# US 12,760,091 B2 · Fibronectin targeting chimeric antigen receptors (CARs)

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- Number: US 12,760,091 B2
- Document ID: US12760091B2
- Status: Granted
- Kind: B2
- Application number: 17/773,431
- Filed: 2020-10-30
- Published: 2022-12-01
- Granted: 2026-10-13
- Earliest claimed: 2019-11-01
- Assignees: The Trustees of the University of Pennsylvania
- Applicants: The Trustees of the University of Pennsylvania
- Inventors: Avery D. Posey, Jr.; Tiffany King
- Web page: https://patentgrep.com/patent/US12760091B2

## CPC

- C07K 16/18: against material from animals or humans
- 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 40/4257: {Mucins, e.g. MUC-1}
- 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 14/7051: {T-cell receptor (TcR)-CD3 complex}
- C07K 14/70517: {CD8}
- C07K 14/70578: {NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 (NGF-receptor C07K14/71, TNF-receptor C07K14/7151)}
- A61K 2239/28: Expressing multiple CARs, TCRs or antigens
- A61K 2239/31: characterized by the route of administration
- A61K 2239/38: characterised by the dose, timing or administration schedule
- A61K 2239/58: Prostate
- 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)

## Abstract

The present disclosure provides compositions and methods comprising chimeric antigen receptors (CARs) capable of binding tumor-specific isoforms of fibronectin.

## Claims

### Claim 1

A chimeric antigen receptor (CAR) comprising an antigen binding domain capable of binding the IIICS domain of fibronectin, a transmembrane domain, and an intracellular domain, wherein the antigen binding domain comprises:
i. at least one heavy chain variable region (HCDR) comprising the nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3; and
ii. at least one light chain variable region (LCDR) comprising the nucleotide sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6.

### Claim 2 (depends on 1)

The CAR of claim 1, wherein the antigen binding domain comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7.

### Claim 3 (depends on 1)

The CAR of claim 1, wherein the antigen binding domain comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8.

### Claim 4 (depends on 1)

The CAR of claim 1, wherein the antigen binding domain is a single-chain variable fragment (scFv) comprising the amino acid sequence of SEQ ID NO: 10 or SEQ ID NO: 11.

### Claim 5 (depends on 1)

A nucleic acid comprising a polynucleotide sequence encoding the chimeric antigen receptor (CAR) of claim 1.

### Claim 6 (depends on 5)

The nucleic acid of claim 5, wherein the antigen binding domain comprises a heavy chain variable region encoded by a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 44 and/or a light chain variable region encoded by a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 45.

### Claim 7 (depends on 5)

The nucleic acid of claim 5, wherein the antigen binding domain is a single-chain variable fragment (scFv) encoded by a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 46 or SEQ ID NO: 47.

### Claim 8 (depends on 5)

The nucleic acid of claim 5, wherein the CAR is encoded by a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 48 or 49.

### Claim 9 (depends on 5)

A vector comprising the nucleic acid of claim 5.

### Claim 10 (depends on 1)

A modified immune cell or precursor cell thereof, comprising the CAR of claim 1.

### Claim 11 (depends on 10)

The modified immune cell or precursor cell thereof of claim 10, wherein the modified cell is a modified T cell.

### Claim 12 (depends on 5)

A method for generating a modified immune cell or precursor cell thereof, comprising introducing into an immune or precursor cell the nucleic acid of claim 5.

### Claim 13 (depends on 12)

The method of claim 12, wherein the nucleic acid is introduced via viral transduction.

### Claim 14 (depends on 13)

The method of claim 13, wherein the viral transduction comprises contacting the immune or precursor cell with a viral vector comprising the nucleic acid encoding a CAR.

### Claim 15 (depends on 14)

The method of claim 14, wherein the viral vector is selected from the group consisting of a retroviral vector, a lentiviral vector, an adenoviral vector, and an adeno-associated viral vector.

### Claim 16 (depends on 15)

The method of claim 15, wherein the viral vector is a lentiviral vector.