ANTI-HER2 GLYCOANTIBODIES AND USES THEREOF

• US 20150344587A1
• Filed: 05/27/2015
• Published: 12/03/2015
• Est. Priority Date: 05/27/2014
• Status: Active Grant

• Current Assignee: Academia Sinica (Government of The Republic of China)
• Original Assignee: Academia Sinica (Government of The Republic of China)
• Inventors: WONG, Chi-Huey, WU, Chung-Yi
• Granted Patent: US 10,005,847 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 31/517 ortho- or peri-condensed with carbocyclic ring systems, e.g. quinazoline, perimidine
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 16/283 against Fc-receptors, e.g. CD16, CD32, CD64 CD23 C07K16/2851
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 2317/14 Specific host cells or culture conditions, e.g. components, pH or temperature
• 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 2317/41 Glycosylation, sialylation, or fucosylation
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C12P 21/005 Glycopeptides, glycoproteins
• CPC Class Codes: C12Y 302/01 Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
• CPC Class Codes: C12Y 302/01051 Alpha-L-fucosidase (3.2.1.51)

Abstract

The present disclosure relates to a novel class of anti-HER2 monoclonal antibodies comprising a homogeneous population of anti-HER2 IgG molecules having the same N-glycan on each of Fc. The antibodies of the invention can be produced from anti-HER2 monoclonal antibodies by Fc glycoengineering. Importantly, the antibodies of the invention have improved therapeutic values with increased ADCC activity and increased Fc receptor binding affinity compared to the corresponding monoclonal antibodies that have not been glycoengineered.

Claims

1. A composition of anti-HER2 glycoantibodies or antigen binding fragments comprising a homogeneous population of anti-HER2 IgG molecules having the same N-glycan on the Fc region of each anti-HER2 IgG molecule.

2. The composition of claim 1, wherein the anti-HER2 IgG molecule comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO:

1, and a light chain having the amino acid sequence set forth in SEQ ID NO;

2.

3. The composition of claim 1, wherein the anti-HER2 IgG molecule comprises a light chain sequence and a heavy chain sequence of Trastuzumab (Herceptin).

4. The composition of claim 1, characterized in that the anti-HER2 glycoantibodies have improved ADCC over Trastuzumab.

5. The composition of claim 1, characterized in that the anti-HER2 glycoantibodies improved binding to Fcγ

RIIIA over Trastuzumab.

6. The composition of claim 1, wherein the N-glycan is attached to the Asn-297 of the Fc region.

7. The composition of claim 1, wherein the N-glycan is free of fucose.

8. The composition of claim 1, wherein the N-glycan comprises at least one α

2-6 terminal sialic acid.

9. The composition of claim 1, wherein the N-glycan comprises two α

2-6 terminal sialic acids.

10. The composition of claim 1, wherein the N-glycan is selected from the group consisting of Sia₂(α

2-6)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia₂(α

2-6)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia₂(α

2-3)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia₂(α

2-3)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia₂(α

2-3/α

2-6)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia₂(α

2-6/α

2-3)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia₂(α

2-31α

2-6)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia₂(α

2-6/α

2-3)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia(α

2-6)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia(α

2-3)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia(α

2-6)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia(α

2-3)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia(α

2-6)GalGlcNAc₂Man₃GlcNAc₂, Sia(α

2-3)GalGlcNAc₂Man₃GlcNAc₂, Sia(α

2-6)GalGlcNAc₃Man₃GlcNAc₂, Sia(α

2-3)GalGlcNAc₃Man₃GlcNAc₂, Gal₂GlcNAc₂Man₃GlcNAc₂, Gal₂GlcNAc₃Man₃GlcNAc₂, GalGlcNAc₂Man₃GlcNAc₂, GalGlcNAc₃Man₃GlcNAc₂, GlcNAc₃Man₃GlcNAc₂, GlcNAc₂Man₃GlcNAc₂, GlcNAcMan₃GlcNAc₂ and Man₃GlcNAc₂.

11. The composition of claim 10, wherein the N-glycan is selected from the group consisting of Sia₂(α

2-6)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia₂(α

2-6)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia₂(α

2-3)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia₂(α

2-3)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia₂(α

2-31α

2-6)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia₂(α

2-6/α

2-3)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia₂(α

2-3/α

2-6)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia₂(α

2-6/α

2-3)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia(α

2-6)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia(α

2-3)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia(α

2-6)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia(α

2-3)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia(α

2-6)GalGlcNAc₂Man₃GlcNAc₂, Sia(α

2-3)GalGlcNAc₂Man₃GlcNAc₂, Sia(α

2-6)GalGlcNAc₃Man₃GlcNAc₂, Sia(α

2-3)GalGlcNAc₃Man₃GlcNAc₂, Gal₂GlcNAc₂Man₃GlcNAc₂ and Gal₂GlcNAc₃Man₃GlcNAc₂.

12. A pharmaceutical formulation comprising a composition of anti-HER2 glycoantibodies or antigen binding fragments according to any one of claims 1-11 and a pharmaceutically acceptable carrier.

13. A method of treating HER2-expressing tumor in a patient in need thereof, the method comprising administering to the patient an effective amount of a pharmaceutical formulation according to claim 12.

14. The method of claim 13, wherein the tumor is a HER2-overexpressing tumor.

15. The method of claim 14, wherein said HER2-overexpressing tumor is breast cancer.

16. The method of claim 13, further comprising administering to the patient an anti-HER2 therapeutic agent.

17. The method of claim 16, wherein the anti-HER2 therapeutic agent is Lapatinib, Trastuzumab or Pertuzumab.

18. The method of claim 13, further comprising administering to the patient a second therapeutic agent as co-administration or as co-formulation.

19. A method of making an anti-HER2 glycoantibody of claim 1, the method comprising:

(a) contacting an anti-HER2 monoclonal antibody with an α

-fucosidase and at least one endoglycosidase, thereby yielding a defucosylated antibody having a single N-acetylglucosamine (GlcNAc), and

(b) adding a carbohydrate moiety to GlcNAc under suitable conditions.

20. The method of claim 23, wherein the anti-HER2 monoclonal antibody is Trastuzumab.

21. The method of claim 19, the anti-HER2 glycoantibody is produced in vitro.

22. The method of claim 19, the anti-HER2 glycoantibody is enzymatically engineered from an antibody obtained by cell culturing.

23. The method of claim 22, wherein the method comprises fed batch culturing of Chinese hamster ovary (CHO) cell line.

24. The method of claim 19, wherein the coupling glycan is selected from the group consisting of Sia₂(α

2-6)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia₂(α

2-6)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia₂(α

2-3)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia₂(α

2-3)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia₂(α

2-3/α

2-6)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia₂(α

2-6/α

2-3)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia₂(α

2-3/α

2-6)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia₂(α

2-6/α

2-3)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia(α

2-6)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia(α

2-3)Gal₂GlcNAc₂Man₃GlcNAc₂, Sia(α

2-6)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia(α

2-3)Gal₂GlcNAc₃Man₃GlcNAc₂, Sia(α

2-6)GalGlcNAc₂Man₃GlcNAc₂, Sia(α

2-3)GalGlcNAc₂Man₃GlcNAc₂, Sia(α

2-6)GalGlcNAc₃Man₃GlcNAc₂, Sia(α

2-3)GalGlcNAc₃Man₃GlcNAc₂, Gal₂GlcNAc₂Man₃GlcNAc₂, Gal₂GlcNAc₃Man₃GlcNAc₂, GalGlcNAc₂Man₃GlcNAc₂, GalGlcNAc₃Man₃GlcNAc₂, GlcNAc₃Man₃GlcNAc₂, GlcNAc₂Man₃GlcNAc₂, GlcNAcMan₃GlcNAc₂ and Man₃GlcNAc₂.

25. The method of claim 24, wherein the coupling glycan is selected from the group consisting of Sia₂(α

2-6)Gal₂GlcNAc₂Man₃GlcNAc, Sia₂(α

2-6)Gal₂GlcNAc₃Man₃GlcNAc, Sia₂(α

2-3)Gal₂GlcNAc₂Man₃GlcNAc, Sia₂(α

2-3)Gal₂GlcNAc₃Man₃GlcNAc, Sia₂(α

2-31α

2-6)Gal₂GlcNAc₂Man₃GlcNAc, Sia₂(α

2-6/α

2-3)Gal₂GlcNAc₂Man₃GlcNAc, Sia₂(α

2-31α

2-6)Gal₂GlcNAc₃Man₃GlcNAc, Sia₂(α

2-6/α

2-3)Gal₂GlcNAc₃Man₃GlcNAc, Sia(α

2-6)Gal₂GlcNAc₂Man₃GlcNAc, Sia(α

2-3)Gal₂GlcNAc₂Man₃GlcNAc, Sia(α

2-6)Gal₂GlcNAc₃Man₃GlcNAc, Sia(α

2-3)Gal₂GlcNAc₃Man₃GlcNAc, Sia(α

2-6)GalGlcNAc₂Man₃GlcNAc, Sia(α

2-3)GalGlcNAc₂Man₃GlcNAc, Sia(α

2-6)GalGlcNAc₃Man₃GlcNAc, Sia(α

2-3)GalGlcNAc₃Man₃GlcNAc, Gal₂GlcNAc₂Man₃GlcNAc and Gal₂GlcNAc₃Man₃GlcNAc.

26. The method of claim 19, wherein the adding in step (b) is performed by a transglycosylase.

27. The method of claim 19, wherein the endoglycosidase is EndoS, EndoS2, EndoH, EndoA, EndoM, EndoF, EndoF2, EndoF3.

28. The method of claim 19, wherein the α

-fucosidase comprises a polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO;

5.

29. The method of claim 19, wherein the α

-fucosidase is a recombinant Bacteroides α

-fucosidase.