Antagonist antibodies against gdf-8 and uses therefor

Abstract

The disclosure provides improved neutralizing anti-GDF-8 antibodies capable of substantially higher levels of expression in host cells compared to previous anti-GDF-8 antibodies. Also provided are methods of using compositions comprising such antibodies to increase muscle mass or strength, and to treat or prevent muscular disorders, neuromuscular disorders, metabolic disorders, adipose tissue disorders or bone disorders.

1 - 61 . (canceled) 
     
     
         62 . A monoclonal antibody, or antigen binding fragment thereof, that specifically binds GDF-8, wherein said antibody specifically binds an epitope comprising residues 25(GLU), 29(TRP), 30(ASP), 31(TRP), 33(ILE), 34(ALA), 35(PRO), 36(LYS), 83(ASN), 84(MET), 85(LEU), 87(PHE), 90(LYS), 91(GLU), 92(GLN), 93(ILE), 94(ILE) and 95(TYR) of the amino acid sequence of SEQ ID NO:1, and wherein said antibody expresses at a level at least 2-fold higher under the same conditions compared to a second antibody comprising heavy chains comprising a VH region defined by the amino acid sequence of SEQ ID NO:7 and a human IgG1 CH region, and light chains comprising a VL region defined by the amino acid sequence of SEQ ID NO:9 and a human kappa CL region. 
     
     
         63 . The monoclonal antibody of  claim 62 , wherein said antibody binds GDF-8 with a Kd of at least 10 −6  M, M, 10 −8  M, 10 −9  M, 10 −10  M, or 10 −11  M. 
     
     
         64 . The monoclonal antibody of  claim 62 , wherein said antibody expresses at a level at least 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 11-fold, or 12-fold higher under the same conditions compared to said second antibody. 
     
     
         65 . The antibody of  claim 64 , wherein said monoclonal antibody expresses at least 12-fold higher than said second antibody when both are expressed in COS cells. 
     
     
         66 . The antibody of  claim 64 , wherein said monoclonal antibody expresses at least 6-fold higher than said second antibody when both are expressed in CHO cells. 
     
     
         67 . The monoclonal antibody of  claim 62 , wherein the VH region of said antibody comprises leucine at Kabat position 108. 
     
     
         68 . The monoclonal antibody of  claim 67 , wherein the fourth framework region (FR4) of said VH region comprises amino acids 106-116 of SEQ ID NO:44. 
     
     
         69 . The monoclonal antibody of  claim 62 , wherein the VH region of said antibody comprises CDRH1, CDRH2, and CDRH3 from the VH region defined by the amino acid sequence of SEQ ID NO:44. 
     
     
         70 . The monoclonal antibody of  claim 69 , wherein CDRH1 comprises the amino acid sequence of SEQ ID NO:10, CDRH2 comprises the amino acid sequence of SEQ ID NO:11, and CDRH3 comprises the amino acid sequence of SEQ ID NO:12. 
     
     
         71 . The monoclonal antibody of  claim 69 , wherein the VH region of said antibody comprises the amino acid sequence of SEQ ID NO:44. 
     
     
         72 . The monoclonal antibody of  claim 62 , wherein the VL region of said antibody comprises glycine or glutamine at Kabat position 100. 
     
     
         73 . The monoclonal antibody of  claim 62 , wherein the fourth framework region of said VL region comprises amino acids 98-107 of SEQ ID NO:46 or amino acids 98-107 of SEQ ID NO:9. 
     
     
         74 . The monoclonal antibody of  claim 62 , wherein the VL region of said antibody comprises CDRL1, CDRL2, and CDRL3 from the VL region defined by the amino acid sequence of SEQ ID NO:46. 
     
     
         75 . The monoclonal antibody of  claim 74 , wherein CDRL1 comprises the amino acid sequence of SEQ ID NO:13, CDRL2 comprises the amino acid sequence of SEQ ID NO:14, and CDRL3 comprises the amino acid sequence of SEQ ID NO:15. 
     
     
         76 . The monoclonal antibody of  claim 74 , wherein the VL region of said antibody comprises the amino acid sequence of SEQ ID NO:46 or the amino acid sequence of SEQ ID NO:9. 
     
     
         77 . The monoclonal antibody of  claim 62 , further comprising an antibody constant heavy (CH) region derived from an antibody subtype selected from the group consisting of IgA, IgG, IgD, IgE, or IgM. 
     
     
         78 . The monoclonal antibody of  claim 77 , wherein said IgG subtype is further selected from the group consisting of IgG1, IgG2, IgG3 and IgG4. 
     
     
         79 . The monoclonal antibody of  claim 78 , wherein said IgG subtype comprises at least one mutation that alters Fc domain function. 
     
     
         80 . The monoclonal antibody of  claim 78 , wherein said IgG subtype is IgG1. 
     
     
         81 . The monoclonal antibody of  claim 62 , wherein said antibody is partially or fully humanized. 
     
     
         82 . The monoclonal antibody of  claim 62 , wherein said antibody is human. 
     
     
         83 . The monoclonal antibody of  claim 62 , wherein the heavy chain of said antibody comprises the amino acid sequence of SEQ ID NO:57. 
     
     
         84 . The monoclonal antibody of  claim 62 , further comprising an antibody constant light (CL) region. 
     
     
         85 . The monoclonal antibody of  claim 84 , wherein said CL region is a kappa or lambda CL region. 
     
     
         86 . The monoclonal antibody of  claim 62 , wherein the light chain of said antibody comprises the amino acid sequence of SEQ ID NO:17. 
     
     
         87 . A pharmaceutical composition comprising the monoclonal antibody of  claim 62  and a pharmaceutically acceptable carrier. 
     
     
         88 . An isolated polynucleotide comprising a nucleic acid sequence encoding the heavy chain of the monoclonal antibody of  claim 62 . 
     
     
         89 . A method of increasing the mass or strength of a muscle of a mammal, comprising administering to a mammal an amount of the monoclonal antibody of  claim 62  effective to increase the mass or strength of a muscle of the mammal. 
     
     
         90 . The method of  claim 89 , wherein the muscle is skeletal muscle. 
     
     
         91 . The method of  claim 90 , wherein the skeletal muscle is active in breathing. 
     
     
         92 . A method of treating a muscular disorder in a mammal characterized by diminished muscle mass or strength, comprising administering to a mammal in need of treatment for a muscular disorder characterized by diminished muscle mass or strength a therapeutically effective amount of the monoclonal antibody of  claim 62 . 
     
     
         93 . The method of  claim 92 , wherein the muscular disorder is selected from the group consisting of muscular dystrophy, muscle atrophy, sarcopenia, cachexia, muscle wasting syndrome, age-related loss of muscle mass or strength, and frailty. 
     
     
         94 . The method of  claim 93 , wherein the muscular disorder is muscular dystrophy. 
     
     
         95 . The method of  claim 94 , wherein the muscular dystrophy is Duchenne muscular dystrophy. 
     
     
         96 . The method of  claim 92 , wherein treatment is effective to increase the subject's performance in a 6 minute walk test. 
     
     
         97 . The method of  claim 92 , wherein said antibody is administered to a subject also being treated with a glucocorticoid.