Therapeutic musk antibodies

Abstract

The present invention relates to antibody-based molecules, including full-length antibodies, antigen-binding domains thereof, and antibody derivatives that are capable of binding to and activating human muscle-specific tyrosine protein kinase (MuSK). The present invention further discloses methods of treating neuromuscular conditions using the aforementioned MuSK antibodies.

1 - 41 . (canceled) 
     
     
         42 . An isolated polynucleotide encoding a heavy chain variable region (VH) and/or a light chain variable region (VL) of an antibody that specifically binds to human muscle-specific tyrosine-protein kinase (MuSK), wherein:
 (a) the VH comprises a CDR-H1, a CDR-H2, and a CDR-H3 of the VH amino acid sequence of SEQ ID NO: 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 131, 133, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, or 263; and   (b) the VL comprises a CDR-L1, a CDR-L2, and a CDR-L3 of the VL amino acid sequence of SEQ ID NO: 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 132, 134, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, or 239.   
     
     
         43 . The isolated polynucleotide of  claim 42 , wherein the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 comprise the amino acid sequences set forth in SEQ ID NOs: 147, 153, 156, 159, 172, and 195; 1, 17, 33, 49, 65, and 81; 2, 18, 34, 50, 66, and 82; 2, 18, 240, 50, 66, and 82; 2, 18, 241, 50, 66, and 82; 2, 18, 242, 50, 66, and 82; 2, 18, 243, 50, 66, and 82; 2, 18, 244, 50, 66, and 82; 2, 18, 245, 50, 66, and 82; 2, 18, 246, 50, 66, and 82; 2, 18, 247, 50, 66, and 82; 3, 19, 35, 51, 67, and 83; 4, 20, 36, 52, 68, and 84; 5, 21, 37, 53, 69, and 85; 6, 22, 38, 54, 70, and 86; 7, 23, 39, 55, 71, and 87; 8, 24, 40, 56, 72, and 88; 9, 25, 41, 57, 73, and 89; 10, 26, 42, 58, 74, and 90; 11, 27, 43, 59, 75, and 91; 12, 28, 44, 60, 76, and 92; 13, 29, 45, 61, 77, and 93; 14, 30, 46, 62, 78, and 94; 15, 31, 47, 63, 79, and 95; 16, 32, 48, 64, 80, and 96; 135, 137, 139, 141, 143, and 145; 135, 137, 248, 141, 143, and 145; 135, 137, 249, 141, 143, and 145; 135, 137, 250, 141, 143, and 145; 135, 137, 251, 141, 143, and 145; 136, 138, 140, 142, 144, and 146; 147, 150, 156, 159, 170, and 180; 147, 150, 156, 159, 171, and 181; 147, 150, 156, 160, 172, and 182; 147, 150, 156, 159, 172, and 183; 147, 150, 156, 159, 171, and 184; 147, 150, 156, 159, 173, and 185; 147, 150, 156, 159, 173, and 186; 148, 151, 157, 161, 174, and 187; 148, 151, 157, 162, 174, and 188; 148, 151, 157, 163, 174, and 188; 148, 151, 157, 164, 174, and 189; 149, 152, 158, 165, 175, and 190; 149, 152, 158, 166, 176, and 191; 149, 152, 158, 167, 177, and 192; 149, 152, 158, 168, 178, and 193; 149, 152, 158, 169, 179, and 194; 147, 153, 156, 159, 172, and 183; 147, 154, 156, 159, 172, and 183; 147, 155, 156, 159, 172, and 183; 147, 154, 156, 159, 172, and 195; or 147, 155, 156, 159, 172, and 195, respectively. 
     
     
         44 . The isolated polynucleotide of  claim 42 , wherein:
 (a) the VH comprises an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 131, 133, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, or 263; and/or   (b) the VL comprises an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 132, 134, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, or 239.   
     
     
         45 . The isolated polynucleotide of  claim 42 , wherein the VH and the VL comprise amino acid sequences that are at least 80% identical to the amino acid sequences of SEQ ID NOs: 234 and 235; 228 and 229; 230 and 231; 232 and 233; 236 and 237; 238 and 239; 97 and 98; 99 and 100; 252 and 100; 253 and 100; 254 and 100; 255 and 100; 256 and 100; 257 and 100; 258 and 100; 259 and 100; 101 and 102; 103 and 104; 105 and 106; 107 and 108; 109 and 110; 111 and 112; 113 and 114; 115 and 116; 117 and 118; 119 and 120; 121 and 122; 123 and 124; 125 and 126; 127 and 128; 131 and 132; 260 and 132; 261 and 132; 262 and 132; 263 and 132; 133 and 134; 196 and 197; 198 and 199; 200 and 201; 202 and 203; 204 and 205; 206 and 207; 208 and 209; 210 and 211; 212 and 213; 214 and 215; 216 and 217; 218 and 219; 220 and 221; 222 and 223; 224 and 225; or 226 and 227, respectively. 
     
     
         46 . The isolated polynucleotide of  claim 42 , wherein the VH and the VL comprise the amino acid sequences of SEQ ID NOs: 234 and 235; 228 and 229; 230 and 231; 232 and 233; 236 and 237; 238 and 239; 97 and 98; 99 and 100; 252 and 100; 253 and 100; 254 and 100; 255 and 100; 256 and 100; 257 and 100; 258 and 100; 259 and 100; 101 and 102; 103 and 104; 105 and 106; 107 and 108; 109 and 110; 111 and 112; 113 and 114; 115 and 116; 117 and 118; 119 and 120; 121 and 122; 123 and 124; 125 and 126; 127 and 128; 131 and 132; 260 and 132; 261 and 132; 262 and 132; 263 and 132; 133 and 134; 196 and 197; 198 and 199; 200 and 201; 202 and 203; 204 and 205; 206 and 207; 208 and 209; 210 and 211; 212 and 213; 214 and 215; 216 and 217; 218 and 219; 220 and 221; 222 and 223; 224 and 225; or 226 and 227, respectively. 
     
     
         47 . The isolated polynucleotide of  claim 42 , wherein the polynucleotide further encodes:
 (a) a human IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM heavy chain constant region, and/or   (b) a human lambda light chain constant region or a human kappa light chain constant region.   
     
     
         48 . The isolated polynucleotide of  claim 42 , wherein the polynucleotide further encodes a human IgG1 heavy chain constant region comprising L234A and L235A mutations. 
     
     
         49 . A vector comprising the isolated polynucleotide of  claim 42 . 
     
     
         50 . A host cell comprising the isolated polynucleotide of  claim 42 . 
     
     
         51 . A method of producing an antibody that specifically binds to human muscle-specific tyrosine-protein kinase (MuSK) comprising culturing the host cell of  claim 50  so that the polynucleotide is expressed and the antibody is produced. 
     
     
         52 . A host cell comprising:
 (a) a first polynucleotide encoding a VH comprising a CDR-H1, a CDR-H2, and a CDR-H3 of the VH amino acid sequence of SEQ ID NO: 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 131, 133, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, or 263; and   (b) a second polynucleotide encoding a VL comprising a CDR-L1, a CDR-L2, and a CDR-L3 of the VL amino acid sequence of SEQ ID NO: 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 132, 134, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, or 239.   
     
     
         53 . The host cell of  claim 52 , wherein the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 comprise the amino acid sequences of SEQ ID NOs: 147, 153, 156, 159, 172, and 195; 1, 17, 33, 49, 65, and 81; 2, 18, 34, 50, 66, and 82; 2, 18, 240, 50, 66, and 82; 2, 18, 241, 50, 66, and 82; 2, 18, 242, 50, 66, and 82; 2, 18, 243, 50, 66, and 82; 2, 18, 244, 50, 66, and 82; 2, 18, 245, 50, 66, and 82; 2, 18, 246, 50, 66, and 82; 2, 18, 247, 50, 66, and 82; 3, 19, 35, 51, 67, and 83; 4, 20, 36, 52, 68, and 84; 5, 21, 37, 53, 69, and 85; 6, 22, 38, 54, 70, and 86; 7, 23, 39, 55, 71, and 87; 8, 24, 40, 56, 72, and 88; 9, 25, 41, 57, 73, and 89; 10, 26, 42, 58, 74, and 90; 11, 27, 43, 59, 75, and 91; 12, 28, 44, 60, 76, and 92; 13, 29, 45, 61, 77, and 93; 14, 30, 46, 62, 78, and 94; 15, 31, 47, 63, 79, and 95; 16, 32, 48, 64, 80, and 96; 135, 137, 139, 141, 143, and 145; 135, 137, 248, 141, 143, and 145; 135, 137, 249, 141, 143, and 145; 135, 137, 250, 141, 143, and 145; 135, 137, 251, 141, 143, and 145; 136, 138, 140, 142, 144, and 146; 147, 150, 156, 159, 170, and 180; 147, 150, 156, 159, 171, and 181; 147, 150, 156, 160, 172, and 182; 147, 150, 156, 159, 172, and 183; 147, 150, 156, 159, 171, and 184; 147, 150, 156, 159, 173, and 185; 147, 150, 156, 159, 173, and 186; 148, 151, 157, 161, 174, and 187; 148, 151, 157, 162, 174, and 188; 148, 151, 157, 163, 174, and 188; 148, 151, 157, 164, 174, and 189; 149, 152, 158, 165, 175, and 190; 149, 152, 158, 166, 176, and 191; 149, 152, 158, 167, 177, and 192; 149, 152, 158, 168, 178, and 193; 149, 152, 158, 169, 179, and 194; 147, 153, 156, 159, 172, and 183; 147, 154, 156, 159, 172, and 183; 147, 155, 156, 159, 172, and 183; 147, 154, 156, 159, 172, and 195; or 147, 155, 156, 159, 172, and 195, respectively. 
     
     
         54 . The host cell of  claim 52 , wherein
 (a) the VH comprises an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 131, 133, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, or 263; and/or   (b) the VL comprises an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 132, 134, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, or 239.   
     
     
         55 . The host cell of  claim 52 , wherein the VH and the VL comprise amino acid sequences that are at least 80% identical to the amino acid sequences of SEQ ID NOs: 234 and 235;
 228 and 229; 230 and 231; 232 and 233; 236 and 237; 238 and 239; 97 and 98; 99 and 100; 252 and 100; 253 and 100; 254 and 100; 255 and 100; 256 and 100; 257 and 100; 258 and 100; 259 and 100; 101 and 102; 103 and 104; 105 and 106; 107 and 108; 109 and 110; 111 and 112; 113 and 114; 115 and 116; 117 and 118; 119 and 120; 121 and 122; 123 and 124; 125 and 126; 127 and 128; 131 and 132; 260 and 132; 261 and 132; 262 and 132; 263 and 132; 133 and 134; 196 and 197; 198 and 199; 200 and 201; 202 and 203; 204 and 205; 206 and 207; 208 and 209; 210 and 211; 212 and 213; 214 and 215; 216 and 217; 218 and 219; 220 and 221; 222 and 223; 224 and 225; or 226 and 227, respectively.   
     
     
         56 . The host cell of  claim 52 , wherein the VH and the VL comprise the amino acid sequences of SEQ ID NOs: 234 and 235; 228 and 229; 230 and 231; 232 and 233; 236 and 237; 238 and 239; 97 and 98; 99 and 100; 252 and 100; 253 and 100; 254 and 100; 255 and 100; 256 and 100; 257 and 100; 258 and 100; 259 and 100; 101 and 102; 103 and 104; 105 and 106; 107 and 108; 109 and 110; 111 and 112; 113 and 114; 115 and 116; 117 and 118; 119 and 120; 121 and 122; 123 and 124; 125 and 126; 127 and 128; 131 and 132; 260 and 132; 261 and 132; 262 and 132; 263 and 132; 133 and 134; 196 and 197; 198 and 199; 200 and 201; 202 and 203; 204 and 205; 206 and 207;
 208 and 209; 210 and 211; 212 and 213; 214 and 215; 216 and 217; 218 and 219; 220 and 221; 222 and 223; 224 and 225; or 226 and 227, respectively.   
     
     
         57 . The host cell of  claim 52 , wherein:
 (a) the first polynucleotide further encodes a human IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM heavy chain constant region, and/or   (b) the second polynucleotide further encodes a human lambda light chain constant region or a human kappa light chain constant region.   
     
     
         58 . The host cell of  claim 52 , wherein the first polynucleotide further encodes a human IgG1 heavy chain constant region comprising L234A and L235A mutations. 
     
     
         59 . The host cell of  claim 52 , wherein the first polynucleotide and the second polynucleotide are comprised within a single vector. 
     
     
         60 . The host cell of  claim 52 , wherein the first polynucleotide and the second polynucleotide are comprised within separate vectors. 
     
     
         61 . A method of producing an antibody that specifically binds to human muscle-specific tyrosine-protein kinase (MuSK) comprising culturing the host cell of  claim 52  so that the polynucleotides are expressed and the antibody is produced.