# US 2019/0169303 A1 · Humanized Anti-OX40 Antibodies and Uses Thereof

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- Number: US 2019/0169303 A1
- Document ID: US20190169303A1
- Status: Published application
- Kind: A1
- Application number: 16/215,263
- Filed: 2018-12-10
- Published: 2019-06-06
- Earliest claimed: 2014-10-10
- Applicants: MedImmune, LLC
- Inventors: Scott A. Hammond; Michael Oberst; Qun Du; Melissa Damschroder
- Web page: https://patentgrep.com/patent/US20190169303A1

## CPC

- C07K 16/2878: {against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95}
- A61P 35/00: Antineoplastic agents
- A61K 2039/505: {comprising antibodies}
- C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
- 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 2317/51: Complete heavy chain or Fd fragment, i.e. VH + CH1
- C07K 2317/515: Complete light chain, i.e. VL + CL
- C07K 2317/73: Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
- C07K 2317/732: Antibody-dependent cellular cytotoxicity [ADCC]
- C07K 2317/734: Complement-dependent cytotoxicity [CDC]
- 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 disclosure provides humanized anti-OX40 antibodies. Also provided are methods of making such antibodies, and methods of use, e.g., treatment of cancer.

## Claims

### Claim at position 1

1-70. (cancelled)

### Claim 71

An antibody or antigen-binding fragment thereof, comprising:
a humanized heavy chain variable region (VH) and a humanized light chain variable region (VL);
wherein the VH comprises an amino acid sequence with the formula:
HFW1-HCDR1-HFW2-HCDR2-HFW3-HCDR3-HFW4,
wherein HFW 1 is SEQ ID NO: 6 or SEQ ID NO: 7, HCDR1 is SEQ ID NO: 8, HFW2 is SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, or SEQ ID NO: 13, HCDR2 is SEQ ID NO: 14, SEQ ID NO: 15 or SEQ ID NO: 16, HFW3 is SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, or SEQ ID NO: 24, HCDR3 is SEQ ID NO: 25, SEQ ID NO: 26, or SEQ ID NO: 27, and HFW4 is SEQ ID NO: 28;
wherein the VL comprises the amino acid sequence SEQ ID NO: 29 or SEQ ID NO: 32; and
wherein the antibody or antigen-binding fragment thereof can specifically bind to human OX40.

### Claim 72 (depends on 71)

The antibody or antigen-binding fragment thereof of claim 71, wherein the VH comprises the amino acid sequence of SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, or SEQ ID NO: 67.

### Claim 73 (depends on 71)

The antibody or antigen-binding fragment thereof of claim 71 further comprising a light chain constant region or fragment thereof fused to the C-terminus of the VL, wherein the light chain constant region is a human kappa constant region.

### Claim 74 (depends on 71)

The antibody or antigen-binding fragment thereof of claim 71, further comprising a heavy chain constant region or fragment thereof fused to the C-terminus of the VH, wherein the heavy chain constant region is a human IgG1 constant region, a human IgG4P constant region, a human IgG1TM constant region, or a murine IgG1 constant region.

### Claim 75 (depends on 71)

The antibody or antigen-binding fragment thereof of claim 71 further comprising the heavy chain amino acid sequence SEQ ID NO: 71 and the light chain amino acid sequence SEQ ID NO: 30.

### Claim 76 (depends on 71)

The antibody or antigen-binding fragment of claim 71, wherein the antigen-binding fragment is an Fv fragment, an Fab fragment, an F(ab′)2 fragment, an Fab′ fragment, a dsFv fragment, an scFv fragment, or an sc(Fv)2 fragment, or any combination thereof.

### Claim 77 (depends on 71)

The antibody or antigen-binding fragment thereof of claim 71, which can specifically bind to OX40 as expressed on Jurkat cells, primary activated CD4+ or CDS+ T cells from human, cynomolgus monkey, rhesus monkey, or any combination thereof and which does not bind to murine or rat OX40.

### Claim 78 (depends on 71)

The antibody or antigen-binding fragment thereof of claim 71, which does not cross react with related Tumor Necrosis Factor Receptor Superfamily (TNFRSF) proteins.

### Claim 79 (depends on 71)

The antibody or antigen-binding fragment thereof of claim 71, which can induce dose-dependent proliferation of activated CD4+ T cells and dose-dependent cytokine release in primary activated human CD4+ T cells in a plate-based assay.

### Claim 80 (depends on 79)

The antibody or antigen-binding fragment thereof of claim 79, wherein the cytokine is IFNγ, TNFα, IL-5, IL-10, IL-2, IL-4, IL-13, IL-8, IL-12 p70, IL-1, or any combination thereof.

### Claim 81 (depends on 71)

The antibody or antigen-binding fragment thereof of claim 71, which can activate the NFKB pathway in OX40 expressing T cells in the presence of FcγR-expressing cells.

### Claim 82 (depends on 71)

The antibody or antigen-binding fragment thereof of claim 71, which can trigger complement-dependent or antibody-dependent cellular cytotoxicity against OX40-expressing cells.

### Claim 83 (depends on 71)

The antibody or antigen-binding fragment thereof of claim 71, wherein administration of an effective dose to a subject in need of cancer treatment can inhibit tumor growth in the subject.

### Claim 84 (depends on 83)

The antibody or antigen-binding fragment thereof of claim 83, wherein the tumor growth inhibition is achieved in the presence of T cells.

### Claim 85 (depends on 84)

The antibody or antigen-binding fragment thereof of claim 84, wherein tumor growth is inhibited by at least 10%, at least 20%, at least 30%, at least 40%, and least 50%, at least 60%, or at least 70% compared to administration of an isotype-matched control antibody or antigen-binding fragment thereof.

### Claim 86 (depends on 71)

A composition comprising the antibody or antigen-binding fragment thereof of claim 71 and a carrier.

### Claim 87 (depends on 71)

A polynucleotide comprising a nucleic acid that encodes the antibody or antigen-binding fragment thereof of claim 71.

### Claim 88 (depends on 87)

The polynucleotide of claim 87, wherein the polynucleotide comprises the nucleic acid of SEQ ID NO: 60, the nucleic acid of SEQ ID NO: 31, the nucleic acid of SEQ ID NO: 72, or any combination thereof.

### Claim 89 (depends on 87)

A vector comprising the polynucleotide of claim 87.

### Claim 90 (depends on 87)

A host cell comprising the polynucleotide of claim 87.

### Claim 91 (depends on 90)

A method of producing an antibody or antigen-binding fragment thereof, comprising culturing the host cell of claim 90 under conditions in which the antibody or antigen-binding fragment thereof encoded by the polynucleotide is expressed; and recovering the antibody or antigen-binding fragment thereof.

### Claim 92 (depends on 71)

A method to promote survival or proliferation of activated T cells, comprising contacting activated T cells with the antibody or antigen-binding fragment thereof of claim 71, wherein the antibody or antigen-binding fragment thereof can specifically bind to OX40 on the surface of the T cells, wherein the activated T cells are activated CD4+ T cells, activated CDS+T cells, or a combination thereof.

### Claim 93 (depends on 71)

A method of inducing cytokine release from activated T cells, comprising contacting activated T cells with the antibody or antigen-binding fragment thereof of claim 71, wherein the antibody or antigen-binding fragment thereof can specifically bind to OX40 on the surface of the T cells; and releasing a cytokine from the activated T cells.

### Claim 94 (depends on 93)

The method of claim 93, wherein the cytokine is IFNγ, TNFα, IL-5, IL-10, IL-2, IL-4, IL-13, IL-8, IL-12 p70, IL-1, or any combination thereof.

### Claim 95 (depends on 71)

A method of promoting T cell activation, comprising contacting T cells with the antibody or antigen-binding fragment thereof of claim 71, wherein the antibody or antigen-binding fragment thereof can specifically bind to OX40 on the surface of the T cells.

### Claim 96 (depends on 95)

The method of claim 95, wherein T cell activation can be measured through stimulation of the NFKB signal transduction pathway.

### Claim 97 (depends on 95)

The method of claim 95, wherein the T cells are activated CD4+ T cells, activated CDS+ T cells, or a combination thereof.

### Claim 98 (depends on 97)

The method of claim 97, wherein the activated CD4+ T cells are human CD4+ T cells, cynomolgus monkey CD4+ T cells, rhesus monkey CD4+ T cells, or a combination thereof.

### Claim 99 (depends on 95)

The method of claim 95, wherein the contacting comprises administering an effective amount of the antibody or antigen-binding fragment thereof to a subject.

### Claim 100 (depends on 86)

A method of treating cancer in a subject, comprising administering to a subject in need of treatment an effective amount of the composition of claim 86.

### Claim 101 (depends on 100)

The method of claim 100, wherein the cancer is a solid tumor.

### Claim 102 (depends on 101)

The method of claim 101, wherein administration of the composition can inhibit tumor growth, can promote tumor reduction, or both.

### Claim 103 (depends on 102)

The method of claim 102, wherein tumor growth inhibition is achieved in the presence of T cells.

### Claim 104 (depends on 71)

A method of enhancing an immune response in a subject comprising administering to a subject in need thereof a therapeutically effective amount of the antibody or antigen-binding fragment thereof of claim 71.

### Claim 105 (depends on 71)

The antibody or antigen-binding fragment thereof of claim 71, which binds to an epitope of human OX40 that falls within amino acids 108 to 146 of SEQ ID NO: 91.

### Claim 106 (depends on 105)

The antibody or antigen-binding fragment thereof of claim 105, wherein the epitope comprises at least amino acids leucine 116 and alanine 126 of SEQ ID NO: 91.

### Claim 107 (depends on 71)

The antibody or antigen-binding fragment thereof of claim 71, which binds to a mouse OX40 variant comprising SEQ ID NO: 92, except for a Q113L mutation and a V124A mutation.

### Claim 108 (depends on 71)

An isolated peptide consisting of 100 or fewer amino acids and comprising amino acids 108 to 146 of SEQ ID NO: 91 except for one, two, three, four, five, or six single amino acid substitutions, deletions, or insertions at any position except L116 and A126, wherein the peptide can be specifically bound by the antibody or antigen-binding fragment thereof of claim 71.