US2020247885A1PendingUtilityA1
P2x4 antibodies & uses thereof
Est. expiryMay 2, 2034(~7.8 yrs left)· nominal 20-yr term from priority
Inventors:Wendy A WilliamsClare JonesJames ButtonJohn LinleyHarm Jan SnijderLing HuangYoko ShibataSudharsan SridharanMaria GrovesClaire Louise Dobson
C07K 2317/92C07K 2317/622C07K 2317/56C07K 2317/21C07K 16/28C07K 2317/24C07K 2317/34C07K 2317/76A61P 25/04A61K 2039/505C07K 2317/565C07K 2317/75
50
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Claims
Abstract
The invention provides antibodies that specifically bind a P2X4 polypeptide and modulate P2X4 channel activity, recombinant P2X4 polypeptides and methods for generating such polypeptides, as well as compositions and methods for generating anti-P2X4 antibodies, and methods of using P2X4 antibodies for the treatment of neuropathic pain and other indications.
Claims
exact text as granted — not AI-modifiedWhat is claimed is:
1 . An antibody or antigen binding fragment thereof that specifically binds a human P2X4 polypeptide and modulates channel activity.
2 . The antibody of claim 1 , wherein the antibody is a P2X4 potentiator.
3 . The antibody of claim 1 , wherein the antibody is a P2X4 antagonist.
4 . The antibody of claim 1 , wherein the antibody binds an epitope comprising human P2X4 amino acids 110-166.
5 . The antibody of claim 4 , wherein the antibody binds an epitope comprising one or more human P2X4 amino acids selected from the group consisting of amino acids 118, 122-139, 145, 159, 180, 183, 184, 231, and 244.
6 . The antibody of claim 1 , wherein the antibody or fragment thereof comprises:
a. a heavy chain variable region CDR1 comprising a sequence:
(SEQ ID NO: 1009)
X 1 X 2 X 3 X 4 X 5 ,
wherein
X 1 is G, N, S, D, or R;
X 2 is Y, A, H, F, or S;
X 3 is A, W, Y, S, G, F, W, E, D, or P;
X 4 is M, I, W, L, I, F, or V;
X 5 is S, G, T, H, or N; and/or
b. a heavy chain variable region CDR2 comprising a sequence:
(SEQ ID NO: 1010)
X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 X 15 X 16 ,
wherein
X 1 is A, R, I, T, E, S, A, V, W, N, G, E, R, or Y;
X 2 is I or M;
X 3 is S, K, Y, D, N, W, or I;
X 4 is S, D, G, H, N, R, Y, or V;
X 5 is G, D, S, F, N, R, F, D, or T;
X 6 is G, S, N, or T;
X 7 is S, T, D, Y, N, A, E, M, F, or D;
X 8 is T, I, K, or A;
X 9 is Y, D, R, N, G, Q, E, H, or K;
X 10 is Y, Q, S, or V;
X 11 is A, S, N, or V;
X 12 is D, A, P, R, or Q;
X 13 is S, P, K, or N;
X 14 is V, F, L, or A;
X 15 is K, Q, or E;
X 16 is G, S, A, or D;
wherein the heavy chain variable region CDR2 optionally comprises an insertion of 1-3 amino acids, X a X b X c between amino acids X 3 and X 4 , wherein
X a is G, S, P, W, Y, E, A, R, or N; and
X b X c are KT, respectively
and/or
c. a heavy chain variable region CDR3 comprising a sequence:
(SEQ ID NO: 1011)
X 1 X 2 X3X 4 X 5 X 6 X 7 X 8 ,
X 1 is E, N, D, R, K, G, S, A, Y, V, P, or H;
X 2 is E, L, R, Q, T, G, F, P, Y, K, A, S, V, or F;
X 3 is R, A, T, G, V, S, M, W, Y, D, H, N, E, L, or I;
X 4 is G, L, R, D, T, G, Y, S, E, F, Q, C, I, M, V, N, K, or P;
X 5 is S, G, Y, D, W, T, S, N, I, D, V, E, or C;
X 6 is Y, A, S, W, T, L, G, E, F, K, V, I, or D;
X 7 is D, E, or G; and
X 8 is Y, S, V, L, M, Q, I, S, I, H, F, or D;
wherein the heavy chain variable region CDR3 optionally comprises an insertion of 1-14 amino acids X a -X n (SEQ ID NO: 1012), wherein
X a is F, R, S, Y, L, D, G, V, I, T, or A;
X b is G, R, Y, F, T, D, S, G, V, M, D, or R;
X c is F, W, A, G, T, I, S, F, Y, C, L, V, R, or N;
X d is S, F, M, G, Y, L, S, A, D, L, R, V, C, or S;
X e is G, Y, S, T, P, F, Y, R, A, E, G, Q, N, or L;
X f is Y, N, G, T, R, F, A, M, W, P, or V;
X g is Y, M, S, V, F, A, P, S, D, R, H, P, E, or R
X h is Y, G, M, F, G, P, V, F, H, T, or G,
X i is T, I G, R, or F;
X j is Y, G, H, or E;
X k is Y, G, F, or N;
X 1 is F, or N;
X m is Y; and
X n is F.
7 . The antibody of claim 1 , wherein the antibody or fragment thereof comprises,
a. a light chain variable region CDR1 comprising a sequence:
(SEQ ID NO: 1013)
X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 ,
wherein
X 1 is T, G, R, S, or Q;
X 2 is G, A, or L;
X 3 is S, T, D, or H;
X 4 is S, N, K, A, Q, T, or V;
X 5 is G, I, L, S, or D;
X 6 is A, G, R, P, I, D, S, E, or T;
X 7 is G, N, M, D, K, S, R, Y, or T;
X 8 is Y, K, F, Q, S, N, Y, D, H, or R;
X 9 is D, N, Y, W, F, M, G, or S;
X 10 is V, A, L, I, G, or P;
X 11 is H, T, S, Y, A, Q, Y, N, or F;
wherein the light chain variable region CDR1 optionally comprises an insertion of between 1 and 3 amino acids X a -X c between X 4 and X 5 , wherein
X a is S or G;
X b is N, D or S; and
X c is I or V;
b. a light chain variable region CDR2 comprising a sequence:
(SEQ ID NO: 1014)
X 1 X 2 X 3 X 4 X 5 X 6 X 7 ,
wherein
X 1 is G, Y, Q, K, N, D, R, A, or E;
X 2 is N, D, K, A, V, G, or T;
X 3 is N, S, T, I, K, Y, or D;
X 4 is N, D, Y, K, E, T, N, S, or Q;
X 5 is R, or L;
X 6 is P, E, A, S, or Q;
X 7 is S, P, or T; and/or
c. a light chain variable region CDR3 comprising a sequence:
(SEQ ID NO: 1015)
X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 ,
wherein
X 1 is Q, N, A, G, D, S, or L;
X 2 is S, V, A, Q, T, L, or H;
X 3 is Y, W, R, A, S, Q, T, or G;
X 4 is D, Y, I, N, M, or H;
X 5 is T, M, S, N, D, R, G, or K;
X 6 is N, T, S, G, F, L, or D;
X 7 is L, T, G, P, A, I, or N;
X 8 is K, W, V, I, P, G, L, R, or Y; and
X 9 is V, L, or T;
wherein the light chain variable region CDR3 optionally comprises an insertion of between 1 and 3 amino acids X a -X c between X 7 and Xs, wherein
X a is D, N, A, T, S, I or H;
X b is H, Y, G, A, R, L, S, or P; and
X c is S.
8 . The antibody of claim 1 , wherein the antibody or fragment thereof comprises a V H comprising:
a. a heavy chain variable region CDR1 comprising the sequence: SX 1 AMS (SEQ ID NO: 1016) where X 1 is Y or F; b. a heavy chain variable region CDR2 comprising the sequence: AISGSG XISTYYADSVKG (SEQ ID NO: 1017) where X 1 is S or G; c. a heavy chain variable region CDR3 comprising the sequence: X 1 X 2 DX 3 WSX 4 X 5 X 6 X 7 X 8 TAFDL (SEQ ID NO: 1018), where X 1 is H, D or Q; X 2 is W, M, F, H, or R; X 3 is W, Y or F; X 4 is T, N, G, or P; X 5 is R, A, S, G, or Y; X 6 is S, P, N or T; X 7 is G, S, R, or K: X 8 is P, M, A or L. optionally in combination with a V L comprising a. a light chain variable region CDR1 comprising the sequence SGDALPRQYAY (SEQ ID NO: 1019) b. a light chain variable region CDR2 comprising the sequence KDSX 1 RPS (SEQ ID NO: 1020) where X 1 is E or F. c. a light chain variable region CDR3 comprising the sequence QSADSSGTYXIV (SEQ ID NO: 1021), where X 11 s V or A.
9 . The antibody of claim 8 , wherein the antibody or fragment thereof comprises a V H comprising:
a. a heavy chain variable region CDR1 comprising the sequence: SFAMS (SEQ ID NO: 813); b. a heavy chain variable region CDR2 comprising the sequence: AISGSG GSTYYADSVKG (SEQ ID NO: 814); c. a heavy chain variable region CDR3 comprising the sequence:
(SEQ ID NO: 815)
QFDYWSTYSGPTAFDL;
in combination with a V L comprising
a. a light chain variable region CDR1 comprising the sequence SGDALPRQYAY (SEQ ID NO: 822)
b. a light chain variable region CDR2 comprising the sequence KDSERPS (SEQ ID NO: 823)
c. a light chain variable region CDR3 comprising the sequence QSADSSGTYVV (SEQ ID NO: 824).
10 . The antibody of claim 1 , wherein the antibody or fragment thereof comprises:
a. a heavy chain variable region CDR1 comprising a sequence:
(SEQ ID NO: 1022)
X 1 X 2 X 3 X 4 X 5 ,
wherein
X 1 is S, N, D, T, or R;
X 2 is G, Y, or F;
X 3 is Y, H, S, G, D, or F;
X 4 is D, V, or I;
X 5 is N, H, C, R, S or is absent;
wherein the heavy chain variable region CDR1 optionally comprises amino acids X 6 and X 7 , which are V and S, respectively;
b. a heavy chain variable region CDR2 comprising a sequence:
(SEQ ID NO: 1023)
X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 X 15 X 16 ,
wherein
X 1 is M, V, L, I, A, G, or T;
X 2 is G or I;
X 3 is Y, W, N, or C;
X 4 is Y, G, D, or W;
X 5 is S, D, or E;
X 6 is G or D;
X 7 is S, Y, N, or I;
X 8 is T, P, or K;
X 9 is N, A, G, D, or V;
X 10 is Y or F;
X 11 is N;
X 12 is P, S, or E;
X 13 is S, A, or N;
X 14 is L or F;
X 15 is K;
X 16 is S, G, or N;
wherein the heavy chain variable region CDR2 optionally comprises an insertion of amino acids X a and X b between X 3 and X 4 , wherein X a is I or P and X b is S;
c. a heavy chain variable region CDR3 comprising a sequence:
(SEQ ID NO: 1024)
X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 ,
wherein
X 1 is G, S, A, or R;
X 2 is M, G, Y, S, L, R, or V;
X 3 is M, D, I, V, H, M, or S;
X 4 is V, Y, M, W, or S;
X 5 is L, Y, S or absent;
X 6 is I, D, V, T, G, S, Y, or absent;
X 7 is P, G, D, S, or A; and
X 8 is N, Y, or T;
wherein the heavy chain variable region CDR3 optionally comprises an insertion of between 1 and 6 amino acids of X a -X f (SEQ ID NO: 1025) between X 6 and X 7 , wherein
X a is G, T, D, or Y;
X b is S, A, G, or F;
X c is Y, V, P, or F;
X d is Y or F;
X e is Y; and
X f is E, F, or G.
11 . The antibody of claim 1 , wherein the antibody or fragment thereof comprises:
a. a light chain variable region CDR1 comprising a sequence:
(SEQ ID NO: 1026)
X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 ,
wherein
X 1 is K, Q, or R;
X 2 is A or T;
X 3 is S, R, or N;
X 4 is K or Q;
X 5 is S, D, R, N, L, or I;
X 6 is I or S;
X 7 is T, G, V, or N;
X 8 is N, S, H, or K;
X 9 is Y or W;
X 10 is L, M, or I;
X 11 is A, S, or Y;
b. a light chain variable region CDR2 comprising a sequence:
(SEQ ID NO: 1027)
X 1 X 2 X 3 X 4 X 5 X 6 X 7 ,
wherein
X 1 is S, D, E, or Y;
X 2 is G, A, or T;
X 3 is S or T;
X 4 is T, S, K, or A;
X 5 is L;
X 6 is Q, A, or V;
X 7 is S or D;
c. a light chain variable region CDR3 comprising a sequence:
(SEQ ID NO: 1028)
X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 ,
wherein
X 1 is Q, L, or H
X 2 is Q or K;
X 3 is Y, A, W, or T;
X 4 is Y, H, S, or D;
X 5 is E, S, R, T, or N;
X 6 is K, N, T, L, or H;
X 7 is P;
X 8 is Y, W, L, N, P, or R;
X 9 is T.
12 . A polynucleotide encoding the antibody or antigen binding fragment thereof of claim 1 .
13 . A host cell comprising a vector comprising the polynucleotide of claim 12 .
14 . A method for treating neuropathic pain, the method comprising administering to a patient in need thereof an effective amount of an antibody of claim 1 or antigen binding fragment thereof.
15 . A method for the large scale production of a recombinant P2X4 polypeptide, the method comprising expressing a human P2X4 protein in an SF9 cell at 27° C. for 72 hours;
extracting the P2X4 protein by solubilizing in a buffer comprising n-Dodecyl-beta-D-Maltoside, n-Dodecyl thio-Maltoside, CHAPS, and the Cholesteryl Hemisuccinate;
then isolating the solubilized protein.
16 . A pharmaceutical composition comprising one or more antibodies or antigen binding fragment thereof according to claim 1 and a pharmaceutically acceptable excipient.
17 . A pharmaceutical composition comprising one or more antibodies or antigen binding fragment thereof according to claim 8 and a pharmaceutically acceptable excipient.
18 . A polynucleotide encoding the antibody or antigen binding fragment thereof of claim 8 .
19 . A host cell comprising a vector comprising the polynucleotide of claim 18 .
20 . A method for treating neuropathic pain, the method comprising administering to a patient in need thereof an effective amount of an antibody of claim 8 or antigen binding fragment thereof.Cited by (0)
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