# US 11,993,653 B2 · Antibodies and methods of use thereof

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- Number: US 11,993,653 B2
- Document ID: US11993653B2
- Status: Granted
- Kind: B2
- Application number: 16/435,175
- Filed: 2019-06-07
- Published: 2020-01-23
- Granted: 2024-05-28
- Earliest claimed: 2016-12-07
- Assignees: Agenus Inc.; Ludwig Institute for Cancer Research; Memorial Sloan Kettering Cancer Center
- Applicants: Agenus Inc.; Ludwig Institute for Cancer Research Ltd.; Memorial Sloan Kettering Cancer Center
- Inventors: Marc Van Dijk; Cornelia Anne Mundt; Gerd Ritter; David Schaer; Jedd David Wolchok; Taha Merghoub; Nicholas Stuart Wilson; David Adam Savitsky; Mark Arthur Findeis; Roberta Zappasodi; Rikke Baek Holmgaard; Jean-Marie Cuillerot; Igor Proscurshim; Olga Shebanova
- Web page: https://patentgrep.com/patent/US11993653B2

## CPC

- C07K 16/2818: {against CD28 or CD152}
- A61K 39/39558: {against tumor tissues, cells, antigens}
- A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P 35/00: Antineoplastic agents
- A61P 35/04: specific for metastasis
- A61P 37/04: Immunostimulants
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- A61K 2039/507: {Comprising a combination of two or more separate antibodies}
- C07K 2317/21: from primates, e.g. man
- C07K 2317/31: multispecific
- C07K 2317/33: Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
- C07K 2317/34: Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
- C07K 2317/565: Complementarity determining region [CDR]
- 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)
- 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)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

Provided are antibodies that specifically bind to CTLA-4 and/or PD-1 and antagonize CTLA-4 and/or PD-1 function. Also provided are pharmaceutical compositions and kits comprising these antibodies, nucleic acids encoding these antibodies, expression vectors and host cells for making these antibodies, and methods of treating a subject using these antibodies either alone or in combination.

## Claims

### Claim 1

A method of increasing T cell activation in a subject that has cancer, the method comprising administering to the subject an effective amount of a therapeutic combination comprising a first isolated antibody that specifically binds to human CTLA-4 and a second isolated antibody that specifically binds to human PD-1, wherein:
(a) the first isolated antibody comprises a heavy chain variable region (VH) comprising the complementarity determining regions CDRH1, CDRH2, and CDRH3 of the VH amino acid sequence of SEQ ID NO: 1 and a light chain variable region (VL) comprising the complementarity determining regions CDRL1, CDRL2, and CDRL3 of the VL amino acid sequence of SEQ ID NO: 14, and
(b) the second isolated antibody comprises a heavy chain variable region (VH) comprising the complementarity determining regions CDRH1, CDRH2 and CDRH3 of the VH amino acid sequence of SEQ ID NO: 66 and a light chain variable region (VL) comprising the complementarity determining regions CDRL1, CDRL2 and CDRL3 of the VL amino acid sequence of SEQ ID NO: 74.

### Claim 2 (depends on 1)

The method of claim 1, wherein CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the first isolated antibody comprise the amino acid sequences set forth in SEQ ID NOs: 20, 22, 24, 27, 30, and 36, respectively.

### Claim 3 (depends on 1)

The method of claim 1, wherein the first isolated antibody comprises
a heavy chain variable region and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 1 and 14, respectively.

### Claim 4 (depends on 1)

The method of claim 1, wherein the first isolated antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 51, and a light chain comprising the amino acid sequence of SEQ ID NO: 59.

### Claim 5 (depends on 1)

The method of claim 1, wherein the first isolated antibody comprises:
(a) a heavy chain constant region selected from the group consisting of human IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2;
(b) a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 60;
(c) an IgG1 heavy chain constant region comprising S239D/I332E mutations, numbered according to the EU numbering system;
(d) a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 61;
(e) an IgG1 heavy chain constant region comprising S239D/A330L/I332E mutations, numbered according to the EU numbering system;
(f) a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 62;
(g) an IgG1 heavy chain constant region comprising L235V/F243L/R292P/Y300L/P396L mutations, numbered according to the EU numbering system;
(h) a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 63;
(i) an IgG1 heavy chain constant region, wherein the IgG1 heavy chain constant region is non-fucosylated;
(j) a light chain constant region selected from the group consisting of human Igκ and Igλ; or
(k) a light chain constant region comprising the amino acid sequence of SEQ ID NO: 64.

### Claim 6 (depends on 1)

The method of claim 1, wherein CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the second isolated antibody comprise the amino acid sequences set forth in SEQ ID NOs: 75, 76, 81, 83, 84, and 85, respectively.

### Claim 7 (depends on 1)

The method of claim 1, wherein the second isolated antibody comprises a heavy chain variable region and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 66 and 74, respectively.

### Claim 8 (depends on 1)

The method of claim 1, wherein the second isolated antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 91 and a light chain comprising the amino acid sequence of SEQ ID NO: 93.

### Claim 9 (depends on 1)

The method of claim 1, wherein:
(a) the first isolated antibody comprises a heavy chain variable region and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 1 and 14, respectively; and
(b) the second isolated antibody comprises a heavy chain variable region and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 66 and 74, respectively.

### Claim 10 (depends on 1)

The method of claim 1, wherein:
(a) the first isolated antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 51 and a light chain comprising the amino acid sequence of SEQ ID NO: 59; and
(b) the second isolated antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 91 and a light chain comprising the amino acid sequence of SEQ ID NO: 93.

### Claim 11 (depends on 1)

The method of claim 1, wherein the second isolated antibody comprises:
(a) a heavy chain constant region selected from the group consisting of human IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2;
(b) an IgG1 heavy chain constant region comprising an N297A mutation, numbered according to the EU numbering system;
(c) a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 94;
(d) an IgG4 heavy chain constant region comprising an S228P mutation, numbered according to the EU numbering system;
(e) a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 95;
(f) a light chain constant region selected from the group consisting of human Igκ and Igλ; or
(g) a light chain constant region comprising the amino acid sequence of SEQ ID NO: 64.

### Claim 12 (depends on 1)

The method of claim 1, wherein:
(a) the first isolated antibody is administered at 0.3 mg/kg or 1 mg/kg;
(b) the second isolated antibody is administered at 1 mg/kg, 3 mg/kg, or 6 mg/kg;
(c) the second isolated antibody is administered at a dose of 200 mg;
(d) the first isolated antibody is administered at 0.3 mg/kg, and the second isolated antibody is administered at 1 mg/kg;
(e) the first isolated antibody is administered at 1 mg/kg, and the second isolated antibody is administered at 1 mg/kg;
(f) the first isolated antibody is administered at 1 mg/kg, and the second isolated antibody is administered at 3 mg/kg;
(g) the first isolated antibody is administered at 1 mg/kg, and the second isolated antibody is administered at 6 mg/kg;
(h) the first isolated antibody is administered every six weeks;
(i) the second isolated antibody is administered every two weeks or every three weeks;
(j) the first isolated antibody is administered at 0.3 mg/kg every six weeks, and the second isolated antibody is administered at 1 mg/kg every two weeks;
(k) the first isolated antibody is administered at 1 mg/kg every six weeks, and the second isolated antibody is administered at 1 mg/kg every two weeks;
(l) the first isolated antibody is administered at 1 mg/kg every six weeks, and the second isolated antibody is administered at 3 mg/kg every two weeks; or
(m) the first isolated antibody is administered at 1 mg/kg every six weeks, and the second isolated antibody is administered at 6 mg/kg every three weeks.

### Claim 13 (depends on 1)

The method of claim 1, wherein:
(a) the cancer is a metastatic or locally advanced solid tumor;
(b) the cancer is a metastatic or locally advanced, unresectable squamous cell carcinoma, adenosquamous carcinoma, or adenocarcinoma of the cervix,
(c) no standard therapy is available for the cancer;
(d) the cancer is refractory to a standard therapy,
(e) the cancer has relapsed after a standard therapy;
(f) the cancer is HPV positive;
(g) the cancer is a non-small cell lung cancer (NSCLC);
(h) the cancer is a Stage IV, metastatic, or locally advanced NSCLC;
(i) the cancer is a non-small cell lung cancer (NSCLC), and wherein the NSCLC has no EGFR or ALK genomic tumor aberrations;
(j) the cancer is a non-small cell lung cancer (NSCLC), and wherein the subject has received no prior systemic chemotherapy treatment for the NSCLC;
(k) the cancer is a cutaneous squamous-cell carcinoma (cSCC);
(l) the cancer is a cutaneous squamous-cell carcinoma (cSCC), and wherein is not curable with radiation therapy; or
(m) the percentage of tumor cells in a sample of the cancer that exhibit detectable membrane expression of PD-L1 is at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, or 90%.

### Claim 14 (depends on 13)

The method of claim 13, wherein:
(a) the first and second isolated antibodies are administered as a first cancer therapy after diagnosis of the cancer;
(b) the first and second isolated antibodies are administered as a first cancer therapy after diagnosis of tumor progression that has occurred despite previous treatment of the cancer with a different cancer therapy; or
(c) the first and second isolated antibodies are administered as a first cancer therapy after diagnosis of toxicity of a different cancer therapy.

### Claim 15

A method of increasing T cell activation in a subject that has cervical cancer, the method comprising administering to the subject an effective amount of a therapeutic combination comprising a first isolated antibody that specifically binds to human CTLA-4 and a second isolated antibody that specifically binds to human PD-1, wherein:
(a) the first isolated antibody comprises a heavy chain variable region (VH) comprising the complementarity determining regions CDRH1, CDRH2, and CDRH3 of the VH amino acid sequence of SEQ ID NO: 1 and a light chain variable region (VL) comprising the complementarity determining regions CDRL1, CDRL2, and CDRL3 of the VL amino acid sequence of SEQ ID NO: 14, and
(b) the second isolated antibody comprises a heavy chain variable region (VH) comprising the complementarity determining regions CDRH1, CDRH2 and CDRH3 of the VH amino acid sequence of SEQ ID NO: 66 and a light chain variable region (VL) comprising the complementarity determining regions CDRL1, CDRL2 and CDRL3 of the VL amino acid sequence of SEQ ID NO: 74.

### Claim 16 (depends on 15)

The method of claim 15, wherein the cancer has relapsed after platinum-based chemotherapy.

### Claim 17 (depends on 15)

The method of claim 15, wherein the cancer is advanced or metastatic.

### Claim 18 (depends on 15)

The method of claim 15, wherein:
(a) the first isolated antibody comprises a heavy chain variable region and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 1 and 14, respectively; and
(b) the second isolated antibody comprises a heavy chain variable region and a light chain variable region comprising the amino acid sequences set forth in SEQ ID NOs: 66 and 74, respectively.

### Claim 19 (depends on 15)

The method of claim 15, wherein:
(a) the first isolated antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 51 and a light chain comprising the amino acid sequence of SEQ ID NO: 59; and
(b) the second isolated antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 91 and a light chain comprising the amino acid sequence of SEQ ID NO: 93.