CD20-Binding polypeptide compositions and methods

• US 20050069545A1
• Filed: 08/13/2004
• Published: 03/31/2005
• Est. Priority Date: 08/14/2003
• Status: Active Grant

• Current Assignee: Merck Patent GmbH (Merck & Co., Inc.)
• Original Assignee: Merck Patent GmbH (Merck & Co., Inc.)
• Inventors: Gillies, Stephen D., Williams, Stephen, Carr, Francis Joseph
• Granted Patent: US 8,147,832 B2
• Time in Patent Office: Days
• US Class Current: 424/144.100
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C07K 16/2887 against CD20
• CPC Class Codes: C07K 16/3076 against structure-related tumour-associated moieties
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/734 Complement-dependent cytotoxicity [CDC]
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: Y10S 424/80 Antibody or fragment thereof whose amino acid sequence is disclosed in whole or in part
• CPC Class Codes: Y10S 424/801 involving antibody or fragment thereof produced by recombinant dna technology

Abstract

A CD20-binding polypeptide composition of the invention comprises at least one polypeptide selected from the group consisting of a polypeptide having the amino acid residue sequence of SEQ ID NO: 1 (Vh of 2B8 antibody), but which includes at least one amino acid residue substitution in SEQ ID NO: 1 selected from the group consisting of V₁₂K, A₁₄P, M₂₀V, I₄₈T, A₆₈T, Q₈₂E, T₈₇R, S₉₁T, and T₁₀₆W; a polypeptide having the amino acid residue sequence of SEQ ID NO: 4 (Vk of 2B8 antibody), but which includes at least one amino acid residue substitution in SEQ ID NO: 4 selected from the group consisting of L₁₁I, S₁₂T, S₂₇T, V₂₉A, G₄₀T, V₅₉S, S₆₉T, L₇₂M, R₇₆S, and V₇₇L; a polypeptide having the amino acid residue sequence of SEQ ID NO: 9 (Vh of Leu16 antibody), but which includes at least one amino acid residue substitution in SEQ ID NO: 9 selected from the group consisting of V₁₂K, M₂₀V, A₆₈T, Q₈₂E, T₈₇R, S₉₁T, D₉₃V, and A₁₁₄T; and a polypeptide having the amino acid residue sequence of SEQ ID NO: 11 (Vk of Leu16 antibody), but which includes at least one amino acid residue substitution in SEQ ID NO: 11 selected from the group consisting of L₁₁I, S₁₂T, A₅₉S, S₆₉T, L₇₂M, R₇₆S, and V₇₇L. The compositions are useful for diagnosis and treatment of autoimmune diseases.

Claims

1. A CD20-binding polypeptide composition comprising at least one polypeptide selected from the group consisting of:

a modified heavy chain variable region polypeptide having the amino acid residue sequence of SEQ ID NO;

1, but which includes at least one amino acid residue substitution in SEQ ID NO;

1 selected from the group consisting of V₁₂K, A₁₄P, M₂₀V, I₄₈T, A₆₈T, Q₈₂E, T₈₇R, S₉₁T, and T₁₀₆W;

a modified light chain variable region polypeptide having the amino acid residue sequence of SEQ ID NO;

4, but which includes at least one amino acid residue substitution in SEQ ID NO;

4 selected from the group consisting of L₁₁I, S₁₂T, S₂₇T, V₂₉A, G₄₀T, V₅₉S, S₆₉T, L₇₂M, R₇₆S, and V₇₇L;

a modified heavy chain variable region polypeptide having the amino acid residue sequence of SEQ ID NO;

9, but which includes at least one amino acid residue substitution in SEQ ID NO;

9 selected from the group consisting of V₁₂K, M₂₀V, A₆₈T, Q₈₂E, T₈₇R, S₉₁T, D₉₃V, and A₁₁₄T; and

a modified light chain variable region polypeptide having the amino acid residue sequence of SEQ ID NO;

11, but which includes at least one amino acid residue substitution in SEQ ID NO;

11 selected from the group consisting of L₁₁I, S₁₂T, A₅₉S, S₆₉T, L₇₂M, R₇₆S, and V₇₇L.

2. The CD20-binding polypeptide composition of claim 1 comprising at least one modified heavy chain variable region polypeptide and at least one modified light chain variable region polypeptide.

3. The CD20-binding polypeptide composition of claim 2 in the form of a chimeric antibody and further including a human heavy chain constant region and a human light chain constant region.

4. The CD20-binding polypeptide composition of claim 3 wherein the human heavy chain constant region is an IgG constant region.

5. The CD20-binding polypeptide composition of claim 4 wherein the IgG constant region is an IgG1 constant region.

6. The CD20-binding polypeptide composition of claim 3 wherein the human light chain constant region is a human kappa light chain constant region.

7. The CD20-binding polypeptide composition of claim 3 in the form of a fusion protein with a cytokine.

8. The CD20-binding polypeptide composition of claim 7 wherein the cytokine is IL-2.

9. The CD20-binding polypeptide composition of claim 1 in the form of a fusion protein with a cytokine.

10. The CD20-binding polypeptide composition of claim 9 wherein the cytokine is IL-2.

11. The CD20-binding polypeptide composition of claim 1 in the form of a Fab antibody fragment, a single-chain Fv antibody fragment, and minibody.

23. A pharmaceutical composition comprising a CD20-binding polypeptide composition of claim 1, together with a pharmaceutically acceptable carrier, excipient, diluent.

24. The pharmaceutical composition of claim 23 further comprising an additional pharmacologically effective drug.

25. A method of treating an autoimmune disease comprising administering to a patient a therapeutically effective amount of a CD20-binding polypeptide composition of claim 1.

26. A method of treating an autoimmune disease comprising administering to a patient a therapeutically effective amount of a pharmaceutical composition of claim 23.

12. A CD20-binding polypeptide composition comprising at least one polypeptide segment having an amino acid residue sequence selected from the group consisting of SEQ ID NO:

2, SEQ ID NO;

3 and SEQ ID NO;

10.

13. A CD20-binding polypeptide composition comprising at least one polypeptide segment having an amino acid residue sequence selected from the group consisting of SEQ ID NO:

5 SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, and SEQ IDNO;

12.

14. A CD20-binding polypeptide composition comprising a polypeptide segment having an amino acid residue sequence selected from the group consisting of SEQ ID NO:

2, SEQ ID NO;

3, and SEQ ID NO;

10; and

a polypeptide segment having an amino acid residue sequence selected from the group consisting of SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, and SEQ ID NO;

12.

15. The CD20-binding polypeptide composition of claim 14 comprising a polypeptide segment having an amino acid residue sequence of SEQ ID NO:

2 and a polypeptide segment having an amino acid residue sequence of SEQ ID NO;

5.

16. The CD20-binding polypeptide composition of claim 14 comprising a polypeptide segment having an amino acid residue sequence of SEQ ID NO:

2 and a polypeptide segment having an amino acid residue sequence of SEQ ID NO;

6.

17. The CD20-binding polypeptide composition of claim 14 comprising a polypeptide segment having an amino acid residue sequence of SEQ ID NO:

2 and a polypeptide segment having an amino acid residue sequence of SEQ ID NO;

7.

18. The CD20-binding polypeptide composition of claim 14 comprising a polypeptide segment having an amino acid residue sequence of SEQ ID NO:

2 and a polypeptide segment having an amino acid residue sequence of SEQ ID NO;

8.

19. The CD20-binding polypeptide composition of claim 14 comprising a polypeptide segment having an amino acid residue sequence of SEQ ID NO:

3 and a polypeptide segment having an amino acid residue sequence of SEQ ID NO;

6.

20. The CD20-binding polypeptide composition of claim 14 comprising a polypeptide segment having an amino acid residue sequence of SEQ ID NO:

3 and a polypeptide segment having an amino acid residue sequence of SEQ ID NO;

8.

21. The CD20-binding polypeptide composition of claim 14 comprising a polypeptide segment having an amino acid residue sequence of SEQ ID NO:

10 and a polypeptide segment having an amino acid residue sequence of SEQ ID NO;

12.

22. A CD20-binding polypeptide composition substantially free from T cell epitopes and comprising a polypeptide having an amino acid residue sequence selected from the group consisting of SEQ ID NO:

15, SEQ ID NO;

16, SEQ ID NO;

17, SEQ ID NO;

18, SEQ ID NO;

19, SEQ ID NO;

20, SEQ ID NO;

21, SEQ ID NO;

22, SEQ ID NO;

23, SEQ ID NO;

24, SEQ ID NO;

25, SEQ ID NO;

26, SEQ ID NO;

27, SEQ ID NO;

28, and mixtures thereof.

27. An isolated nucleic acid encoding at least one polypeptide segment selected from the group consisting of:

a modified heavy chain variable region polypeptide having the amino acid residue sequence of SEQ ID NO;

1, but which includes at least one amino acid residue substitution in SEQ ID NO;

1 selected from the group consisting of V₁₂K, A₁₄P, M₂₀V, I₄₈T, A₆₈T, Q₈₂E, T₈₇R, S₉₁T, and T₁₀₆W;

a modified light chain variable region polypeptide having the amino acid residue sequence of SEQ ID NO;

4, but which includes at least one amino acid residue substitution in SEQ ID NO;

4 selected from the group consisting of L₁₁I, S₁₂T, S₂₇T, V₂₉A, G₄₀T, V₅₉S, S₆₉T, L₇₂M, R₇₆S, and V₇₇L;

a modified heavy chain variable region polypeptide having the amino acid residue sequence of SEQ ID NO;

9, but which includes at least one amino acid residue substitution in SEQ ID NO;

9 selected from the group consisting of V₁₂K, M₂₀V, A₆₈T, Q₈₂E, T₈₇R, S₉₁T, D₉₃V, and A₁₁₄T; and

a modified light chain variable region polypeptide having the amino acid residue sequence of SEQ ID NO;

11, but which includes at least one amino acid residue substitution in SEQ ID NO;

11 selected from the group consisting of L₁₁I, S₁₂T, A₅₉S, S₆₉T, L₇₂M, R₇₆S, and V₇₇L.