Antibody and antibody fragment introduced new modification sites

• US 9,150,639 B2
• Filed: 03/28/2011
• Issued: 10/06/2015
• Est. Priority Date: 03/26/2010
• Status: Active Grant

• Current Assignee: Kyowa Kirin Co., Ltd. (Kirin Holdings Company Limited)
• Original Assignee: Kyowa Hakko Kirin Company Limited (Kirin Holdings Company Limited)
• Inventors: Shiraishi, Yasuhisa, Yamasaki, Motoo
• Primary Examiner(s): HUYNH, PHUONG N
• Application Number:
  US13/072,924
• Publication Number:
  US 20120009621A1
• Time in Patent Office: 1,653 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 47/50 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/61 the organic macromolecular compound being a polysaccharide or a derivative thereof
• CPC Class Codes: A61K 47/68 the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
• CPC Class Codes: A61K 47/6803 Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
• CPC Class Codes: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• 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 2317/14 Specific host cells or culture conditions, e.g. components, pH or temperature
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/522 CH1 domain

Abstract

The present invention relates to an antibody or antibody fragment comprising novel Cys residue, to which a hydrophilic macromolecular group or amphipathic macromolecular group can be bound at a high efficiency. In addition, the present invention relates to a monoclonal antibody modified product or an antibody fragment modified product in which cysteine residue is chemically modified.

Claims

1. A monoclonal IgG antibody or antigen-binding fragment thereof, wherein said monoclonal IgG antibody or antigen-binding fragment thereof comprises a human constant region in which one or more amino acids having a solvent accessible surface area ratio of 20% or less are substituted with a cysteine residue, wherein said one or more amino acids that are substituted are selected from the group consisting of (1) to (5):

(1) the amino acid at position 124 of human IgG light chain region in Kabat numbering,

(2) the amino acid at position 198 of human IgG light chain region in Kabat numbering,

(3) the amino acid at position 201 of human IgG light chain region in Kabat numbering,

(4) the amino acid at position 147 of human IgG heavy chain region in EU numbering, and

(5) the amino acid at position 183 of human IgG heavy chain region in EU numbering.

3. The monoclonal IgG antibody or antigen-binding fragment thereof according to claim 1, wherein the amino acid at position 124 of human IgG light chain region in Kabat numbering is substituted with a cysteine residue.

4. The monoclonal IgG antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein one or more amino acids in a CH1 region and/or a light chain constant region of said monoclonal IgG antibody or antigen-binding fragment are substituted with a cysteine residue.

5. The monoclonal IgG antibody or antigen-binding fragment thereof according to claim 1, wherein at least one of said substituted cysteine residues is chemically modified.

10. The monoclonal IgG antibody or antigen-binding fragment thereof according to claim 1 or 2, which has a cytotoxicity.

11. The monoclonal IgG antibody or antigen-binding fragment thereof according to claim 10, wherein the cytotoxicity is an antibody-dependent cellular cytotoxicity or a complement-dependent cytotoxicity.

12. The monoclonal IgG antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein the antigen-binding fragment thereof is an antibody fragment selected from the group consisting of a Fab, a Fab′

and a F(ab′

)₂.

13. The monoclonal IgG antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein the monoclonal IgG antibody is a recombinant antibody.

14. The monoclonal IgG antibody or antigen-binding fragment thereof according to claim 13, wherein the recombinant antibody is a chimeric antibody, a humanized antibody or a human antibody.

2. A monoclonal IgG antibody or antigen-binding fragment thereof, wherein said monoclonal IgG antibody or antigen-binding fragment thereof comprises a human constant region in which an amino acid having a solvent accessible surface area ratio of 20% or less is substituted with a cysteine residue, wherein said amino acid having a solvent accessible surface area ratio of 20% or less is position 140 of human IgG heavy chain region in EU numbering, and wherein said substituted cysteine residue is chemically modified.

6. The monoclonal IgG antibody or antigen-binding fragment thereof according to claim 2 or 5,

(A) wherein the substituted cysteine residue is chemically modified by a chemical modification reaction under non-reducing conditions;

(B) wherein 40% or more of said substituted cysteine residues are chemically modified;

(C) wherein the chemical modification is binding of a thiol group of the cysteine residue with a modification group comprising a hydrophilic macromolecule or amphipathic macromolecule, wherein the hydrophilic macromolecule or amphipathic macromolecule is a polyoxyalkylene, polyol or polysaccharide; and

/or

(D) wherein said chemical modification is binding of a thiol group of the cysteine residue with a modification group comprising a functional molecule, wherein the functional molecule is a drug, a biologically active peptide, a biologically active protein, a nucleic acid, a radiolabeled compound, a sugar chain, a lipid or a fluorescent compound.

7. The monoclonal IgG antibody or antigen-binding fragment thereof according to claim 6, wherein the functional molecule is a nucleic acid.

8. The monoclonal IgG antibody or antigen-binding fragment thereof according to claim 6, wherein the drug is an antitumor agent, an antibiotic or an antiviral agent.

9. The monoclonal IgG antibody or antigen-binding fragment thereof according to claim 6, wherein said modification group has a molecular weight of 500 Da to 100 kDa.