overlapping sequence motifs selected from the amino acid sequences of Table 2. 
     
     
         35 . The fusion protein of  claim 33 , wherein no three contiguous amino acids of the XTEN sequence are identical unless the amino acid is serine, in which case no more than three contiguous amino acids are serine residues. 
     
     
         36 . The fusion protein of  claim 28 , further comprising a second XTEN sequence that is linked to the growth hormone, wherein the XTEN sequence exhibits at least about 95% sequence identity to a first XTEN selected from Table 3, and the second XTEN sequence exhibits at least about 95% sequence identity to the first XTEN or a second XTEN selected from Table 3. 
     
     
         37 . The fusion protein of  claim 28 , wherein the XTEN polypeptide is linked at the growth hormone N-terminal, wherein the XTEN polypeptide comprises a sequence that exhibits at least about 80% sequence identity to the amino acid sequence selected from the group consisting of AEPAGSPTSTEEGTPGSGTASSSPGSSTPSGATGSPGASPGTSSTGS and AEPAGSPTSTEEGASPGTSSTGSPGSSTPSGATGSPGSSTPSGATGS. 
     
     
         38 . The fusion protein of  claim 28  or  claim 29 , wherein the growth hormone is a human growth hormone. 
     
     
         39 . The fusion protein of  claim 28  or  claim 29 , wherein at least about 90% of the XTEN sequence consists of non-overlapping sequence motifs, and wherein the content of any one amino acid type in the full-length XTEN does not exceed 30%. 
     
     
         40 . A pharmaceutical composition comprising the fusion protein of  claim 28 . 
     
     
         41 . A method of treating a growth hormone-related condition in a subject, the method comprising administering to the subject a therapeutically effective amount of the fusion protein of  claims 28 , wherein the growth hormone-related condition is selected from growth-hormone deficiency, Turner's Syndrome, Prader-Willi Syndrome, chronic renal failure, intrauterine growth retardation, idiopathic short stature, AIDS wasting, obesity, multiple sclerosis, fibromyalgia, Crohn's disease, ulcerative colitis, muscular dystrophy, low muscle mass, or low bone density. 
     
     
         42 . A method of producing a fusion protein comprising growth hormone fused to one or more extended recombinant polypeptides (XTEN), the method comprising:
 (a) providing a host cell that comprises a recombinant polynucleotide molecule that encodes the fusion protein;   (b) culturing the host cell such that the host cell expresses the fusion protein, wherein the fusion protein is the fusion protein of 28; and   (c) recovering the fusion protein.   
     
     
         43 . An isolated nucleic acid comprising a polynucleotide sequence encoding the fusion protein of  claim 28 .