# US 2023/0054718 A1 · Anti-galectin-9 antibody and uses thereof

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- Number: US 2023/0054718 A1
- Document ID: US20230054718A1
- Status: Published application
- Kind: A1
- Application number: 17/789,961
- Filed: 2021-01-06
- Published: 2023-02-23
- Earliest claimed: 2020-01-07
- Assignees: Hifibio (Hk) Limited
- Applicants: HiFiBio (HK) Limited
- Inventors: Germain MARGALL DUCOS; Rachel PACHERIE; Nicola Arturo Aldo BELTRAMINELLI; Francisco ADRIAN; Liang SCHWEIZER; Muriel DAVID; Yun-Yueh LU
- Web page: https://patentgrep.com/patent/US20230054718A1

## CPC

- C07K 16/2851: {against the lectin superfamily, e.g. CD23, CD72}
- A61P 35/00: Antineoplastic agents
- C07K 16/2818: {against CD28 or CD152}
- A61K 2039/505: {comprising antibodies}
- A61K 2039/507: {Comprising a combination of two or more separate antibodies}
- C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
- C07K 2317/55: Fab or Fab'
- C07K 2317/565: Complementarity determining region [CDR]
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding
- C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

The invention provides humanized monoclonal antibodies against Gal9, and method of using the antibodies to treat cancer, including combination therapy with antagonists of the PD-1/PD-L1 immune checkpoint.

## Claims

### Claim 1

An isolated monoclonal antibody, or an antigen-binding fragment thereof, wherein, said monoclonal antibody or antigen-binding fragment thereof is specific for Galectin-9, and wherein said monoclonal antibody comprises:
(1a) a heavy chain variable region (HCVR), comprising a HCVR CDR1 sequence of SEQ ID NO: 2, a HCVR CDR2 sequence of SEQ ID NO: 4, and a HCVR CDR3 sequence of SEQ ID NO: 6; and,
(1b) a light chain variable region (LCVR), comprising a LCVR CDR1 sequence of SEQ ID NO: 10, a LCVR CDR2 sequence of SEQ ID NO: 12, and a LCVR CDR3 sequence of SEQ ID NO: 14; or
(2a) a heavy chain variable region (HCVR), comprising a HCVR CDR1 sequence of SEQ ID NO: 18, a HCVR CDR2 sequence of SEQ ID NO: 20, and a HCVR CDR3 sequence of SEQ ID NO: 22; and,
(2b) a light chain variable region (LCVR), comprising a LCVR CDR1 sequence of SEQ ID NO: 26, a LCVR CDR2 sequence of SEQ ID NO: 28, and a LCVR CDR3 sequence of SEQ ID NO: 30; or
(3a) a heavy chain variable region (HCVR), comprising a HCVR CDR1 sequence of SEQ ID NO: 34, a HCVR CDR2 sequence of SEQ ID NO: 36, and a HCVR CDR3 sequence of SEQ ID NO: 38; and,
(3b) a light chain variable region (LCVR), comprising a LCVR CDR1 sequence of SEQ ID NO: 42, a LCVR CDR2 sequence of SEQ ID NO: 44, and a LCVR CDR3 sequence of SEQ ID NO: 46; or
(4a) a heavy chain variable region (HCVR), comprising a HCVR CDR1 sequence of SEQ ID NO: 50, a HCVR CDR2 sequence of SEQ ID NO: 52, and a HCVR CDR3 sequence of SEQ ID NO: 54; and,
(4b) a light chain variable region (LCVR), comprising a LCVR CDR1 sequence of SEQ ID NO: 58, a LCVR CDR2 sequence of SEQ ID NO: 60, and a LCVR CDR3 sequence of SEQ ID NO: 62; or
(5a) a heavy chain variable region (HCVR), comprising a HCVR CDR1 sequence of SEQ ID NO: 66, a HCVR CDR2 sequence of SEQ ID NO: 68, and a HCVR CDR3 sequence of SEQ ID NO: 70; and,
(5b) a light chain variable region (LCVR), comprising a LCVR CDR1 sequence of SEQ ID NO: 74, a LCVR CDR2 sequence of SEQ ID NO: 76, and a LCVR CDR3 sequence of SEQ ID NO: 78; or
(6a) a heavy chain variable region (HCVR), comprising a HCVR CDR1 sequence of SEQ ID NO: 82, a HCVR CDR2 sequence of SEQ ID NO: 84, and a HCVR CDR3 sequence of SEQ ID NO: 86; and,
(6b) a light chain variable region (LCVR), comprising a LCVR CDR1 sequence of SEQ ID NO: 90, a LCVR CDR2 sequence of SEQ ID NO: 92, and a LCVR CDR3 sequence of SEQ ID NO: 94; or
(7a) a heavy chain variable region (HCVR), comprising a HCVR CDR1 sequence of SEQ ID NO: 98, a HCVR CDR2 sequence of SEQ ID NO: 100, and a HCVR CDR3 sequence of SEQ ID NO: 102; and,
(7b) a light chain variable region (LCVR), comprising a LCVR CDR1 sequence of SEQ ID NO: 106, a LCVR CDR2 sequence of SEQ ID NO: 108, and a LCVR CDR3 sequence of SEQ ID NO: 110; or
(8a) a heavy chain variable region (HCVR), comprising a HCVR CDR1 sequence of SEQ ID NO: 114, a HCVR CDR2 sequence of SEQ ID NO: 116, and a HCVR CDR3 sequence of SEQ ID NO: 118; and,
(8b) a light chain variable region (LCVR), comprising a LCVR CDR1 sequence of SEQ ID NO: 122, a LCVR CDR2 sequence of SEQ ID NO: 124, and a LCVR CDR3 sequence of SEQ ID NO: 128.

### Claim 2 (depends on 1)

The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, wherein:
(1c) the antibody or antigen-binding fragment thereof of (1a) and (1b) further comprises a HFR3 sequence of SEQ ID NO: 5, and optionally further comprises a HFR1 sequence of SEQ ID NO: 1; or
(2c) the antibody or antigen-binding fragment thereof of (2a) and (2b) further comprises a HFR3 sequence of SEQ ID NO: 21, and optionally further comprises a HFR1 sequence of SEQ ID NO: 17; or
(3c) the antibody or antigen-binding fragment thereof of (3a) and (3b) further comprises a HFR3 sequence of SEQ ID NO: 37, and optionally further comprises a HFR1 sequence of SEQ ID NO: 33; or
(4c) the antibody or antigen-binding fragment thereof of (4a) and (4b) further comprises a HFR3 sequence of SEQ ID NO: 53, and optionally further comprises a HFR1 sequence of SEQ ID NO: 49; or
(5c) the antibody or antigen-binding fragment thereof of (5a) and (5b) further comprises a HFR3 sequence of SEQ ID NO: 69, and optionally further comprises a HFR1 sequence of SEQ ID NO: 65; or
(6c) the antibody or antigen-binding fragment thereof of (6a) and (6b) further comprises a HFR3 sequence of SEQ ID NO: 85, and optionally further comprises a HFR1 sequence of SEQ ID NO: 81; or
(7c) the antibody or antigen-binding fragment thereof of (7a) and (7b) further comprises a HFR3 sequence of SEQ ID NO: 101, and optionally further comprises a HFR1 sequence of SEQ ID NO: 97; or
(8c) the antibody or antigen-binding fragment thereof of (8a) and (8b) further comprises a HFR3 sequence of SEQ ID NO: 117, and optionally further comprises a HFR1 sequence of SEQ ID NO: 113.

### Claim 3 (depends on 1)

The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, wherein:
(1A) the HCVR sequence is SEQ ID NO: 8; and/or,
(1B) the LCVR sequence is SEQ ID NO: 16, or,
(2A) the HCVR sequence is SEQ ID NO: 24; and/or,
(2B) the LCVR sequence is SEQ ID NO: 32, or,
(3A) the HCVR sequence is SEQ ID NO: 40; and/or,
(3B) the LCVR sequence is SEQ ID NO: 48, or,
(4A) the HCVR sequence is SEQ ID NO: 56; and/or,
(4B) the LCVR sequence is SEQ ID NO: 64, or,
(5A) the HCVR sequence is SEQ ID NO: 72; and/or,
(5B) the LCVR sequence is SEQ ID NO: 80, or,
(6A) the HCVR sequence is SEQ ID NO: 88; and/or,
(6B) the LCVR sequence is SEQ ID NO: 96, or
(7A) the HCVR sequence is SEQ ID NO: 104; and/or,
(7B) the LCVR sequence is SEQ ID NO: 112, or
(8A) the HCVR sequence is SEQ ID NO: 120; and/or,
(8B) the LCVR sequence is SEQ ID NO: 128.

### Claim 4 (depends on 1)

The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1, which is a humanized antibody, and comprises:
(1) the HCVR sequence of SEQ ID NO: 8 and the LCVR sequence of SEQ ID NO: 16; or,
(2) the HCVR sequence of SEQ ID NO: 72 and the LCVR sequence of SEQ ID NO: 80.

### Claim 5 (depends on 1)

The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein said antigen-binding fragment thereof is an Fab, Fab′, F(ab′)2, Fd, single chain Fv or scFv, disulfide linked Fv, V-NAR domain, IgNar, intrabody, IgGΔCH2, minibody, F(ab′)3, tetrabody, triabody, diabody, single-domain antibody, DVD-Ig, Fcab, mAb2, (scFv)2, or scFv-Fc.

### Claim 6 (depends on 1)

The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, wherein said monoclonal antibody or antigen-binding fragment thereof cross-reacts with mouse Gal9.

### Claim 7 (depends on 1)

The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, wherein said monoclonal antibody or antigen-binding fragment thereof binds to human Gal9 with an EC50 of about 0.1-0.2 nM, and/or binds to mouse Gal9 with an EC50 of about 0.5-1.0 nM.

### Claim 8 (depends on 1)

The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, wherein said monoclonal antibody or antigen-binding fragment thereof binds human Gal9 with a Kd of less than about 25 nM, 20 nM, 15 nM, 10 nM, 5 nM, 2 nM, or 1 nM.

### Claim 9 (depends on 1)

The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, which binds to Gal9 and inhibits Gal9 binding to a Gal9 receptor (e.g., TIM3 or CD44).

### Claim 10 (depends on 1)

The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, which neutralizes Gal-9-induced Th1 apoptosis of T cells (such as CD4+ T cells).

### Claim 11 (depends on 1)

The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, which suppresses Gal9-induced Treg expansion.

### Claim 12 (depends on 1)

The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, which synergistically inhibits tumor growth in vivo and/or prolongs survival in a mouse with a xenograph tumor with an antagonist of an immune checkpoint.

### Claim 13 (depends on 1)

The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, wherein the antagonist of the immune checkpoint is an antibody or antigen-binding fragment thereof specific for PD-1 or PD-L1.

### Claim 14 (depends on 1)

A method of treating cancer in a patient in need thereof, the method comprising administering to the patient an effective amount of the isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, and an antagonist of an immune checkpoint.

### Claim 15 (depends on 14)

The method of claim 14, wherein the immune checkpoint is PD-1/PD-L1 immune checkpoint, and optionally wherein the antagonist of the immune checkpoint is an antibody or antigen-binding fragment thereof specific for PD-1 or PD-L1; a peptide inhibitor of PD-1/PD-L1; a small molecule inhibitor of PD-L1; a macrocyclic peptide; or any combination thereof.

### Claim 16 (depends on 1)

The method of claim 1, wherein the cancer is a hematological cancer (such as AML and DLBCL), or a solid tumor (such as breast cancer, head and neck cancer, lung cancer, melanoma (including uveal melanoma), colon cancer, renal carcinoma, ovarian cancer, liver cancer, and prostate cancer).

### Claim 17 (depends on 1)

The method of claim 1, further comprising administering to the patient a chemotherapeutic agent, an anti-angiogenesis agent, a growth inhibitory agent, an immune-oncology agent, and/or an anti-neoplastic composition.

### Claim 18 (depends on 1)

A polynucleotide encoding the heavy chain or the light chain or the antigen-binding portion thereof of the isolated monoclonal antibody of claim 1.

### Claim 19 (depends on 18)

The polynucleotide of claim 18, which is codon optimized for expression in a human cell.

### Claim 20 (depends on 18)

A vector comprising the polynucleotide of claim 18, wherein the vector is an expression vector selected from (a mammalian expression vector, a yeast expression vector, an insect expression vector, or a bacterial expression vector).

### Claim 21 (depends on 1)

A method of rescuing or promoting effector T cells proliferation and/or enhancing effector T cell activity in a patient diagnosed with cancer, in risk of developing cancer or having cancer relapse, or a method of identifying and treating a patient having cancer, the method comprising: administering to the patient an effective amount of the isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, upon identifying the patient as having a level of Galectin-9 in a sample from the patient higher than a reference level of Galectin-9 in a healthy or a control individual.

### Claim 22 (depends on 21)

The method of claim 21, further comprising identifying the patient as having the level of Galectin-9 in the sample higher than the reference level, by comparing the level of Galectin-9 in the sample to the reference level.

### Claim 23 (depends on 21)

The method of claim 21, further comprising administering to the patient an antagonist of an immune checkpoint, and optionally wherein the immune checkpoint is PD-1/PD-L1 immune checkpoint.

### Claim 24 (depends on 23)

The method of claim 23, wherein the antagonist of the immune checkpoint is an antibody or antigen-binding fragment thereof specific for PD-1 or PD-L1; a (non-antibody) peptide inhibitor of PD-1/PD-L1; a small molecule inhibitor of PD-L1; a macrocyclic peptide; or any combination thereof.

### Claim 25 (depends on 21)

The method of claim 21, wherein the cancer is a hematological cancer (such as AML and DLBCL), or a solid tumor (such as breast cancer, head and neck cancer, lung cancer, melanoma (including uveal melanoma), colon cancer, renal carcinoma, ovarian cancer, liver cancer, and prostate cancer).

### Claim 26 (depends on 25)

The method of claim 25, wherein the patient is an FAB M0, M1, M4 or M5 AML patient, or wherein the patient is not an FAB M2 or M3 AML patient.

### Claim 27 (depends on 21)

The method of claim 21, wherein the sample is a blood sample, a plasma sample, or a serum sample.

### Claim 28 (depends on 1)

A method of rescuing or promoting effector T cells proliferation and/or enhancing effector T cell activity in a patient diagnosed with AML, in risk of developing AML or having AML relapse, or a method of identifying and treating a patient having AML, the method comprising: administering to the patient an effective amount of the isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, upon identifying the patient as having a level of Galectin-9-encoding mRNA in a bone-marrow (BM)-derived mononuclear cell (MNC) sample from the patient statistically significantly higher or lower than a reference level in BM-derived MNC or CD34+ cells in a healthy or a control individual.

### Claim 29 (depends on 28)

The method of claim 28, wherein (1) the level of Galectin-9-encoding mRNA in the BM-derived MNC sample from the patient is significantly higher than the reference level when the patient is an FAB M0, M1, M2, M4, or M5 AML patient, or (2) the level of Galectin-9-encoding mRNA in the BM-derived MNC sample from the patient is significantly lower than the reference level when the patient is an FAB M3 AML patient.

### Claim 30

An antibody, or an antigen-binding portion thereof, directed against or specific for Galectin-9, for use in the treatment of cancer, wherein said antibody or antigen-binding portion thereof rescues effector T cell proliferation, and/or enhances effector T cell activity, and optionally wherein the effector T cell is a Th1 cell.

### Claim 31 (depends on 1)

A method of rescuing or promoting effector T cell proliferation, and/or enhancing effector T cell activity, comprising contacting said effector T cell with the isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, and optionally wherein the effector T cell is a Th1 cell.

### Claim 32 (depends on 1)

A method of inducing or promoting immune memory in a subject, comprising administering to the subject an effective amount of a composition comprising the isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, wherein the immune memory is effective in inhibiting or reducing tumor progression or recurrence, or cancer cell proliferation in the subject.