Homogenous antibody drug conjugates via enzymatic methods

• US 10,357,472 B2
• Filed: 06/11/2015
• Issued: 07/23/2019
• Est. Priority Date: 06/12/2014
• Status: Active Grant

• Current Assignee: CSPC Megalith Biopharmaceutical Company Limited
• Original Assignee: CSPC Dophen Corporation (CSPC Pharmaceutical Group Limited)
• Inventors: Hu, Sean, Allen, Lisha
• Primary Examiner(s): Alstrum-Acevedo, James H
• Assistant Examiner(s): D'Ambrosio, Thea
• Application Number:
  US15/317,907
• Publication Number:
  US 20170106096A1
• Time in Patent Office: 1,503 Days
• Field of Search: None
• CPC Class Codes: A61K 31/255 of sulfoxy acids or sulfur analogues thereof
• CPC Class Codes: A61K 38/16 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof enzyme inhibitors A61K38/005
• CPC Class Codes: A61K 38/164 from bacteria
• CPC Class Codes: A61K 38/45 Transferases (2)
• CPC Class Codes: A61K 47/6803 Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
• CPC Class Codes: A61K 47/6855 the tumour determinant being from breast cancer cell
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 1/00 General methods for the preparation of peptides , i.e. processes for the organic chemical preparation of peptides or proteins of any length
• CPC Class Codes: C07K 14/195 from bacteria
• 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 16/30 from tumour cells
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/40 characterized by post-translational modification
• CPC Class Codes: C07K 2317/41 Glycosylation, sialylation, or fucosylation
• CPC Class Codes: C07K 2317/524 CH2 domain
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C12N 9/1044 Protein-glutamine gamma-glutamyltransferase (2.3.2.13), i.e. transglutaminase or factor XIII
• CPC Class Codes: C12Y 203/02013 Protein-glutamine gamma-glutamyltransferase (2.3.2.13), i.e. transglutaminase or factor XIII

Abstract

The present application in one aspect provide Fc-containing polypeptide conjugates comprising an Fc-containing polypeptide conjugated to a conjugate moiety, wherein the Fc-containing polypeptide comprises an N-glycosylated Fc region comprising an acceptor glutamine residue flanked by an N-glycosylation site and wherein the conjugate moiety is conjugated to the Fc-containing polypeptide via the acceptor glutamine residue. Also provided are methods of making such Fc-containing polypeptide conjugates by using a wildtype or engineered transglutaminases. Further provided are engineered transglutaminases specifically designed for carrying out such reactions.

Claims

1. An antibody drug conjugate comprising an antibody conjugated to a conjugation moiety via an endogenous acceptor glutamine residue on the antibody, wherein the antibody drug conjugate is glycosylated in the Fc region, wherein the conjugation moiety comprises a cleavable linker, and wherein the conjugation moiety is:

2. The antibody drug conjugate of claim 1, wherein the antibody drug conjugate is N-glycosylated in the Fc region.

3. The antibody drug conjugate of claim 1, wherein the antibody is a human antibody.

4. The antibody drug conjugate of claim 1, wherein the antibody is a humanized antibody.

5. The antibody drug conjugate of claim 1, wherein both heavy chains of the antibody are conjugated to the conjugation moiety.

6. The antibody drug conjugate of claim 1, wherein the antibody is an anti-Her2 antibody.

7. The antibody drug conjugate of claim 6, wherein the antibody is trastuzumab.

8. The antibody drug conjugate of claim 6, wherein the anti-Her2 antibody is N-glycosylated in the Fc region.

9. The antibody drug conjugate of claim 8, wherein the anti-Her2 antibody is N-glycosylated at position 297, and wherein the anti-Her2 antibody is conjugated to a conjugation moiety via an endogenous acceptor glutamine residue at position 295.

10. A composition comprising the antibody drug conjugate of claim 1, wherein at least about 50% of antibody drug conjugates in the composition are glycosylated in the Fc region.

11. The composition of claim 10, wherein at least 50% of the antibody drug conjugates in the composition has a molar ratio between the antibody and the conjugation moiety of 1:

1 or 1;

2.

12. A composition comprising the antibody drug conjugate of claim 9, wherein at least about 50% of antibody drug conjugates in the composition are glycosylated in the Fc region.

13. The composition of claim 12, wherein at least 50% of the antibody drug conjugates in the composition has a molar ratio between the antibody and the conjugation moiety of about 1:

2.

14. A pharmaceutical composition comprising the antibody drug conjugate of claim 1.

15. A pharmaceutical composition comprising the antibody drug conjugate of claim 9.