LTBP complex-specific inhibitors of TGF-beta 1 and uses thereof

Patent metadata

Field	Value
Publication number	US-12358992-B2
Application number	US-201816634336-A
Country	US
Kind code	B2
Filing date	Jul 27, 2018
Priority date	Jul 28, 2017
Publication date	Jul 15, 2025
Grant date	Jul 15, 2025

Abstract

Disclosed herein are inhibitors, such as antibodies, and antigen binding portions thereof, that selectively bind complexes of LTBP1-TGFβ1 and/or LTBP3-TGFβ1. The application also provides methods of use of these inhibitors for, for example, inhibiting TGFβ1 activation, and treating subjects suffering from TGFβ1-related disorders, such as fibrotic conditions. Methods of selecting a context-dependent or context-independent isoform-specific TGFβ1 inhibitor for a subject in need thereof are also provided.

First claim

What is claimed is:

1. A method for making a composition comprising an antibody, or an antigen-binding fragment thereof, that specifically binds a human LTBP1-proTGFβ1 complex, and does not bind a GARP-proTGFβ1, LRRC33-proTGFβ1, mature TGFβ1, GARP-proTGFβ2, LRRC33-proTGFβ2, mature TGFβ2, GARP-proTGFβ3, LRRC33-proTGFβ3 and mature TGFβ3; wherein the antibody, or the antigen-binding fragment thereof, inhibits TGFβ1 but does not inhibit TGFβ2 or TGFβ3, the method comprising steps of:
i) providing an antigen comprising human LTBP1-proTGFβ1,
ii) selecting antibodies, or antigen-binding fragments thereof, that specifically bind the antigen of step (i), and do not bind GARP-proTGFβ1, LRRC33-proTGFβ1, mature TGFβ1, GARP-proTGFβ2, LRRC33-proTGFβ2, mature TGFβ2, GARP-proTGFβ3, LRRC33-proTGFβ3, and mature TGFβ3, so as to provide specific binders of human LTBP1-proTGFβ1;
iii) selecting from the antibodies, or the antigen-binding fragments thereof, of step ii), antibodies, or antigen-binding fragments thereof, that inhibit activation of TGFβ1, so as to identify specific inhibitors of TGFβ1 activation, wherein the selection step comprises a cell-based assay to measure TGFβ activation, wherein the cell-based assay comprises:
transfecting αVβ integrin-expressing cells with a plasmid encoding human LTBP1 and a plasmid encoding proTGFβ1;
coating an assay plate with fibronectin;
plating the transfected cells expressing human LTBP1 and proTGFβ1 on the assay plate coated with fibronectin, wherein the expressed human LTBP1-proTGFβ1 complex binds to the fibronectin;
incubating in the presence of the antibody or the antigen-binding fragment thereof;
adding reporter cells expressing a TGFβ-responsive promoter element, wherein the reporter cells further comprise a reporter gene under the control of the TGFβ-responsive promoter element, wherein the reporter gene comprises a luciferase gene;
reading the results from the reporter cells, wherein the data generated reflects levels of TGFβ activation; and
identifying the antibodies or the antigen-binding fragments thereof that inhibit TGFβ1 activation; and

iv) formulating an antibody or an antigen-binding fragment thereof, selected from the previous step into a pharmaceutical composition,

thereby making the composition comprising the antibody, or the antigen-binding fragment thereof.

2. The method of claim 1 , wherein the reporter cells are CAGA12 cells.

3. A method for making a composition comprising an antibody, or an antigen-binding fragment thereof, that specifically binds a human LTBP3-proTGFβ1 complex, and does not bind a GARP-proTGFβ1, LRRC33-proTGFβ1, mature TGFβ1, GARP-proTGFβ2, LRRC33-proTGFβ2, mature TGFβ2, GARP-proTGFβ3, LRRC33-proTGFβ3 and mature TGFβ3; wherein the antibody, or the antigen-binding fragment thereof, inhibits TGFβ1 but does not inhibit TGFβ2 or TGFβ3, the method comprising steps of:
i) providing an antigen comprising human LTBP3-proTGFβ1,
ii) selecting antibodies, or antigen-binding fragments thereof, that specifically bind the antigen of step (i), and do not bind GARP-proTGFβ1, LRRC33-proTGFβ1, mature TGFβ1, GARP-proTGFβ2, LRRC33-proTGFβ2, mature TGFβ2, GARP-proTGFβ3, LRRC33-proTGFβ3, and mature TGFβ3, so as to provide specific binders of human LTBP3-proTGFβ1;
iii) selecting from the antibodies, or the antigen-binding fragments thereof, of step ii), antibodies, or antigen-binding fragments thereof, that inhibit activation of TGFβ1, so as to identify specific inhibitors of TGFβ1 activation, wherein the selection step comprises a cell-based assay to measure TGFβ activation, wherein the cell-based assay comprises:
transfecting αVβ integrin-expressing cells with a plasmid encoding human LTBP3 and a plasmid encoding proTGFβ1;
coating an assay plate with fibronectin;
plating the transfected cells expressing human LTBP3 and proTGFβ1 on the assay plate coated with fibronectin, wherein the expressed human LTBP3-proTGFβ1 complex binds to the fibronectin;
incubating in the presence of the antibody or the antigen-binding fragment thereof;
adding reporter cells expressing a TGFβ-responsive promoter element, wherein the reporter cells further comprise a reporter gene under the control of the TGFβ-responsive promoter element, wherein the reporter gene comprises a luciferase gene;
reading the results from the reporter cells, wherein the data generated reflects levels of TGFβ activation; and
identifying the antibodies or the antigen-binding fragments thereof that inhibit TGFβ1 activation; and

iv) formulating an antibody or an antigen-binding fragment thereof, selected from the previous step into a pharmaceutical composition,

thereby making the composition comprising the antibody, or the antigen-binding fragment thereof.

4. The method of claim 3 , wherein the reporter cells are CAGA12 cells.

Assignees

• Scholar Rock Inc

Inventors

• Schurpf Thomas
• Littlefield Christopher
• Carven Gregory J
• Datta Abhishek

Classifications

• C07K2317/10

  characterized by their source of isolation or production · CPC title

• G01N33/531

  Production of immunochemical test materials · CPC title

• G01N33/537

  with separation of immune complex from unbound antigen or antibody · CPC title

• C07K2317/76

  Antagonist effect on antigen, e.g. neutralization or inhibition of binding · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title