Fibronectin based scaffold domain proteins that bind to myostatin

Patent metadata

Field	Value
Publication number	US-10245302-B2
Application number	US-201615282277-A
Country	US
Kind code	B2
Filing date	Sep 30, 2016
Priority date	Sep 13, 2012
Publication date	Apr 2, 2019
Grant date	Apr 2, 2019

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 nucleic acid encoding a polypeptide comprising a fibronectin type III tenth ( 10 Fn3) domain which binds to myostatin and (i) comprises AB, BC, CD, DE, EF, and FG loops, wherein at least one loop of the BC, DE, and FG loops of the 10 Fn3 domain has 0, 1, 2, or 3 amino acid substitutions relative to the respective BC, DE, and FG loops of SEQ ID NOs: 34, 39, and 75.

2. The nucleic acid of claim 1 , wherein at least one loop of the BC, DE, and FG loops of the 10 Fn3 domain has 1 amino acid substitution relative to the respective BC, DE, and FG loops of SEQ ID NOs: 34, 39, and 75.

3. The nucleic acid of claim 1 , wherein:
(i) the serine at position 3 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of A, C, D, F, H, I, K, L, N, Q, R, T, V, W, or Y;
(ii) the leucine at position 4 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from M or V;
(iii) the proline at position 5 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of A, C, D, E, I, K, L, M, N, Q, R, S, T, V, or Y;
(iv) the histidine at position 6 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of A, C, D, E, F, G, I, K, L, M, N, Q, R, S, T, V, W, or Y;
(v) the glutamine at position 7 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, or Y;
(vi) the glycine at position 8 of the BC loop (SEQ ID NO: 34) is substituted with the amino acid S;
(vii) the lysine at position 9 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of A, C, D, E, F, G, H, I, L, M, N, Q, R, S, T, V, W, or Y;
(viii) the alanine at position 10 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of C, G, L, M, S, or T; or
(ix) the asparagine at position 11 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of A, C, F, H, P, Q, R, S, or Y.

4. The nucleic acid of claim 3 , wherein:
(i) the serine at position 3 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of C, F, I, V, W, or Y;
(ii) the histidine at position 6 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of C, D, E, F, G, I, K, L, M, N, Q, R, S, T, V, W, or Y;
(iii) the lysine at position 9 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of A, C, G, H, I, L, M, N, Q, R, S, V, W, or Y;
(iv) the alanine at position 10 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of G, L, M, or S; or
(v) the asparagine at position 11 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of C, H, Q, S, or Y.

5. The nucleic acid of claim 3 , wherein:
(i) the serine at position 3 of the BC loop (SEQ ID NO: 34) is substituted with the amino acid F or W;
(ii) the histidine at position 6 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of C, F, G, I, K, L, M, N, R, S, T, V, W, or Y;
(iii) the glutamine at position 7 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of A, C, E, F, H, I, K, L, M, P, R, S, T, V, or Y;
(iv) the lysine at position 9 of the BC loop (SEQ ID NO: 34) is substituted with an amino acid selected from the group consisting of A, C, H, L, M, N, R, V, W, or Y;
(v) the alanine at position 10 of the BC loop (SEQ ID NO: 34) is substituted with the amino acid G or L; or
(vi) the asparagine at position 11 of the BC loop (SEQ ID NO: 34) is substituted with the amino acid H or Q.

6. The nucleic acid of claim 1 , wherein the valine at position 5 of the DE loop (SEQ ID NO: 39) is substituted with an amino acid selected from the group consisting of A, C, D, E, F, I, K, L, M, N, Q, S, or T.

7. The nucleic acid of claim 6 , wherein the valine at position 5 of the DE loop (SEQ ID NO: 39) is substituted with an amino acid selected from the group consisting of C, E, I, L, M, Q, or T.

8. The nucleic acid of claim 6 , wherein the valine at position 5 of the DE loop (SEQ ID NO: 39) is substituted with an amino acid selected from the group consisting of C, E, I, L, or M.

9. The nucleic acid of claim 1 , wherein:
(i) the valine at position 2 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selected from the group consisting of A, C, F, I, L, M, Q, T, W, or Y;
(ii) the threonine at position 3 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selected from the group consisting of A, C, F, G, H, I, K, L, M, N, Q, R, S, V, W, or Y;
(iii) the aspartic acid at position 4 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selected from the group consisting of A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, or Y;
(iv) the threonine at position 5 of the FG loop (SEQ ID NO: 75) is substituted to with an amino acid selected from the group consisting of A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, or Y;
(v) the glycine at position 6 of the FG loop (SEQ ID NO: 75) is substituted to with an amino acid selected from the group consisting of A, C, D, E, F, H, I, K, L, M, N, Q, R, S, T, V, W, or Y;
(vi) the tyrosine at position 7 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selected from the group consisting of A, C, F, H, I, L, M, N, P, S, T, V, or W;
(vii) the leucine at position 8 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selected from the group consisting of A, C, E, F, H, I, K, M, N, Q, R, S, T, V, W, or Y;
(viii) the lysine at position 9 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selected from the group consisting of A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, or Y;
(ix) the tyrosine at position 10 of the FG loop (SEQ ID NO: 75) is substituted with the amino acid F or W; or
(x) the lysine at position 11 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selected from the group consisting of A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, or Y.

10. The nucleic acid of claim 9 , wherein:
(i) the valine at position 2 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selected from the group consisting of A, C, I, L, or M;
(ii) the threonine at position 3 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selected from the group consisting of C, F, H, I, L, M, Q, R, S, V, W, or Y;
(iii) the aspartic acid at position 4 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selected from the group consisting of A, C, E, F, G, H, I, L, M, N, P, Q, S, T, V, W, or Y;
(iv) the threonine at position 5 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selected from the group consisting of A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, V, W, or Y;
(v) the glycine at position 6 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selected from the group consisting of A, D, E, F, H, I, L, M, N, Q, S, T, V, W, or Y;
(vi) the tyrosine at position 7 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selected from the group consisting of C, F, I, L, M, P, T, V, or W;
(vii) the leucine at position 8 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selected from the group consisting of C, F, H, I, K, M, N, Q, R, T, V, W, or Y;
(viii) the lysine at position 9 of the FG loop (SEQ ID NO: 75) is substituted with an amino acid selec

Assignees

• Bristol Myers Squibb Co

Inventors

• Cload Sharon
• Engle Linda
• Lipovsek Dasa
• Madireddi Malavi
• Rakestraw Ginger Chao
• Swain Joanna
• Zhao Wenjun
• Wei Hui
• Yamniuk Aaron P
• Ramamurthy Vidhyashankar
• Kozhich Alexander T
• Corbett Martin J
• Krystek Jr Stanley Richard

Classifications

• 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

• A61P7/12

  Antidiuretics, e.g. drugs for diabetes insipidus (ADH A61P5/10) · CPC title

• 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