# US 11,077,182 B2 · Methods of preparing anti-human papillomavirus antigen T cells

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- Number: US 11,077,182 B2
- Document ID: US11077182B2
- Status: Granted
- Kind: B2
- Application number: 17/113,570
- Filed: 2020-12-07
- Published: 2021-04-29
- Granted: 2021-08-03
- Earliest claimed: 2013-07-15
- Assignees: The United States of Americans represented by the Secretary, Department of Health and Human Services
- Applicants: The United States of America, as represented by the Secretary, Department of Health and Human Services
- Inventors: Christian S. Hinrichs; Steven A. Rosenberg
- Web page: https://patentgrep.com/patent/US11077182B2

## CPC

- A61K 39/12: Viral antigens
- A61K 35/17: Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes (when activated by a specific antigen A61K39/00)
- A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
- A61K 40/4273: {Glycoprotein 100 [Gp100]}
- A61K 40/46: Viral antigens
- A61P 1/04: for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
- A61P 11/04: for throat disorders
- A61P 15/00: Drugs for genital or sexual disorders (for disorders of sex hormones A61P5/24); Contraceptives
- A61P 15/02: for disorders of the vagina
- A61P 35/00: Antineoplastic agents
- C12N 5/0636: {T lymphocytes}
- C12N 5/0638: {Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK]}
- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- A61K 2039/5158: {Antigen-pulsed cells, e.g. T-cells}
- A61K 2039/572: {cytotoxic response}
- A61K 2039/585: {wherein the target is cancer}
- C12N 2501/2302: Interleukin-2 (IL-2)
- C12N 2502/30: tumour cells
- C12N 2710/20011: Papillomaviridae
- 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
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Disclosed are methods of preparing an isolated population of human papillomavirus (HPV)-specific T cells comprise dividing an HPV-positive tumor sample into multiple fragments; separately culturing the multiple fragments; obtaining T cells from the cultured multiple fragments; testing the T cells for specific autologous HPV-positive tumor recognition; selecting the T cells that exhibit specific autologous HPV-positive tumor recognition; and expanding the number of selected T cells to produce a population of HPV-specific T cells for adoptive cell therapy. Related methods of treating or preventing cancer using the T cells are also disclosed.

## Claims

### Claim 1

A method of treating or preventing HPV+ cancer in a patient, the method comprising:
(a) culturing multiple fragments of an HPV+ tumor sample from the patient in the presence of at least one cytokine;
(b) obtaining T-cells from the cultured fragments;
(c) expanding the number of T-cells to produce an expanded population of HPV-specific T-cells using one or both of (i) irradiated allogenic feeder cells and (ii) irradiated autologous feeder cells; and one or both of (iii) OKT3 antibody and (iv) interleukin-2 wherein the T-cells have not been depleted of CD4+ cells;
(d) optionally, adding a second culturing step following (c); and
(e) administering the expanded population of HPV-specific T-cells to the patient in an amount effective to treat or prevent the HPV+ cancer in the patient.

### Claim 2 (depends on 1)

The method of claim 1, wherein a second culturing step is added after step (c) to further expand the number of selected T-cells.

### Claim 3 (depends on 1)

The method of claim 1, wherein the cytokine in step (a) is IL-2.

### Claim 4 (depends on 3)

The method of claim 3, wherein the IL-2 in step (a) is present at 6000 IU.

### Claim 5 (depends on 1)

The method of claim 1, wherein the IL-2 in step (c) is present at 6000 IU.

### Claim 6 (depends on 1)

The method of claim 1, further comprising, prior to step (a), obtaining an HPV+ tumor sample from the patient and fragmenting said tumor samples.

### Claim 7 (depends on 6)

The method of claim 6, wherein the tumor samples are fragmented mechanically or enzymatically.

### Claim 8 (depends on 1)

The method of claim 1, wherein the expanded population of HPV-specific T-cells recognizes HPV 16-positive cervical cancer cells.

### Claim 9 (depends on 1)

The method of claim 1, wherein the expanded population of HPV-specific cells recognizes an HPV antigen selected from the group consisting of HPV 16 E6 and HPV 16 E7.

### Claim 10 (depends on 1)

The method of claim 1, wherein the expanded population of HPV-specific cells recognizes an HPV antigen selected from the group consisting of HPV 18 E6 and HPV 18 E7.

### Claim 11 (depends on 1)

The method of claim 1, further comprising administering to the patient nonmyleoablative lymphodepleting chemotherapy prior to step (e).

### Claim 12 (depends on 1)

The method of claim 1, wherein the expanded population of cells obtained in (d) comprises multiple T-cells having different HPV specificities.

### Claim 13 (depends on 1)

The method of claim 1, wherein the expanded population of cells obtained in (d) secretes at least about 200 pg/mL of interferon-y.

### Claim 14 (depends on 1)

The method of claim 1, wherein the HPV+ cancer is selected from head and neck squamous cell carcinoma (HNSCC) and cervical cancer.

### Claim 15 (depends on 14)

The method of claim 14, wherein the HPV+ cancer is cervical cancer.

### Claim 16 (depends on 14)

The method of claim 14, wherein the HPV+ cancer is head and neck squamous cell carcinoma.

### Claim 17 (depends on 16)

The method of claim 16, wherein the head and neck squamous cell carcinoma is selected from laryngeal cancer, hypopharyngeal cancer, nasal cavity cancer, nasopharyngeal cancer, oral cancer, oropharyngeal cancer, and salivary gland cancer.

### Claim 18 (depends on 1)

The method of claim 1, wherein the expansion step occurs over about 10 to about 14 days.

### Claim 19 (depends on 18)

The method of claim 18, wherein the population of HPV-specific T-cells is increased at least about 1000-fold to about 3000-fold.

### Claim 20 (depends on 1)

The method of claim 1, wherein the expanded population of HPV-specific T-cells is administered at a dose of from about 1×1010 to about 13.7×1010.

### Claim 21 (depends on 1)

The method of claim 1, wherein the expanded population of HPV-specific T-cells is administered with a pharmaceutically acceptable carrier.

### Claim 22 (depends on 21)

The method of claim 21, wherein the pharmaceutically acceptable carrier is saline, 5% dextrose in water, Ringer's lactate, an electrolyte solution, or PLASMA-LYTE A.

### Claim 23 (depends on 1)

The method of claim 1, wherein the expanded population of HPV-specific T-cells is enriched for CD4+ T-cells.

### Claim 24 (depends on 1)

The method of claim 1, wherein the culturing step (a) is performed for at least about 12 days.