Serum albumin binding molecules

Patent metadata

Field	Value
Publication number	US-10934572-B2
Application number	US-201916244921-A
Country	US
Kind code	B2
Filing date	Jan 10, 2019
Priority date	May 3, 2010
Publication date	Mar 2, 2021
Grant date	Mar 2, 2021

Abstract

The present invention relates to an antibody-like protein based on the tenth fibronectin type III domain (10Fn3) that binds to serum albumin. The invention further relates to fusion molecules comprising a serum albumin-binding 10Fn3 joined to a heterologous protein for use in diagnostic and therapeutic applications.

First claim

We claim:

1. A method of increasing the serum half-life of a therapeutic moiety, the method comprising conjugating to the therapeutic moiety a fibronectin type III tenth ( 10 Fn3) domain which binds to domain 1 or 2 of human serum albumin with a K D of 1 μM or less, wherein the 10 Fn3 domain comprises a modified amino acid sequence in one or more of the BC, DE, and FG loops relative to the wild-type human 10 Fn3 domain (SEQ ID NO: 1), and wherein the polypeptide comprises an amino acid sequence which is at least 75% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 8, 12, 16, 20, and 24-44.

2. The method of claim 1 , wherein the BC, DE, and FG loops of the 10 Fn3 domain are selected from the group consisting of:
(a) a BC loop comprising the amino acid sequence set forth in SEQ ID NO: 5, a DE loop comprising the amino acid sequence set forth in SEQ ID NO: 6, and an FG loop comprising the amino acid sequence set forth in SEQ ID NO: 7,
(b) a BC loop comprising the amino acid sequence set forth in SEQ ID NO: 9, a DE loop comprising the amino acid sequence set forth in SEQ ID NO: 10, and an FG loop comprising the amino acid sequence set forth in SEQ ID NO: 11,
(c) a BC loop comprising the amino acid sequence set forth in SEQ ID NO: 13, a DE loop comprising the amino acid sequence set forth in SEQ ID NO: 14, and an FG loop comprising the amino acid sequence set forth in SEQ ID NO: 15,
(d) a BC loop comprising the amino acid sequence set forth in SEQ ID NO: 17, a DE loop comprising the amino acid sequence set forth in SEQ ID NO: 18, and an FG loop comprising the amino acid sequence set forth in SEQ ID NO: 19, and
(e) a BC loop comprising the amino acid sequence set forth in SEQ ID NO: 21, a DE loop comprising the amino acid sequence set forth in SEQ ID NO: 22, and an FG loop comprising the amino acid sequence set forth in SEQ ID NO: 23.

3. The method of claim 1 , wherein the 10 Fn3 domain comprises an amino acid sequence which is at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 8, 12, 16, 20, and 24-44.

4. The method of claim 1 , wherein the 10 Fn3 domain comprises an amino sequence selected from the group consisting of SEQ ID NOs: 4, 8, 12, 16, 20, and 24-44.

5. The method of claim 1 , wherein the 10 Fn3 domain further comprises an N-terminal extension sequence selected from the group consisting of SEQ ID NOs: 45-53.

6. The method of claim 1 , wherein the 10 Fn3 domain further comprises a C-terminal extension sequence selected from the group consisting of SEQ ID NOs: 54-64 and 215.

7. The method of claim 1 , wherein the therapeutic moiety is a small organic molecule, a nucleic acid, or a protein.

8. The method of claim 1 , wherein the therapeutic moiety targets receptors, receptor ligands, viral coat proteins, immune system proteins, hormones, enzymes, antigens, or cell signaling proteins.

9. A method of treating a disease condition in a subject, the method comprising administering to the subject a therapeutically effective amount of a fusion polypeptide comprising a fibronectin type III tenth ( 10 Fn3) domain and a therapeutic moiety, wherein the 10 Fn3 domain binds to domain 1 or 2 of human serum albumin with a K D of 1 μM or less, wherein the 10 Fn3 domain comprises a modified amino acid sequence in one or more of the BC, DE, and FG loops relative to the wild-type human 10 Fn3 domain (SEQ ID NO: 1), wherein the polypeptide comprises an amino acid sequence which is at least 75% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 8, 12, 16, 20, and 24-44, and wherein the disease condition responds to the biological activity of the therapeutic moiety contained in the fusion polypeptide.

10. The method of claim 9 , wherein the BC, DE, and FG loops of the 10 Fn3 domain are selected from the group consisting of:
(a) a BC loop comprising the amino acid sequence set forth in SEQ ID NO: 5, a DE loop comprising the amino acid sequence set forth in SEQ ID NO: 6, and an FG loop comprising the amino acid sequence set forth in SEQ ID NO: 7,
(b) a BC loop comprising the amino acid sequence set forth in SEQ ID NO: 9, a DE loop comprising the amino acid sequence set forth in SEQ ID NO: 10, and an FG loop comprising the amino acid sequence set forth in SEQ ID NO: 11,
(c) a BC loop comprising the amino acid sequence set forth in SEQ ID NO: 13, a DE loop comprising the amino acid sequence set forth in SEQ ID NO: 14, and an FG loop comprising the amino acid sequence set forth in SEQ ID NO: 15,
(d) a BC loop comprising the amino acid sequence set forth in SEQ ID NO: 17, a DE loop comprising the amino acid sequence set forth in SEQ ID NO: 18, and an FG loop comprising the amino acid sequence set forth in SEQ ID NO: 19, and
(e) a BC loop comprising the amino acid sequence set forth in SEQ ID NO: 21, a DE loop comprising the amino acid sequence set forth in SEQ ID NO: 22, and an FG loop comprising the amino acid sequence set forth in SEQ ID NO: 23.

11. The method of claim 9 , wherein the 10 Fn3 domain comprises an amino acid sequence which is at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 8, 12, 16, 20, and 24-44.

12. The method of claim 9 , wherein the 10 Fn3 domain comprises an amino sequence selected from the group consisting of SEQ ID NOs: 4, 8, 12, 16, 20, and 24-44.

13. The method of claim 9 , wherein the 10 Fn3 domain further comprises a C-terminal extension sequence selected from the group consisting of SEQ ID NOs: 54-64 and 215.

14. The method of claim 9 , wherein the 10 Fn3 domain further comprises an N-terminal extension sequence selected from the group consisting of SEQ ID NOs: 45-53.

15. The method of claim 9 , wherein the therapeutic moiety is a small organic molecule, a nucleic acid, or a protein.

16. The method of claim 9 , wherein the therapeutic moiety targets receptors, receptor ligands, viral coat proteins, immune system proteins, hormones, enzymes, antigens, or cell signaling proteins.

Assignees

• Bristol Myers Squibb Co

Inventors

• Gosselin Michael L
• Fabrizio David
• Swain Joanna F
• Mitchell Tracy
• Camphausen Ray
• Cload Sharon T
• Furfine Eric
• Morin Paul E
• Mukherjee Ranjan
• Taylor Simeon I

Classifications

• C07K14/78 Primary

  Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG] · CPC title

• A61K38/39

  Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG] · CPC title

• C07K19/00

  Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes} · CPC title

• C12N15/62

  DNA sequences coding for fusion proteins · CPC title

• A61K47/62 Primary

  the modifying agent being a protein, peptide or polyamino acid · CPC title