Anti-PACAP antibodies
The present invention is directed to antagonistic antibodies and antigen binding fragments thereof having binding specificity for PACAP. These antibodies inhibit, block or neutralize at least one biological effect associated with PACAP, e.g., vasodilation. In exemplary embodiments these antibodies and antigen binding fragments thereof may comprise specific V H , V L , and CDR polypeptides described herein. In some embodiments these antibodies and antigen binding fragments thereof bind to and/or compete for binding to specific epitope(s) on human PACAP. The invention is further directed to using these antagonistic anti-PACAP antibodies, and binding fragments thereof, for the diagnosis, assessment, and treatment of diseases and disorders associated with PACAP and conditions where antagonism of PACAP-related activities, such as vasodilation, mast cell degranulation, and/or neuronal activation, are therapeutically beneficial, e.g., headache and migraine indications.
1. An antibody or antigen-binding fragment having binding specificity to Pituitary Adenylate-Cyclase-Activating Polypeptide (PACAP) which is selected from:
(A) an antibody or antigen-binding fragment that comprises:
(i) a variable heavy chain polypeptide comprising complementarity determining region 1, 2 and 3 (CDR1, CDR2 and CDR3) polypeptides respectively comprising the amino sequences of SEQ ID NO: 604; SEQ ID NO: 606; and SEQ ID NO: 608; and
(ii) a variable light chain polypeptide comprising CDR1, CDR2 and CDR3 polypeptides respectively comprising the amino sequences of SEQ ID NO: 624; SEQ ID NO: 626; and SEQ ID NO: 628; or
(B) an antibody or antigen-binding fragment that comprises:
(i) a variable heavy chain polypeptide comprising CDR1, CDR2 and CDR3 polypeptides respectively comprising the amino sequences of SEQ ID NO: 564; and SEQ ID NO: 566; and SEQ ID NO: 568, and
(ii) a variable light chain polypeptide comprising CDR1, CDR2 and CDR3 polypeptides respectively comprising the amino sequences of SEQ ID NO: 584; SEQ ID NO: 586; and SEQ ID NO: 588.
2. The antibody or antigen-binding fragment of claim 1 (A) which comprises a variable heavy chain polypeptide comprising an amino acid sequence possessing at least 90% sequence identity to SEQ ID NO: 602 and a variable light chain polypeptide comprising an amino acid sequence possessing at least 90% sequence identity to SEQ ID NO: 622.
3. The antibody or antigen-binding fragment of claim 1 (B) which comprises a variable heavy chain polypeptide comprising an amino acid sequence possessing at least 90% sequence identity to SEQ ID NO: 562 and a variable light chain polypeptide comprising an amino acid sequence possessing at least 90% sequence identity to SEQ ID NO: 582.
4. The antibody or antigen-binding fragment of claim 1 (A) which comprises a variable heavy chain polypeptide comprising an amino acid sequence possessing at least 95% sequence identity to SEQ ID NO: 602 and a variable light chain polypeptide comprising an amino acid sequence possessing at least 95% sequence identity to SEQ ID NO: 622.
5. The antibody or antigen-binding fragment of claim 1 (B) which comprises a variable heavy chain polypeptide comprising an amino acid sequence possessing at least 95% sequence identity to SEQ ID NO: 562 and a variable light chain polypeptide comprising an amino acid sequence possessing at least 95% sequence identity to SEQ ID NO: 582.
6. The antibody or antigen-binding fragment of claim 1 (A) which comprises a variable heavy chain polypeptide possessing the amino acid sequence of SEQ ID NO: 602 and a variable light chain polypeptide possessing the amino acid sequence of SEQ ID NO: 622.
7. The antibody or antigen-binding fragment of claim 1 (B) which comprises a variable heavy chain polypeptide possessing the amino acid sequence of SEQ ID NO: 562 and a variable light chain polypeptide possessing the amino acid sequence of SEQ ID NO: 582.
8. The antibody or antigen-binding fragment of claim 1 (A) which comprises a heavy chain polypeptide possessing the amino acid sequence of SEQ ID NO: 601 and a light chain polypeptide possessing the amino acid sequence of SEQ ID NO: 621.
9. The antibody or antigen-binding fragment of claim 1 (B) which comprises a heavy chain polypeptide possessing the amino acid sequence of SEQ ID NO: 561 and a light chain polypeptide possessing the amino acid sequence of SEQ ID NO: 562.
10. The antibody or antigen-binding fragment of claim 1 (A) or (B), which is human, humanized or chimerized.
11. The antibody or antigen-binding fragment of claim 1 (A) or (B), which comprises a human constant domain.
12. The antibody or antigen-binding fragment of claim 1 (A) or (B), which comprises a human constant domain selected from IgG1, IgG2, IgG3 and IgG4.
13. The antibody or antigen-binding fragment of claim 1 (A) or (B), which comprises a human constant domain which is mutated to impair or increase at least one effector function or glycosylation.
14. The antigen binding-fragment of claim 1 (A) or (B), which is selected from the group consisting of an scFv, camelbody, nanobody, Immunoglobulin New Antigen Receptor (“IgNAR”), fragment antigen binding (“Fab”) fragment, Fab′ fragment, monovalent antibody fragment, and a F(ab′) 2 fragment.
15. The antigen-binding fragment of claim 1 (A) or (B), which substantially or entirely lacks N-glycosylation and/or O-glycosylation and/or comprising an Fc region that contains one or more mutations that alters or eliminates N- and/or O-glycosylation.
16. The antigen-binding fragment of claim 1 (A) or (B), comprising an Fc region that has been modified to alter at least one effector function, half-life, proteolysis, or glycosylation.
17. The antibody or antigen-binding fragment of claim 1 (A) or (B), which binds to PACAP with a binding affinity (K D ) of less than or equal to 5×10 −5 M, 10 −5 M, 5×10 −6 M, 10 −6 M, 5×10 −7 M, 10 −7 M, 5×10 −8 M, 10 −8 M, 5×10 −9 M, 10 −9 M, 5×10 −10 M, 10 −10 M, 5×10 −11 M, 10 −11 M, 5×10 −12 M, 10 −12 M, 5×10 −13 M, or 10 −13 M, optionally as determined by ELISA, bio-layer interferometry (“BLI”), kinetic exclusion assay or surface plasmon resonance at 25° or 37° C.
18. The antibody or antigen-binding fragment of claim 1 (A) or (B), which binds to PACAP with an off-rate (k off ) of less than or equal to 5×10 −4 s −1 , 10 −4 s −1 , 5×10 −5 s −1 , or 10 −5 s −1 .
19. The antibody or antigen-binding fragment of claim 1 (A) or (B), which
(i) when administered to a subject inhibits or neutralizes at least one biological effect elicited by PACAP;
(ii) neutralizes or inhibits PACAP activation of at least one of pituitary adenylate cyclase-activating 1 receptor (PAC1-R), vasoactive intestinal peptide rector type 1 (VPAC1-R), or vasoactive intestinal peptide rector type 2 (VPAC2-R);
(iii) neutralizes or inhibits PACAP activation of each of PAC1-R, VPAC1-R, and VPAC2-R;
(iv) neutralizes or inhibits PACAP activation of PAC1-R;
(v) when administered is capable of inhibiting or preventing PACAP binding to at least one of PAC1-R, VPAC1-R, or VPAC2-R;
(vi) when administered is capable of inhibiting or preventing PACAP binding to each of PAC1-R, VPAC1-R, and VPAC2-R;
(vii) when administered is capable of inhibiting or preventing PACAP binding to PAC1-R-expressing cells, VPAC1-R-expressing cells, and/or VPAC2-R-expressing cells;
(viii) inhibits or blocks PACAP-induced cyclic AMP (cAMP) production;
(ix) when administered reduces PACAP-induced vasodilation, mast cell degranulation and/or neuronal activation;
(x) has a stronger affinity for PACAP as compared to vasoactive intestinal peptide (VIP) and/or does not bind to VIP, wherein the affinity of said antibody or antibody fragment to PACAP optionally is at least 10-fold, 30-fold, 100-fold, 300-fold, 1000-fold, 3000-fold, 10000-fold, 30000-fold, 100000-fold, 300000-fold, 1000000-fold, 3000000-fold, 10000000-fold, 30000000-fold or more stronger than the affinity of said antibody or antibody fragment to VIP; or
(xi) any combination of the foregoing.
20. A composition comprising at least one anti-PACAP antibody or antigen-binding fragment according to claim 1 and a pharmaceutically acceptable carrier.