# US 12,005,078 B2 · Genetically engineered drug resistant T cells and methods of using the same

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- Number: US 12,005,078 B2
- Document ID: US12005078B2
- Status: Granted
- Kind: B2
- Application number: 15/756,937
- Filed: 2016-09-06
- Published: 2018-09-06
- Granted: 2024-06-11
- Earliest claimed: 2015-09-03
- Assignees: The UAB Research Foundation; Emory University; Children's Healthcare of Atlanta, Inc.
- Applicants: The UAB Research Foundation; Emory University; Children's Healthcare of Atlanta, Inc.
- Inventors: Lawrence S. Lamb; H. Trent Spencer; G. Yancey Gillespie
- Web page: https://patentgrep.com/patent/US12005078B2

## CPC

- A61K 31/495: having six-membered rings with two {or more} nitrogen atoms as the only ring heteroatoms, e.g. piperazine {or tetrazines}(A61K31/48 takes precedence {; with three nitrogen atoms A61K31/53})
- A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
- A61K 40/15: Natural-killer [NK] cells; Natural-killer T [NKT] cells
- A61K 40/30: characterised by the recombinant expression of specific molecules in the cells of the immune system
- A61K 40/31: Chimeric antigen receptors [CAR]
- A61K 40/4204: {Epidermal growth factor receptors [EGFR]}
- A61K 40/4217: {Receptors for interleukins [IL]}
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61P 35/00: Antineoplastic agents
- C07K 14/7051: {T-cell receptor (TcR)-CD3 complex}
- C07K 14/7155: {for interleukins [IL]}
- C07K 16/30: from tumour cells
- C12N 5/0636: {T lymphocytes}
- C12N 5/0638: {Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK]}
- C12N 5/0646: {Natural killers cells [NK], NKT cells}
- C12N 9/12: transferring phosphorus containing groups, e.g. kinases (2.7)
- C12Y 207/10001: Receptor protein-tyrosine kinase (2.7.10.1)
- A61K 2239/31: characterized by the route of administration
- A61K 2239/38: characterised by the dose, timing or administration schedule
- A61K 2239/47: Brain; Nervous system
- A61K 2300/00: Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
- C07K 2319/70: containing domain for protein-protein interaction
- C12N 2501/72: Transferases [EC 2.]
- 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)
- C12Y: ENZYMES

## Abstract

The present disclosure provides novel cell compositions engineered to express at least a chimeric antigen receptor and a survival factor. Methods of using such cell compositions are also described.

## Claims

### Claim 1

A method of treating a subject suffering from a tumor, the method comprising:
a. administering to the subject a cell composition comprising γδ T cells expressing at least NKG2D, a chimeric antigen receptor (CAR) directed to a tumor antigen,, and a polypeptide that confers resistance to a chemotherapeutic agent, wherein the chemotherapeutic agent increases the expression of a NKG2D ligand on the tumor; and
b. administering to the subject the chemotherapeutic agent, wherein the chemotherapeutic agent is administered either before administration of the cell composition to the subject, after administration of the cell composition to the subject, concurrently with administration of the cell composition to the subject or any combination of the foregoing;
wherein the CAR comprises an endodomain, and the endodomain consists of a CD3-zeta signaling domain or one costimulatory domain.

### Claim 2 (depends on 1)

The method of claim 1 further comprising at least one additional administration of the cell composition to the subject.

### Claim 3 (depends on 1)

The method of claim 1, wherein the chemotherapeutic agent is administered before administration of the cell composition to the subject, or concurrently with administration of the cell composition to the subject, or a combination of the foregoing.

### Claim 4 (depends on 1)

The method of claim 1, wherein the cell composition is administered to the subject on day X and the chemotherapeutic agent is administered to the subject 12 to 72 hours prior to day X, 12 to 72 hours after day X or both 12 to 72 hours prior to and after day X.

### Claim 5 (depends on 1)

The method of claim 1, wherein the cell composition is administered to the subject on day X and the chemotherapeutic agent is administered to the subject 12 to 72 hours prior to day X, 12 to 72 hours after day X or both 12 to 72 hours prior to and after day X, followed by an additional administration of the cell composition to the subject on day Y, with optional administration of the chemotherapeutic agent to the subject 12 to 72 hours prior to day Y, 12 to 72 hours after day Y or both 12 to 72 hours prior to and after day Y.

### Claim 6 (depends on 1)

The method of claim 1, wherein the cancer is selected from the group consisting of brain cancer, breast cancer, prostate cancer, lung cancer, colon cancer, epithelial cancer, head and neck cancer, skin cancer, cancers of the genito-urinary tract, ovarian cancer, endometrial cancer, cervical cancer, kidney cancer, gastric cancer, cancer of the small intestine, liver cancer, pancreatic cancer, gall bladder cancer, cancers of the bile duct, esophageal cancer, cancer of the salivary glands, thyroid cancer, and a hematologic malignancy.

### Claim 7 (depends on 1)

The method of claim 1, wherein the cancer is selected from the group consisting of pineal tumors, pituitary tumors, PNET, schwannoma, lymphoma, medulloblastoma, meningioma, metastatic brain cancer, neurofibroma, neuronal & mixed neuronal-glial tumors, oligoastrocytoma, oligodendroglioma, astrocytoma, atypical teratoid rhaboid tumor (ATRT), chondrosarcoma, choroid plexus tumors, craniopharyngioma, ependymoma, germ cell tumor, neuroblastoma, glioblastoma and glioma.

### Claim 8 (depends on 1)

The method of claim 1, wherein the cancer is sensitive to the chemotherapeutic agent.

### Claim 9 (depends on 1)

The method of claim 1, wherein the cancer is resistant to the chemotherapeutic agent.

### Claim 10 (depends on 1)

The method of claim 1, wherein the tumor antigen is selected from the group consisting of EphA2, B cell maturation antigen (BCMA), B7-H3, B7-H6, CALX, CA9, CD22, CD19, CD20, ROR1, kappa or light chain, carcinoembryonic antigen, alpha-fetoprotein, CA-125, Glypican-3, epithelial tumor antigen, melanoma-associated antigen, EGP2, EGP40, EPCAM, ERBB3, ERBB4, ErbB3/4, PAP, FAR, FBP, fetal AchR, Folate Receptor α, mutated p53, mutated ras, HER2, ERBB2, HER3, folate binding protein, HIV-1 envelope glycoprotein gp120, HIV-1 envelope glycoprotein gp41, 5T4, 8H9, GD2, CD123, CD23, CD33, CD30, CD38, CD56, c-Met, fap, mesothelin, GD3, HERV-K, IL-11Ra, IL-13Ra, CSPG4, Lewis-Y, MCSP, Mucl, Muc16, NCAM, NKG2D ligands, NY-ESO-1, PRAME, PSCA, PSC1, PSMA, EGFR, Sp17, SURVIVIN, TAG72, TEM1, TEM8, EGFRvIII and VEGFR2.

### Claim 11 (depends on 1)

The method of claim 1, wherein the cancer is glioblastoma and the tumor antigen is selected from the group consisting of ULBP-1, ULBP-2, ULBP-3, ULBP-4, ULBP-5, ULBP-6, MIC-A, MIC-B, EGFRvIII and IL13Rα.

### Claim 12 (depends on 1)

The method of claim 1, wherein the cancer is glioblastoma and the tumor antigen is EGFRvIII or IL13Rα.

### Claim 13 (depends on 1)

The method of claim 1, wherein the cancer is neuroblastoma and the tumor antigen is selected from the group consisting of ULBP-1, ULBP-2, ULBP-3, ULBP-4, ULBP-5, ULBP-6, MIC-A, MIC-B and GD2.

### Claim 14 (depends on 1)

The method of claim 1, wherein the cancer is neuroblastoma and the tumor antigen is GD2.

### Claim 15 (depends on 1)

The method of claim 1, wherein the cell composition further comprises at least one of αβ T cells and NK cells.

### Claim 16 (depends on 1)

The method of claim 1, wherein the cell composition further comprises αβ T cells and NK cells.

### Claim 17 (depends on 1)

The method of claim 1, wherein the γδ T cells are present at greater than or equal to 60% of the total cell population, αβ T cells are present at less than or equal to 5% of the total cell population and NK cells are present at less than or equal to 25% of the total cell population, as measured by flow cytometry.

### Claim 18 (depends on 1)

The method of claim 1, wherein the chemotherapeutic agent is a nucleoside-analog, alkylating agent, antimetabolite, antibiotic, topoisomerase inhibitor, mitotic inhibitor, differentiating agent, or hormone therapy agent.

### Claim 19 (depends on 1)

The method of claim 1, wherein the polypeptide is O6-Methylguanine-DNA methyltransferase, multidrug resistance protein 1 or 5′ nucleotidase II.

### Claim 20 (depends on 1)

The method of claim 1, wherein the chemotherapeutic agent is an alkylating agent and the polypeptide is O6-Methylguanine-DNA methyltransferase.

### Claim 21 (depends on 1)

The method of claim 1, wherein the chemotherapeutic agent is temozolomide and the polypeptide is O6-Methylguanine-DNA methyltransferase.

### Claim 22 (depends on 1)

The method of claim 1, wherein the γδ T cells are engineered to express the CAR directed to a tumor antigen and the polypeptide, and the receptor for NKG2D is naturally present on the γδ T cells.

### Claim 23 (depends on 1)

The method of claim 1, wherein the γδ T cells further express a suicide gene.

### Claim 24 (depends on 1)

The method of claim 1, wherein the chemotherapeutic agent increases the expression of a NKG2D ligand selected from the group consisting of ULBP-1, ULBP-2, ULBP-3, ULBP-4, ULBP-5, ULBP-6, MIC-A and MIC-B.

### Claim 25 (depends on 1)

The method of claim 1, wherein the γδ T cells are obtained from the subject and optionally expanded ex vivo.

### Claim 26 (depends on 1)

The method of claim 1, wherein the cancer is glioblastoma and the tumor antigen is a NKG2D ligand.

### Claim 27 (depends on 1)

The method of claim 1, wherein the endodomain consists of the CD3-zeta signaling domain.

### Claim 28 (depends on 1)

The method of claim 1, wherein the endodomain consists of one co-stimulatory domain.

### Claim 29 (depends on 28)

The method of claim 28, wherein the co-stimulatory domain is selected from the group consisting of CD28, 41BB, DAP10, OX40 and ICOS.

### Claim 30 (depends on 6)

The method of claim 6, wherein the hematologic malignancy is selected from the group consisting of leukemia, lymphoma, multiple myeloma, and myelodysplastic syndrome.