Antigen Binding Proteins

• US 20120237506A1
• Filed: 02/27/2012
• Published: 09/20/2012
• Est. Priority Date: 02/28/2011
• Status: Active Grant

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: Bossenmaier, Birgit, Kettenberger, Hubert, Klein, Christian, Schaefer, Wolfgang, Schwaiger, Manfred, Sustmann, Claudio, Kuenkele, Klaus-Peter, Regula, Joerg-Thomas
• Granted Patent: US 9,982,036 B2
• Time in Patent Office: Days
• US Class Current: 424/133.1
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• 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 16/2863 against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 2317/41 Glycosylation, sialylation, or fucosylation
• CPC Class Codes: C07K 2317/64 comprising a combination of variable region and constant region components
• CPC Class Codes: C07K 2317/66 comprising a swap of domains, e.g. CH3-CH2, VH-CL or VL-CH1
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/77 Internalization into the cell
• 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
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: G01N 21/553 and using surface plasmons fluorescence excitation G01N21/648; enhanced Raman G01N21/658

Abstract

The present invention relates to antigen binding proteins comprising two Fc parts, methods for their production, pharmaceutical compositions containing said antigen binding proteins, and uses thereof.

Claims

1. An antigen binding protein comprising

a) two modified heavy chains of an antibody which specifically binds to an antigen, wherein VH of each heavy chain is replaced by the VL of said antibody, said modified heavy chains being associated with each other via their CH3 domains of the Fc part;

b) two modified heavy chains of said antibody wherein CH1 of each heavy chain is replaced by CL of said antibody, said modified heavy chains being associated with each other via their CH3 domains of the Fc part;

and wherein the VL domains of the heavy chains of a) are associated with the VH domains of the heavy chains of b), and the CH1 domains of the heavy chains of a) are associated with the CL domains of the heavy chains of b).

2. The antigen binding protein according to claim 1, comprising

the CH3 domains of the Fc part of the modified heavy chains of a) and the CH3 domains of the Fc part of the modified heavy chains of b) are of the same isotype

3. The antigen binding protein according to claim 2, comprising

the CH2 and CH3 domains of the Fc part of the modified heavy chains of a) and the CH2 and CH3 domains of the Fc part of the modified heavy chains of b) are of the same isotype.

4. The antigen binding protein according to claim 3, comprising

the CH2 and CH3 domains of the Fc part of the modified heavy chains of a) and the CH2 and CH3 domains of the Fc part of the modified heavy chains of b) are of the IgG isotype.

5. The antigen binding protein according to claim 4, comprising

the CH2 and CH3 domains of the Fc part of the modified heavy chains of a) and the CH2 and CH3 domains of the Fc part of the modified heavy chains of b) are of the IgG1 isotype.

6. The antigen binding protein according to claim 1, characterized in comprising

a) two modified heavy chains comprising the amino acid sequence of SEQ ID NO:

1; and

b) two modified heavy chains comprising the amino acid sequence of SEQ ID NO;

2;

a) two modified heavy chains comprising the amino acid sequence of SEQ ID NO;

3; and

b) two modified heavy chains comprising the amino acid sequence of SEQ ID NO;

4;

or

a) two modified heavy chains comprising the amino acid sequence of SEQ ID NO;

5; and

b) two modified heavy chains comprising the amino acid sequence of SEQ ID NO;

6.

7. The antigen binding protein according to claim 5, comprising

either the two modified heavy chains of a),

or the two modified heavy chains of b),

are further modified by the amino acid substitutions S364G, L368F, D399K and K409D (wherein the amino acid positions are numbered according to the EU Index of Kabat).

8. The antigen binding protein according to claim 1, characterized in comprising

a) two modified heavy chains comprising the amino acid sequence of SEQ ID NO:

1; and

b) two modified heavy chains comprising the amino acid sequence of SEQ ID NO;

2;

wherein either the two modified heavy chains of a),

or the two modified heavy chains of b),

are further modified by the amino acid substitutions S364G, L368F, D399K and K409D (wherein the amino acid positions are numbered according to the EU Index of Kabat);

a) two modified heavy chains comprising the amino acid sequence of SEQ ID NO;

3; and

b) two modified heavy chains comprising the amino acid sequence of SEQ ID NO;

4;

wherein either the two modified heavy chains of a),

or the two modified heavy chains of b),

are further modified by the amino acid substitutions S364G, L368F, D399K and K409D (wherein the amino acid positions are numbered according to the EU Index of Kabat);

or

a) two modified heavy chains comprising the amino acid sequence of SEQ ID NO;

5; and

b) two modified heavy chains comprising the amino acid sequence of SEQ ID NO;

6

wherein either the two modified heavy chains of a),

or the two modified heavy chains of b),

are further modified by the amino acid substitutions S364G, L368F, D399K and K409D (wherein the amino acid positions are numbered according to the EU Index of Kabat).

9. The antigen binding protein according to claim 1, comprising

the CH3 domains of the Fc part of the modified heavy chains of a) and the CH3 domains of the Fc part of the modified heavy chains of b) are of a different isotype.

10. The antigen binding protein according to claim 9, comprising

the CH3 domains of the Fc part of the modified heavy chains of a) are of the IgG1 isotype;

and the CH3 domains of the Fc part of the modified heavy chains of b) are of the IgA isotype.

11. The antigen binding protein according to claim 10, characterized in comprising

a) two modified heavy chains comprising the amino acid sequence of SEQ ID NO:

7; and

b) two modified heavy chains comprising the amino acid sequence of SEQ ID NO;

4.

12. The antigen binding protein according to claim 10, comprising

The CH2 and CH3 domains of the Fc part of the modified heavy chains of a) are of the IgG1 isotype;

and the CH2 and CH3 domains of the Fc part of the modified heavy chains of b) are of the IgA isotype.

13. The monovalent antigen binding protein according to claims 1 to 11 comprising the CH2 domain of the Fc parts of a) and b) are of IgG1 isotype, and the antigen binding protein is afucosylated with an amount of fucose of 80% or less of the total amount of oligosaccharides (sugars) at Asn297 is of human IgG1 isotype.

14. A pharmaceutical composition comprising an antigen binding protein according to claims 1 to 13.

15. The antigen binding protein according to claims 1 to 13 for use in the treatment of cancer.

16. Use of antigen binding protein according to claims 1 to 13 for the manufacture of a medicament for the treatment of cancer.

17. A method for the treatment of a patient in need of therapy, characterized by administering to the patient a therapeutically effective amount of an antigen binding protein according to claims 1 to 13.

18. A method for the preparation of an antigen binding protein according to claims 1 to 13

comprising the steps of

a) transforming a host cell with vectors comprising nucleic acid molecules encoding an antigen binding protein according to claims 1 to 13,

b) culturing the host cell under conditions that allow synthesis of said antigen binding protein molecule;

and

c) recovering said antigen binding protein molecule from said culture.

19. A nucleic acid encoding an antigen binding protein according to claims 1 to 13.

20. A vector comprising nucleic acid according to claim 19.

21. A host cell comprising the vector according to claim 20.