Anti-CGRP compositions and use thereof
The present invention is directed to antibodies and fragments thereof having binding specificity for CGRP. Another embodiment of this invention relates to the antibodies described herein, and binding fragments thereof, comprising the sequences of the V H , V L and CDR polypeptides described herein, and the polynucleotides encoding them. The invention also contemplates conjugates of anti-CGRP antibodies and binding fragments thereof conjugated to one or more functional or detectable moieties. The invention also contemplates methods of making said anti-CGRP antibodies and binding fragments thereof. Embodiments of the invention also pertain to the use of anti-CGRP antibodies, and binding fragments thereof, for the diagnosis, assessment and treatment of diseases and disorders associated with CGRP.
1. An isolated polynucleotide or polynucleotides which encode an anti-human calcitonin gene related peptide (CGRP) antibody or antibody fragment, wherein said antibody or antibody fragment comprises (i) a variable light (V L ) chain polypeptide that comprises the V L complementarity determining region (CDR) polypeptides CDR1, CDR2, and CDR3, respectively, of SEQ ID NO: 55, SEQ ID NO: 56, and SEQ ID NO: 57, and (ii) a variable heavy (V H ) chain polypeptide that comprises the V H CDR polypeptides CDR1, CDR2, and CDR3, respectively, of SEQ ID NO: 58, SEQ ID NO: 59, and SEQ ID NO: 60.
2. The isolated polynucleotide or polynucleotides of claim 1 , wherein said polynucleotide or polynucleotides comprise (i) a polynucleotide sequence that encodes the V L chain polypeptide of said anti-human CGRP antibody or antibody fragment which polypeptide comprises an amino acid sequence which possesses at least 90% sequence identity to SEQ ID NO: 51, SEQ ID NO: 41, or SEQ ID NO: 31, and (ii) a polynucleotide which encodes the V H chain polypeptide of said anti-human CGRP antibody or antibody fragment which polypeptide comprises an amino acid sequence which possesses at least 90% sequence identity to SEQ ID NO: 53, SEQ ID NO: 43, or SEQ ID NO: 33.
3. The isolated polynucleotide or polynucleotides of claim 1 , wherein said polynucleotide or polynucleotides comprise (i) a polynucleotide that encodes the V L chain polypeptide of said anti-human CGRP antibody or antibody fragment which polynucleotide comprises a sequence which possesses at least 90% sequence identity to SEQ ID NO: 191, SEQ ID NO: 181, or SEQ ID NO: 171, and (ii) a polynucleotide that encodes the V H chain polypeptide of said anti-human CGRP antibody or antibody fragment which polynucleotide comprises a sequence which possesses at least 90% sequence identity to SEQ ID NO: 193, SEQ ID NO: 183, or SEQ ID NO: 173.
4. The isolated polynucleotide or polynucleotides of claim 1 , wherein said polynucleotide or polynucleotides comprise (i) a polynucleotide that encodes the V L chain polypeptide of SEQ ID NO: 51, SEQ ID NO: 41, or SEQ ID NO: 31, and (ii) a polynucleotide which encodes the V H chain polypeptide of SEQ ID NO: 53, SEQ ID NO: 43, or SEQ ID NO: 33.
5. The isolated polynucleotide or polynucleotides of claim 1 , wherein said polynucleotide or polynucleotides comprise (i) a polynucleotide that encodes a V L chain polypeptide wherein said polynucleotide comprises the sequence of SEQ ID NO: 191, SEQ ID NO: 181, or SEQ ID NO: 171, and (ii) a polynucleotide that encodes a V H chain polypeptide wherein said polynucleotide comprises the sequence of SEQ ID NO: 193, SEQ ID NO: 183, or SEQ ID NO: 173.
6. The isolated polynucleotide or polynucleotides of claim 1 , wherein said polynucleotide or polynucleotides comprise (i) a polynucleotide that encodes a light chain polypeptide having an amino acid sequence which possesses at least 90% sequence identity to SEQ ID NO: 52, SEQ ID NO: 42, or SEQ ID NO: 32, and (ii) a polynucleotide that encodes a heavy chain polypeptide having an amino acid sequence which possesses at least 90% sequence identity to SEQ ID NO: 54, SEQ ID NO: 44, or SEQ ID NO: 34.
7. The isolated polynucleotide or polynucleotides of claim 1 , which comprise (i) a sequence that encodes a light chain polypeptide having a sequence which possesses at least 90% sequence identity to SEQ ID NO: 192, SEQ ID NO: 182, or SEQ ID NO: 172, and (ii) a polynucleotide that encodes a heavy chain polypeptide having a sequence which possesses at least 90% sequence identity to SEQ ID NO: 194, SEQ ID NO: 184, or SEQ ID NO: 174.
8. The isolated polynucleotide or polynucleotides of claim 1 , which comprise (i) a polynucleotide that encodes a light chain polypeptide having the sequence of SEQ ID NO: 52, SEQ ID NO: 42, or SEQ ID NO: 32, and (ii) a polynucleotide that encodes a heavy chain polypeptide having the sequence of SEQ ID NO: 54, SEQ ID NO: 44, or SEQ ID NO: 34.
9. The isolated polynucleotide or polynucleotides of claim 1 , which comprise (i) a polynucleotide that encodes a light chain polypeptide which polynucleotide comprises the sequence of SEQ ID NO: 192, SEQ ID NO: 182, or SEQ ID NO: 172, and (ii) a polynucleotide that encodes a heavy chain polypeptide, which polynucleotide comprises the sequence of SEQ ID NO: 194, SEQ ID NO: 184, or SEQ ID NO: 174.
10. A host cell comprising said polynucleotide or polynucleotides of claim 1 .
11. The cell of claim 10 , which is a yeast cell belonging to the genus Pichia.
12. The cell of claim 11 , wherein the Pichia is Pichia pastoris.
13. The cell of claim 10 , which is a CHO cell.
14. The isolated polynucleotide or polynucleotides of claim 1 , wherein said polynucleotide or polynucleotides are comprised of yeast preferred codons or human preferred codons.
15. The isolated polynucleotide or polynucleotides of claim 1 , wherein the anti-CGRP antibody or antibody fragment encoded by said isolated polynucleotide or polynucleotides further comprises a human IgG1, IgG2, IgG3, or IgG4 constant region or fragment thereof.
16. The isolated polynucleotide or polynucleotides of claim 15 , wherein the anti-CGRP antibody or antibody fragment encoded thereby comprises a human IgG1 constant region or fragment thereof.
17. The isolated polynucleotide or polynucleotides of claim 1 , wherein the anti-CGRP antibody or antibody fragment encoded thereby binds CGRP with an off-rate (K off ) of less than or equal to 10 −4 S −1 , 5×10 −5 S −1 , 10 −5 S −1 , 5×10 −6 S −1 , 10 −6 S −1 , 5×10 −7 S −1 , or 10 −7 S −1 .
18. The isolated polynucleotide or polynucleotides of claim 1 , wherein said anti-CGRP antibody or antibody fragment encoded thereby contains an Fc region having a mutation that alters effector function, half-life, proteolysis, and/or glycosylation.
19. The isolated polynucleotide or polynucleotides of claim 1 , wherein said anti-CGRP antibody or antibody fragment encoded thereby is a humanized, single chain, or chimeric antibody.
20. The isolated polynucleotide or polynucleotides of claim 1 , wherein said anti-CGRP antibody or antibody fragment encoded thereby is humanized.
21. The isolated polynucleotide or polynucleotides of claim 1 , wherein said encoded anti-CGRP antibody fragment is selected from a Fab fragment, a Fab′ fragment, and a F(ab′) 2 fragment.
22. An expression vector comprising the polynucleotide or polynucleotides of claim 1 .
23. An expression vector comprising the polynucleotide or polynucleotides of claim 2 .
24. An expression vector comprising the polynucleotide or polynucleotides of claim 3 .
25. An expression vector comprising the polynucleotide or polynucleotides of claim 4 .
26. An expression vector comprising the polynucleotide or polynucleotides of claim 5 .
27. An expression vector comprising the polynucleotide or polynucleotides of claim 6 .
28. An expression vector comprising the polynucleotide or polynucleotides of claim 7 .
29. An expression vector comprising the polynucleotide or polynucleotides of claim 8 .
30. An expression vector comprising the polynucleotide or polynucleotides of claim 9 .