HIGH-AFFINITY, ISOFORM-SELECTIVE TGF[beta]1 INHIBITORS AND USE THEREOF

Abstract

Disclosed herein are monoclonal antibodies and antigen-binding fragments thereof capable of selectively inhibiting TGF[beta]1 with high potency. Related compositions, methods and therapeutic use are also disclosed.

Description

[0001] related application

[0002] This application claims the benefit of and priority to: U.S. Provisional Patent Application No. 62 / 696,752, filed July 11, 2018; U.S. Provisional Patent Application No. 62 / 718,196, filed August 13, 2018; U.S. Provisional Patent Application No. 62 / 737,534 filed on November 9, 2018; U.S. Provisional Patent Application No. 62 / 810,263 filed on February 25, 2019 and April 2019 US Provisional Patent Application No. 62 / 827,552, filed on 1st, each entitled "High Affinity, Context-Independent TGFβ1 Inhibitors and Uses Thereof," each of which is expressly incorporated herein by reference in its entirety.

[0003] sequence listing

[0004] This application contains a Sequence Listing, which has been filed electronically in ASCII format, and is hereby incorporated by reference in its entirety. Said ASCII copy, created on July 11, 2019, is named 127036-03520_SL.txt and is 261,252 bytes in size. Background technique

[0005] Transforming growth factor ...

Examples

preparation example Construction

[0554] The preparation method of the pharmaceutical composition comprising such antibodies or fragments thereof may include steps of purification, formulation, sterile filtration, packaging and the like. For example, certain steps, such as sterile filtration, are performed according to guidelines established by relevant regulatory agencies, such as the FDA. Such compositions are available in single-use containers, such as pre-filled syringes, or in multidose containers, such as vials.

[0555] modify

[0556] Antibodies of the present disclosure, or antigen-binding portions thereof, can be modified with detectable labels or detectable moieties, including but not limited to: enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, radioactive materials, positron-emitting metals, non- Radioactive paramagnetic metal ions and affinity labels for the detection and isolation of GARP-TGFβ1 complexes, LTBP1-TGFβ1 complexes, LTBP3-TGFβ1 compl...

Embodiment approach

[0969] 1. An antibody or antigen-binding fragment thereof that binds to the following antigen complex with a KD of ≤ 10 nM (optionally ≤ 5 nM) as measured by an assay based on solution equilibrium titration each of:

[0970] i) hLTBP1-proTGFβ1;

[0971] ii) hLTBP3-proTGFβ1;

[0972] iii) hGARP-proTGFβ1; and,

[0973] iv) hLRRC33-proTGFβ1;

[0974] Wherein the antibody or fragment thereof is a fully human or humanized antibody or fragment thereof.

[0975] 2. The antibody or antigen-binding fragment of embodiment 1, which binds to said i) hLTBP1-proTGFβ1 and said ii) each of the hLTBP3-proTGFβ1 complexes, wherein optionally, the antibody or fragment binds each of the complexes with a KD of ≤ 1 nM as measured by an assay based on solution equilibrium titration

[0976] 3. An antibody or antigen-binding fragment thereof that binds to an antigen complex with a KD of ≤ 200 pM (optionally ≤ 100 pM) as measured by a solution equilibrium titration-based assay each of:

[0977] ...

Embodiment 1

[1183] Example 1: In vitro binding properties

[1184] 1) BLI-based assay:

[1185] pass Assay Affinity of Ab4, Ab5, Ab6 and Ab3 was measured on human proTGFβ1 cells while activity was measured by CAGA12 reporter cells detecting inhibition of human proTGFβ1. A protocol for measuring the affinity of antibodies for the complexes provided herein is summarized in Table 19 below, and a summarized list of affinity properties for exemplary antibodies of the present disclosure is provided in Table 8 herein.

[1186] Table 19: For performing Exemplary Protocol for Binding Assays

[1187]

[1188]

[1189]For example, the binding of Ab6 to the TGFβ antigen was measured using biolayer interferometry on a FortéBio Octet Red 384 using polystyrene 96-well black half-area plates (Greiner Bio-One). Ab6 and human mature TGFβ1, TGFβ2 were performed after coupling the antigen to the Amine Responsive Second Generation (AR2G) biosensor (FortéBio) using the Amine Responsive Second Gener...

[0001] related application

[0002] This application claims the benefit of and priority to: U.S. Provisional Patent Application No. 62 / 696,752, filed July 11, 2018; U.S. Provisional Patent Application No. 62 / 718,196, filed August 13, 2018; U.S. Provisional Patent Application No. 62 / 737,534 filed on November 9, 2018; U.S. Provisional Patent Application No. 62 / 810,263 filed on February 25, 2019 and April 2019 US Provisional Patent Application No. 62 / 827,552, filed on 1st, each entitled "High Affinity, Context-Independent TGFβ1 Inhibitors and Uses Thereof," each of which is expressly incorporated herein by reference in its entirety.

[0003] sequence listing

[0004] This application contains a Sequence Listing, which has been filed electronically in ASCII format, and is hereby incorporated by reference in its entirety. Said ASCII copy, created on July 11, 2019, is named 127036-03520_SL.txt and is 261,252 bytes in size. Background technique

[0005] Transforming growth factor ...

Examples

preparation example Construction

[0554] The preparation method of the pharmaceutical composition comprising such antibodies or fragments thereof may include steps of purification, formulation, sterile filtration, packaging and the like. For example, certain steps, such as sterile filtration, are performed according to guidelines established by relevant regulatory agencies, such as the FDA. Such compositions are available in single-use containers, such as pre-filled syringes, or in multidose containers, such as vials.

[0555] modify

[0556] Antibodies of the present disclosure, or antigen-binding portions thereof, can be modified with detectable labels or detectable moieties, including but not limited to: enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, radioactive materials, positron-emitting metals, non- Radioactive paramagnetic metal ions and affinity labels for the detection and isolation of GARP-TGFβ1 complexes, LTBP1-TGFβ1 complexes, LTBP3-TGFβ1 compl...

Embodiment approach

[0969] 1. An antibody or antigen-binding fragment thereof that binds to the following antigen complex with a KD of ≤ 10 nM (optionally ≤ 5 nM) as measured by an assay based on solution equilibrium titration each of:

[0970] i) hLTBP1-proTGFβ1;

[0971] ii) hLTBP3-proTGFβ1;

[0972] iii) hGARP-proTGFβ1; and,

[0973] iv) hLRRC33-proTGFβ1;

[0974] Wherein the antibody or fragment thereof is a fully human or humanized antibody or fragment thereof.

[0975] 2. The antibody or antigen-binding fragment of embodiment 1, which binds to said i) hLTBP1-proTGFβ1 and said ii) each of the hLTBP3-proTGFβ1 complexes, wherein optionally, the antibody or fragment binds each of the complexes with a KD of ≤ 1 nM as measured by an assay based on solution equilibrium titration

[0976] 3. An antibody or antigen-binding fragment thereof that binds to an antigen complex with a KD of ≤ 200 pM (optionally ≤ 100 pM) as measured by a solution equilibrium titration-based assay each of:

[0977] ...

Embodiment 1

[1183] Example 1: In vitro binding properties

[1184] 1) BLI-based assay:

[1185] pass Assay Affinity of Ab4, Ab5, Ab6 and Ab3 was measured on human proTGFβ1 cells while activity was measured by CAGA12 reporter cells detecting inhibition of human proTGFβ1. A protocol for measuring the affinity of antibodies for the complexes provided herein is summarized in Table 19 below, and a summarized list of affinity properties for exemplary antibodies of the present disclosure is provided in Table 8 herein.

[1186] Table 19: For performing Exemplary Protocol for Binding Assays

[1187]

[1188]

[1189]For example, the binding of Ab6 to the TGFβ antigen was measured using biolayer interferometry on a FortéBio Octet Red 384 using polystyrene 96-well black half-area plates (Greiner Bio-One). Ab6 and human mature TGFβ1, TGFβ2 were performed after coupling the antigen to the Amine Responsive Second Generation (AR2G) biosensor (FortéBio) using the Amine Responsive Second Gener...

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