Methods for controlling blood pharmacokinetics of antibodies

Patent metadata

Field	Value
Publication number	US-11046784-B2
Application number	US-29503907-A
Country	US
Kind code	B2
Filing date	Mar 30, 2007
Priority date	Mar 31, 2006
Publication date	Jun 29, 2021
Grant date	Jun 29, 2021

Abstract

The present inventors discovered that the half-life in blood of an IgG antibody which is a polypeptide comprising an FcRn-binding domain can be controlled by controlling the surface charge through modification of residues exposed on the surface among residues in the variable regions of the IgG antibody. Antibodies whose half-life in blood had been controlled by the methods of the present invention were confirmed to actually retain the original activity. The methods of the present invention are widely applicable to polypeptides comprising an FcRn-binding domain, such as IgG antibodies, which are recycled via the FcRn salvage pathway regardless of the type of target antigen.

First claim

The invention claimed is:

1. A method for producing a nucleic acid, the method comprising:
(a) providing a nucleotide sequence encoding a first antibody heavy chain of a first IgG antibody, the first heavy chain comprising an Fc domain;
(b) producing a nucleic acid encoding a second antibody heavy chain,
wherein the second heavy chain is identical to the first heavy chain except at one or more amino acid positions in the first heavy chain selected from the group consisting of heavy chain framework region positions H1, H8, H10, H12, H13, H15, H16, H19, H23, H26, H39, H42, H43, H46, H68, H72, H76, H81, H83, H85, and H86 (numbered according to the Kabat numbering system), wherein the amino acid residue at at least one of the one or more positions in the second heavy chain is a different amino acid residue of a different charge than the amino acid residue at the corresponding position in the first heavy chain, and
wherein a second IgG antibody comprising two copies of the second heavy chain binds to the same antigen as the first IgG antibody and has altered pharmacokinetics in blood compared to the first IgG antibody; and
(c) determining that the second IgG antibody has a pI shift compared to the first IgG antibody,
wherein either
(i) the pI of the second IgG antibody is decreased compared to the pI of the first IgG antibody, and the second IgG antibody has a prolonged half-life in blood, a prolonged mean residence time in blood, or a decreased blood clearance compared to the first IgG antibody; or
(ii) the pI of the second IgG antibody is increased compared to the pI of the first IgG antibody.

2. The method of claim 1 , wherein the altered pharmacokinetics in blood comprises a change in any one or more of the following parameters: half-life in blood, mean residence time in blood, or blood clearance.

3. The method of claim 1 , further comprising:
expressing the nucleic acid, thereby producing the second heavy chain.

4. A nucleic acid produced by the method of claim 1 .

5. The method of claim 1 , wherein the second IgG antibody is a humanized IgG antibody or a chimeric IgG antibody.

6. The method of claim 1 , wherein the one or more amino acid positions are selected from the group consisting of heavy chain framework region positions H10, H12, H23, H39, and H43, numbered according to the Kabat numbering system.

7. The method of claim 6 , wherein each different amino acid residue is selected from the group consisting of glutamic acid, aspartic acid, lysine, arginine, and histidine.

8. The method of claim 1 , wherein the pI shift is a decrease in pI of at least 0.3, and the altered pharmacokinetics in blood comprises increased half-life in blood or decreased clearance from blood.

9. The method of claim 1 , wherein the pI shift is an increase in pI of at least 0.3, and the altered pharmacokinetics in blood comprises decreased half-life in blood or increased clearance from blood.

10. The method of claim 1 , wherein the one or more positions are at least two positions.

11. The method of claim 1 , wherein the antigen-binding activity of the second IgG antibody is not reduced compared to the antigen-binding activity of the first IgG antibody.

12. A method for producing a nucleic acid, the method comprising:
producing a desired nucleic acid encoding a desired antibody heavy chain, the desired heavy chain having been previously designed by a method comprising:
(a) providing a nucleotide sequence encoding a first heavy chain of a first IgG antibody, the first heavy chain comprising an Fc domain;
(b) selecting one or more amino acid positions in the first heavy chain from the group consisting of heavy chain framework region positions H1, H8, H10, H12, H13, H15, H16, H19, H23, H26, H39, H42, H43, H46, H68, H72, H76, H81, H83, H85, and H86 (numbered according to the Kabat numbering system);
(c) designing the desired heavy chain to comprise an amino acid sequence that differs from the sequence of the first heavy chain in that the amino acid residue at at least one of the one or more amino acid positions selected in (b) is substituted with a different amino acid residue of a different charge;
wherein a second IgG antibody comprising two copies of the desired heavy chain binds to the same antigen as the first IgG antibody and has altered pharmacokinetics in blood compared to the first IgG antibody; and
(d) determining that the second IgG antibody has a pI shift compared to the first IgG antibody,
wherein either
(i) the pI of the second IgG antibody is decreased compared to the pI of the first IgG antibody, and the second IgG antibody has a prolonged half-life in blood, a prolonged mean residence time in blood, or a decreased blood clearance compared to the first IgG antibody; or
(ii) the pI of the second IgG antibody is increased compared to the pI of the first IgG antibody.

13. A method for producing a nucleic acid, the method comprising:
(a) providing a nucleotide sequence encoding a first heavy chain of a first IgG antibody that is not human or humanized, the heavy chain comprising an Fc domain;
(b) producing a nucleic acid encoding a second antibody heavy chain, the second heavy chain differing from the first heavy chain in that (i) the second heavy chain is humanized, and (ii) the second heavy chain has a different amino acid residue of a different charge than in the first heavy chain at one or more amino acid positions selected from the group consisting of heavy chain framework region positions H1, H8, H10, H12, H13, H15, H16, H19, H23, H26, H39, H42, H43, H46, H68, H72, H76, H81, H83, H85, and H86 (numbered according to the Kabat numbering system),
wherein a second IgG antibody comprising two copies of the second heavy chain binds to the same antigen as the first IgG antibody and has altered pharmacokinetics in blood compared to the first IgG antibody; and
(c) determining that the second IgG antibody has a pI shift compared to the first IgG antibody,
wherein either
(1) the pI of the second IgG antibody is decreased compared to the pI of the first IgG antibody, and the second IgG antibody has a prolonged half-life in blood, a prolonged mean residence time in blood, or a decreased blood clearance compared to the first IgG antibody; or
(2) the pI of the second IgG antibody is increased compared to the pI of the first IgG antibody.

14. The method of claim 13 , wherein the antigen-binding activity of the second IgG antibody is not reduced compared to the antigen-binding activity of the first IgG antibody.

15. The method of claim 13 , wherein the pI shift is a decrease in pI of at least 0.3, and the altered pharmacokinetics in blood comprises increased half-life in blood or decreased clearance from blood.

16. The method of claim 13 , wherein the pI shift is an increase in pI of at least 0.3, and the altered pharmacokinetics in blood comprises decreased half-life in blood or increased clearance from blood.

17. The method of claim 13 , wherein the one or more positions are at least two positions.

18. A method for producing a nucleic acid, the method comprising:
producing a desired nucleic acid encoding a desired antibody heavy chain, the desired heavy chain having been previously designed by a method comprising:
(a) providing a nucleotide sequence encoding a first heavy chain of a first IgG antibody that is not human or humanized, the first heavy chain comprising an Fc domain;
(b) selecting one or more amino acid positions in the first heavy chain from the group consisting of heavy chain framework region positions H1, H8, H10, H12, H13, H15, H16, H19,

Assignees

• Igawa Tomoyuki
• Tsunoda Hiroyuki
• Tachibana Tatsuhiko
• Chugai Pharmaceutical Co Ltd

Inventors

• Igawa Tomoyuki
• Tsunoda Hiroyuki
• Tachibana Tatsuhiko

Classifications

• C12N15/09 Primary

  Recombinant DNA-technology · CPC title

• C12N1/20

  Bacteria; Culture media therefor · CPC title

• C12N1/14

  Fungi (culture of mushrooms A01G18/00; as new plants A01H15/00); Culture media therefor · CPC title

• A61K39/395

  Antibodies (agglutinins A61K38/36 {; as drug carriers A61K47/50}); Immunoglobulins; Immune serum, e.g. antilymphocytic serum · CPC title

• C07K16/40 Primary

  against enzymes · CPC title