COMPOSITIONS AND METHODS FOR GROWTH FACTOR MODULATION

• US 20200131259A1
• Filed: 01/07/2020
• Published: 04/30/2020
• Est. Priority Date: 05/06/2013
• Status: Active Application

• 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
• Application Number:
  US16/736,207
• Publication Number:
  US 20200131259A1
• Time in Patent Office: Days
• 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. A method for manufacturing a pharmaceutical composition that comprises an antibody, or an antigen-binding portion thereof, that inhibits TGF-β

1 signaling, the method comprising the steps of;

(i) providing an antigen comprising a protein complex that comprises;

a) a homodimer comprising prodomains of transforming growth factor beta (TGF-β

1), or prodomain fragments thereof that comprise a straightjacket region that includes at least one cysteine residue present within about the first 4, 5, 6 or 7 N-terminal region; and

,

b) a protein known to associate with a pro-protein of TGF-β

1, selected from the group consisting of;

latent TGF-β

binding protein 1 (LTBP1), latent TGF-β

binding protein 3 (LTBP3), glycoprotein A repetitions predominant (GARP), leucine rich repeat containing 33 (LRRC33), and fragments or variants thereof that comprise two cysteine residues that form disulfide bonds with the straightjacket region of the prodomains, wherein the variants are at least 90% identical to a native sequence;

(ii) selecting for one or more antibodies, or antigen-binding portions thereof, for the ability to bind the antigen of step (i) and inhibit TGF-β

1 signaling; and

,

(iii) expressing the antibody, or the antigen-binding portion thereof, selected in step (ii) in mammalian cells capable of large scale translation.

2. The method of claim 1, further comprising a step of:

(iv) purifying an antibody or antigen-binding portion thereof expressed in step (iii) from the culture medium.

3. The method of claim 1, further comprising a step of:

(iv) formulating the antibody or antigen-binding portion thereof purified in step (iv) into a pharmaceutical composition, wherein the antibody, or antigen-binding portion thereof, is a human or humanized antibody or antigen-binding portion thereof.

4. The method of claim 1, further comprising a step of:

immunizing a host animal with the antigen of step (i).

5. The method of claim 4, further comprising a step of:

collecting lymphocytes from the host animal that bind the antigen of step (i).

6. The method of claim 1, wherein step (ii) comprises screening a library.

7. The method of claim 6, wherein the library is an antibody display library.

8. The method of claim 7, wherein the display library is an antibody fragment display library.

9. The method of claim 8, wherein the antibody fragment display library comprises Fab fragments or single-chain variable fragments (scFvs).

10. The method of claim 7, wherein the display library is expressed in a host selected from the group consisting of yeast, bacteriophage, bacteria and retroviruses.

11. The method of claim 1, further comprising a step of preparing hybridoma cells.

12. The method of claim 1, further comprising a step of:

carrying out negative selection, wherein the antibodies and antigen-binding portions thereof, that bind a mature TGFβ

1 growth factor and/or pro-proteins of related growth factors are removed from the pool.

13. The method of claim 1, further comprising a step of:

carrying out affinity maturation and/or optimization.

15. The method of claim 1, wherein the antigen is a cell-based antigen.

16. The method of claim 1, wherein the antibody binds a latency loop of the prodomain.

17. The method of claim 1, wherein each of the prodomains of TGF-β

1 comprises at least about 50 amino acid residues.

18. The method of claim 1, wherein the protein known to associate with the pro-protein of TGF-β

1, or fragments thereof, comprises at least about 50 amino acid residues.

19. The method of claim 1, wherein the antibodies, or antigen-binding portions thereof, of step (ii) inhibit TGF-β

1 signaling by binding to the protein complex and blocking release of mature TGF-β

1.

20. The method of claim 1, wherein the cysteine residues that form disulfide bonds with the straightjacket region of the prodomains are native cysteine residues.

14. The method of 1, wherein the antibody, or antigen-binding portion thereof, is an IgA, IgD, IgE, IgG or IgM antibody.

21. A method for manufacturing a pharmaceutical composition that comprises an antibody, or an antigen-binding portion thereof, that inhibits TGF-β

1 signaling, the method comprising the steps of;

(i) providing an antigen comprising a protein complex that comprises;

a) a homodimer comprising prodomains of transforming growth factor beta (TGF-β

1), or prodomain fragments thereof that comprise a straightjacket region that includes at least one cysteine residue present within about the first 4, 5, 6 or 7 N-terminal region; and

,

b) a protein known to associate with a pro-protein of TGF-β

1, selected from the group consisting of;

latent TGF-β

binding protein 1 (LTBP1), latent TGF-β

binding protein 3 (LTBP3), glycoprotein A repetitions predominant (GARP), leucine rich repeat containing 33 (LRRC33), and fragments or variants thereof that comprise two cysteine residues that form disulfide bonds with the straightjacket region of the prodomains, wherein the variants are at least 90% identical to a native sequence;

(ii) selecting for one or more antibodies, or antigen-binding portions thereof, for the ability to bind to the protein complex and block release of mature TGF-β

1;

(iii) screening the antibody, or the antigen-binding fragment thereof, that binds to the protein complex in step (ii) for inhibitors of TGF-β

1 signaling;

(iv) expressing the antibody, or the antigen-binding portion thereof, selected in step (iii) in mammalian cells capable of large scale translation.

22. The method of claim 21, further comprising a step of:

(v) purifying an antibody or antigen-binding portion thereof expressed in step (iv) from culture medium.

23. The method of claim 21, further comprising a step of:

(v) formulating the antibody or antigen-binding portion thereof purified in step (iv) into a pharmaceutical composition, wherein the antibody, or antigen-binding portion thereof, is a human or humanized antibody or antigen-binding portion thereof.

24. The method of claim 21, wherein the cysteine residues that form disulfide bonds with the straightjacket region of the prodomains are native cysteine residues.

25. The method of claim 21, wherein the antibody, or the antigen-binding fragment thereof, inhibits TGF-β

1 signaling in step (iii) by

blocking release of mature TGF-β

1 growth factor from pro/latent TGF-β

1; and

/or

blocking release of mature TGF-β

1 growth factor from the protein complex.