COVALENTLY LINKED ANTIGEN-ANTIBODY CONJUGATES

• US 20150258209A1
• Filed: 12/19/2014
• Published: 09/17/2015
• Est. Priority Date: 07/04/2012
• Status: Active Grant

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: BRINKMANN, ULRICH, GEORGES, GUY, GROTE, MICHAEL, HAAS, ALEXANDER, HOFFMANN, EIKE, BENZ, JOERG, DENGL, STEFAN, DZIADEK, SEBASTIAN
• Granted Patent: US 10,517,945 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/6056 Antibodies
• CPC Class Codes: A61K 2039/627 characterised by the linker
• CPC Class Codes: A61K 39/385 Haptens or antigens, bound to carriers
• CPC Class Codes: A61K 47/6835 the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
• CPC Class Codes: A61K 47/6879 the immunoglobulin having two or more different antigen-binding sites, e.g. bispecific or multispecific immunoglobulin
• CPC Class Codes: A61K 47/6897 Pre-targeting systems with two or three steps using antibody conjugates; Ligand-antiligand therapies
• 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 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• 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 16/16 against material from plants
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 16/44 against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids
• CPC Class Codes: C07K 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/40 : characterized by post-translational modification
• CPC Class Codes: C07K 2317/55 : Fab or Fab'
• CPC Class Codes: C07K 2317/565 : Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/624 : Disulfide-stabilized antibody (dsFv)
• CPC Class Codes: C07K 2317/94 : Stability, e.g. half-life, pH, temperature or enzyme-resistance
• CPC Class Codes: G01N 33/6857 : Antibody fragments

Abstract

Herein is reported a conjugate comprising an antigen and an antibody that specifically binds to the antigen wherein a covalent bond is formed between the antigen and an amino acid residue in the CDR2 of the antibody, an antibody comprising a cysteine residue at amino acid position 53 (according to Kabat) in the heavy chain CDR2 and uses thereof.

Claims

1. A conjugate comprising an antigen and an antibody that specifically binds to the antigen wherein a covalent bond is between the antigen and an amino acid residue in a variable region of the antibody.

2. The conjugate of claim 1, wherein the antigen is a hapten and wherein a covalent bond is between the hapten and an amino acid residue in a variable region of the antibody.

3. The conjugate according to claim 1, wherein the amino acid residue in a variable region is in the heavy chain CDR2.

4. The conjugate according to claim 1, wherein the covalent bond is between a cysteine residue in the heavy chain CDR2 of the antibody and the antigen.

5. The conjugate according to claim 4, wherein the cysteine residue in the heavy chain CDR2 of the antibody is at position 52b or at position 53 according to the heavy chain variable domain numbering of Kabat.

6. The conjugate according to claim 1, wherein the covalent bond is a disulfide bond.

7. The conjugate according to claim 1, wherein the antibody is a bispecific antibody comprising a first binding specificity to a first antigen and a second binding specificity to a second antigen.

8. The conjugate according to claim 7, wherein the first binding specificity is to a hapten and a second binding specificity is to a non-hapten antigen.

9. The conjugate according to claim 1, wherein the antibody is a full length antibody.

10. The conjugate according to claim 1, wherein the antibody is a humanized antibody or a human antibody.

11. The conjugate according to claim 1, wherein the conjugate further comprises a therapeutic or detectable moiety.

12. The conjugate according to claim 2, wherein the hapten is selected from the group of haptens consisting of biotin, theophylline, digoxigenin, carborane, and fluorescein.

15. A pharmaceutical formulation comprising the conjugate according to claim 1 and a pharmaceutically acceptable carrier.

17. The conjugate of claim 1, wherein the antigen is a haptenylated compound comprising a hapten, a linker, and a payload.

18. The conjugate of claim 7, wherein the bispecific antibody comprises a first binding site that specifically binds to a haptenylated compound and a second binding specific that specifically binds to a cell surface marker.

19. A method of producing the conjugate of claim 1, comprising combining in solution an antibody that specifically binds to a hapten and comprises a reactive group at one amino acid residue in the CDR2 with a haptenylated compound comprising a reactive group, wherein the haptenylated compound comprises a payload, and recovering the conjugate from the solution.

20. A method of treating a disease in an individual comprising administering an effective amount of the conjugate of claim 1 to an individual in need thereof.

25. A method of detection comprising contacting a biological sample with the conjugate of claim 1 under conditions permissive for binding of the antibody or the conjugate to the target, and detecting whether a complex is formed between the conjugate and the target.

26. The method of claim 19, wherein the payload is a peptide consisting of 5 to 60 amino acids or a detectable label.

13. An antibody that has in the light and/or the heavy chain a cysteine residue in the CDR2.

14. The antibody according to claim 13, wherein the cysteine residue in the heavy chain CDR2 of the antibody is at position 52b or at position 53 according to the heavy chain variable domain numbering of Kabat.

16. A pharmaceutical formulation comprising the antibody according to claim 13 and a pharmaceutically acceptable carrier.

21. A method of producing the antibody of claim 13 comprising

(a) cultivating a cell comprising a nucleic acid encoding the antibody, and

(b) recovering the antibody from the cell or the culture medium.

23. A nucleic acid encoding the antibody of claim 13.

24. A host cell comprising nucleic acid encoding the antibody of claim 13.

22. A method for identifying a position in an antibody CDR2 that can be mutated to a cysteine for the formation of a covalent bond between the residue in the antibody CDR2 and a bound antigen or haptenylated compound comprising

(a) providing a crystal structure of the non-covalent complex of the antibody and the antigen or haptenylated compound, whereby the antigen or haptenylated compound comprises a linker sequence, and

(b) identifying an amino acid residue in the CDR2 of the antibody and in the linker sequence with a distance of 10 to 11 Angstroms between the alpha carbon atoms of the amino acid residue in the antibody CDR2 and the atom in the linker is fused wherein the identified position is the position in an antibody CDR2 that can be mutated to cysteine for the formation of a covalent bond between the residue in the antibody CDR2 and the bound antigen or haptenylated compound.