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 canine is provided. Also provided are caninised antibodies which specifically bind to canine neuronal growth factor (NGF) and neutralise the ability of canine NGF to bind to the p75 or TrkA canine NGF receptor. The invention extends to nucleic acids encoding same and to methods of treating pain and arthritis in a canine using said antibodies and/or nucleic acids.

1 .- 90 . (canceled) 
     
     
         91 . A caninized antibody or a caninized antigen binding fragment thereof which is capable of specifically binding to canine nerve growth factor (NGF) and inhibiting the ability of canine NGF to bind to the p75 canine NGF receptor and/or TrkA canine NGF receptor, wherein the antibody or antigen binding fragment comprises:
 (a) a light chain comprising a light chain variable region comprising the amino acid sequence selected from the group consisting of SEQ ID NO: 1 and substitution variants thereof differing from SEQ ID NO: 1 only by one or more amino acid substitutions selected from:   Q3V;   T5M, T51;   S7T;   A9L, A9P;   S12A;   L13V;   S14A, S14T;   Q15P, Q15R;   E16G, E16D;   E17D;   K18T, K18P, K18A, K18E, K18L;   V19A;   T20S;   T22S, T22Y;   C23Y;   Y36F, Y361, Y36L;   Q37R, Q371, Q37L;   Q38H;   K39R;   P40A, P40S;   G41D;   A43S, A43P, A43T;   K45Q, K45E, K45R;   L46R, L46A, L46G, L46P, L46S;   148L;   Y49E, Y49F, Y49N, Y9S, Y49V;   G47A;   V58A;   P59S;   S60D;   F62L, F62V;   S631;   G64A;   T69A;   E70D;   Y71F;   S72T, S72R;   L73F;   T74K, T74R;   N76S;   L78E;   S80P, S80A;   E81D, E81G, E811, E81N;   V83A, V83G, V83S, V83T;   A84G;   V851, V85L;   F87Y;   G97S;   A100P, A100Q, A100T;   G101E;   T102P;   K103Q, K103S;   V104L, V104W;   E105D, E105R; and   L1061; and   (b) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence selected from the group consisting of SEQ ID NO:2 and substitution variants thereof differing from SEQ ID NO: 2 only by one or more amino acid substitutions selected from:   E1D, E1G;   V2E, V2G, V21, V2L, V2M;   Q3A, Q3E, Q3H, Q3K, Q3L, Q3P, Q3R, Q3S, Q3V;   L4P, L4V;   V5A V5E, V5L, V5M;   E6A, E6Q;   S7F, S7L, S7T;   G9E;   D10G, D10A, D10E, D10N, D10T;   L11Q, L11R, L11V, L11W;   V12A, V121, V12M;   N13Q, N13K, N13R;   P14S, P14T;   G15T, G15A, G15E;   G16E, G16A;   T17S, T17P;   L18R;   T19R, T19G, T19K, T19V;   L20I, L20V;   S21Y;   V23A, V23E, V231, V23L;   V24A, V241, V24G, V24S, V24T;   S25G, S25P, S25T;   G26P, G26R, G26T;   F27P, F271, F27L, F27S, F27T, F27V;   S28T, S28A, S28D, S281, S28L, S28M, S28N, S28P, S28R;   L29F, L291, L29M, L29V;   T30S, T30D, T30G, T30H, T30I, T30K, T30N, T30R, T30V;   W36C;   V37A, V37F, V371, V37L;   Q39H, Q39L;   A40D, A40G, A40P, A40S, A40T, A40V;   L41P;   G42E, G42L, G42R;   R43K, R43A, R43C, R43G, R43M, R43Q;   G44D, G44E, G44R, G44T, G44V;   L45F, L45M, L45P;   E46Q, E46D, E46H, E46L, E46P, E46R;   W47C, W47F, W47L, W47M, W47S, W47Y;   V48F, V481, V48L;   G49A, G49L, G49S, G49T;   R66Q;   L67F, L77V;   T68A, T681, T68S;   169L, I69M, 169T, 169V;   T70S, T70A, T70F;   R71K;   D72E, D72N;   T73N, T73D, T73G, T731, T73S;   S74A, S74D, S74G, S74P, S74T, S74V;   K75E, K75G, K75M, K75N, K75Q, K75R;   S76N, S76D, S76H, S76K, S76R;   T77A, 1771, T77M, T77S;   V78L, V78A, V781, V78M;   F79Y, F79H, F79S, F79T;   L80I;   K81Q, K81A, K81D, K81H, K81E, K81R;   M82L;   H83N, H83D, H83K, H83P, H83R, H83S, H83T;   S84D, S84G, S84N, S84R, S84T;   L85V;   Q86R, Q86G, Q86I, Q86K, Q86S, Q86T;   S87A, S87D, S87G, S87P, S87T, S87V;   E88A, E88D, E88V;   T90A, T90M, T90S;   A91G, A91V;   T92V, T92F, T921, T92L, T92K, T92L, T92M, T92Q;   Y93H;   Y94F, Y94H;   A96C, A96G, A96L, A96M, A96R, A96S, A96T, A96V;   R97K, R97A, R97D, R97E, R97G, R971, R97L, R97M, R97N, R97P, R97Q, R97S, R97T, R97V;   W112L;   G113A, G13S;   Q114D, Q114H, Q114P, Q114R;   T116A, T16I, T116N, T116S;   L117S, L117P, L117Q, L117R;   V1181, V118L, V118P;   T119A, T119F, T1191, T119L, T119P, T119S, T119Y;   V120A;   S121A, S121C, S121P, S121T; and   S122L, S122P.   
     
     
         92 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the light chain variable region comprises a substitution variant of SEQ ID NO: 1 having at least 85% identity to SEQ ID NO: 1. 
     
     
         93 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 1. 
     
     
         94 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the heavy chain variable region comprises a substitution variant of SEQ ID NO: 2 having at least 85% identity to SEQ ID NO: 2. 
     
     
         95 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 2. 
     
     
         96 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the light chain variable region comprises a substitution variant of SEQ ID NO: 1 having at least 85% identity to SEQ ID NO: 1, and the heavy chain variable region comprises a substitution variant of SEQ 1D NO: 2 having at least 85% identity to SEQ ID NO: 2. 
     
     
         97 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 1 and the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 2. 
     
     
         98 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the light chain variable region comprises a substitution variant of SEQ ID NO: 1 differing from SEQ ID NO: 1 by substitutions Q3V, Y71F, N76S, S80P, and F87Y, and the heavy chain variable region comprises a substitution variant of SEQ ID NO: 2 differing from SEQ ID NO: 2 by substitutions K81Q and Q86R. 
     
     
         99 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 3 and the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 4. 
     
     
         100 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the light chain comprises the amino acid sequence selected from SEQ ID NO: 5 and SEQ ID NO: 10. 
     
     
         101 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the heavy chain comprises the amino acid sequence selected from SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 9. 
     
     
         102 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the heavy chain comprises the amino acid sequence selected from SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13 and SEQ ID NO: 14. 
     
     
         103 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the heavy chain comprises the amino acid sequence selected from SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17 and SEQ ID NO: 18. 
     
     
         104 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the heavy chain comprises the amino acid sequence selected from SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21 and SEQ ID NO: 22. 
     
     
         105 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the heavy chain comprises the amino acid sequence selected from SEQ ID NO: 6, SEQ ID NO: 9, SEQ ID NO: 15 and SEQ ID NO: 18. 
     
     
         106 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , wherein the heavy chain comprises the amino acid sequence selected from SEQ ID NO: 11, SEQ ID NO: 14, SEQ ID NO: 19, and SEQ ID NO: 22. 
     
     
         107 . The caninized antibody or caninized antigen binding fragment thereof as claimed in  claim 91 , comprising a type HCA or HCD canine heavy chain constant domain or an aglycosylated type HCB, HCA or HCD canine heavy chain constant domain.