Compositions and methods for growth factor modulation

Patent metadata

Field	Value
Publication number	US-12577296-B2
Application number	US-202016736207-A
Country	US
Kind code	B2
Filing date	Jan 7, 2020
Priority date	May 6, 2013
Publication date	Mar 17, 2026
Grant date	Mar 17, 2026

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

What is claimed is:

1 . A method for manufacturing a pharmaceutical composition that comprises an antibody, or an antigen-binding portion thereof, that inhibits transforming growth factor beta 1 (TGF-β1) signaling, the method comprising the steps of:
(i) screening an antibody display library or hybridoma cell lines for an antibody, or an antigen-binding portion thereof, capable of binding an antigen comprising a transforming growth factor beta 1 prodomain complex (TGF-β1 GPC), that comprises:
a) a latency associated peptide (LAP) dimer, wherein each polypeptide of the dimer has an amino acid sequence comprising SEQ ID NO: 38 or SEQ ID NO: 99; and
b) a growth factor dimer, wherein each polypeptide of the dimer has an amino acid sequence comprising SEQ ID NO: 44; and
selecting an antibody, or antigen-binding portion thereof, that binds to the TGF-β1 GPC;

(ii) providing a growth factor activity assay to measure inhibition of TGF-β1 signaling comprising:
a) contacting the TGF-β1 GPC with the antibody, or antigen-binding portion thereof, selected in step (i), a cell expressing αvβ6 integrin and/or αvβ8 integrin, and a responsive cell comprising a reporter gene comprising a TGF-β1 growth factor responsive promoter operably linked to a sequence encoding a detectable gene product;
b) obtaining TGF-β1 signaling activity data by detecting expression of the detectable gene product, and
c) selecting an antibody, or an antigen-binding portion thereof, that inhibits TGF-β1 signaling;

(iii) preparing expression vectors comprising the coding sequences of the antibody, or the antigen-binding portions thereof, selected in step (ii);
(iv) transfecting the vectors prepared in step (iii) into a cell in culture medium for expression of the antibody, or antigen-binding portion thereof;
(v) purifying the antibody, or the antigen-binding portion thereof, expressed in step (iv) from the culture medium; and
(vi) formulating the antibody, or the antigen-binding portion thereof, purified in step (v) into a pharmaceutical composition.

2 . The method of claim 1 , wherein the antibody display library is an antibody fragment display library.

3 . The method of claim 2 , wherein the antibody fragment display library comprises Fab fragments or single-chain variable fragments (scFvs).

4 . The method of claim 1 , wherein the antibody display library is expressed in a host selected from the group consisting of yeast, bacteriophage, bacteria and retroviruses.

5 . The method of claim 1 , wherein the antibody display library screened in step (i) comprises human antibodies or antigen-binding portions thereof.

6 . A method for manufacturing a pharmaceutical composition that comprises an antibody, or an antigen-binding portion thereof, that inhibits transforming growth factor beta 1 (TGF-β1) signaling, the method comprising the steps of:
(i) screening an antibody display library or hybridoma cell lines for an antibody, or an antigen-binding portion thereof, capable of binding an antigen comprising a transforming growth factor beta 1 prodomain complex (TGF-β1 GPC), that comprises:
a) a latency associated peptide (LAP) dimer, wherein each polypeptide of the dimer has an amino acid sequence comprising SEQ ID NO: 38 or SEQ ID NO: 99; and
b) a growth factor dimer, wherein each polypeptide of the dimer has an amino acid sequence comprising SEQ ID NO: 44; and
selecting an antibody, or antigen-binding portion thereof, that binds to the TGF-β1 GPC;

(ii) providing a growth factor activity assay to measure inhibition of TGF-β1 signaling comprising:
a) contacting the TGF-β1 GPC with the antibody, or antigen-binding portion thereof, selected in step (i), a cell expressing αvβ6 integrin and/or αvβ8 integrin, and a responsive cell comprising a reporter gene comprising a TGF-β1 growth factor responsive promoter operably linked to a sequence encoding a detectable gene product;
b) obtaining TGF-β1 signaling activity data by detecting expression of the detectable gene product, and
c) selecting an antibody, or an antigen-binding portion thereof, that inhibits TGF-β1 signaling;
d) carrying out affinity maturation and/or optimization of the antibody, or antigen-binding portion thereof selected in step (ii) c); or
humanizing the antibody, or the antigen-binding portion thereof selected in step (ii) c); or
selecting an antibody, or antigen-binding portion thereof, from step (ii) c) that cross-reacts with human or mouse forms of the antigen; or
performing an in vitro binding assay for the antibody, or antigen-binding portion thereof, selected in step (ii) c), and selecting an antibody, or antigen-binding portion thereof, that does not bind mature TGF-β1 growth factor or empty LAP dimer;

(iii) preparing expression vectors comprising the coding sequences of the antibody, or the antigen-binding portions thereof, of step (ii) d);
(iv) transfecting the vectors prepared in step (iii) into a cell in culture medium for expression of the antibody, or antigen-binding portion thereof;
(v) purifying the antibody, or the antigen-binding portion thereof, expressed in step (iv) from the culture medium; and
(vi) formulating the antibody, or the antigen-binding portion thereof, purified in step (v) into a pharmaceutical composition.

7 . The method of claim 6 , wherein step (ii) d) is carrying out affinity maturation and/or optimization of the antibody, or antigen-binding portion thereof selected in step (ii) c).

8 . The method of claim 6 , wherein step (ii) d) is humanizing the antibody, or the antigen-binding portion thereof selected in step (ii) c).

9 . The method of claim 6 , wherein step (ii) d) is selecting an antibody, or antigen-binding portion thereof, from step (ii) c) that cross-reacts with human or mouse forms of the antigen.

10 . The method of claim 6 , wherein step (ii) d) is performing an in vitro binding assay for the antibody, or antigen-binding portion thereof, selected in step (ii) c), and selecting an antibody, or antigen-binding portion thereof, that does not bind mature TGF-β1 growth factor or empty LAP dimer.

11 . The method of claim 10 , wherein the in vitro binding assay is an Enzyme-Linked Immunosorbent Assay (ELISA), Surface Plasmon Resonance assay and/or a Fluorescence-Associated Cell Sorting (FACS) assay.

Assignees

• Scholar Rock Inc

Inventors

• Schurpf Thomas
• Mahanthappa Nagesh K
• Straub Michelle Marie

Classifications

• C07K2317/32

  specific for a neo-epitope on a complex, e.g. antibody-antigen or ligand-receptor · CPC title

• C07K16/28

  against receptors, cell surface antigens or cell surface determinants · CPC title

• C07K2317/55

  Fab or Fab' · CPC title

• C12N15/1037

  Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display · CPC title

• C07K2317/21

  from primates, e.g. man · CPC title