MULTISPECIFIC ANTIBODIES

• US 20150166670A1
• Filed: 11/24/2014
• Published: 06/18/2015
• Est. Priority Date: 05/24/2012
• Status: Abandoned Application

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: HAAS, ALEXANDER, KLEIN, CHRISTIAN, SCHAEFER, WOLFGANG, SUSTMANN, CLAUDIO, CASTOLDI, RAFFAELLA
• Application Number:
  US14/551,957
• Publication Number:
  US 20150166670A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• 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 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/41 Glycosylation, sialylation, or fucosylation
• CPC Class Codes: C07K 2317/51 Complete heavy chain or Fd fragment, i.e. VH + CH1
• CPC Class Codes: C07K 2317/515 Complete light chain, i.e. VL + CL
• CPC Class Codes: C07K 2317/66 comprising a swap of domains, e.g. CH3-CH2, VH-CL or VL-CH1
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2319/70 containing domain for protein-protein interaction

Abstract

The present invention relates to bivalent, multispecific antibodies, especially bivalent, trispecific antibodies which bind to human HER1, human HER2, and human HER3, their manufacture and use.

Claims

1. A multispecific antibody, comprising:

A) a) the light chain and heavy chain of a full length antibody which specifically binds to a first antigen and second antigen; and

b) the modified light chain and modified heavy chain of a full length antibody which specifically binds to a third antigen, wherein the variable domains VL and VH are replaced by each other, and/or wherein the constant domains CL and CH1 are replaced by each other;

or

B) a) the light chain and heavy chain of a full length antibody which specifically binds to a first antigen; and

b) the modified light chain and modified heavy chain of a full length antibody which specifically binds to a second antigen and third antigen, wherein the variable domains VL and VH are replaced by each other, and/or wherein the constant domains CL and CH1 are replaced by each other;

or

C) a) the light chain and heavy chain of a full length antibody which specifically binds to a first antigen and a second antigen; and

b) the modified light chain and modified heavy chain of a full length antibody which specifically binds to a third antigen and fourth antigen, wherein the variable domains VL and VH are replaced by each other, and/or wherein the constant domains CL and CH1 are replaced by each other; and

wherein the antibody is a bivalent, tri- or tetraspecific antibody.

2. The multispecific antibody according to claim 1, wherein the antibody is trispecific and comprises

A) a) the light chain and heavy chain of a full length antibody which specifically binds to a first antigen and second antigen;

and

b) the modified light chain and modified heavy chain of a full length antibody which specifically binds to a third antigen, wherein the variable domains VL and VH are replaced by each other, and/or wherein the constant domains CL and CH1 are replaced by each other;

or

B) a) the light chain and heavy chain of a full length antibody which specifically binds to a first antigen; and

b) the modified light chain and modified heavy chain of a full length antibody which specifically binds to a second antigen and third antigen, wherein the variable domains VL and VH are replaced by each other, and/or wherein the constant domains CL and CH1 are replaced by each other.

3. The multispecific antibody according to claim 2, wherein

under A) the first antigen is human HER1, the second antigen human HER3 and the third antigen is human HER2;

or

under B) the first antigen is human HER2, the second antigen human HER1 and the third antigen is human HER3.

4. The multispecific antibody according to claim 1, wherein the antibody is a bivalent, trispecific antibody and comprises a

a) the light chain and heavy chain of a full length antibody which specifically binds to human HER1 and human HER3;

and

b) the modified light chain and modified heavy chain of a full length antibody which specifically binds to human HER2, wherein the variable domains VL and VH are replaced by each other, and/or wherein the constant domains CL and CH1 are replaced by each other.

5. The multispecific antibody according to claim 4, wherein the antibody comprises the amino acid sequences of SEQ ID NOs:

4, 9, 13 and 18.

6. The multispecific antibody according to claim 3, wherein

under A) the first antigen is human HER1, the second antigen human HER3 and the third antigen is human cMET;

or

under B) the first antigen is human cMET, the second antigen human HER1 and the third antigen is human HER3.

7. The multispecific antibody according to claim 1, wherein the antibody is tetraspecific and comprises

a) the light chain and heavy chain of a full length antibody which specifically binds to a first antigen and a second antigen;

and

b) the modified light chain and modified heavy chain of a full length antibody which specifically binds to a third antigen and fourth antigen, wherein the variable domains VL and VH are replaced by each other, and/or wherein the constant domains CL and CH1 are replaced by each other.

8. The multispecific antibody according to claim 1, wherein in the modified light chain and modified heavy chain under b) the variable domains VL and VH are replaced by each other;

and wherein the constant domains CL and CH1 are replaced by each other.

9. The multispecific antibody according to claim 1, wherein in the modified light chain and modified heavy chain under b) (only) the variable domains VL and VH are replaced by each other.

10. The multispecific antibody according to claim 1, wherein in the modified light chain and modified heavy chain under b) (only) the constant domains CL and CH1 are replaced by each other.

11. The multispecific antibody according to claim 1, comprising

the CH3 domain of the heavy chain of the full length antibody of a) and

the CH3 domain of the modified heavy chain of the full length antibody of b) each meet at an interface which comprises an original interface between the antibody CH3 domains;

wherein said interface is altered to promote the formation of the trispecific or tetraspecific antibody, wherein the alteration is characterized in that;

i) the CH3 domain of one heavy chain is altered, so that within the original interface the CH3 domain of one heavy chain that meets the original interface of the CH3 domain of the other heavy chain within the tri- or tetraspecific antibody,

an amino acid residue is replaced with an amino acid residue having a larger side chain volume, thereby generating a protuberance within the interface of the CH3 domain of one heavy chain which is positionable in a cavity within the interface of the CH3 domain of the other heavy chain

and

ii) the CH3 domain of the other heavy chain is altered, so that within the original interface of the second CH3 domain that meets the original interface of the first CH3 domain within the tri- or tetraspecific antibody

an amino acid residue is replaced with an amino acid residue having a smaller side chain volume, thereby generating a cavity within the interface of the second CH3 domain within which a protuberance within the interface of the first CH3 domain is positionable.

12. The multispecific antibody according to claim 11, wherein the said amino acid residue having a larger side chain volume is selected from the group consisting of arginine (R), phenylalanine (F), tyrosine (Y), tryptophan (W) and said amino acid residue having a smaller side chain volume is selected from the group consisting of alanine (A), serine (S), threonine (T), valine (V).

13. The multispecific antibody according to claim 11, wherein both CH3 domains are further altered by the introduction of cysteine (C) as amino acid in the corresponding positions of each CH3 domain such that a disulfide bridge between both CH3 domains can be formed.

14. A method for the preparation of a multispecific antibody according to claim 1 comprising the steps of

a) transforming a host cell with vectors comprising nucleic acid molecules encoding

A) a) the light chain and heavy chain of a full length antibody which specifically binds to a first antigen and second antigen;

and

b) the modified light chain and modified heavy chain of a full length antibody which specifically binds to a third antigen, wherein the variable domains VL and VH are replaced by each other, and/or wherein the constant domains CL and CH1 are replaced by each other;

or

B) a) the light chain and heavy chain of a full length antibody which specifically binds to a first antigen; and

b) the modified light chain and modified heavy chain of a full length antibody which specifically binds to a second antigen and third antigen, wherein the variable domains VL and VH are replaced by each other, and/or wherein the constant domains CL and CH1 are replaced by each other;

or

C) a) the light chain and heavy chain of a full length antibody which specifically binds to a first antigen and a second antigen; and

b) the modified light chain and modified heavy chain of a full length antibody which specifically binds to a third antigen and fourth antigen, wherein the variable domains VL and VH are replaced by each other, and/or wherein the constant domains CL and CH1 are replaced by each other.

b) culturing the host cell under conditions that allow synthesis of said antibody molecule; and

c) recovering said antibody molecule from said culture.

15. Nucleic acid encoding the multispecific antibody according to claim 1.

16. Vectors comprising nucleic acid encoding the multispecific antibody according to claim 1.

17. A host cell comprising the vectors according to claim 16.

19. A pharmaceutical composition comprising an antibody according to claim 1 and at least one pharmaceutically acceptable excipient.

20. An antibody according to claim 1 for use in the treatment of cancer.

22. A method for the treatment of a patient in need of therapy, comprising administering to the patient a therapeutically effective amount of an antibody according to claim 1.

23. A method for the treatment of a patient suffering from cancer, comprising administering to the patient a therapeutically effective amount of an antibody according to claim 1.

18. (canceled)

21. (canceled)