Anti-PACAP antibodies and uses thereof
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. A nucleic acid or nucleic acids which encode for an anti-human Pituitary Adenylate Cyclase-Activating Polypeptide (“PACAP”) antibody or antigen-binding fragment thereof that antagonizes, inhibits, neutralizes or blocks at least one biological effect associated with human PACAP, wherein said anti-human PACAP antibody or antigen-binding fragment thereof comprises:
(A)
(i) a variable heavy chain polypeptide comprising complementarity determining region 1, 2 and 3 (CDR1, CDR2 and CDR3) polypeptides respectively comprising the amino acid 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 acid sequences of SEQ ID NO: 624; SEQ ID NO: 626; and SEQ ID NO: 628; or
(B)
(i) a variable heavy chain polypeptide comprising CDR1, CDR2 and CDR3 polypeptides respectively comprising the amino acid 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 acid sequences of SEQ ID NO: 584; SEQ ID NO: 586; and SEQ ID NO: 588.
2. A nucleic acid or nucleic acids according to claim 1 , which encode an antibody or antigen-binding fragment having binding specificity to Pituitary Adenylate-Cyclase-Activating Polypeptide (PACAP) which comprises:
(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 acid 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 acid sequences of SEQ ID NO: 624; SEQ ID NO: 626; and SEQ ID NO: 628.
3. The nucleic acid or nucleic acids of claim 1 ,
which is embodiment (A) wherein the encoded antibody or antigen-binding fragment 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; or
which is embodiment (B) wherein the encoded antibody or antigen-binding fragment 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 nucleic acid or nucleic acids of claim 1 ,
which is embodiment (A) wherein the encoded antibody or antigen-binding fragment 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; or
which is embodiment (B) wherein the encoded antibody or antigen-binding fragment 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.
5. The nucleic acid or nucleic acids of claim 1 ,
which is embodiment (A) wherein the encoded antibody or antigen-binding fragment 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; or
which is embodiment (B) wherein the encoded antibody or antigen-binding fragment 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.
6. The nucleic acid or nucleic acids of claim 1 ,
which is embodiment (A) wherein the encoded antibody or antigen-binding fragment 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; or
which is embodiment (B) wherein the encoded antibody or antigen-binding fragment 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.
7. The nucleic acid or nucleic acids of claim 1 , which is embodiment (A) or (B), wherein the encoded antibody or antigen-binding fragment is human, humanized or chimerized.
8. The nucleic acid or nucleic acids of claim 1 , which is embodiment (A) or (B), wherein the encoded antibody or antigen-binding fragment comprises a human constant domain.
9. The nucleic acid or nucleic acids of claim 1 , which is embodiment (A) or (B), wherein the encoded antibody or antigen-binding fragment comprises a human constant domain selected from IgG1, IgG2, IgG3 and IgG4.
10. The nucleic acid or nucleic acids of claim 1 , which is embodiment (A) or (B), wherein the encoded antibody or antigen-binding fragment comprises a human constant domain which is mutated to impair or increase at least one effector function or glycosylation.
11. The nucleic acid or nucleic acids of claim 1 , which is embodiment (B), which comprises:
a heavy chain variable region coding sequence at least 90, 95 or 100% identical to that of SEQ ID NO: 572 and a light chain variable region coding sequence at least 90, 95 or 100% identical to that of SEQ ID NO: 582; and/or
a heavy chain coding sequence at least 90, 95 or 100% identical to that of SEQ ID NO: 571 and a light chain variable region coding sequence at least 90, 95 or 100% identical to that of SEQ ID NO: 591.
12. The nucleic acid or nucleic acids of claim 1 , which is embodiment (A), which comprises:
a heavy chain variable region coding sequence at least 90, 95 or 100% identical to that of SEQ ID NO: 612 and a light chain variable region coding sequence at least 90, 95 or 100% identical to that of SEQ ID NO: 632; and/or
a heavy chain coding sequence at least 90, 95 or 100% identical to that of SEQ ID NO: 611 and a light chain variable region coding sequence at least 90, 95 or 100% identical to that of SEQ ID NO: 631.
13. A vector or vectors comprising the nucleic acid or nucleic acids of claim 1 , which is embodiment (A) or (B).
14. An isolated or recombinant host cell comprising the nucleic acid or nucleic acids of claim 1 , which is embodiment (A) or (B), or a vector or vectors comprising said nucleic acid or nucleic acids.
15. The isolated or recombinant host cell of claim 14 , which is a mammalian, bacterial, fungal, yeast, avian, amphibian, plant or insect cell.
16. The isolated or recombinant host cell of claim 15 which is a CHO cell or a Pichia pastoris cell.
17. A method of expressing an anti-PACAP antibody or antigen-binding fragment thereof comprising culturing a host cell according to claim 15 under conditions that provide for expression of said antibody or antigen-binding fragment, optionally a Pichia pastoris yeast cell or CHO cell.
18. The antigen-binding fragment of claim 1 , which is embodiment (A) or (B), which is selected from the group consisting of an scFv, nanobody, fragment antigen binding (“Fab”) fragment, Fab′ fragment, monovalent antibody fragment, and a F(ab′)2 fragment.