# US 11,992,503 B2 · Prostate-specific membrane antigen cars and methods of use thereof

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

- Number: US 11,992,503 B2
- Document ID: US11992503B2
- Status: Granted
- Kind: B2
- Application number: 16/933,728
- Filed: 2020-07-20
- Published: 2020-11-05
- Granted: 2024-05-28
- Earliest claimed: 2018-03-06
- Assignees: The Trustees of the University of Pennsylvania
- Applicants: The Trustees of the University of Pennsylvania
- Inventors: Yangbing Zhao; Szu Hua Sharon Lin; Xiaojun Liu; Anne Chew
- Web page: https://patentgrep.com/patent/US11992503B2

## CPC

- A61K 31/661: Phosphorus acids or esters thereof not having P—C bonds, e.g. fosfosal, dichlorvos, malathion {or mevinphos}
- A61K 31/675: having nitrogen as a ring hetero atom, e.g. pyridoxal phosphate
- A61K 31/7076: containing purines, e.g. adenosine, adenylic acid
- 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 40/421: {Immunoglobulin superfamily}
- A61K 40/4229: {Transforming growth factor [TGF]}
- A61K 40/4276: {Prostate specific membrane antigen [PSMA]}
- A61P 35/00: Antineoplastic agents
- A61P 35/04: specific for metastasis
- C07K 14/7051: {T-cell receptor (TcR)-CD3 complex}
- C07K 14/70517: {CD8}
- C07K 14/70521: {CD28, CD152}
- C07K 14/70575: {NGF/TNF-superfamily, e.g. CD70, CD95L, CD153, CD154 (NGF C07K14/48, TNF C07K14/525)}
- C07K 14/70578: {NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 (NGF-receptor C07K14/71, TNF-receptor C07K14/7151)}
- C07K 16/2818: {against CD28 or CD152}
- C07K 16/2827: {against B7 molecules, e.g. CD80, CD86}
- C07K 16/2863: {against receptors for growth factors, growth regulators}
- C07K 16/3069: {Reproductive system, e.g. ovaria, uterus, testes, prostate}
- C12N 5/0636: {T lymphocytes}
- C12N 5/0638: {Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK]}
- 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 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 2317/31: multispecific
- C07K 2317/55: Fab or Fab'
- C07K 2317/569: Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2319/02: containing a signal sequence
- C07K 2319/03: containing a transmembrane segment
- C07K 2319/32: fusions with soluble part of a cell surface receptor, "decoy receptors"
- C07K 2319/33: fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
- 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
- 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)
- 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 modified immune cells (e.g., modified T cells) comprising a chimeric antigen receptor (CAR) having affinity for a prostate-specific membrane antigen (PSMA) (e.g., human PSMA). The present disclosure provides modified immune cells (e.g., modified T cells) comprising a CAR having affinity for PSMA and a dominant negative receptor and/or a switch receptor. The present disclosure provides modified immune cells (e.g., modified T cells) comprising a CAR having affinity for PSMA and a dominant negative receptor and/or a switch receptor, wherein the modified cell is capable of expressing and secreting a bispecific antibody.

## Claims

### Claim 1

A method of treating a cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition comprising a modified immune cell comprising:
(a) a chimeric antigen receptor (CAR) having affinity for a prostate specific membrane antigen (PSMA) on a target cell, wherein the CAR comprises the amino acid sequence set forth in SEQ ID NO:105; and
(b) a dominant negative receptor consisting of the amino acid sequence set forth in SEQ ID NO:115.

### Claim 2

A method of treating a cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition comprising a modified immune cell, wherein the modified immune cell comprises:
(a) a chimeric antigen receptor (CAR) having affinity for a prostate specific membrane antigen (PSMA) on a target cell, wherein the CAR comprises an antigen binding domain, a transmembrane domain, and an intracellular domain, and wherein:
(i) the antigen binding domain comprises a heavy chain variable region and a light chain variable region selected from the group consisting of:
(a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 21 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 16;
(b) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 28 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 33;
(c) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 40 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 45;
(d) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 52 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 57;
(e) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 64 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 69; or
(f) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 191 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 192; and
(ii) the intracellular domain comprises a 4-1BB signaling domain, an ICOS signaling domain, and a CD3 zeta signaling domain; and
(b) a dominant negative TGF-β receptor type II comprising the amino acid sequence set forth in SEQ ID NO:115.

### Claim 3 (depends on 2)

The method of claim 2, wherein the VH comprises the sequence of SEQ ID NO:191 and the VL comprises the sequence of SEQ ID NO:192.

### Claim 4

A method of treating a cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition comprising a modified immune cell, wherein the modified immune cell comprises:
(a) a chimeric antigen receptor (CAR) having affinity for a prostate specific membrane antigen (PSMA) on a target cell, wherein the CAR comprises an antigen binding domain, a transmembrane domain, and an intracellular domain, and wherein the antigen binding domain comprises an antibody or an antigen-binding fragment thereof comprising:
(i) a heavy chain variable region (VH) that comprises the consensus sequence of SEQ ID NO:194, and/or
(ii) a light chain variable region (VL) that comprises the consensus sequence of SEQ ID NO: 195; and
(b) a dominant negative receptor and/or switch receptor.

### Claim 5 (depends on 4)

The method of claim 4, wherein:
(a) the VH comprises the sequence of SEQ ID NO: 198 and the VL comprises the sequence of SEQ ID NO: 199;
(b) the VH comprises the sequence of SEQ ID NO: 200 and the VL comprises the sequence of SEQ ID NO: 201;
(c) the VH comprises the sequence of SEQ ID NO: 202 and the VL comprises the sequence of SEQ ID NO: 199; and
(d) the VH comprises the sequence of SEQ ID NO: 202 and the VL comprises the sequence of SEQ ID NO: 201.

### Claim 6 (depends on 4)

The method of claim 4, wherein the antigen-binding fragment is selected from the group consisting of a Fab, a single-chain variable fragment (scFv), and a single-domain antibody.

### Claim 7 (depends on 4)

The method of claim 4, wherein the transmembrane domain comprises a transmembrane region derived from CD8.

### Claim 8 (depends on 7)

The method of claim 7, wherein the transmembrane region comprises the amino acid sequence of SEQ ID NO:88.

### Claim 9 (depends on 4)

The method of claim 4, wherein the transmembrane domain further comprises a hinge region derived from CD8.

### Claim 10 (depends on 9)

The method of claim 9, wherein the hinge region comprises the amino acid sequence of SEQ ID NO:86.

### Claim 11 (depends on 4)

The method of claim 4, wherein the intracellular domain comprises two signaling domains selected from the group consisting of a 4-1BB signaling domain, a CD3 zeta signaling domain, and an ICOS signaling domain.

### Claim 12 (depends on 4)

The method of claim 4, wherein the intracellular domain comprises:
(a) two signaling domains selected from the group consisting of a 4-1BB signaling domain comprising the amino acid sequence of SEQ ID NO:92,
(b) a CD3 zeta signaling domain comprising the amino acid sequence of SEQ ID NO:97 or SEQ ID NO:100, and
(c) an ICOS signaling domain comprising the amino acid sequence of SEQ ID NO:203.

### Claim 13 (depends on 4)

The method of claim 4, wherein the dominant negative receptor is a dominant negative TGF-β receptor type II.

### Claim 14 (depends on 13)

The method of claim 13, wherein the dominant negative TGF-β receptor type II comprises the amino acid sequence set forth in SEQ ID NO:115.

### Claim 15 (depends on 4)

The method of claim 4, wherein the modified immune cell is a T cell.

### Claim 16 (depends on 15)

The method of claim 15, wherein the T cell is an autologous cell or an allogeneic cell.

### Claim 17 (depends on 4)

The method of claim 4, wherein the modified cell is derived from a human cell.

### Claim 18 (depends on 4)

The method of claim 4, further comprising administering to the subject a lymphodepleting chemotherapy, wherein the lymphodepleting chemotherapy comprises administering to the subject a therapeutically effective amount of cyclophosphamide at about 200 mg/m2/day to about 2000 mg/m2/day, and/or fludarabine at about 20 mg/m2/day to about 900 mg/m2/day.

### Claim 19 (depends on 4)

The method of claim 4, wherein the cancer is a prostate cancer.

### Claim 20 (depends on 19)

The method of claim 19, wherein the prostate cancer is a castrate-resistant prostate cancer.

### Claim 21 (depends on 20)

The method of claim 20, wherein the castrate-resistant prostate cancer is selected from an advanced castrate-resistant prostate cancer, or a metastatic castrate-resistant prostate cancer.