Anti-nerve growth factor antibodies and methods of preparing and using the same

Abstract

A method of preparing an antibody suitable for use in a feline is provided. Also provided are chimeric and felinised antibodies which specifically bind to feline neuronal growth factor (NGF) and neutralise the ability of feline NGF to bind to the p75 or TrkA feline NGF receptor. The invention extends to nucleic acids encoding same and to methods of treating pain and arthritis in a feline using said antibodies and/or nucleic acids.

1 .- 128 . (canceled) 
     
     
         129 . A method of preparing an anti-nerve growth factor antibody suitable for use in a feline comprising:
 comparing (i) sequences of framework regions of a donor antibody from a species other than a feline, wherein the donor antibody has binding specificity for feline nerve growth factor (NGF) with (ii) sequences of framework regions of one or more feline antibodies, and   modifying the framework regions of the donor antibody to substitute amino acid residues that are different at a corresponding Kabat position in the one or more feline antibodies with amino acid residues present at the corresponding Kabat position in the one or more feline antibodies to produce a modified antibody,   wherein the modified antibody does not contain any amino acid in any position within the framework regions which would be foreign at the corresponding Kabat position in the one or more feline antibodies; and   wherein the modified antibody is capable of specifically binding to feline NGF and inhibiting the ability of feline NGF to bind to the p75 feline NGF receptor and/or TrkA feline NGF receptor.   
     
     
         130 . The method of  claim 129 , wherein one or more of the one or more amino acid residues present at the corresponding Kabat position in the one or more feline antibodies used to modify the framework regions of the donor antibody have the highest homology to the substituted amino acid residue of the donor antibody. 
     
     
         131 . The method of  claim 129 , further comprising replacing at least one constant domain of the heavy and/or light chain of the donor antibody with a constant domain of a heavy and/or light chain of a feline antibody. 
     
     
         132 . The method of  claim 131 , wherein the constant domain of the heavy chain of the donor antibody is replaced with a feline heavy chain constant domain as shown in Genbank Accession No. BAA32229.1 (SEQ ID NO:34). 
     
     
         133 . The method of  claim 129 , wherein the modified antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 or an amino acid sequence which has a sequence identity of at least 85% thereto and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:4 or a sequence which has an amino acid identity of at least 85% thereto. 
     
     
         134 . The method of  claim 129 , wherein the modified antibody comprises a light chain variable region comprising:
 an FR1 framework region comprising the amino acid sequence of SEQ ID NO:26 or an amino acid sequence which has a sequence identity of at least 85% thereto;   an FR2 framework region comprising the amino acid sequence of SEQ ID NO:27 or an amino acid sequence which has a sequence identity of at least 85% thereto;   an FR3 framework region comprising the amino acid sequence of SEQ ID NO:28 or an amino acid sequence which has a sequence identity of at least 85% thereto; and   an FR4 framework region comprising the amino acid sequence of SEQ ID NO:29 or an amino acid sequence which has a sequence identity of at least 85% thereto;   and a heavy chain variable region comprising:   an FR1 framework region comprising the amino acid sequence of SEQ ID NO:30 or an amino acid sequence which has a sequence identity of at least 85% thereto;   an FR2 framework region comprising the amino acid sequence of SEQ ID NO:31 or an amino acid sequence which has a sequence identity of at least 85% thereto;   an FR3 framework region comprising the amino acid sequence of SEQ ID NO:32 or an amino acid sequence which has a sequence identity of at least 85% thereto; and   an FR4 framework region comprising the amino acid sequence of SEQ ID NO:33 or an amino acid sequence which has a sequence identity of at least 85% thereto.   
     
     
         135 . A method for treating, inhibiting or ameliorating pain in a feline subject, or for the treatment of arthritis or an arthritic condition in a feline subject, or for the treatment of a condition caused by, associated with or resulting in the increased expression of feline NGF or increased sensitivity to NGF in a feline subject, or for the treatment of a tumour induced to proliferate by NGF in a feline and conditions associated therewith, comprising administering to a feline in need thereof a modified antibody prepared by the method of  claim 129 . 
     
     
         136 . A kit for the treatment of pain in felines, or for the treatment of a condition associated with pain, or for the treatment, amelioration or inhibition of pain associated with osteoarthritis, inflammation, pruritis, immune mediated polyarthritis or rheumatoid arthritis comprising a modified antibody prepared by a method of  claim 129  and instructions for use of the same.