# US 12,595,312 B2 · CD133-binding agents and uses thereof

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- Number: US 12,595,312 B2
- Document ID: US12595312B2
- Status: Granted
- Kind: B2
- Application number: 17/511,420
- Filed: 2021-10-26
- Published: 2022-03-10
- Granted: 2026-04-07
- Earliest claimed: 2016-10-19
- Assignees: The Governing Council of the University of Toronto; McMaster University
- Applicants: The Governing Council of the University of Toronto; McMaster University
- Inventors: Jason Moffat; Jarrett Adams; Guohua Pan; Sachdev Sidhu; Rashida Williams; Xiaoyu Zhang; Sheila K. Singh; Parvez Vora; Chitra Venugopal
- Web page: https://patentgrep.com/patent/US12595312B2

## CPC

- C07K 16/2896: {against molecules with a "CD"-designation, not provided for elsewhere}
- 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 40/421: {Immunoglobulin superfamily}
- A61K 40/4224: {Molecules with a "CD" designation not provided for elsewhere}
- A61K 47/6849: {the antibody targeting a receptor, a cell surface antigen or a cell surface determinant}
- 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 14/70596: {Molecules with a "CD"-designation not provided for elsewhere}
- C07K 16/2809: {against the T-cell receptor (TcR)-CD3 complex}
- C07K 16/468: {Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies}
- G01N 33/57557: {of other specific parts of the body, e.g. brain}
- A61K 2039/505: {comprising antibodies}
- A61K 2239/47: Brain; Nervous system
- C07K 2317/21: from primates, e.g. man
- C07K 2317/31: multispecific
- C07K 2317/33: Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
- C07K 2317/52: Constant or Fc region; Isotype
- C07K 2317/55: Fab or Fab'
- C07K 2317/565: Complementarity determining region [CDR]
- C07K 2317/567: Framework region [FR]
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2317/73: Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
- C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
- 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: 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)

## Abstract

This disclosure is directed to novel CD133-binding agents. The disclosure is also directed to uses of novel CD133-binding agents for detecting CD133-expressing cells and/or quantitating levels of cellular CD133 expression, for targeting CD133-expressing cells, for decreasing levels of CD133 in CD133-expressing cells and for treating or preventing cancer.

## Claims

### Claim 1

An antibody or an antigen binding fragment thereof that specifically binds to a CD133 polypeptide, wherein said antibody or antigen binding fragment thereof comprises a light chain variable domain comprising complementarity determining regions (CDRs) comprising SEQ ID NOs: 6, 7, and 8 and a heavy chain variable domain comprising CDRs comprising SEQ ID NOs: 9, 10, and 11.

### Claim 2 (depends on 1)

The antibody or antigen binding fragment thereof according to claim 1, wherein said antibody is a single chain variable fragment (scFv) antibody or a single chain Fab fragment (scFab) antibody.

### Claim 3 (depends on 1)

The antibody or antigen binding fragment thereof according to claim 1, wherein said antigen binding fragment is selected from a Fab fragment, a Fab′ fragment, and an Fv fragment.

### Claim 4 (depends on 1)

The antibody or antigen binding fragment thereof according to claim 1, wherein said antibody is a humanized antibody or a chimeric antibody.

### Claim 5 (depends on 1)

The antibody or antigen binding fragment thereof according to claim 1, wherein the light chain variable domain comprises an amino acid sequence that is at least 70% identical to SEQ ID NO: 2 and/or wherein the heavy chain variable domain comprises an amino acid sequence that is at least 70% identical to SEQ ID NO: 3.

### Claim 6 (depends on 1)

The antibody or antigen binding fragment thereof according to claim 1, wherein the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 2 and/or wherein the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 3.

### Claim 7 (depends on 1)

The antibody or antigen binding fragment thereof according to claim 1, wherein the CD133 polypeptide is a human CD133 polypeptide.

### Claim 8 (depends on 7)

The antibody or antigen binding fragment thereof according to claim 7, wherein the human CD133 polypeptide comprises SEQ ID NO: 1.

### Claim 9 (depends on 1)

The antibody or antigen binding fragment thereof according to claim 1, wherein the CD133 polypeptide is denatured.

### Claim 10 (depends on 1)

The antibody or antigen binding fragment thereof according to claim 1, wherein the CD133 polypeptide is on the surface of a cell.

### Claim 11 (depends on 1)

The antibody or antigen binding fragment thereof according to claim 1, wherein said antibody or antigen binding fragment thereof comprises human constant regions.

### Claim 12 (depends on 1)

The antibody or antigen binding fragment thereof according to claim 1, wherein said antibody is an IgG antibody.

### Claim 13 (depends on 1)

The antibody or antigen binding fragment thereof according to claim 1, wherein said antibody is an IgA1, IgA2, IgD, IgG1, IgG2, IgG3, IgG4, IgE or IgM antibody.

### Claim 14 (depends on 1)

The antibody or antigen binding fragment thereof according to claim 1, wherein said antibody is a bispecific antibody.

### Claim 15 (depends on 14)

The antibody or antigen binding fragment thereof according to claim 14, wherein said bispecific antibody binds to a CD3 polypeptide.

### Claim 16 (depends on 14)

The antibody or antigen binding fragment thereof according to claim 14, wherein said bispecific antibody comprises:
(a) a polypeptide comprising SEQ ID NO: 22 and a polypeptide comprising SEQ ID NO: 23;
(b) a polypeptide comprising SEQ ID NO: 24 and a polypeptide comprising SEQ ID NO: 25;
(c) a polypeptide comprising SEQ ID NO: 26; or
(d) a polypeptide comprising SEQ ID NO: 27.

### Claim 17 (depends on 1)

A chimeric antigen receptor (CAR) that specifically binds to a CD133 polypeptide, wherein said CAR comprises an extracellular antigen binding domain comprising the antibody or antigen binding fragment thereof according to claim 1.

### Claim 18 (depends on 1)

The antibody or antigen binding fragment thereof according to claim 1, wherein said antibody or antigen binding fragment thereof is detectably labeled.

### Claim 19 (depends on 1)

An immunoconjugate comprising the antibody or antigen binding fragment thereof according to claim 1 and an effector agent.

### Claim 20 (depends on 19)

The immunoconjugate according to claim 19, wherein said effector agent is an anti-neoplastic agent or a toxin.

### Claim 21 (depends on 1)

A pharmaceutical composition comprising the antibody or antigen binding fragment thereof according to claim 1 and a carrier.

### Claim 22 (depends on 1)

A method for treating cancer expressing CD133 in a subject, said method comprising administering to the subject an effective amount of the antibody or antigen binding fragment thereof according to claim 1.

### Claim 23 (depends on 22)

The method according to claim 22, wherein said cancer is selected from a metastatic melanoma, a brain cancer, a prostate cancer, a pancreatic cancer, and a colon cancer.

### Claim 24 (depends on 23)

The method according to claim 23, wherein said brain cancer is a glioblastoma or a medulloblastoma.

### Claim 25 (depends on 14)

A method for treating cancer expressing CD133 in a subject, said method comprising administering to the subject an effective amount of the bispecific antibody according to claim 14.

### Claim 26 (depends on 17)

A method for treating cancer expressing CD133 in a subject, said method comprising administering to the subject an effective amount of T cells expressing a CAR according to claim 17.

### Claim 27 (depends on 1)

A method for detecting CD133 and/or quantifying the levels of CD133 expressed by cells, said method comprising contacting a sample suspected of comprising CD133 or cells expressing CD133 with the antibody or antigen binding fragment thereof according to claim 1.