# US 2014/0193358 A1 · Treating cancer stem cells using targeted cargo proteins

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- Number: US 2014/0193358 A1
- Document ID: US20140193358A1
- Status: Published application
- Kind: A1
- Application number: 14/045,958
- Filed: 2013-10-04
- Published: 2014-07-10
- Earliest claimed: 2008-09-19
- Assignees: Protox Therapeutics, Inc.
- Inventors: Fahar Merchant
- Web page: https://patentgrep.com/patent/US20140193358A1

## CPC

- A61K 38/45: Transferases (2)
- A61K 9/0019: {Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00)}
- A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61K 47/6415: {Toxins or lectins, e.g. clostridial toxins or Pseudomonas exotoxins}
- A61K 47/642: {the peptide or protein in the drug conjugate being a cytokine, e.g. IL2, chemokine, growth factors or interferons being the inactive part of the conjugate}
- A61K 47/68: the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K 47/6829: {Bacterial toxins, e.g. diphteria toxins or Pseudomonas exotoxin A}
- A61K 47/6851: {the antibody targeting a determinant of a tumour cell}
- A61N 5/10: X-ray therapy; Gamma-ray therapy; Particle-irradiation therapy (A61N5/01 takes precedence)
- A61P 35/00: Antineoplastic agents
- C07K 14/4747: {Apoptosis related proteins}
- C07K 16/30: from tumour cells
- A61K 2035/124: {the cells being hematopoietic, bone marrow derived or blood cells}
- C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
- C07K 2319/33: fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
- C07K 2319/55: containing a fusion with a toxin, e.g. diphteria toxin
- C07K 2319/74: containing a fusion for binding to a cell surface receptor
- C12Y 204/02036: NAD(+)--diphthamide ADP-ribosyltransferase (2.4.2.36)
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- 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)
- A61N: ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY (measurement of bioelectric currents A61B; surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body A61B18/00; anaesthetic apparatus in general A61M; incandescent lamps H01K; infrared radiators for heating H05B)
- 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)
- C12Y: ENZYMES
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

The disclosure provides targeted cargo proteins that are useful for targeting cancer stem cells, and methods of their use in treating cancer.

## Claims

### Claim 29

A method of treating a cancer stem cell in a subject, comprising:
administering to the subject a targeted cargo protein, wherein the targeted cargo protein comprises:
(a) one or more cargo moieties; and
(b) one or more targeting moieties that bind to a target displayed by a cancer stem cell, wherein the targeting moiety is derived from a natural ligand to the target, thereby treating the cancer stem cell in the subject.

### Claim 30 (depends on 29)

The method of claim 29, wherein the cargo moiety comprises a toxin.

### Claim 31 (depends on 30)

The method of claim 30, wherein the toxin comprises a bacterial toxin, animal toxin, or plant toxin.

### Claim 32 (depends on 30)

The method of claim 30, wherein the toxin comprises a pore-forming toxin.

### Claim 33 (depends on 32)

The method of claim 32, wherein the pore-forming toxin comprises aerolysin or proaerolysin.

### Claim 34 (depends on 31)

The method of claim 31, wherein the plant toxin comprises bouganin or ricin.

### Claim 35 (depends on 31)

The method of claim 31, wherein the bacterial toxin comprises Pseudomonas exotoxin, cholera toxin, or diphtheria toxin.

### Claim 36 (depends on 29)

The method of claim 29, wherein the cargo moiety comprises a pro-apoptosis member of the BCL-2 family selected from BAX, BAD, BAT, BAK, BIK, BOK, BID BIM, BMF and BOK.

### Claim 37

A method of treating a cancer stem cell in a subject, comprising:
administering to the subject a targeted cargo protein, wherein the targeted cargo protein comprises:
(a) one or more cargo moieties selected from aerolysin, proaerolysin, bouganin, ricin, Pseudomonas exotoxin, cholera toxin, diphtheria toxin, and BAD; and
(b) one or more targeting moieties that bind to a target displayed by a cancer stem cell, thereby treating the cancer stem cell in the subject.

### Claim 38 (depends on 37)

The method according to claim 37, wherein the one or more targeting moieties is selected from an antibody, ligand or ligand variant.

### Claim 39 (depends on 29)

The method of claim 29, wherein the target displayed by the cancer stem cell comprises a receptor selected from the group consisting of: IL-4, IL-3, IL-2, EGF, and GMCSF or an antigen comprising EpCAM, mesothelin, or CD22.

### Claim 40 (depends on 37)

The method of claim 37, wherein the targeting moiety comprises a humanized antibody.

### Claim 41 (depends on 29)

The method of claim 29, wherein the targeted cargo protein comprises a human cargo moiety selected from the group consisting of RNase A and perform.

### Claim 42 (depends on 29)

The method of claim 29, wherein the targeted cargo protein comprises a fusion protein.

### Claim 43 (depends on 29)

The method of claim 29, wherein the targeted cargo protein is present in a pharmaceutically acceptable carrier.

### Claim 44 (depends on 29)

The method of claim 29, wherein the subject has a recurrent cancer or a newly diagnosed cancer.

### Claim 45 (depends on 29)

The method of claim 29, wherein the subject is refractory.

### Claim 46 (depends on 29)

The method of claim 29, further comprising:
determining whether the subject is refractory to radiation or chemotherapy; wherein if the subject is refractory it indicates that they will benefit from administration of the targeted cargo protein.

### Claim 47 (depends on 29)

The method of claim 29, further comprising administering chemotherapy or radiation therapy to the subject after administering the targeted cargo protein, or surgically removing at least part of a tumor after administering the targeted cargo protein.

### Claim 48 (depends on 29)

The method of claim 29, further comprising administering chemotherapy or radiation therapy to the subject before administering the targeted cargo protein, or surgically removing at least part of a tumor before administering the targeted cargo protein.

### Claim 49 (depends on 29)

The method of claim 29, further comprising administering chemotherapy or radiation therapy to the subject during treatment with the targeted cargo protein, or administering the targeted cargo protein during surgical removal of least part of a tumor in the subject.

### Claim 50 (depends on 29)

The method of claim 29, further comprising:
administering to the subject an agonist that sensitizes the cancer stem cells prior to administering the targeted cargo protein.

### Claim 51 (depends on 29)

The method of claim 29, wherein the targeted cargo protein is administered intratumorally.

### Claim 52 (depends on 29)

The method of claim 29, wherein the targeted cargo protein comprises Pseudomonas exotoxin linked to circularly permuted IL-4, IL-2 linked to aerolysin, IL-2 linked to proaerolysin, IL4 linked to BAD, GMCSF linked to BAD, EGF linked to proaerolysin, anti-EpCAM antibody linked to Pseudomonas exotoxin, anti-EpCAM antibody linked to bouganin, anti-mesothelin antibody linked to PE, anti-CD22 antibody linked to PE, anti-CD22 antibody linked to RNase A, and anti-PSMA antibody linked to thapsigargin.

### Claim 53 (depends on 29)

The method of claim 29, further comprising removing hematopoietic stem cells from the subject prior to administering the targeted cargo protein.

### Claim 54 (depends on 53)

The method of claim 53, further comprising re-introducing to the subject the removed hematopoietic stem cells.

### Claim 55 (depends on 29)

The method of claim 29, wherein the method further treats a bulk tumor in the subject.

### Claim 56 (depends on 29)

The method of claim 29, wherein the targeted cargo protein further comprises a polymer, for example to increase stability, increase circulating half life or reduce immunogenicity of the targeted cargo protein.