Compositions and methods for growth factor modulation

• US 9,758,577 B2
• Filed: 06/20/2016
• Issued: 09/12/2017
• Est. Priority Date: 05/06/2013
• Status: Active Grant

• Current Assignee: Scholar Rock, Inc. (Scholar Rock Holding Corp.)
• Original Assignee: Scholar Rock, Inc. (Scholar Rock Holding Corp.)
• Inventors: Schurpf, Thomas, Mahanthappa, Nagesh K., Straub, Michelle Marie
• Primary Examiner(s): KEMMERER, ELIZABETH
• Application Number:
  US15/187,278
• Publication Number:
  US 20160289318A1
• Time in Patent Office: 449 Days
• Field of Search: None
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 39/001134 Transforming growth factor [TGF]
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/02 Muscle relaxants, e.g. for tetanus or cramps
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/495 : Transforming growth factor [TGF]
• CPC Class Codes: C07K 16/005 : constructed by phage libraries
• CPC Class Codes: C07K 16/22 : against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/28 : against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/2863 : against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 2317/21 : from primates, e.g. man
• CPC Class Codes: C07K 2317/24 : containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/31 : multispecific
• CPC Class Codes: C07K 2317/32 : specific for a neo-epitope on a complex, e.g. antibody-antigen or ligand-receptor
• CPC Class Codes: C07K 2317/33 : Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/55 : Fab or Fab'
• CPC Class Codes: C07K 2317/565 : Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/76 : Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2317/94 : Stability, e.g. half-life, pH, temperature or enzyme-resistance
• CPC Class Codes: C12N 15/1037 : Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display
• CPC Class Codes: G01N 2333/495 : Transforming growth factor [TGF]
• CPC Class Codes: G01N 2333/71 : for growth factors; for growth regulators

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.

Claims

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-β

1, 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.