Nucleic acids encoding inhibitory anti-factor XII/XIIa monoclonal antibodies

Patent metadata

Field	Value
Publication number	US-9856325-B2
Application number	US-201615343401-A
Country	US
Kind code	B2
Filing date	Nov 4, 2016
Priority date	Jul 22, 2011
Publication date	Jan 2, 2018
Grant date	Jan 2, 2018

Abstract

The invention relates to inhibitory anti-factor XII/FXIIa antibodies and methods of their use.

First claim

The invention claimed is:

1. A nucleic acid encoding an anti-Factor XII/XIIa monoclonal antibody or antigen-binding fragment thereof comprising an immunoglobulin heavy chain variable (vH) region and an immunoglobulin light chain variable (vL) region,
wherein the vH region comprises:
a heavy chain CDR1 consisting of the sequence of SEQ ID NO: 6;
a heavy chain CDR2 consisting of the sequence of GIX 1 X 2 X 3 X 4 X 5 X 6 TVYADSVKG (SEQ ID NO: 8), wherein X 1 is R, N, or D; X 2 is P, V, I, or M; X 3 is S, P, or A; X 4 is G, L, V, or T; X 5 can be any amino acid; and X 6 is T, G, or S; and
a heavy chain CDR3 consisting of the sequence of ALPRSGYLX 1 X 2 X 3 X 4 YYYYALDV (SEQ ID NO: 10), wherein X 1 is I, M, or V; X 2 is S or K; X 3 is P, K, T, or H; and X 4 is H, N, G, or Q; and
wherein the vL region comprises:
a light chain CDR1 consisting of the sequence set forth in any one of SEQ ID NOs: 11 and 44-51;
a light chain CDR2 consisting of the sequence of SEQ ID NO: 12; and
a light chain CDR3 consisting of the sequence of AX 1 WX 2 X 3 X 4 X 5 RX 6 X 7 (SEQ ID NO: 14), wherein X 1 is A or S; X 5 is L or V; X 6 is G, L, or K; and X 2 , X 3 , X 4 and X 7 can be any amino acid.

2. A vector comprising the nucleic acid of claim 1 , operably linked to a promoter sequence.

3. A cell comprising the vector of claim 2 .

4. A method of producing an anti-Factor XII/XIIa monoclonal antibody or antigen-binding fragment thereof, the method comprising (1) culturing the cell according to claim 3 under conditions to express the anti-Factor XII/XIIa monoclonal antibody or antigen-binding fragment thereof; and (2) purifying the anti-Factor XII/XIIa monoclonal antibody or antigen-binding fragment thereof from the culture supernatant.

5. The nucleic acid of claim 1 , wherein the vH region comprises an amino acid sequence more than 85% identical to the sequence of SEQ ID NO: 4.

6. The nucleic acid of claim 1 , wherein the vL region comprises an amino acid sequence more than 85% identical to the sequence of SEQ ID NO: 5.

7. The nucleic acid of claim 1 , wherein the heavy chain CDR2 consists of the sequence set forth in any one of SEQ ID NOs: 7 and 29-38.

8. The nucleic acid of claim 1 , wherein the heavy chain CDR3 consists of the sequence set forth in any one of SEQ ID NOs: 9 and 39-43.

9. The nucleic acid of claim 1 , wherein
X 5 is G, Y, Q, K, R, N, or M in the heavy chain CDR2;
X 2 is D, Y, E, T, W, E, or S in the light chain CDR3;
X 3 is A, N, I, L, V, P, Q, or E in the light chain CDR3;
X 4 is S, D, P, E, Q, or R In the light chain CDR3; and/or
X 7 is V, A, D, T, M, or G in the light chain CDR3.

10. The nucleic acid of claim 1 , wherein the light chain CDR3 consists of the sequence set forth in any one of SEQ ID NOs: 13 and 52-63.

11. The nucleic acid of claim 1 , wherein the vH region comprises a heavy chain CDR1 consisting of the sequence of SEQ ID NO: 6, a heavy chain CDR2 consisting of the sequence of SEQ ID NO: 7, a heavy chain CDR3 consisting of the sequence of SEQ ID NO: 9, and wherein the vL region comprises a light chain CDR1 consisting of the sequence of SEQ ID NO: 11, a light chain CDR2 consisting of the sequence of SEQ ID NO: 12, and a light chain CDR3 consisting of the sequence of SEQ ID NO: 13.

12. The nucleic acid of claim 1 , wherein the vH region consists of the sequence of SEQ ID NO: 4, and wherein the vL region consists of the sequence of SEQ ID NO: 5.

13. The nucleic acid of claim 1 , wherein the vH region comprises a heavy chain CDR1 consisting of the sequence of SEQ ID NO: 6, a heavy chain CDR2 consisting of the sequence of SEQ ID NO: 29, a heavy chain CDR3 consisting of the sequence of SEQ ID NO: 9, and wherein the vL region comprises a light chain CDR1 consisting of the sequence of SEQ ID NO: 11, a light chain CDR2 consisting of the sequence of SEQ ID NO: 12, and a light chain CDR3 consisting of the sequence of SEQ ID NO: 13.

14. The nucleic acid of claim 1 , wherein the vH region comprises a heavy chain CDR1 consisting of the sequence of SEQ ID NO: 6, a heavy chain CDR2 consisting of the sequence of SEQ ID NO: 30, a heavy chain CDR3 consisting of the sequence of SEQ ID NO: 9, and wherein the vL region comprises a light chain CDR1 consisting of the sequence of SEQ ID NO: 11, a light chain CDR2 consisting of the sequence of SEQ ID NO: 12, and a light chain CDR3 consisting of the sequence of SEQ ID NO: 13.

15. The nucleic acid of claim 1 , wherein the vH region comprises a heavy chain CDR1 consisting of the sequence of SEQ ID NO: 6, a heavy chain CDR2 consisting of the sequence of SEQ ID NO: 31, a heavy chain CDR3 consisting of the sequence of SEQ ID NO: 9, and wherein the vL region comprises a light chain CDR1 consisting of the sequence of SEQ ID NO: 11, a light chain CDR2 consisting of the sequence of SEQ ID NO: 12, and a light chain CDR3 consisting of the sequence of SEQ ID NO: 13.

16. The nucleic acid of claim 1 , wherein the vH region comprises a heavy chain CDR1 consisting of the sequence of SEQ ID NO: 6, a heavy chain CDR2 consisting of the sequence of SEQ ID NO: 32, a heavy chain CDR3 consisting of the sequence of SEQ ID NO: 9, and wherein the vL region comprises a light chain CDR1 consisting of the sequence of SEQ ID NO: 11, a light chain CDR2 consisting of the sequence of SEQ ID NO: 12, and a light chain CDR3 consisting of the sequence of SEQ ID NO: 13.

17. The nucleic acid of claim 1 , wherein the vH region comprises a heavy chain CDR1 consisting of the sequence of SEQ ID NO: 6, a heavy chain CDR2 consisting of the sequence of SEQ ID NO: 7, a heavy chain CDR3 consisting of the sequence of SEQ ID NO: 9, and wherein the vL region comprises a light chain CDR1 consisting of the sequence of SEQ ID NO: 44, a light chain CDR2 consisting of the sequence of SEQ ID NO: 12, and a light chain CDR3 consisting of the sequence of SEQ ID NO: 13.

18. The nucleic acid of claim 1 , wherein the antibody or antigen-binding fragment thereof has a more than 2 fold higher binding affinity to human Factor XIIa-beta than to human Factor XII and is capable of inhibiting the amidolytic activity of human Factor XIIa at a concentration of 100 nM or lower in an in vitro amidolytic activity assay by 80% or more.

19. The nucleic acid of claim 1 , wherein the antibody or antigen-binding fragment thereof inhibits the amidolytic activity of Factor XIIa-alpha by more than 50% in an in vitro amidolytic activity assay when used at a molar ratio of FXIIa-alpha to antibody of 1:0.2.

20. The nucleic acid of claim 1 , wherein the antibody or antigen-binding fragment thereof binds murine FXII/FXIIa; and wherein the antibody or antigen-binding fragment thereof binds to a polypeptide comprising the sequence of SEQ ID NO: 2 in which (a) the asparagine residue at position 398 of SEQ ID NO: 2 is substituted for lysine; or (b) the isoleucine residue at position 438 of SEQ ID NO: 2 is substituted for alanine, and wherein the affinity of the antibody or antigen-binding fragment thereof for the polypeptide in (a) or (b) is lower than the affinity of the antibody or antigen-binding fragment thereof for a polypeptide comprising SEQ ID NO: 2 without the corresponding substitution.

21. The nucleic acid of claim 1 , wherein the antibody or antigen-binding fragment thereof binds human Factor XIIa-beta with a K D of better than 10 −7 M.

22. The nucleic acid of claim 1 , wherein the antibody or antigen-binding fragment thereof competes with Infestin for binding to human Factor XIIa-beta.

Assignees

• Csl Behring Gmbh
• Csl Ltd

Inventors

• Panousis Con
• Rayzman Veronika
• Nash Andrew
• Wilson Michael
• Schmidbauer Stefan
• Nolte Marc

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title

• A61P7/10

  Antioedematous agents; Diuretics · CPC title

• A61P7/02

  Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title