Compositions and methods for growth factor modulation

Patent metadata

Field	Value
Publication number	US-9758577-B2
Application number	US-201615187278-A
Country	US
Kind code	B2
Filing date	Jun 20, 2016
Priority date	May 6, 2013
Publication date	Sep 12, 2017
Grant date	Sep 12, 2017

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

The invention claimed is:

1. An antibody produced by a method comprising the steps of:
i. expressing mammalian proGDF-8;
ii. subjecting the expressed mammalian proGDF-8 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), so as to form a growth factor prodomain complex (GPC);
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 that bind to the GPC formed by enzymatic cleavage and excluding phage particles that bind to isolated GDF-8 growth factor or isolated GDF-8 prodomain; and
v. producing recombinant antibodies having complementarity determining regions (CDRs) of an antibody fragment 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 to remove 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 comprising an isotype selected from the group consisting of IgG1, IgG2, IgG3, IgG4, IgA, IgGA2, IgD, IgE, and IgM.

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

16. The antibody of claim 1 , wherein said mammalian proGDF-8 comprises the amino acid sequence of SEQ ID NO: 5.

Assignees

• Scholar Rock Inc

Inventors

• Schurpf Thomas
• Mahanthappa Nagesh K
• Straub Michelle Marie

Classifications

• A61P7/00

  Drugs for disorders of the blood or the extracellular fluid · CPC title

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title

• A61P37/02

  Immunomodulators · CPC title

• A61P43/00

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