IP Library › Granted Patent US 8,969,289
Granted Patent B2
US 8,969,289 · App. 13/098,851 · Granted Mar 3, 2015

Serum albumin binding molecules

Inventors: Michael L. Gosselin (Boston, MA); David Fabrizio (Swampscott, MA); Joanna F. Swain (Concord, MA); Tracy Mitchell (Billerica, MA); Ray Camphausen (Wayland, MA); Sharon T. Cload (Cambridge, MA); Eric Furfine (Concord, MA); Paul E. Morin (Pennington, NJ); Ranjan Mukherjee (Churchville, PA); Simeon I. Taylor (Skillman, NJ)
Assignee: Bristol-Myers Squibb Company
C07K14/78A61K47/48238C12N15/62A61K38/00A61K47/48284
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 8,969,289
App. No.
13/098,851
Granted
Mar 3, 2015
Kind
B2
Abstract

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

Claims (38)

1. A polypeptide comprising a fibronectin type III tenth ( 10 Fn3) domain, wherein the 10 Fn3 domain binds to 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 at least 75% identical to an amino acid sequence selected from SEQ ID NOs: 4, 8, 12, 16, 20, and 24-44.

2. The polypeptide of claim 1 , wherein the 10 Fn3 domain binds to domain 1 or 2 of human serum albumin with a KD of 1 μM or less.

3. The polypeptide of claim 1 , wherein the 10 Fn3 domain binds to serum albumin at a pH range of 5.5 to 7.4.

4. The polypeptide claim 1 , wherein the 10 Fn3 domain binds to HSA with a K D of 200 nM or less at pH 5.5.

5. The polypeptide of claim 1 , wherein the 10 Fn3 domain also binds to one or more of rhesus serum albumin (RhSA), cynomolgous monkey serum albumin (CySA), or murine serum albumin (MuSA).

6. The polypeptide of claim 1 , wherein the serum half-life of the polypeptide in the presence of serum albumin is at least 2 hours.

7. A fusion polypeptide comprising a fibronectin type III tenth ( 10 Fn3) domain and a heterologous protein, wherein the 10 Fn3 domain binds to human serum albumin with a K D of 1 μM or less and comprises an amino acid sequence 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.

8. The fusion polypeptide of claim 7 , wherein the 10 Fn3 domain comprises an amino acid sequence selected from SEQ ID NOs: 4, 8, 12, 16, 20, and 24-44.

9. The fusion polypeptide of claim 7 , wherein the 10 Fn3 domain comprises a BC loop having the amino acid sequence set forth in SEQ ID NO: 5, a DE loop having the amino acid sequence set forth in SEQ ID NO: 6, and an FG loop having the amino acid sequence set forth in SEQ ID NO: 7.

10. The fusion polypeptide of claim 7 , wherein the heterologous protein is selected from fibroblast growth factor 21 (FGF21), insulin, insulin receptor peptide, (bone morphogenetic protein 9 (BMP-9), amylin, peptide YY (PYY3-36), pancreatic polypeptide (PP), interleukin 21 (IL-21), glucagon-like peptide 1 (GLP-1), Plectasin, Progranulin, Atstrrin, Osteocalcin (OCN), Apelin, or a polypeptide comprising a 10 Fn3 domain.

11. The fusion polypeptide of claim 10 , wherein the heterologous protein is FGF21.

12. A polypeptide comprising a fibronectin type III tenth ( 10 Fn3) domain, wherein the 10 Fn3 domain (i) comprises a modified amino acid sequence in one or more of the AB, BC, CD, DE, EF and FG loops relative to the wild-type 10 Fn3 domain, (ii) binds to a target molecule not bound by the wild-type 10 Fn3 domain, and (iii) comprises a C-terminal tail having a sequence (ED)n, wherein n is an integer from 3 to 7.

13. A pharmaceutical composition comprising the polypeptide of claim 1 .

14. The polypeptide 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.

15. The polypeptide of claim 1 , wherein the 10 Fn3 domain comprises an amino acid sequence at least 80% identical to an amino acid sequence selected from SEQ ID NOs: 4, 8, 12, 16, 20, and 24-44.

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

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

18. The polypeptide of claim 1 , wherein the serum half-life of the polypeptide in the presence of albumin is at least 5-fold greater than the serum half-life of the polypeptide in the absence of serum albumin.

19. The polypeptide of claim 1 , wherein the 10 Fn3 domain further comprises an N-terminal extension sequence selected from any one of the amino acid sequences set forth in SEQ ID NOs: 45-53.

20. The polypeptide of claim 1 , wherein the 10 Fn3 domain further comprises a C-terminal extension sequence selected from any one of the amino acid sequences set forth in SEQ ID NOs: 54-64 and 215.

21. The polypeptide of claim 18 , wherein the 10 Fn3 domain comprises an amino acid sequence selected from SEQ ID NOs: 89-116 and 222-225.

22. The polypeptide of claim 1 , wherein the 10 Fn3 domain comprises 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.

23. The polypeptide of claim 1 , wherein the 10 Fn3 domain comprises 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.

24. The polypeptide of claim 1 , wherein the 10 Fn3 domain comprises 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.

25. The polypeptide of claim 1 , wherein the 10 Fn3 domain comprises 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.

26. The polypeptide of claim 1 , wherein the 10 Fn3 domain comprises 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.

27. The polypeptide of claim 1 , wherein the 10 Fn3 domain comprises an amino acid sequence at least 75% identical to the amino acid sequence of SEQ ID NO: 4.

28. The polypeptide of claim 15 , wherein the 10 Fn3 domain comprises an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 4.

29. The polypeptide of claim 16 , wherein the 10 Fn3 domain comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 4.

30. The polypeptide of claim 17 , wherein the 10 Fn3 domain comprises the amino sequence of SEQ ID NO: 4.

31. The polypeptide of claim 7 , wherein the 10 Fn3 domain comprises an amino acid sequence at least 75% identical to the amino acid sequence of SEQ ID NO: 4.

32. The polypeptide of claim 8 , wherein the 10 Fn3 domain comprises the amino acid sequence of SEQ ID NO: 4.

33. The polypeptide of claim 21 , wherein the 10 Fn3 domain comprises the amino acid sequence of SEQ ID NO: 90.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jul 28, 2011
From: GOSSELIN, MICHAEL L.; FABRIZIO, DAVID; SWAIN, JOANNA F.; MITCHELL, TRACY; CAMPHAUSEN, RAY; CLOAD, SHARON T.; FURFINE, ERIC; MORIN, PAUL; MUKHERJEE, RANJAN; TAYLOR, SIMEON I.
To: BRISTOL-MYERS SQUIBB COMPANY
Reel/Frame 026665/0533 →
Continuity (2)
Provisional Application 61330672 · May 3, 2010
Related Publication 20110305663A1 · Dec 15, 2011