# US 12,692,298 B2 · Screening and antitumor use of KRAS mutation specific T cell receptor

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- Number: US 12,692,298 B2
- Document ID: US12692298B2
- Status: Granted
- Kind: B2
- Application number: 18/247,018
- Filed: 2021-09-29
- Published: 2023-11-23
- Granted: 2026-07-28
- Earliest claimed: 2020-09-29
- Assignees: Institute of Microbiology, Chinese Academy of Sciences
- Applicants: Institute of Microbiology, Chinese Academy of Sciences
- Inventors: Fu Gao; Shuguang Tan; Dan Lu
- Web page: https://patentgrep.com/patent/US12692298B2

## CPC

- C07K 14/7051: {T-cell receptor (TcR)-CD3 complex}
- A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
- A61K 40/32: T-cell receptors [TCR]
- A61K 40/4253: {GTPases, e.g. Ras or Rho}
- A61P 35/00: Antineoplastic agents
- G01N 33/5758: {involving compounds serving as markers for tumours, cancers or neoplasias, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides or metabolites}
- C12N 2740/16043: viral genome or elements thereof as genetic vector
- G01N 2333/7051: T-cell receptor (TcR)-CD3 complex
- 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
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- 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 disclosure provides two specific T cell receptors targeting a G12V or G12C mutant epitope of a KRAS gene, and an anti-tumor use thereof. Each of the two T cell receptors consists of an α peptide chain and a β peptide chain. Further provided are an antigen binding fragment of the T cell receptors, a nucleic acid encoding the T cell receptors or an antigen binding fragment thereof, a vector comprising the nucleic acid, and a host cell comprising the vector. Further provided is a method for preparing a specific T cell receptor for a KRAS G12V mutation or an antigen-binding fragment thereof. The specific T cell receptor and antigen-binding fragment thereof can be used as an immune activator to stimulate an immune response of an organism, thereby generating an effect against tumors and other diseases.

## Claims

### Claim 1

A T cell receptor (TCR) or antigen-binding fragment thereof, capable of binding to a complex of KRAS-G12V8-16 epitope of SEQ ID NO: 35 and HLA-A11, a complex of KRAS-G12C8-16 epitope of SEQ ID NO: 37 and HLA-A11, or a complex of KRAS-G12V8-16 epitope of SEQ ID NO: 35 and HLA-A03 and comprising an α-chain variable region and a β-chain variable region, wherein the TCR or antigen-binding fragment thereof comprises the following α-chain complementarity determining regions (CDRs) and β-chain complementarity determining regions (CDRs):
an α-chain complementarity determining region CDR1 comprising SEQ ID NO: 3;
an α-chain complementarity determining region CDR2 comprising SEQ ID NO: 4;
an α-chain complementarity determining region CDR3 comprising SEQ ID NO: 5;
a β-chain complementarity determining region CDR1 comprising SEQ ID NO: 8;
a β-chain complementarity determining region CDR2 comprising SEQ ID NO: 9; and
a β-chain complementarity determining region CDR3 comprising SEQ ID NO: 10;
or
an α-chain complementarity determining region CDR1 comprising SEQ ID NO: 13;
an α-chain complementarity determining region CDR2 comprising SEQ ID NO: 14;
an α-chain complementarity determining region CDR3 comprising SEQ ID NO: 15;
a β-chain complementarity determining region CDR1 comprising SEQ ID NO: 18;
a β-chain complementarity determining region CDR2 comprising SEQ ID NO: 19; and
a β-chain complementarity determining region CDR3 comprising SEQ ID NO: 20.

### Claim 2 (depends on 1)

The T cell receptor (TCR) or antigen-binding fragment thereof of claim 1, which comprises:
an α-chain variable region comprising SEQ ID NO: 2, and
a β-chain variable region comprising SEQ ID NO: 7;
or
an α-chain variable region as set forth in SEQ ID NO: 12, and
a β-chain variable region comprising SEQ ID NO: 17.

### Claim 3 (depends on 1)

The TCR or antigen-binding fragment thereof according to claim 1, wherein the TCR is a murine TCR, a human-mouse chimeric TCR or a humanized TCR.

### Claim 4 (depends on 1)

A polynucleotide encoding the TCR or antigen-binding fragment thereof of claim 1, which is one or more sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:6, SEQ ID NO:11, and SEQ ID NO:16.

### Claim 5 (depends on 4)

An expression vector comprising the polynucleotide of claim 4, wherein the expression vector is a lentivirus vector.

### Claim 6 (depends on 5)

A host cell comprising the expression vector of claim 5.

### Claim 7 (depends on 1)

A method for preparing the TCR or antigen-binding fragment thereof of claim 1, comprising:
i) culturing a host cell comprising an expression vector, wherein the expression vector comprises a polynucleotide, and the polynucleotide comprises one or more sequences selected from the group consisting of SEQ ID NO:1, SEQ ID NO:6, SEQ ID NO:11, and SEQ ID NO:16;
ii) recovering the TCR or antigen-binding fragment thereof of claim 1 from the host cell or a culture medium thereof.

### Claim 8 (depends on 1)

A pharmaceutical composition, which comprises the TCR or antigen-binding fragment thereof according to claim 1, and a pharmaceutically acceptable carrier.

### Claim 9 (depends on 1)

A method for increasing interferon (IFN)-γ cytokine level secreted by a T cell, comprising contacting a T cell expressing the TCR or antigen-binding fragment thereof of claim 1 with a polypeptide comprising a KRAS-G12V8-16 mutation of SEQ ID NO: 35 or a KRAS-G12C8-16 mutation of SEQ ID NO: 37, thereby increasing IFN-γ cytokine level secreted by the T cell.

### Claim 10 (depends on 1)

A method for detecting a tumor cell expressing a KRAS-G12V8-16 mutation of SEQ ID NO: 35 or a KRAS-G12C8-16 mutation of SEQ ID NO: 37, comprising contacting a protein comprising the TCR or antigen-binding fragment thereof of claim 1 with the tumor cell to be detected, thereby detecting presence or absence of the tumor cell expressing the KRAS-G12V8-16 mutation or the KRAS-G12C8-16 mutation.

### Claim 11 (depends on 1)

A method for treating a tumor with a KRAS-G12V8-16 mutation of SEQ ID NO: 35 and/or a KRAS-G12C8-16 mutation of SEQ ID NO: 37 of a KRAS gene in a patient, comprising administering to the patient a T cell expressing the TCR or antigen-binding fragment thereof of claim 1, thereby treating the patient.

### Claim 12 (depends on 11)

The method of claim 11, wherein the tumor is selected from pancreas cancer, colorectal cancer, or lung cancer.

### Claim 13 (depends on 12)

The method of claim 12, wherein the tumor is non-small cell lung cancer.

### Claim 14 (depends on 11)

The method of claim 11, wherein the TCR or antigen-binding fragment thereof comprises:
an α-chain variable region comprising SEQ ID NO: 2, and
a β-chain variable region comprising SEQ ID NO: 7;
or
an α-chain variable region comprising SEQ ID NO: 12, and
a β-chain variable region comprising SEQ ID NO: 17.

### Claim 15 (depends on 11)

The method of claim 11, wherein the TCR is a murine TCR, a human-mouse chimeric TCR or a humanized TCR.