STAPHYLOCOCCUS AUREUS SPECIFIC ANTIBODIES AND USES THEREOF

• US 20130164308A1
• Filed: 12/18/2012
• Published: 06/27/2013
• Est. Priority Date: 12/22/2011
• Status: Active Grant

• Current Assignee: Rinat Neuroscience Corp. (Pfizer Inc.)
• Original Assignee: Rinat Neuroscience Corp. (Pfizer Inc.)
• Inventors: SHELTON, David Louis, STROP, Pavel, FOLETTI, Davide Luciano, CHAPARRO RIGGERS, Javier Fernando, GLANVILLE, Jacob Eli Gunn, SHAUGHNESSY, Lee Mary Beth, ZHAI, Wenwu
• Granted Patent: US 8,715,673 B2
• Time in Patent Office: Days
• US Class Current: 424/165.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 39/40 bacterial
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: C07K 16/1271 from Micrococcaceae (F), e.g. Staphylococcus
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2317/94 Stability, e.g. half-life, pH, temperature or enzyme-resistance

Abstract

The present invention provides antagonizing antibodies that bind to S. aureus alpha-toxin. The invention further provides a method of obtaining such antibodies and antibody encoding nucleic acids. The invention further relates to therapeutic methods for use of these antibodies for the treatment and/or prevention of staphylococcal disease, including, for example, pneumonia, bacteremia, sepsis, eye infection, and abscess.

Claims

1. An isolated antagonist antibody that specifically binds to S. aureus alpha-toxin and comprises:

a heavy chain variable region (VH) comprising a VH complementarity determining region one (CDR1), VH CDR2, and VH CDR3 of the VH sequence of SEQ ID NO;

35; and

a light chain variable region (VL) comprising a VL CDR1, VL CDR2, and VL CDR3.

2. The isolated antagonist antibody of claim 1, wherein the VL comprises a VL CDR1, VL CDR2, and VL CDR3 of the VL sequence of SEQ ID NO:

3.

3. The isolated antagonist antibody of claim 1, wherein the VH CDR1 comprises the amino acid sequence of SEQ ID NO:

5, 6 or 7, the VH CDR2 comprises the amino acid sequence of SEQ ID NO;

8 or 9, the VH CDR3 comprises the amino acid sequence of SEQ ID NO;

54, the VL CDR1 comprises the amino acid sequence of SEQ ID NO;

15, the VL CDR2 comprises the amino acid sequence of SEQ ID NO;

16, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO;

17.

4. The isolated antagonist antibody of claim 3, wherein the VH comprises the amino acid sequence of SEQ ID NO:

35 or a variant thereof with one or several conservative amino acid substitutions in residues that are not within a CDR, and the VL comprises the amino acid sequence of SEQ ID NO;

3 or a variant thereof with one or several amino acid substitutions in amino acids that are not within a CDR.

5. The isolated antagonist antibody of claim 3, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:

72 or 73 and a light chain comprising the amino acid sequence of SEQ ID NO;

23.

6. The isolated antagonist antibody of claim 1, wherein the antibody further comprises an immunologically inert constant region.

7. The isolated antagonist antibody of claim 6, wherein the antibody has an isotype that is selected from the group consisting of IgG₂, IgG_2Δ

a, IgG₄, IgG<sub>4Δ

b</sub>, IgG<sub>4Δ

c</sub>, IgG₄ S228P, IgG<sub>4Δ

b</sub> S228P and IgG<sub>4Δ

c</sub> S228P.

8. The isolated antagonist antibody of claim 6, wherein the constant region is aglycosylated Fc.

17. A pharmaceutical composition comprising the antagonist antibody of claim 1.

18. A cell line that recombinantly produces the antagonist antibody of claim 1.

19. A nucleic acid encoding the antagonist antibody of claim 1.

20. A method of protecting an individual from a staphylococcal disease, the method comprising administering to an individual suffering from or at risk for a staphylococcal disease an effective amount of the antagonist antibody of claim 1.

21. The method of claim 20, wherein the individual is suffering from a staphylococcal disease and/or a staphylococcal infection.

22. The method of claim 20, wherein the individual is suffering from a staphylococcal infection.

23. The method of claim 20, wherein the staphylococcal disease is selected from the group consisting of staphylococcal abscess, staphylococcal bacteremia, staphylococcal sepsis, staphylococcal eye infection, staphylococcal skin infection, staphylococcal soft tissue infection, staphylococcal osteomyelitis, staphylococcal mastitis, and staphylococcal lung disease.

24. The method of claim 20, wherein the method further comprises administering an effective amount of an antiinfective agent, an antibiotic agent, and an antimicrobial agent.

9. An isolated antagonist antibody that specifically binds to S. aureus alpha-toxin and binds to an epitope that is the same as or overlaps with the epitope on S. aureus alpha-toxin recognized by DF1.1.

10. The isolated antagonist antibody of claim 9, wherein the antibody recognizes an epitope comprising amino acid residues 30-32, 64-72, 78-79, 205, 207-208, 210-213, 253, 274, and 276 of the alpha-toxin amino acid sequence shown in SEQ ID NO:

74.

11. The isolated antagonist antibody of claim 9, wherein the antibody blocks alpha-toxin from binding to a cell membrane.

12. The isolated antagonist antibody of claim 9, wherein the antibody binds to alpha-toxin with an equilibrium dissociation constant of about 20 pM or less.

13. The isolated antagonist antibody of claim 9, wherein the antibody comprises a heavy chain variable region (VH) complementarity determining region three (CDR3) sequence of SEQ ID NO:

54 and variants thereof wherein the amino acid at position 1 is D, E or G;

the amino acid at position 2 is H, L, M, R, V or Y;

the amino acid at position 3 is E, G, L or Y;

the amino acid at position 4 is A, E, G, V or Y;

the amino acid at position 5 is A, I, N, R, S, T or Y;

the amino acid at position 6 is G, R, S or Y;

the amino acid at position 7 is D, G, Q or R;

the amino acid at position 8 is C, D, G, N, R, S, or W;

the amino acid at position 9 is G, Q, R, S or Y;

the amino acid at position 10 is G, L, P or R;

the amino acid at position 11 is A, H, P, Q or T;

the amino acid at position 12 is D, E, G, H, K, N or S;

the amino acid at position 13 is A, L, P or V;

the amino acid at position 14 is F or L;

the amino acid at position 15 is D, E or S; and

the amino acid at position 16 is F, I, L or V.

14. The isolated antagonist antibody of claim 13, wherein the antibody comprises:

a VH CDR1 comprising the amino acid sequence of SEQ ID NO;

5, 6 or 7; and

a VH CDR2 comprising the amino acid sequence of SEQ ID NO;

8 or 9.

15. The isolated antagonist antibody of claim 13, wherein the antibody comprises a light chain variable region (VL) derived from a human germline V_κ

sequence selected from the group consisting of IGKV3-20, IGKV3-15, IGKV1-39, IGKV3-11, IGKV2-28, IGKV1-12, IGKV1-9, IGKV4-1, IGKV3D-20, IGLV1-40, IGKV1-5, IGKV3-NL5, IGKV1-17, IGKV3-NL1, IGKV1-6, IGKV2-40, IGKV1-33, IGKV1-27, IGLV2-14, IGLV6-57, IGLV1-51, IGKV6-21, IGLV1-47, IGLV3-21, IGLV1-44, IGKV1D-16, IGKV1-13, IGKV3-NL4, IGKV2-24, and IGKV1-8.

16. The isolated antagonist antibody of claim 13, wherein the antibody comprises:

a light chain variable region (VL) complementarity determining region three (CDR3) sequence of SEQ ID NO;

17 and variants thereof wherein the amino acid at position 1 is E, H, L, M or Q;

the amino acid at position 2 is E, H, N, Q, R or S;

the amino acid at position 3 is A, C, D, F, G, H, L, P, R, S, T, V or Y;

the amino acid at position 4 is A, D, E, F, G, H, I, K, L, M, N, R, S, T, V or Y;

the amino acid at position 5 is A, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W or Y;

the amino acid at position 6 is A, D, F, G, I, K, L, M, N, P, Q, S, T, V, W or Y;

the amino acid at position 7 is A, D, E, F, G, H, I, L, M, N, P, Q, R, S, T or V;

the amino acid at position 8 is A, E, F, G, H, I,K, L, M, N, P, Q, R, S, T, V, W or Y; and

the amino acid at position 9 is A, G, I, K, L, N, P, S, T or Y.