Compositions and methods for growth factor modulation

Patent metadata

Field	Value
Publication number	US-2016289318-A1
Application number	US-201615187278-A
Country	US
Kind code	A1
Filing date	Jun 20, 2016
Priority date	May 6, 2013
Publication date	Oct 6, 2016
Grant date	—

Abstract

Provided herein are proteins, antibodies, assays and methods useful for modulating growth factor levels and/or activities. In some embodiments, such growth factors are members of the TGF-β superfamily of proteins.

First claim

1 . An antibody produced by a method comprising the steps of:
i. expressing proGDF-8 (SEQ ID NO: 5); ii. forming a growth factor prodomain complex (GPC) by subjecting the expressed proGDF-8 (SEQ ID NO: 5) to enzymatic cleavage with one or more of furin, bone morphogenetic protein-1 (BMP-1), mammalian tolloid protein (mTLD), mammalian tolloid-like 1 (mTLL1), and mammalian tolloid-like 2 (mTLL2); iii. carrying out solid-phase or solution-phase enrichment with an antibody fragment phage display library, wherein the GPC formed by enzymatic cleavage is used as a target antigen; iv. selecting phage particles bound to the GPC formed by enzymatic cleavage; and v. producing recombinant antibodies having complementarity determining region (CDR) amino acid sequences derived from the antibody fragments expressed at the surface of the selected phage particles.

2 . The antibody of claim 1 , wherein said antibody is human or humanized.

3 . The antibody of claim 1 , wherein said antibody is subjected to affinity maturation.

4 . The antibody of claim 1 , wherein said method further comprises:
vi. screening the recombinant antibodies and selecting those which inhibit release of GDF-8 growth factor from GDF-8 GPCs.

5 . The antibody of claim 4 , wherein antibodies that inhibit GDF-8 activity are selected.

6 . The antibody of claim 1 , wherein said method further comprises:
vi. conducting a negative selection binding assay to exclude antibodies that bind to one or more undesired antigens.

7 . The antibody of claim 6 , wherein said one or more undesired antigens are selected from the group consisting of GDF-8 prodomain, GDF-8 growth factor, murine proGDF-8, proGDF-11, proTGF-β1, and a protein with an amino acid sequence selected from the group consisting of SEQ ID NOs: 207-230.

8 . The antibody of claim 7 , wherein said antibody is human or humanized.

9 . The antibody of claim 7 , wherein said antibody is subjected to affinity maturation.

10 . The antibody of claim 1 , wherein phage particle binding is determined using one or more binding assays selected from the group consisting of enzyme-linked immunosorbent assays, surface plasmon resonance assays, and flow cytometry assays.

11 . The antibody of claim 1 , wherein said enzymatic cleavage comprises sequential enzymatic cleavage with at least two enzymes.

12 . The antibody of claim 11 , wherein said antibody is human or humanized.

13 . The antibody of claim 11 , wherein said antibody is subjected to affinity maturation.

14 . The antibody of claim 1 , wherein at least one CDR amino acid sequence has at least one amino acid deletion, addition, or substitution relative to an amino acid sequence of said antibody fragments expressed at the surface of the selected phage particles.

15 . The antibody of claim 1 comprising an isotype selected from the group consisting of IgG1, IgG2, IgG3, IgG4, IgA, IgGA2, IgD, IgE, and IgM.

16 . The antibody of claim 1 comprising a bispecific antibody.

Assignees

• Scholar Rock Inc

Inventors

• Schurpf Thomas
• Mahanthappa Nagesh K
• Straub Michelle Marie

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P7/00

  Drugs for disorders of the blood or the extracellular fluid · 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

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title