Antibodies to lrp6

Abstract

Anti-LRP6 antibodies and antigen-binding fragments thereof, as well as pharmaceutical compositions comprising such antibodies and antigen-binding fragments are described. These anti-LRP6 antibodies can be used to enhance Wnt activity and/or antagonize Dkk1 activity. Also described are methods of therapy using such antibodies and antigen-binding regions to bind modulate Wnt/LRP6 signaling to promote tissue homeostasis, regeneration and repair in diseases such as, but not limited to, bone disorders, such as osteoporosis, rheumatoid arthritis, and osteolytic lesions caused by osteoarthritis and multiple myeloma, gastrointestinal disease and wound healing.

We claim: 
     
         1 . An isolated antibody or antigen-binding fragment thereof, comprising:
 a) a heavy chain, comprising a first variable region, comprising an amino acid sequence as set forth in SEQ ID NO: 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 72, 74, 78, 82, 86, 90, 94, 98, or 102; and   b) a light chain comprising a second variable region, comprising an amino acid sequence as set forth in SEQ ID NO: 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 76, 80, 84, 88, 92, 96, 100, or 104.   
     
     
         2 . An isolated antibody or antigen-binding fragment thereof, comprising:
 a) a heavy chain, comprising a first variable region, comprising a sequence that has at least 90% identity to an amino acid sequence as set forth in SEQ ID NO: 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 72, 74, 78, 82, 86, 90, 94, 98, or 102; and   b) a light chain comprising a second variable region, comprising a sequence that has at least 90% identity to an amino acid sequence as set forth in SEQ ID NO: 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 76, 80, 84, 88, 92, 96, 100, or 104, wherein the antibody binds LRP6 with high affinity.   
     
     
         3 . An isolated antibody or antigen-binding fragment thereof, comprising:
 a) a heavy chain, comprising a first variable region, comprising a sequence that has at least 95% identity to an amino acid sequence as set forth in SEQ ID NO: 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 72, 74, 78, 82, 86, 90, 94, 98, or 102; and   b) a light chain comprising a second variable region, comprising a sequence that has at least 95% identity to an amino acid sequence as set forth in SEQ ID NO: 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 76, 80, 84, 88, 92, 96, 100, or 104, wherein the antibody binds LRP6 with high affinity.   
     
     
         4 . An isolated antibody or antigen-binding fragment thereof, comprising:
 a) a heavy chain, comprising a first variable region, comprising a sequence that has at least 99% identity to an amino acid sequence as set forth in SEQ ID NO: 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 72, 74, 78, 82, 86, 90, 94, 98, or 102; and   b) a light chain, comprising a second variable region, comprising a sequence that has at least 99% identity to an amino acid sequence as set forth in SEQ ID NO: 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 76, 80, 84, 88, 92, 96, 100, or 104, wherein the antibody binds to LRP6 with high affinity.   
     
     
         5 . The antibody of any one of  claims 1 - 4 , wherein the antibody stimulates Wnt activity. 
     
     
         6 . The antibody of any one of  claims 1 - 4 , wherein the antibody antagonizes Dkk1 activity. 
     
     
         7 . The antibody of  claim 1 , wherein the heavy chain and the light chain are connected by a flexible linker to form a single chain antibody. 
     
     
         8 . The antibody of  claim 7 , which is a single-Fv antibody. 
     
     
         9 . The antibody of  claim 1 , which is selected from the group consisting of an Fab antibody, an Fab′ antibody, an (Fab′)2 antibody, a fully human antibody, a humanized antibody, and a chimeric antibody. 
     
     
         10 . A nucleic acid molecule encoding the antibody of any one of  claims 1 - 6 . 
     
     
         11 . A host cell transformed with the nucleic acid of  claim 10 . 
     
     
         12 . A pharmaceutical composition comprising an antibody of  claim 1 , further comprising a suitable carrier. 
     
     
         13 . A pharmaceutical composition comprising an antibody of  claim 1 , further comprising at least one therapeutic agent. 
     
     
         14 . A method of treating a bone disorder in a patient comprising administering the pharmaceutical agent of  claim 12  or  13 . 
     
     
         15 . The method of  claim 14 , wherein the bone disorder is selected from the group consisting of rheumatoid arthritis, osteoarthritis, ankylosing spondylosis, osteoporosis, and osteolytic lesions associated with multiple myeloma. 
     
     
         16 . A method of healing wounds in a patient comprising administering the pharmaceutical agent of  claim 12  or  13 . 
     
     
         17 . A method of treating gastrointestinal diseases in a patient comprising administering the pharmaceutical agent of  claim 12  or  13 . 
     
     
         18 . The method of  claim 17 , wherein the gastrointestinal disease is inflammatory bowel disease. 
     
     
         19 . A method of detecting the level of LRP6 in a biological sample, comprising contacting the sample with the antibody or antigen-binding fragment of  claim 1 . 
     
     
         20 . A selective binding agent comprising any of SEQ ID NO: 108-110, 114-116, 120-122, 126-128, 132-134, 138-140, 144-146, 150-152, 156-158, 162-164, 168-170, 174-176, 180-182, 186-188, 192-194, 198-200, 204-206, 210-212, 216-218, 222-224, 228-230, 234-236, 240-242, 246-248, 252-254, 258-260, 264-266, 270-272, 276-278, 282-284, 288-290, 294-296, 300-302, 306-308, 312-314, 318-320, 324-326, 330-332, 336-338, 342-344, 348-350, 354-356, 360-362, or 366-368. 
     
     
         21 . A nucleic acid molecule encoding the selective binding agent of  claim 20 . 
     
     
         22 . A host cell transformed with the nucleic acid molecule of  claim 21 . 
     
     
         23 . The selective binding agent of  claim 20 , wherein the heavy chain and the light chain are connected by a flexible linker to form a single chain antibody. 
     
     
         24 . The selective binding agent of  claim 23 , which is a single-Fv antibody. 
     
     
         25 . The selective binding agent of  claim 20 , which is selected from the group consisting of an Fab antibody, an (Fab′)2 antibody, a fully human antibody, a humanized antibody, and a chimeric antibody. 
     
     
         26 . A pharmaceutical composition comprising the selective binding agent of  claim 20 , further comprising a suitable carrier. 
     
     
         27 . A pharmaceutical composition comprising the selective binding agent of  claim 20 , further comprising at least one therapeutic agent. 
     
     
         28 . A method of treating a bone disorder in a patient, comprising administering the pharmaceutical composition of  claim 26  or  27 . 
     
     
         29 . The method of  claim 28 , wherein the bone disorder is selected from the group consisting of rheumatoid arthritis, osteoarthritis, ankylosing spondylosis, osteoporosis, and osteolytic lesions associated with multiple myeloma. 
     
     
         30 . A method of detecting the level of LRP6 in a biological sample, comprising contacting the sample with the selective binding agent of  claim 20 . 
     
     
         31 . An antibody or antigen-binding fragment thereof according to any of  claims 1 - 6 , wherein the antibody or antigen-binding fragment has at least one property selected from the group consisting of:
 a) competes for binding to LRP6 with antibody selected from the group consisting of 77.2, 135.16, 213.7, 240.8, 413.1, 421.1, 498.3, 537.2, 606.4, 620.1, 856.6, 923.3, 931.1, 993.9, 995.5, 1115.3, 1213.2, 1253.12, 1281.1, 1293.11, 1433.8, 1470.2, and 1903.1;   b) binds to the same epitope of LRP6 as an antibody selected from 77.2, 135.16, 213.7, 240.8, 413.1, 421.1, 498.3, 537.2, 606.4, 620.1, 856.6, 923.3, 931.1, 993.9, 995.5, 1115.3, 1213.2, 1253.12, 1281.1, 1293.11, 1433.8, 1470.2, and 1903.1; or   c) binds to the same antigen as that bound by an antibody selected from the group consisting of 77.2, 135.16, 213.7, 240.8, 413.1, 421.1, 498.3, 537.2, 606.4, 620.1, 856.6, 923.3, 931.1, 993.9, 995.5, 1115.3, 1213.2, 1253.12, 1281.1, 1293.11, 1433.8, 1470.2, and 1903.1.   
     
     
         32 . A selective binding agent according to any of  claim 35 ,  36 , or  37 , wherein the selective binding agent has at least one property selected from the group consisting of:
 a) competes for binding to LRP6 with an antibody selected from 77.2, 135.16, 213.7, 240.8, 413.1, 421.1, 498.3, 537.2, 606.4, 620.1, 856.6, 923.3, 931.1, 993.9, 995.5, 1115.3, 1213.2, 1253.12, 1281.1, 1293.11, 1433.8, 1470.2, and 1903.1;   b) binds to the same epitope of LRP6 as an antibody selected from the group consisting of 77.2, 135.16, 213.7, 240.8, 413.1, 421.1, 498.3, 537.2, 606.4, 620.1, 856.6, 923.3, 931.1, 993.9, 995.5, 1115.3, 1213.2, 1253.12, 1281.1, 1293.11, 1433.8, 1470.2, and 1903.1; or   c) binds to the same antigen as that bound by an antibody selected from the group consisting of 77.2, 135.16, 213.7, 240.8, 413.1, 421.1, 498.3, 537.2, 606.4, 620.1, 856.6, 923.3, 931.1, 993.9, 995.5, 1115.3, 1213.2, 1253.12, 1281.1, 1293.11, 1433.8, 1470.2, and 1903.1.   
     
     
         33 . An isolated antibody or antigen-binding fragment thereof that binds to an epitope within residues 263-283 of an LRP6 polypeptide comprising SEQ ID NO: 2. 
     
     
         34 . The antibody of  claim 33 , wherein the antibody binds to the LRP6 polypeptide of SEQ ID NO: 2 with a KD of less than or equal to 10<9>M. 
     
     
         35 . An isolated cell line that produces an antibody according to any one of  claims 1 - 6 . 
     
     
         36 . An isolated cell line that produces a specific binding agent according to  claim 20 . 
     
     
         37 . An isolated cell line that produces an antibody selected from the group of 77.2, 135.16, 213.7, 240.8, 413.1, 421.1, 498.3, 537.2, 606.4, 620.1, 856.6, 923.3, 931.1, 993.9, 995.5, 1115.3, 1213.2, 1253.12, 1281.1, 1293.11, 1433.8, 1470.2, and 1903.1. 
     
     
         38 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 77.2. 
     
     
         39 . The antibody of  claim 38 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         40 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 135.16. 
     
     
         41 . The antibody of  claim 40 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         42 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 213.7. 
     
     
         43 . The antibody of  claim 42 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         44 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 240.8. 
     
     
         45 . The antibody of  claim 44 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         46 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 413.1. 
     
     
         47 . The antibody of  claim 46 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         48 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 421.1. 
     
     
         49 . The antibody of  claim 48 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         50 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 498.3. 
     
     
         51 . The antibody of  claim 50 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         52 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 537.2. 
     
     
         53 . The antibody of  claim 52 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         54 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 606.4. 
     
     
         55 . The antibody of  claim 54 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         56 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 620.1. 
     
     
         57 . The antibody of  claim 56 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         58 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 856.6. 
     
     
         59 . The antibody of  claim 58 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         60 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 923.3. 
     
     
         61 . The antibody of  claim 60 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         62 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 931.1. 
     
     
         63 . The antibody of  claim 62 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         64 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 993.9. 
     
     
         65 . The antibody of  claim 64 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         66 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 995.5. 
     
     
         67 . The antibody of  claim 66 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         68 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 1115.3. 
     
     
         69 . The antibody of  claim 68 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         70 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 1213.2. 
     
     
         71 . The antibody of  claim 70 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         72 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 1253.12. 
     
     
         73 . The antibody of  claim 72 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         74 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 1281.1. 
     
     
         75 . The antibody of  claim 74 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         76 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 1293.11. 
     
     
         77 . The antibody of  claim 76 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         78 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 1433.8. 
     
     
         79 . The antibody of  claim 78 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         80 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 1470.2. 
     
     
         81 . The antibody of  claim 80 , wherein said antibody is a humanized or chimeric form thereof. 
     
     
         82 . The antibody of  claim 1 , wherein said antibody is murine anti-LRP6 monoclonal antibody 1903.1. 
     
     
         83 . The antibody of  claim 82 , wherein said antibody is a humanized or chimeric form thereof.