Methods of determining whether anti-PACAP antibodies inhibit PACAP-associated photophobia or light aversion
This invention relates to methods of screening for anti-PACAP antibodies, or anti-PACAP receptor antibodies, and antigen binding fragments thereof, for potential use in treating or preventing PACAP-associated photophobia or light aversion, and therapeutic compositions containing and methods of using anti-PACAP antibodies, or anti-PACAP receptor antibodies, and antigen binding fragments thereof.
1. A method of determining whether an antibody or antigen binding fragment inhibits Pituitary Adenylate Cyclase-Activating Peptide (PACAP)-associated photophobia or light aversion, which comprises:
(i) providing at least one first test subject and at least one second test subject;
(ii) administering PACAP to the at least one first test subject and the at least one second test subject;
(iii) after step (ii) further administering to the at least one first test subject an anti-PACAP antibody or antigen binding fragment comprising
(a) a heavy chain variable region (VH) comprising a complementarity determining region 1 (CDR1) sequence consisting of the amino acid sequence of SEQ ID NO: 444, a complementarity determining region 2 (CDR2) sequence consisting of the amino acid sequence of SEQ ID NO: 446, and a complementarity determining region 3 (CDR3) sequence consisting of the amino acid sequence of SEQ ID NO: 448 and
(b) a light chain variable region (VL) comprising a CDR1 sequence consisting of the amino acid sequence of SEQ ID NO: 464, a CDR2 sequence consisting of the amino acid sequence of SEQ ID NO: 466, and a CDR3 sequence consisting of the amino acid sequence of SEQ ID NO: 468;
(iv) comparing the response of the at least one first test subject and at least one second test subject to light; and
(v) based on the comparison in (iv), identifying whether the anti-PACAP antibody or antigen binding fragment yields a decreased light aversion or decreased photophobia in the at least one first test subject as compared with the at least one second test subject, thereby determining whether the anti-PACAP antibody or antigen binding fragment inhibits PACAP-associated photophobia or light aversion.
2. A method of determining whether an antibody or antigen binding fragment inhibits Pituitary Adenylate Cyclase-Activating Peptide (PACAP)-associated photophobia or light aversion, which comprises:
(i) providing at least one first test subject and at least one second test subject;
(ii) administering to the at least one first test subject an anti-PACAP antibody or antigen binding fragment comprising
(a) a VH comprising a CDR1 sequence consisting of the amino acid sequence of SEQ ID NO: 444, a CDR2 sequence consisting of the amino acid sequence of SEQ ID NO: 446, and CDR3 sequence consisting of the amino acid sequence of SEQ ID NO: 448 and
(b) a VL comprising a CDR1 sequence consisting of the amino acid sequence of SEQ ID NO: 464, a CDR2 sequence consisting of the amino acid sequence of SEQ ID NO: 466, and a CDR3 sequence consisting of the amino acid sequence of SEQ ID NO: 468;
(iii) after step (ii) further administering PACAP to the at least one first test subject and the at least one second test subject;
(iv) comparing the response of the at least one first test subject and at least one second test subject to light; and
(v) based on the comparison in (iv), identifying whether the anti-PACAP antibody or antigen binding fragment yields a decreased light aversion or decreased photophobia in the at least one first test subject as compared with the at least one second test subject, thereby determining whether the anti-PACAP antibody or antigen binding fragment inhibits PACAP-associated photophobia or light aversion.
3. The method of claim 1 or 2 , wherein:
(I) the VH comprises the amino acid sequence of SEQ ID NO: 442;
(II) the VL comprises the amino acid sequence of SEQ ID NO: 462; or
(III) the VH comprises the amino acid sequence of SEQ ID NO: 442 and the VL comprises the amino acid sequence of SEQ ID NO: 462.
4. The method of claim 1 or 2 , wherein said anti-PACAP antibody or antigen binding fragment comprises:
(I) a heavy chain polypeptide comprising the amino acid sequence of SEQ ID NO: 441;
(II) a light chain polypeptide comprising the amino acid sequence of SEQ ID NO: 461; or
(III) a heavy chain polypeptide comprising the amino acid sequence of SEQ ID NO: 441 and a light chain polypeptide comprising the amino acid sequence of SEQ ID NO: 461.
5. The method of claim 1 or 2 , wherein said anti-PACAP antibody or antigen binding fragment is humanized or chimeric.
6. The method of claim 1 or 2 , wherein said anti-PACAP antibody or antigen binding fragment comprises an antigen binding fragment selected from a single chain antibody, fragment antigen binding (“Fab”) fragment, Fab′ fragment, monovalent antibody fragment, and a F(ab′)2 fragment of the antibody.
7. The method of claim 1 or 2 , wherein said anti-PACAP antibody or antigen binding fragment comprises—(a) a VH comprising a CDRI sequence consisting of the amino acid sequence of SEQ ID NO: 444, a CDR2 sequence consisting of the amino acid sequence of SEQ ID NO: 446, and a CDR3 sequence consisting of the amino acid sequence of SEQ ID NO: 448; (b) a VL comprising a CDRI sequence consisting of the amino acid sequence of SEQ ID NO: 464, a CDR2 sequence consisting of the amino acid sequence of SEQ ID NO: 466, and a CDR3 sequence consisting of the amino acid sequence of SEQ ID NO: 468; and (c) a human Fc region of a human IgG which is selected from a human IgG1, IgG2, IgG3, or IgG4.
8. The method of claim 7 , wherein the amino acid sequence of the Fc region of the human IgG selected from the human IgG1, IgG2, IgG3, or IgG4 comprises a mutation to eliminate: (i) at least one N-glycosylation; (ii) at least one O-glycosylation; or (iii) at least one N-glycosylation and at least O-glycosylation.
9. The method of claim 1 or 2 , wherein said anti-PACAP antibody or antigen binding fragment entirely lacks:
(i) N-glycosylation;
(ii) O-glycosylation; or
(iii) N-glycosylation and O-glycosylation.