Humanized anti-CD3 specific antibodies

• US 6,706,265 B1
• Filed: 06/07/1995
• Issued: 03/16/2004
• Est. Priority Date: 03/24/1992
• Status: Expired due to Fees

• Current Assignee: BTG International Limited (Boston Scientific Corporation)
• Original Assignee: BTG International Limited (Boston Scientific Corporation)
• Inventors: Waldmann, Herman, Clark, Michael R., Gorman, Scott D., Routledge, Edward G., Bolt, Sarah L.
• Primary Examiner(s): Chan, Christina
• Assistant Examiner(s): DiBrino, Marianne
• Application Number:
  US08/478,684
• Time in Patent Office: 3,205 Days
• Field of Search: 530/387.3, 530/388.75, 530/387.1, 530/388.2, 424/154.1, 424/133.1
• US Class Current: 424/133.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/41 Glycosylation, sialylation, or fucosylation
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/74 Inducing cell proliferation

Abstract

Novel aglycosylated antibodies having a binding affinity for the CD3 antigen complex are of value for use in therapy, particularly in immunosuppression.

Claims

1. An aglycosylated IgG antibody which specifically binds CD3, having a heavy chain with three CDRs comprising the amino acid sequences (a) Ser-Phe-Pro-Met-Ala (SEQ ID NO:

1), (b) Thr-Ile-Ser-Thr-Ser-Gly-Gly-Arg-Thr-Tyr-Tyr-Arg-Asp-Ser-Val-Lys-Gly (SEQ ID NO;

2), (c) Phe-Arg-Gln-Tyr-Ser-Gly-Gly-Phe-Asp-Tyr (SEQ ID NO;

3), or an antigen binding fragment thereof, and a light chain with three CDRs comprising the amino acid sequences;

(d) Thr-Leu-Ser-Ser-Gly-Asn-Ile-Glu-Asn-Asn-Tyr-Val-His (SEQ ID NO;

4),

(e) Asp-Asp-Asp-Lys-Arg-Pro-Asp (SEQ ID NO;

5),

(f) His-Ser-Tyr-Val-Ser-Ser-Phe-Asn-Val (SEQ ID NO;

6),

or an antigen binding fragment thereof, the heavy chain CDRs being arranged in the order (a), (b), (c) in the leader to constant domain direction and the light chain CDRs being arranged in the order (d), (e), (f) in the leader to constant domain direction.

2. An aglycosylated antibody according to claim 1, which specifically binds human CD3.

3. An aglycosylated antibody according to claim 1, in which the variable domain framework regions are of or are derived from those of rat or mouse origin.

4. An aglycosylated antibody according to claim 1, in which the CDRs are of different origin than the variable framework region.

5. An aglycosylated antibody according to claim 1, in which the variable domain framework regions are of or are derived from those of human origin.

6. An aglycosylated antibody according to claim 1 having a heavy chain variable domain which comprises

Glu-Valn-Leu-Leu-Glu-Ser-Gly-Gly-Gly-Leu-Val-Gln-Pro-Gly-Gly-Ser-Leu-Arg-Leu-Ser-Cys-Ala-Ala-Ser-Gly-Phe-Thr-Phe-Ser-Ser-Phe-Pro-Met-Ala-Trp-Val-Arg-Gln-Ala-Pro-Gly-Lys-Gly-Leu-Glu-Trp-Val-Ser-Thr-Ile-Ser-Thr-Ser-Gly-Gly-Arg-Thr-Tyr-Tyr-Arg-Asp-Ser-Val-Lys-Gly-Arg-Phe-Thr-Ile-Ser-Arg-Asp-Asn-Ser-Lys-Asn-Thr-Leu-Tyr-Leu-Gln-Met-Asn-Ser-Leu-Arg-Ala-Glu-Asp-Thr-Ala-Val-Tyr-Tyr-Cys-Ala-Lys-Phe-Arg-Gln-Tyr-Ser-Gly-Gly-Phe-Asp-Tyr-Trp-Gly-Gln-Gly-Thr-Leu-Val-Thr-Val-Ser-Ser (SEQ ID NO: 11).

7. An aglycosylated antibody according to claim 1 having a light chain variable domain which comprises Asp-Phe-Met-Leu-Thr-Gln-Pro-His-Ser-Val-Ser-Glu-Ser-Pro-Gly-Lys-Thr-Val-Ile-Ile-Ser-Cys-Thr-Leu-Ser-Ser-Gly-Asn-Ile-Glu-Asn-Asn-Tyr-Val-His-Trp-Tyr-Gln-Gln-Arg-Pro-Gly-Arg-Ala-Pro-Thr-Thr-Val-Ile-Phe-Asp-Asp-Asp-Lys-Arg-Pro-Asp-Gly-Val-Pro-Asp-Arg-Phe-Ser-Gly-Ser-Ile-Asp-Arg-Ser-Ser-Asn-Ser-Ala-Ser-Leu-Thr-Ile-Ser-Gly-Leu-Gln-Thr-Glu-Asp-Glu-Ala-Asp-Tyr-Tyr-Cys-His-Ser-Tyr-Val-Ser-Ser-Phe-Asn-Val-Phe-Gly-Gly-Gly-Thr-Lys-Leu-Thr-Val-Leu (SEQ ID NO:

16).

8. An aglycosylated antibody according to claim 1, in which the constant domains are of or are derived from those of rat or mouse origin.

9. An aglycosylated antibody according to claim 1, in which the CDRs are of different origin than the constant region.

10. An aglycosylated antibody according to claim 1, in which the constant domains are of or are derived from those of human origin.

11. An aglycosylated antibody according to claim 1, in which the constant region is of an IgG isotype.

12. An aglycosylated antibody according to claim 10, in which the constant region is of an IgG1 isotype.

13. An aglycosylated antibody according to claim 10, in which asparagine residue at position 297 of each constant region heavy chain is replaced by an alternative amino acid residue.

14. An aglycosylated antibody according to claim 13, in which the asparagine residue is replaced by an alanine residue.

15. An aglycosylated antibody according to claim 1, in which only one of the arms thereof has an affinity for CD3.

16. An aglycosylated antibody according to claim 1 which is monovalent.

17. An aglycosylated antibody according to claim 16, in which one half of the antibody consists of a complete heavy chain and light chain and the other half consists of a similar but truncated heavy chain lacking the binding site for the light chain.

18. A composition comprising an aglycosylated antibody as claimed in claim 1 together with a pharmaceutically acceptable diluent or carrier.

19. A method of immunosuppressing graft rejection, comprising administering to a patient in need thereof an immunosuppressive effective amount of an aglycosylated antibody as claimed in claim 1.

20. An aglycosylated antibody according to claim 6 having a light chain variable domain which comprises

Ase-Phe-Met-Leu-Thr-Gln-Pro-His-Ser-Val-Ser-Glu-Ser-Pro-Gly-Lys-Thr-Val-Ile-Ile-Ser-Cys-Thr-Leu-Ser-Ser-Gly-Asn-Ile-Glu-Asn-Asn-Tyr-Val-His-Trp-Tyr-Gln-Gln-Arg-Pro-Gly-Arg-Ala-Pro-Thr-Thr-Val-Ile-Phe-Asp-Asp-Asp-Lys-Arg-Pro-Asp-Gly-Val-Pro-Asp-Arg-Phe-Ser-Gly-Ser-Ile-Asp-Arg-Ser-Ser-Asn-Ser-Ala-Ser-Leu-Thr-Ile-Ser-Gly-Leu-Gln-Thr-Glu-Asp-Glu-Ala-Asp-Tyr-Tyr-Cys-His-Ser-Tyr-Val-Ser-Ser-Phe-Asn-Val-Phe-Gly-Gly-Gly-Thr-Lys-Leu-Thr-Val-Leu (SEQ ID NO.16).