Deimmunized serum-binding domains and their use in extending serum half-life

Patent metadata

Field	Value
Publication number	US-9376495-B2
Application number	US-201214118516-A
Country	US
Kind code	B2
Filing date	May 16, 2012
Priority date	May 21, 2011
Publication date	Jun 28, 2016
Grant date	Jun 28, 2016

Abstract

The present invention is directed to a polypeptide (for example, an antigen-binding molecule) that comprises a polypeptide portion of a deimmunized serum-binding protein capable of binding to said serum protein. The presence of the serum-binding protein extends the serum half-life of the polypeptide, relative to the serum half-life of the polypeptide if lacking the polypeptide portion of the deimmunized serum-binding protein. The invention also pertains to methods and uses that employ such molecules.

First claim

What is claimed is:

1. A polypeptide that comprises a portion of a deimmunized albumin-binding protein capable of binding to serum albumin; wherein said deimmunized albumin-binding protein portion is a variant of a wild-type albumin-binding domain (ABD) of a Streptococcal Protein G, said wild-type ABD having the amino acid sequence of SEQ ID NO: 304; wherein said variant ABD has an amino acid sequence that differs from that of SEQ ID NO: 304 in comprising:
(A) a valine to alanine variation at position 31 of SEQ ID NO:304; a valine to alanine variation at position 34 of SEQ ID NO:304; or an aspartate to alanine variation at position 39 of SEQ ID NO:304;
(B) a leucine to alanine variation at position 24 of SEQ ID NO:304; an isoleucine to alanine variation at, position 25 of SEQ ID NO:304; and a valine to alanine variation at position 25 of SEQ ID NO:304; or
(C) an asparagine to aspartic acid variation at position 26 of SEQ ID NO:304; a threonine to serine substitution at position 30 of SEQ ID NO:304; and a valine to alanine variation at position 31 of SEQ ID NO:304;
wherein said deimmunized albumin-binding protein portion extends the serum half-life of said polypeptide, relative to the serum half-life of said polypeptide if lacking said portion of said deimmunized albumin-binding protein.

2. The polypeptide of claim 1 , wherein said polypeptide comprises an additional portion of a deimmunized albumin-binding protein, wherein said portions of said deimmunized albumin-binding protein are both capable of binding to said serum albumin.

3. The polypeptide of claim 1 , wherein said variant ABD comprises said valine to alanine variation at position 31 of SEQ ID NO:304; said valine to alanine variation at position 34 of SEQ ID NO:304; or said aspartate to alanine variation at position 39 of SEQ ID NO:304.

4. The polypeptide of claim 1 , wherein said variant ABD comprises said leucine to alanine variation at position 24 of SEQ ID NO:304; said isoleucine to alanine variation at position 25 of SEQ ID NO:304; and said valine to alanine variation at position 31 of SEQ ID NO:304.

5. The polypeptide of claim 1 , wherein said variant ABD comprises said asparagine to aspartic acid variation at position 26 of SEQ ID NO:304; said threonine to serine substitution at position 30 of SEQ ID NO:304; and said valine to alanine variation at position 31 of SEQ ID NO:304.

6. The polypeptide of claim 1 , wherein said variant ABD has the sequence of SEQ ID NO: 323, SEQ ID NO: 324, SEQ ID NO: 326, SEQ ID NO: 327, SEQ ID NO: 328, or SEQ ID NO: 329.

7. The polypeptide of claim 1 , wherein said polypeptide comprises an antigen-binding molecule.

8. The polypeptide of claim 7 , wherein said antigen-binding molecule is a diabody composed of at least a first and a second polypeptide chain which interact with one another to form two antigen-binding sites, wherein at least one of said polypeptide chains comprises said portion of said deimmunized albumin-binding protein that is capable of binding to said serum albumin.

9. The polypeptide of claim 8 , wherein both said first and said second polypeptide chains comprise said portion of said deimmunized albumin-binding protein capable of binding to said serum albumin.

10. The polypeptide of claim 8 , wherein said first and said second polypeptide chains are covalently linked to one another.

11. The polypeptide of claim 8 , wherein said diabody binds to:
(A) the Natural Killer Group 2D (NKG2D) receptor or the T-cell receptor (TCR); and
(B) a tumor-associated antigen.

12. The polypeptide of claim 7 , wherein said antigen is a breast cancer antigen, an ovarian cancer antigen, a prostate cancer antigen, a cervical cancer antigen, a pancreatic carcinoma antigen, a lung cancer antigen, a bladder cancer antigen, a colon cancer antigen, a testicular cancer antigen, a glioblastoma cancer antigen, an antigen associated with a B cell malignancy, an antigen associated with multiple myeloma, an antigen associated with non-Hodgkin's lymphoma, or an antigen associated with chronic lymphocytic leukemia.

13. A method for extending the serum half-life of a polypeptide, which comprises covalently linking said polypeptide to a polypeptide portion of a deimmunized albumin-binding protein, said extension of serum half-life being relative to the serum half-life of said polypeptide if lacking said albumin-binding protein; wherein said deimmunized albumin-binding protein portion is a variant of a wild-type albumin-binding domain (ABD) of a Streptococcal Protein G, said wild-type ABD having the amino acid sequence of SEQ ID NO: 304; wherein said variant ABD has an amino acid sequence that differs from that of SEQ ID NO: 304 in comprising:
(A) a valine to alanine variation at position 31 of SEQ ID NO:304; a valine to alanine variation at position 34 of SEQ ID NO:304; or an aspartate to alanine variation at position 39 of SEQ ID NO:304;
(B) a leucine to alanine variation at position 24 of SEQ ID NO:304; an isoleucine to alanine variation at, position 25 of SEQ ID NO:304; and a valine to alanine variation at position 31 of SEQ ID NO:304; or
(C) an asparagine to aspartic acid variation at position 26 of SEQ ID NO:304; a threonine to serine substitution at position 30 of SEQ ID NO:304; and a valine to alanine variation at position 31 of SEQ ID NO:304.

14. The method of claim 13 , wherein said polypeptide comprises an additional portion of a deimmunized albumin-binding protein, wherein said portions of said deimmunized albumin-binding protein are both capable of binding to said serum albumin.

15. The method of claim 13 , wherein said variant ABD comprises said valine to alanine variation at position 31 of SEQ ID NO:304; said valine to alanine variation at position 34 of SEQ ID NO:304; or said aspartate to alanine variation at position 39 of SEQ ID NO:304.

16. The method of claim 13 , wherein said variant ABD comprises said leucine to alanine variation at position 24 of SEQ ID NO:304; said isoleucine to alanine variation at position 25 of SEQ ID NO:304; and said valine to alanine variation at position 31 of SEQ ID NO:304.

17. The method of claim 16 , wherein said variant ABD comprises said asparagine to aspartic acid variation at position 26 of SEQ ID NO:304; said threonine to serine substitution at position 30 of SEQ ID NO:304; and said valine to alanine variation at position 31 of SEQ ID NO:304.

18. The method of claim 13 , wherein said variant ABD has the sequence of SEQ ID NO: 323, SEQ ID NO: 324, SEQ ID NO: 326, SEQ ID NO: 327, SEQ ID NO: 328, or SEQ ID NO: 329.

19. The method of claim 13 , wherein said polypeptide comprises an antigen-binding molecule.

20. The method of claim 19 , wherein said antigen-binding molecule is a diabody composed of at least a first and a second polypeptide chain which interact with one another to form two antigen-binding sites, wherein at least one of said polypeptide chains comprises said portion of said deimmunized albumin-binding protein that is capable of binding to said serum albumin.

21. The method of claim 20 , wherein both said first and said second polypeptide chains comprise said portion of said deimmunized albumin-binding protein capable of binding to said serum albumin.

22. The method of claim 20 , wherein said first and said second polypeptide chains are covalently linked to one another.

23. The method of claim 20 , wherein said diabody binds to:
(A) the Natural Killer Group 2D (NKG2D) receptor or the T-cell receptor (TCR); and
(B) a tumor-a

Assignees

• Bonvini Ezio
• Barat Bhaswati
• Huang Ling
• Johnson Leslie S
• Macrogenics Inc

Inventors

• Bonvini Ezio
• Barat Bhaswati
• Huang Ling
• Johnson Leslie S

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• C07K2317/24

  containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered · CPC title

• C07K2319/73

  containing coiled-coiled motif (leucine zippers) · CPC title

• C07K2317/31

  multispecific · CPC title

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title