Fibronectin based scaffold domain proteins that bind to myostatin

Patent metadata

Field	Value
Publication number	US-9662373-B2
Application number	US-201514632436-A
Country	US
Kind code	B2
Filing date	Feb 26, 2015
Priority date	Sep 13, 2012
Publication date	May 30, 2017
Grant date	May 30, 2017

Abstract

The present invention relates to fibronectin-based scaffold domain proteins that bind to myostatin. The invention also relates to the use of these proteins in therapeutic applications to treat muscular dystrophy, cachexia, sarcopenia, osteoarthritis, osteoporosis, diabetes, obesity, COPD, chronic kidney disease, heart failure, myocardial infarction, and fibrosis. The invention further relates to cells comprising such proteins, polynucleotides encoding such proteins or fragments thereof, and to vectors comprising the polynucleotides encoding the proteins.

First claim

We claim:

1. A method of increasing muscle mass in a subject comprising administering to the subject a sufficient amount of a polypeptide comprising a fibronectin type III tenth ( 10 Fn3) domain which binds to myostatin and comprises the amino acid sequence of SEQ ID NO: 6, wherein (Z) x , (Z) y and (Z) z comprise:
(a) the amino acid sequences of SEQ ID NOs: 7, 39 and 46, respectively;
(b) the amino acid sequences of SEQ ID NOs: 8, 40 and 47, respectively;
(c) the amino acid sequences of SEQ ID NOs: 12, 42 and 51, respectively;
(d) the amino acid sequences of SEQ ID NOs: 17, 39 and 56, respectively;
(e) the amino acid sequences of SEQ ID NOs: 18, 39 and 57, respectively;
(f) the amino acid sequences of SEQ ID NOs: 17, 39 and 62, respectively;
(g) the amino acid sequences of SEQ ID NOs: 32, 44 and 73, respectively;
(h) the amino acid sequences of SEQ ID NOs: 33, 45 and 74, respectively;
(i) the amino acid sequences of SEQ ID NOs: 34, 39 and 75, respectively:
(j) the amino acid sequences of SEQ ID NOs: 35, 39 and 75, respectively; or
(k) the amino acid sequences of SEQ ID NOs: 38, 39 and 79.

2. The method of claim 1 , wherein (Z) x , (Z) y and (Z) z comprise the amino acid sequences of SEQ ID NOs: 34, 39 and 75, respectively.

3. The method of claim 1 , wherein the subject has a disorder associated with muscle wasting and/or muscle atrophy.

4. The method of claim 3 , wherein the subject has a myopathy.

5. The method of claim 4 , wherein the subject has a neuromuscular disorder.

6. The method of claim 5 , wherein the neuromuscular disorder is muscular dystrophy.

7. The method of claim 1 , wherein the subject has Duchenne's muscular dystrophy.

8. The method of claim 1 , wherein muscle mass is increased by about 10% or more.

9. The method of claim 8 , wherein muscle mass is increased by about 20% or more.

10. The method of claim 9 , wherein muscle mass is increased by about 30% or more.

11. The method of claim 1 , wherein the polypeptide binds to myostatin with a K D of less than 100 nM.

12. The method of claim 11 , wherein the polypeptide binds to myostatin with a K D of less than 10 nM.

13. The method of claim 1 , wherein the 10 Fn3 domain of the polypeptide further comprises an N-terminal extension sequence comprising the amino acid sequence of SEQ ID NO: 307.

14. The method of claim 1 , wherein the 10 Fn3 domain of the polypeptide further comprises a C-terminal extension sequence comprising amino acid residues EI.

15. The method of claim 1 , wherein the 10 Fn3 domain of the polypeptide further comprises an N-terminal extension sequence comprising the amino acid sequence of SEQ ID NO: 307, and a C-terminal extension sequence comprising amino acid residues EI.

16. The method of claim 1 , wherein the polypeptide further comprises one or more pharmacokinetic (PK) moieties selected from the group consisting of polyethylene glycol, sialic acid, Fc, Fc fragment, transferrin, serum albumin, a serum albumin binding protein, and a serum immunoglobulin binding protein.

17. The method of claim 16 , wherein the PK moiety and the polypeptide are linked via a linker comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 181-187.

18. The method of claim 16 , wherein the PK moiety is an Fc.

19. The method of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 273.

20. The method of claim 15 , wherein the 10 Fn3 domain comprises the amino acid sequence of SEQ ID NO: 281.

21. The method of claim 18 , wherein the Fc comprises IgG1 CH2 and CH3 domains.

22. The method of claim 21 , wherein the Fc comprises SEQ ID NO: 276.

23. A method of increasing muscle mass in a subject with Duchenne's muscular dystrophy comprising administering to the subject a sufficient amount of a polypeptide comprising a fibronectin type III tenth ( 10 Fn3) domain which binds to myostatin and comprises the amino acid sequence of SEQ ID NO: 6, wherein (Z) x , (Z) y and (Z) z comprise SEQ ID NOs: 34, 39 and 75, respectively.

24. The method of claim 23 , wherein the 10 Fn3 domain comprises the amino acid sequence of SEQ ID NO: 281.

25. The method of claim 23 , wherein the polypeptide further comprises one or more pharmacokinetic (PK) moieties selected from the group consisting of polyethylene glycol, sialic acid, Fc, Fc fragment, transferrin, serum albumin, a serum albumin binding protein, and a serum immunoglobulin binding protein.

26. The method of claim 25 , wherein the PK moiety and the polypeptide are linked via a linker comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 181-187.

27. The method of claim 25 , wherein the PK moiety is an Fc.

28. The method of claim 27 , wherein the Fc comprises IgG1 CH2 and CH3 domains.

29. The method of claim 27 , wherein the Fc comprises SEQ ID NO: 276.

30. The method of claim 23 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 273.

31. A method of increasing muscle mass in a subject with Duchenne's muscular dystrophy comprising administering to the subject a sufficient amount of a polypeptide comprising a fibronectin type III tenth ( 10 Fn3) domain which binds to myostatin and comprises the amino acid sequence of SEQ ID NO: 273.

Assignees

• Bristol Myers Squibb Co

Inventors

• Cload Sharon
• Engle Linda
• Lipovsek Dasa
• Madireddi Malavi
• Rakestraw Ginger Chao
• Swain Joanna
• Zhao Wenjun
• Kozhich Alexander T
• Corbett Martin J

Classifications

• A61P5/50

  for increasing or potentiating the activity of insulin · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P3/06

  Antihyperlipidemics · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title