# US 11,896,617 B2 · Polynucleotides encoding rituximab-resistant chimeric antigen receptors

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- Number: US 11,896,617 B2
- Document ID: US11896617B2
- Status: Granted
- Kind: B2
- Application number: 16/857,573
- Filed: 2020-04-24
- Published: 2020-12-10
- Granted: 2024-02-13
- Earliest claimed: 2019-04-26
- Assignees: Allogene Therapeutics, Inc.
- Applicants: Allogene Therapeutics, Inc.
- Inventors: Thomas Charles Pertel; Barbra Johnson Sasu; Mark W. Leonard
- Web page: https://patentgrep.com/patent/US11896617B2

## CPC

- 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)
- C07K 14/7051: {T-cell receptor (TcR)-CD3 complex}
- A61K 31/7088: Compounds having three or more nucleosides or nucleotides
- 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 40/4211: {CD19 or B4}
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61P 35/00: Antineoplastic agents
- A61P 35/02: specific for leukemia
- C07K 14/70517: {CD8}
- C07K 14/70596: {Molecules with a "CD"-designation not provided for elsewhere}
- C07K 16/2803: {against the immunoglobulin superfamily}
- C12N 5/0636: {T lymphocytes}
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- A61K 2039/505: {comprising antibodies}
- A61K 2239/38: characterised by the dose, timing or administration schedule
- A61K 2239/48: Blood cells, e.g. leukemia or lymphoma
- C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding
- C07K 2319/33: fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
- C12N 2510/00: Genetically modified cells
- C12N 2740/15043: viral genome or elements thereof as genetic vector
- 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)
- 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
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Provided herein are polynucleotides encoding chimeric antigen receptors (CARs) comprising a CD19 antigen binding domain that specifically binds to CD19 and is resistant to rituximab binding; and immune cells comprising these CD19-specific CARs, e.g., CAR-T cells. Also provided are methods of making and using these CD19-specific CARs, and immune cells comprising these CD19-specific CARs.

## Claims

### Claim 1

An isolated polynucleotide encoding a polypeptide comprising an anti-CD19 chimeric antigen receptor (CAR) that comprises the amino acid sequence of SEQ ID NO: 9, wherein the polypeptide does not comprise a rituximab binding site, wherein the polynucleotide comprises a nucleic acid sequence having at least 90% identity to SEQ ID NO: 3 and a short EF1a promoter that is capable of expressing the anti-CD19 chimeric antigen receptor (CAR) in a mammalian T cell, and wherein the short EF1a promoter comprises the nucleic acid sequence of SEQ ID NO:16 and does not comprise the nucleic acid sequence of SEQ ID NO:39.

### Claim 2 (depends on 1)

The isolated polynucleotide of claim 1, wherein the polypeptide further comprises a safety switch.

### Claim 3 (depends on 2)

The isolated polynucleotide of claim 2, wherein the safety switch is linked to the CD19 CAR using a linker peptide.

### Claim 4 (depends on 2)

The isolated polynucleotide of claim 2, wherein the safety switch is linked to the anti-CD19 CAR using a T2A linker.

### Claim 5 (depends on 2)

The isolated polynucleotide of claim 2, wherein the safety switch comprises an antibody binding site.

### Claim 6 (depends on 2)

The isolated polynucleotide of claim 2, wherein the polypeptide comprises a CD34 epitope.

### Claim 7 (depends on 6)

The isolated polynucleotide of claim 6, wherein the CD34 epitope is a QBEND-10 epitope.

### Claim 8 (depends on 1)

The isolated polynucleotide of claim 1, wherein the polypeptide further comprises a CD8 hinge/transmembrane domain.

### Claim 9 (depends on 1)

The isolated polynucleotide of claim 1, wherein the polynucleotide encodes a polypeptide that comprises any one of the amino acid sequences of SEQ ID NOs: 11-14.

### Claim 10 (depends on 1)

A vector comprising the isolated polynucleotide of claim 1.

### Claim 11 (depends on 10)

The vector of claim 10, wherein the vector is a retroviral vector, a DNA vector, a plasmid, an RNA vector, an adenoviral vector, an adenovirus associated vector, a lentiviral vector, or any combination thereof.

### Claim 12 (depends on 1)

An engineered immune cell comprising the isolated polynucleotide of claim 1, wherein the engineered immune cell does not express a rituximab binding site.

### Claim 13 (depends on 10)

An engineered immune cell comprising the vector of claim 10, wherein the engineered immune cell does not express a rituximab binding site.

### Claim 14 (depends on 12)

The engineered immune cell of claim 12, wherein the immune cell is a T cell, tumor infiltrating lymphocyte (TIL), NK cell, TCR-expressing cell, dendritic cell, or NK-T cell.

### Claim 15 (depends on 14)

The engineered immune cell of claim 14, wherein the cell is an autologous T cell.

### Claim 16 (depends on 14)

The engineered immune cell of claim 14, wherein the cell is an allogeneic T cell.

### Claim 17 (depends on 12)

The engineered immune cell of claim 12, wherein the cell is resistant to rituximab.

### Claim 18 (depends on 12)

A pharmaceutical composition comprising the engineered immune cell of claim 12 and a pharmaceutically acceptable excipient.

### Claim 19 (depends on 1)

The isolated polynucleotide of claim 1, wherein the polynucleotide comprises a nucleic acid sequence having at least 95% identity to SEQ ID NO: 3.

### Claim 20 (depends on 1)

The isolated polynucleotide of claim 1, wherein the polynucleotide comprises a nucleic acid sequence having at least 96% identity to SEQ ID NO: 3.

### Claim 21 (depends on 1)

The isolated polynucleotide of claim 1, wherein the polynucleotide comprises a nucleic acid sequence having at least 98% identity to SEQ ID NO: 3.

### Claim 22 (depends on 1)

The isolated polynucleotide of claim 1, wherein the polynucleotide comprises a nucleic acid sequence having at least 99% identity to SEQ ID NO: 3.

### Claim 23 (depends on 1)

The isolated polynucleotide of claim 1, wherein the polynucleotide comprises the nucleic acid sequence shown in SEQ ID NO: 3.