# US 10,869,889 B2 · Method and compositions for cellular immunotherapy

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,869,889 B2
- Document ID: US10869889B2
- Status: Granted
- Kind: B2
- Application number: 16/657,666
- Filed: 2019-10-18
- Published: 2020-03-12
- Granted: 2020-12-22
- Earliest claimed: 2012-08-20
- Assignees: Fred Hutchinson Cancer Research Center; Seattle Children's Hospital
- Applicants: Fred Hutchinson Cancer Research Center; Seattle Children's Hospital
- Inventors: Michael C. Jensen; Stanley R. Riddell; Michael Hudecek
- Web page: https://patentgrep.com/patent/US10869889B2

## CPC

- C07K 14/705: Receptors; Cell surface antigens; Cell surface determinants {(tumour specific antigens C07K14/4748)}
- 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 40/4202: {Receptors, cell surface antigens or cell surface determinants}
- A61K 40/4205: {Her-2/neu/ErbB2, Her-3/ErbB3 or Her 4/ ErbB4}
- A61K 40/4211: {CD19 or B4}
- A61P 1/00: Drugs for disorders of the alimentary tract or the digestive system
- A61P 1/04: for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
- A61P 1/18: for pancreatic disorders, e.g. pancreatic enzymes
- A61P 11/00: Drugs for disorders of the respiratory system
- A61P 13/08: of the prostate
- A61P 13/12: of the kidneys
- A61P 15/00: Drugs for genital or sexual disorders (for disorders of sex hormones A61P5/24); Contraceptives
- A61P 31/12: Antivirals
- A61P 35/00: Antineoplastic agents
- A61P 35/02: specific for leukemia
- A61P 37/02: Immunomodulators
- C07K 14/7051: {T-cell receptor (TcR)-CD3 complex}
- C07K 14/70521: {CD28, CD152}
- C07K 14/7151: {for tumor necrosis factor [TNF], for lymphotoxin [LT]}
- C07K 16/2803: {against the immunoglobulin superfamily}
- C07K 16/32: against translation products of oncogenes
- C07K 16/40: against enzymes
- A61K 2239/17: Hinge-spacer domain
- A61K 2239/31: characterized by the route of administration
- A61K 2239/48: Blood cells, e.g. leukemia or lymphoma
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2317/64: comprising a combination of variable region and constant region components
- C07K 2319/03: containing a transmembrane segment
- C07K 2319/30: Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
- C07K 2319/70: containing domain for protein-protein interaction
- 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)
- 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

## Abstract

The present invention provides nucleic acids, vectors, host cells, methods and compositions to confer and/or augment immune responses mediated by cellular immunotherapy, such as by adoptively transferring CD8+ central memory T cells or combinations of central memory T cells with CD4+ T cells that are genetically modified to express a chimeric receptor. In embodiments the genetically modified host cell comprises a nucleic acid comprising a polynucleotide coding for a ligand binding domain, a polynucleotide comprising a customized spacer region, a polynucleotide comprising a transmembrane domain, and a polynucleotide comprising an intracellular signaling domain. It has been surprisingly found that the length of the spacer region can affects the ability of chimeric receptor modified T cells to recognize target cells in vitro and affects in vivo efficacy of the chimeric receptor modified T cells. Pharmaceutical formulations produced by the method, and methods of using the same, are also described.

## Claims

### Claim 1

A chimeric receptor, the chimeric receptor comprising an extracellular binding region consisting of an scFv and a polypeptide spacer, a transmembrane domain and an intracellular signaling domain, wherein:
(a) the scFv binds to an epitope in the extracellular region of CD19, wherein the scFv comprises a variable heavy (VH) chain domain comprising the sequences DYGVS (SEQ ID NO:74), VIWGSETTYYNSALKS (SEQ ID NO:60) and YAMDYWG (SEQ ID NO:61), and a variable light (VL) chain domain comprising the sequences RASQDISKYLN (SEQ ID NO:57), SRLHSGV (SEQ ID NO:58) and GNTLPYTFG (SEQ ID NO:59);
(b) the polypeptide spacer is located between the scFv and the transmembrane domain, wherein the polypeptide spacer consists of the amino acid sequence of SEQ ID NO:21; and
(c) the intracellular signaling domain comprises the signaling domain of CD3ζ and a costimulatory domain.

### Claim 2

A chimeric receptor, the chimeric receptor comprising:
(a) an scFv that binds to an epitope in the extracellular region of CD19, wherein the scFv comprises the variable heavy (VH) chain domain and the variable light (VL) chain domain encoded by SEQ ID NO:3;
(b) a transmembrane domain;
(c) a polypeptide spacer located between the scFv and the transmembrane domain, wherein the polypeptide spacer consists of the amino acid sequence of SEQ ID NO:21; and
(d) an intracellular signaling domain that comprises the signaling domain of CD3ζ and a costimulatory domain.

### Claim 3 (depends on 2)

The chimeric receptor of claim 2, wherein the scFv is encoded by SEQ ID NO:3.

### Claim 4 (depends on 1)

The chimeric receptor of claim 1, wherein:
the transmembrane domain consists of the transmembrane domain of CD8 or of CD28; and
the costimulatory domain comprises the signaling domain of 4-1BB or of CD28.

### Claim 5 (depends on 2)

The chimeric receptor of claim 2, wherein:
the transmembrane domain consists of the transmembrane domain of CD8 or of CD28; and
the costimulatory domain comprises the signaling domain of 4-1BB or of CD28.

### Claim 6 (depends on 1)

The chimeric receptor of claim 1, wherein:
the transmembrane domain consists of the amino acid sequence encoded by the polynucleotide sequence of SEQ ID NO:5; and
the intracellular signaling domain comprises the amino acid sequence encoded by the polynucleotide sequence of SEQ ID NO:6 and the amino acid sequence encoded by SEQ ID NO:7.

### Claim 7 (depends on 2)

The chimeric receptor of claim 2, wherein:
the transmembrane domain consists of the amino acid sequence encoded by the polynucleotide sequence of SEQ ID NO:5; and
the intracellular signaling domain comprises the amino acid sequence encoded by the polynucleotide sequence of SEQ ID NO:6 and the amino acid sequence encoded by SEQ ID NO:7.

### Claim 8 (depends on 1)

The chimeric receptor of claim 1, wherein the chimeric receptor comprises:
the anti-CD19 scFv encoded by SEQ ID NO:3;
the polypeptide spacer encoded by SEQ ID NO:4;
the transmembrane domain encoded by SEQ ID NO:5;
the costimulatory domain encoded by SEQ ID NO:6; and
the CD3ζ signaling domain encoded by SEQ ID NO:7.

### Claim 9 (depends on 1)

A T cell comprising the chimeric receptor of claim 1.

### Claim 10 (depends on 9)

A composition, comprising the T cell of claim 9 in a pharmaceutically acceptable excipient.

### Claim 11 (depends on 8)

A T cell comprising the chimeric receptor of claim 8.

### Claim 12 (depends on 11)

A composition, comprising the T cell of claim 11 in a pharmaceutically acceptable excipient.

### Claim 13 (depends on 2)

A T cell comprising the chimeric receptor of claim 2.

### Claim 14 (depends on 13)

A composition, comprising the T cell of claim 13 in a pharmaceutically acceptable excipient.

### Claim 15 (depends on 6)

A T cell comprising the chimeric receptor of claim 6.

### Claim 16 (depends on 15)

A composition, comprising the T cell of claim 15 in a pharmaceutically acceptable excipient.

### Claim 17 (depends on 7)

A T cell comprising the chimeric receptor of claim 7.

### Claim 18 (depends on 17)

A composition, comprising the T cell of claim 17 in a pharmaceutically acceptable excipient.