# US 2023/0065784 A1 · Composition and method for reducing expression of checkpoint inhibitors in t cells expressing a car or ctl

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

- Number: US 2023/0065784 A1
- Document ID: US20230065784A1
- Status: Published application
- Kind: A1
- Application number: 17/440,832
- Filed: 2020-03-20
- Published: 2023-03-02
- Earliest claimed: 2019-03-21
- Assignees: City of Hope
- Applicants: City of Hope
- Inventors: Saul J. Priceman; John C. Burnett; Yukiko Yamaguchi; Elizabeth Epps
- Web page: https://patentgrep.com/patent/US20230065784A1

## CPC

- C07K 14/7051: {T-cell receptor (TcR)-CD3 complex}
- C07K 16/3069: {Reproductive system, e.g. ovaria, uterus, testes, prostate}
- A61K 39/3955: {against proteinaceous materials, e.g. enzymes, hormones, lymphokines}
- A61K 39/39558: {against tumor tissues, cells, antigens}
- 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 40/36: Immune checkpoint inhibitors
- A61K 40/4274: {Prostate associated antigens e.g. Prostate stem cell antigen [PSCA]; Prostate carcinoma tumor antigen [PCTA]; Prostatic acid phosphatase [PAP]; Prostate-specific G-protein-coupled receptor [PSGR]}
- C07K 16/2818: {against CD28 or CD152}
- C12N 15/113: Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)}
- C12N 15/63: Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- A61K 2039/585: {wherein the target is cancer}
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2319/03: containing a transmembrane segment
- C07K 2319/33: fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
- C12N 2310/14: interfering nucleic acids [NA]
- C12N 2310/531: Stem-loop; Hairpin
- C12N 2320/31: Combination therapy
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Described herein is an approach for targeting multiple critical checkpoint genes involved in T cell exhaustion, for example, PD-1, TIM3, and LAG-3 in a manner that allows knock-down of their expression in T cells that also express a T cell receptor targeted to cancer cells, e.g., a CAR targeted to a cancer antigen or a T cell receptor (TCR).

## Claims

### Claim 1

A nucleic acid vector comprising an shRNA sequence targeted to an mRNA encoding a checkpoint inhibitor selected from the group consisting of PD-1, TIM3 and LAG3 and a nucleic acid sequence encoding a chimeric antigen receptor (CAR).

### Claim 2 (depends on 1)

The nucleic acid vector of claim 1, wherein the CAR targets an antigen selected from the group consisting of IL-13Ra, HER2, CD19 and PSCA.

### Claim 3 (depends on 1)

The nucleic acid vector of claim 1, wherein each shRNA sequence is operably linked to a promoter.

### Claim 4 (depends on 3)

The nucleic acid vector of claim 3, wherein each shRNA sequence is operably linked to the same promoter.

### Claim 5 (depends on 1)

The nucleic acid vector of claim 1, wherein the vector comprises two or more different shRNA nucleic acid sequences targeted to the same mRNA encoding a checkpoint inhibitor.

### Claim 6 (depends on 1)

The nucleic acid vector of claim 1, wherein each shRNA sequence is targeted to a different mRNA encoding a checkpoint inhibitor.

### Claim 7 (depends on 1)

The nucleic acid vector of claim 1, wherein the vector comprises an shRNA sequence targeted to an mRNA encoding PD-1 and an shRNA sequence targeted to an mRNA encoding TIM3.

### Claim 8 (depends on 1)

The nucleic acid vector of claim 1, wherein the vector comprises an shRNA sequence targeted to an mRNA encoding PD-1 and an shRNA sequence targeted to an mRNA encoding LAG3.

### Claim 9 (depends on 1)

The nucleic acid vector of claim 1, wherein the vector comprises an shRNA sequence targeted to an mRNA encoding TIM3 and an shRNA sequence targeted to an mRNA encoding LAG3.

### Claim 10 (depends on 3)

The nucleic acid vector of claim 3, wherein each shRNA sequence is operably linked to a different promoter.

### Claim 11 (depends on 3)

The nucleic acid vector of claim 3, comprising two or more different shRNA sequences that are operably linked to the same promoter.

### Claim 12 (depends on 3)

The nucleic acid vector of claim 3, wherein the promoter is selected from the group consisting of an H1 DNA POL III promoter, an U6 DNA POL III promoter, and a 75K DNA POL III.

### Claim 13 (depends on 1)

The nucleic acid vector of claim 1, wherein the shRNA targeted to an mRNA encoding PD-1 is targeted to SEQ ID NO:25.

### Claim 14 (depends on 1)

The nucleic acid vector of claim 1, wherein the shRNA targeted to an mRNA encoding TIM3 is targeted to SEQ ID NO:26.

### Claim 15 (depends on 1)

The nucleic acid vector of claim 1, wherein the shRNA targeted to an mRNA encoding LAG3 is targeted to SEQ ID NO:27.

### Claim 16 (depends on 1)

The nucleic acid vector of claim 1, wherein the shRNA targeted to PD-1 comprises a nucleic acid sequence selected from the group consisting of SEQ ID Nos 12-14.

### Claim 17 (depends on 1)

The nucleic acid vector of claim 1, wherein the shRNA targeted to TIM3 comprises a nucleic acid sequence selected from the group consisting of SEQ ID Nos: 15-17.

### Claim 18 (depends on 1)

The nucleic acid vector of claim 1, wherein the shRNA targeted to LAG3 comprises a nucleic acid sequence selected from the group consisting of SEQ ID Nos: 18-20.

### Claim 19 (depends on 1)

The nucleic acid vector of claim 1, wherein each shRNA sequence comprises:
sense sequence and an antisense sequence, wherein the sense and the antisense sequence, wherein the sense sequence comprises a nucleotide sequence identical to a target sequence in an mRNA encoding a checkpoint inhibitor.

### Claim 20 (depends on 1)

The nucleic acid vector of claim 1, wherein the vector is a lentiviral vector.

### Claim 21

A nucleic acid vector comprising an H1 DNA POL III promoter, an U6 DNA POL III promoter, and a 75K DNA POL III promoter, where each promoter is operably linked to an shRNA nucleic acid sequence targeted to an mRNA encoding a checkpoint inhibitor.

### Claim 22 (depends on 21)

The nucleic acid vector of claim 21, wherein each shRNA nucleic acid sequence is targeted to the same mRNA encoding a checkpoint inhibitor.

### Claim 23 (depends on 21)

The nucleic acid vector of claim 21, wherein each shRNA is targeted to a different mRNA encoding a checkpoint inhibitor.

### Claim 24 (depends on 21)

The nucleic acid vector of claim 21, wherein the checkpoint inhibitor is selected from the group consisting of PD-1, TIM3 and LAG3.

### Claim 25 (depends on 21)

The nucleic acid vector of claim 21, wherein the shRNA targeted to an mRNA encoding PD-1 is targeted to SEQ ID NO:25.

### Claim 26 (depends on 21)

The nucleic acid vector of claim 21, wherein the shRNA targeted to an mRNA encoding TIM3 is targeted to SEQ ID NO:26.

### Claim 27 (depends on 21)

The nucleic acid vector of claim 21, wherein the shRNA targeted to an mRNA encoding LAG3 is targeted to SEQ ID NO:27.

### Claim 28 (depends on 25)

The nucleic acid vector of claim 25, wherein the shRNA targeted to PD-1 comprises a nucleic acid sequence selected from the group consisting of SEQ ID Nos 12-14.

### Claim 29 (depends on 26)

The nucleic acid vector of claim 26, wherein the shRNA targeted to TIM3 comprises a nucleic acid sequence selected from the group consisting of SEQ ID Nos: 15-17.

### Claim 30 (depends on 27)

The nucleic acid vector of claim 27, wherein the shRNA targeted to LAG3 comprises a nucleic acid sequence selected from the group consisting of SEQ ID Nos: 18-20.

### Claim 31 (depends on 1)

A nucleic acid vector of claim 1, wherein the vector comprises the nucleotide sequence of SEQ ID NO:24.

### Claim 32 (depends on 1)

The nucleic acid vector of claim 1, wherein each shRNA sequence comprises:
sense sequence and an antisense sequence, wherein the sense and the antisense sequence, wherein the sense sequence comprises a nucleotide sequence identical to a target sequence in an mRNA encoding a checkpoint inhibitor.

### Claim 33 (depends on 1)

The nucleic acid vector of claim 1, wherein the vector is a lentiviral vector.

### Claim 34 (depends on 21)

The nucleic acid vector of claim 21, wherein the vector further comprises a nucleotide sequence encoding a chimeric antigen receptor.

### Claim 35 (depends on 34)

The nucleic acid vector of claim 34, wherein the CAR targets an antigen selected from the group consisting of: PSCA, HER2 and CD19.

### Claim 36 (depends on 44)

The nucleic acid vector of claim 44, wherein the CAR comprises, from amino to carboxy terminus: a targeting domain for targeting an antigen; a spacer domain; a transmembrane domain; a co-stimulatory domain; and a CD3ζ signaling domain.

### Claim 37 (depends on 1)

A T cell harboring the nucleic acid vector of claim 1.

### Claim 38 (depends on 1)

A method for reducing the expression of a checkpoint inhibitor in T cell, the method comprising introducing the nucleic acid vector of claim 1 into the T cell.