# US 12,005,103 B2 · Universal anti-tag chimeric antigen receptor-expressing t cells and methods of treating cancer

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- Number: US 12,005,103 B2
- Document ID: US12005103B2
- Status: Granted
- Kind: B2
- Application number: 17/228,001
- Filed: 2021-04-12
- Published: 2021-10-28
- Granted: 2024-06-11
- Earliest claimed: 2010-12-14
- Assignees: University of Maryland
- Applicants: University of Maryland, Baltimore
- Inventors: Eduardo Davila; Koji Tamada
- Web page: https://patentgrep.com/patent/US12005103B2

## CPC

- A61K 39/0011: {Cancer 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 39/39558: {against tumor tissues, cells, antigens}
- A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
- A61K 40/31: Chimeric antigen receptors [CAR]
- A61K 40/4204: {Epidermal growth factor receptors [EGFR]}
- A61K 40/4205: {Her-2/neu/ErbB2, Her-3/ErbB3 or Her 4/ ErbB4}
- A61K 47/555: {pre-targeting systems involving an organic compound, other than a peptide, protein or antibody, for targeting specific cells}
- A61K 47/6855: {the tumour determinant being from breast cancer cell}
- A61K 47/6859: {the tumour determinant being from liver or pancreas cancer cell}
- A61K 47/6863: {the tumour determinant being from stomach or intestines cancer cell}
- A61K 47/6897: {Pre-targeting systems with two or three steps using antibody conjugates; Ligand-antiligand therapies}
- A61P 35/00: Antineoplastic agents
- C07K 14/7051: {T-cell receptor (TcR)-CD3 complex}
- C07K 16/2863: {against receptors for growth factors, growth regulators}
- C07K 16/44: against material not provided for elsewhere {, e.g. haptens, metals, DNA, RNA, amino acids}
- C12N 5/0636: {T lymphocytes}
- A61K 2039/5156: {expressing foreign proteins}
- A61K 2039/572: {cytotoxic response}
- A61K 2039/585: {wherein the target is cancer}
- A61K 2239/31: characterized by the route of administration
- A61K 2239/38: characterised by the dose, timing or administration schedule
- A61K 2239/50: Colon
- A61K 2239/54: Pancreas
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2319/00: Fusion polypeptide
- C07K 2319/03: containing a transmembrane segment
- C12N 2501/515: CD3, T-cell receptor complex
- C12N 2510/00: Genetically modified cells
- 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
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention provides a universal, yet adaptable, anti-tag chimeric antigen receptor (AT-CAR) system which provides T cells with the ability and specificity to recognize and kill target cells, such as tumor cells, that have been marked by tagged antibodies. As an example, αFITC-CAR-expressing T cells have been developed that specifically recognize various human cancer cells when those cells are bound by cancer-reactive FITC-labeled antibodies. The activation of αFITC-CAR-expressing T cells is shown to induce efficient target lysis, T cell proliferation, and cytokine/chemokine production. The system can be used to treating subjects having cancer.

## Claims

### Claim 1

A method of treating cancer in a subject, comprising:
(a) administering a formulation of tagged proteins to a subject in need of treatment, wherein the tagged proteins bind a cancer cell in the subject, and
(b) administering a therapeutically-effective population of anti-tag chimeric receptor (AT-CAR)-expressing effector cells to the subject, wherein the AT-CAR comprises a tag-binding domain, a transmembrane domain, and a T cell activation domain, where the T cell activation domain consists of one or more of the cytoplasmic region of OX40, the cytoplasmic region of HVEM, and FcRε, where the T cell activation domain does not comprise CD3ζ, and wherein the AT-CAR-expressing effector cells bind the tagged proteins and induce cancer cell death, thereby treating cancer in a subject.

### Claim 2

A method of treating cancer in a subject, comprising:
(a) administering one or more formulations of tagged proteins to a subject in need of treatment, wherein the tagged proteins bind a cancer cell in the subject, and
(b) administering one or more therapeutically-effective populations of AT-CAR-expressing effector cells to the subject, wherein the AT-CAR comprises a tag-binding domain, a transmembrane domain, and a T cell activation domain, where the T cell activation domain consists of one or more of the cytoplasmic region of OX40, the cytoplasmic region of HVEM, and FcRε, where the T cell activation domain does not comprise CD3ζ, and wherein the AT-CAR-expressing effector cells bind the tagged proteins and induce cancer cell death, thereby treating cancer in a subject.

### Claim 3

A method of treating cancer in a subject, comprising:
(a) administering at least two formulations of tagged proteins to a subject in need of treatment, wherein the tagged proteins bind a cancer cell in the subject, and
(b) administering at least two therapeutically-effective populations of AT-CAR-expressing effector cells to the subject, wherein the AT-CAR comprises a tag-binding domain, a transmembrane domain, and a T cell activation domain, where the T cell activation domain consists of one or more of the cytoplasmic region of OX40, the cytoplasmic region of HVEM, and FcRε, where the T cell activation domain does not comprise CD3ζ, and wherein the AT-CAR-expressing effector cells bind the tagged proteins and induce cancer cell death, thereby treating cancer in a subject.

### Claim 4 (depends on 1)

The method of claim 1, wherein the tagged proteins are tagged with a tag selected from the group consisting of fluorescein isothiocyanate (FITC), streptavidin, biotin, dinitrophenol, peridinin chlorophyll protein complex, green fluorescent protein, phycoerythrin (PE), horse radish peroxidase, palmitoylation, nitrosylation, alkalanine phosphatase, glucose oxidase, and maltose binding protein.

### Claim 5 (depends on 1)

The method of claim 1, wherein the protein of the tagged proteins is an antibody or an antigen-binding fragment thereof.

### Claim 6 (depends on 5)

The method of claim 5, wherein the antibody or antigen-binding fragment thereof is cetuximab, nimotuzumab, panitumumab, retuximab, omalizumab, tositumomab, trastuzumab, gemtuzumab, or alemtuzumab, or an antigen-binding fragment of any one thereof.

### Claim 7 (depends on 1)

The method of claim 1, wherein the tag-binding domain is an antibody or an antigen-binding fragment thereof.

### Claim 8 (depends on 1)

The method of claim 1, wherein the tag-binding domain specifically binds FITC, biotin, PE, or streptavidin.

### Claim 9 (depends on 7)

The method of claim 7, wherein the antigen-binding fragment is a single chain variable fragment (scFv).

### Claim 10 (depends on 7)

The method of claim 7, wherein the antigen-binding fragment is a single chain variable fragment (scFv) that specifically binds FITC, biotin, PE, or streptavidin.

### Claim 11 (depends on 7)

The method of claim 7, wherein the transmembrane domain is the hinge and transmembrane regions of the human CD8α chain.

### Claim 12 (depends on 1)

The method of claim 1, wherein the effector cells of the population of AT-CAR-expressing effector cells comprises CD3+CD8+ T cells, CD3+CD4+ T cells, gamma delta T cells, cytokine induced killer (CIK) cells, natural killer (NK) cells, natural killer T (NKT) cells, tumor infiltrating lymphocytes (TILs), macrophages, or microglia.

### Claim 13 (depends on 1)

The method of claim 1, wherein the formulation of tagged proteins is administered to the subject prior to administration of the therapeutically-effective population of AT-CAR-expressing effector cells.

### Claim 14 (depends on 1)

The method of claim 1, wherein the formulation of tagged proteins are administered to the subject concurrently with administration of the therapeutically-effective population of AT-CAR-expressing effector cells.

### Claim 15 (depends on 1)

The method of claim 1, wherein the formulation of tagged proteins are administered to the subject after administration of the therapeutically-effective population of AT-CAR-expressing effector cells.

### Claim 16 (depends on 1)

The method of claim 1, wherein the formulation of tagged proteins and the therapeutically-effective population of AT-CAR-expressing effector cells are administered to the subject in any order.

### Claim 17 (depends on 1)

The method of claim 1, wherein AT-CAR-expressing effector cell binding to the tagged proteins, which are bound to a cancer cell, induces cytolytic activation of the effector cells.

### Claim 18 (depends on 1)

The method of claim 1, wherein the subject is a human.