# US 12,187,808 B2 · GD2-based chimeric antigen receptor and application thereof

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- Number: US 12,187,808 B2
- Document ID: US12187808B2
- Status: Granted
- Kind: B2
- Application number: 17/262,487
- Filed: 2019-07-24
- Published: 2021-10-07
- Granted: 2025-01-07
- Earliest claimed: 2018-07-24
- Assignees: Beijing Meikang Geno-Immune Biotechnology Co., Ltd.
- Applicants: Beijing Meikang Geno-Immune Biotechnology Co. Ltd.
- Inventors: Yu Chen Liu
- Web page: https://patentgrep.com/patent/US12187808B2

## CPC

- C07K 16/3084: {against tumour-associated gangliosides}
- 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 40/4258: {Gangliosides, e.g. GM2, GD2 or GD3}
- A61P 35/00: Antineoplastic agents
- C07K 14/7051: {T-cell receptor (TcR)-CD3 complex}
- C07K 14/70517: {CD8}
- C07K 14/70521: {CD28, CD152}
- C07K 14/70578: {NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 (NGF-receptor C07K14/71, TNF-receptor C07K14/7151)}
- C12N 5/0636: {T lymphocytes}
- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- C12N 9/6472: {Cysteine endopeptidases (3.4.22)}
- C12N 15/86: Viral vectors
- C12Y 304/22062: Caspase-9 (3.4.22.62)
- 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 2239/31: characterized by the route of administration
- A61K 2239/38: characterised by the dose, timing or administration schedule
- C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2317/73: Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
- C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding
- C07K 2319/02: containing a signal sequence
- 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 2319/33: fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
- C12N 2510/00: Genetically modified cells
- C12N 2740/15021: Viruses as such, e.g. new isolates, mutants or their genomic sequences
- C12N 2740/15043: viral genome or elements thereof as genetic vector
- C12N 2740/16043: viral genome or elements thereof as genetic vector
- 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
- 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 application relates to a GD2-based chimeric antigen receptor comprising an antigen-binding domain, a transmembrane domain, a costimulatory signaling domain, a CD3ζ signaling domain, and a self-destructive domain in tandem arrangement; wherein the antigen-binding domain binds to a tumor surface antigen, wherein the tumor surface antigen is GD2, and the antigen-binding domain is a single-chain antibody against the tumor surface antigen GD2, wherein the self-destructive domain is a caspase 9 domain.

## Claims

### Claim 1

A chimeric antigen receptor, wherein
the chimeric antigen receptor comprises a Secretory signal peptide, GD2 antigen-binding domain, CD8α and/or CD28 transmembrane domain, CD28 signaling domain, 4-1BB signaling domain, CD3ζ signaling domain, 2A sequence, and caspase 9 domain in tandem arrangement;
the chimeric antigen receptor is Secretory-GD2-CD28-4-1BB-CD3ζ-2A-FBKP.Casp9; and
the chimeric antigen receptor Secretory-GD2-CD28-4-1BB-CD3ζ-2A-FBKP.Casp9 comprises the amino acid sequence as shown in SEQ ID NOs: 13-14.

### Claim 2 (depends on 1)

A viral vector encoding the chimeric antigen receptor according to claim 1.

### Claim 3 (depends on 2)

The viral vector according to claim 2, wherein the viral vector is a lentiviral vector and/or a retroviral vector.

### Claim 4 (depends on 2)

A method of obtaining recombinant lentivirus, comprising co-transfection of a mammalian cell with the viral vector according to claim 2 and packaging helper plasmids pNHP and pHEF-VSVG.

### Claim 5 (depends on 4)

The method according to claim 4, wherein the mammalian cell is selected from the group consisting of a 293 cell, a 293T cell, a TE671 cell, and the combination thereof.

### Claim 6 (depends on 1)

A T cell which is transfected with a nucleic acid sequence encoding the chimeric antigen receptor according to claim 1 for expression.

### Claim 7 (depends on 6)

The T cell according to claim 6, wherein the T cell is transfected via a viral vector and/or a eukaryotic expression plasmid.

### Claim 8 (depends on 1)

A method for treating a tumor, comprising administrating to a patient in need thereof an effective amount of chimeric antigen receptor T cells or immune cells or tumor therapeutics comprising the chimeric antigen receptor according to claim 1.

### Claim 9 (depends on 8)

The method according to claim 8, wherein the tumor is a cancer in which the GD2 antigen is specifically expressed.

### Claim 10 (depends on 9)

The method according to claim 9, wherein the tumor in which the GD2 antigen is specifically expressed is neuroblastoma.