# US 12,110,331 B2 · Canine monoclonal antibodies against canine cytotoxic T lymphocyte associated protein 4 (CTLA-4)

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- Number: US 12,110,331 B2
- Document ID: US12110331B2
- Status: Granted
- Kind: B2
- Application number: 18/491,182
- Filed: 2023-10-20
- Published: 2024-04-04
- Granted: 2024-10-08
- Earliest claimed: 2021-04-21
- Assignees: The Trustees of the University of Pennsylvania; Vetigenics, Inc.
- Applicants: The Trustees of the University of Pennsylvania; Vetigenics, Inc.
- Inventors: Nicola J. Mason; Donald L. Siegel; Nicholas Chester
- Web page: https://patentgrep.com/patent/US12110331B2

## CPC

- C07K 16/2818: {against CD28 or CD152}
- A61P 35/00: Antineoplastic agents
- C07K 2317/20: characterized by taxonomic origin
- C07K 2317/565: Complementarity determining region [CDR]
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2317/734: Complement-dependent cytotoxicity [CDC]
- 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

## Abstract

The present invention relates to canine antibodies, binding polypeptides, and scFvs specific for canine cytotoxic T lymphocyte associated protein 4 (CTLA-4).

## Claims

### Claim 1

An antibody or antigen-binding fragment thereof comprising an antigen-binding domain that specifically binds to an epitope of canine cytotoxic T lymphocyte-associated protein 4 (CTLA-4), wherein the antigen-binding domain comprises:
(i) a heavy chain variable region comprising an amino acid sequence having at least 80% identity to the amino acid sequence of the heavy chain variable region set forth in SEQ ID NO: 6; and
(ii) a light chain variable region comprising an amino acid sequence having at least 80% identity to the amino acid sequence set forth in SEQ ID NO: 8;
wherein the heavy chain variable region comprises three heavy chain complementarity determining regions (HCDRs), wherein HCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 1, HCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 2, and HCDR3 comprises the amino acid sequence set forth in of SEQ ID NO: 3; and
wherein the light chain variable region comprises three light chain complementarity determining regions (LCDRs), wherein LCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 4, LCDR2 comprises the amino acid sequence of VDG, and LCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5.

### Claim 2 (depends on 1)

The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody or antigen-binding fragment thereof is a full-length antibody.

### Claim 3 (depends on 2)

The antibody or antigen-binding fragment thereof of claim 2, wherein the full-length antibody is a canine antibody.

### Claim 4 (depends on 1)

The antibody or antigen-binding fragment thereof of claim 1, wherein the antigen binding domain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 6.

### Claim 5 (depends on 1)

The antibody or antigen-binding fragment thereof of claim 1, wherein the antigen binding domain comprises a heavy chain variable region comprising an amino acid sequence having at least 95% identity to the amino acid sequence set forth in SEQ ID NO: 6.

### Claim 6 (depends on 1)

The antibody or antigen-binding fragment thereof of claim 1, wherein the antigen binding domain comprises a light chain variable region comprising an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 8.

### Claim 7 (depends on 1)

The antibody or antigen-binding fragment thereof of claim 1, wherein the antigen binding domain comprises a light chain variable region comprising an amino acid sequence having at least 95% identity to the amino acid sequence set forth in SEQ ID NO: 8.

### Claim 8 (depends on 2)

The antibody or antigen-binding fragment thereof of claim 2, wherein the antigen binding domain comprises (i) a heavy chain variable region comprising an amino acid sequence having at least 95% identity to the amino acid sequence set forth in SEQ ID NO: 6; and (ii) a light chain variable region comprising an amino acid sequence having at least 95% identity to the amino acid sequence set forth in SEQ ID NO: 8.

### Claim 9

A full-length antibody comprising an antigen-binding domain that specifically binds to an epitope of canine cytotoxic T lymphocyte associated protein 4 (CTLA-4), wherein the antigen-binding domain comprises:
(i) a heavy chain variable region that comprises three heavy chain complementarity determining regions (HCDRs), wherein HCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 1, HCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 2, and HCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 3; and
(ii) a light chain variable region that comprises three light chain complementarity determining regions (LCDRs), wherein LCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 4, LCDR2 comprises the amino acid sequence of VDG, and LCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5.

### Claim 10 (depends on 9)

The full-length antibody of claim 9, wherein:
(i) the heavy chain variable region comprises an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 6; and
(ii) the light chain variable region comprises an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 8.

### Claim 11 (depends on 9)

The full-length antibody of claim 9, comprising a heavy chain amino acid sequence comprising an amino acid sequence having at least 90% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 10, 12, 14, and 16 and a light chain comprising an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 18.

### Claim 12 (depends on 9)

The full-length antibody of claim 9, wherein the full-length antibody is a canine antibody.

### Claim 13 (depends on 1)

An isolated nucleic acid encoding the antibody or antigen-binding fragment thereof of claim 1.

### Claim 14 (depends on 13)

A vector comprising the isolated nucleic acid of claim 13.

### Claim 15 (depends on 14)

A host cell comprising the vector of claim 14.

### Claim 16 (depends on 15)

A method of producing an antibody or antigen-binding fragment thereof that binds to canine cytotoxic T lymphocyte associated protein 4 (CTLA-4), the method comprising culturing the host cell of claim 15.

### Claim 17 (depends on 1)

A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof of claim 1, and a pharmaceutically acceptable carrier.

### Claim 18 (depends on 1)

A method for treating a cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the antibody or antigen-binding fragment thereof of claim 1, thereby treating the cancer in the subject.

### Claim 19 (depends on 18)

The method of claim 18, wherein the cancer is associated with cytotoxic T lymphocyte associated protein 4 (CTLA-4).

### Claim 20 (depends on 18)

The method of claim 18, wherein the subject is canine.