# US 11,110,124 B2 · Cell derived extracellular vesicles for the treatment of diseases

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- Number: US 11,110,124 B2
- Document ID: US11110124B2
- Status: Granted
- Kind: B2
- Application number: 16/049,136
- Filed: 2018-07-30
- Published: 2019-01-31
- Granted: 2021-09-07
- Earliest claimed: 2017-07-31
- Assignees: Rheinische Friedrich-Wilhelms-Universitaet Bonn
- Applicants: Rheinische Friedrich-Wilhelms-Universitaet Bonn
- Inventors: Christoph Coch; Juliane Dassler-Plenker; Gunther Hartmann
- Web page: https://patentgrep.com/patent/US11110124B2

## CPC

- A61K 35/17: Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes (when activated by a specific antigen A61K39/00)
- A61K 35/12: Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells (vaccines or medicinal preparations containing antigens or antibodies A61K39/00)
- A61K 35/13: Tumour cells, irrespective of tissue of origin (tumour vaccines A61K39/00)
- A61K 35/33: Fibroblasts
- A61K 39/0011: {Cancer antigens}
- A61P 35/00: Antineoplastic agents
- A61P 37/04: Immunostimulants
- C07K 14/705: Receptors; Cell surface antigens; Cell surface determinants {(tumour specific antigens C07K14/4748)}
- C12N 5/0693: {Tumour cells; Cancer cells}
- C12N 15/11: DNA or RNA fragments; Modified forms thereof (DNA or RNA not used in recombinant technology, C07H21/00); {Non-coding nucleic acids having a biological activity}
- A61K 2035/122: {for inducing tolerance or supression of immune responses}
- A61K 2039/55555: {Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers}
- C12N 2501/65: MicroRNA
- C12N 2502/11: blood or immune system cells
- C12N 2502/1157: Monocytes, macrophages
- C12N 2502/1164: NK cells
- C12N 2502/1178: Spleen cells
- C12N 2510/00: Genetically modified cells
- 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 invention pertains to extracellular vesicles (exosomes) that are produced by culturing tumor cells in the presence of various ligands or activators of innate immunity receptors and subsequently isolating and purifying the vesicles in the culture supernatant. The invention provides also pharmaceutical compositions comprising the inventive exosomes. The exosomes of the invention are provided as medicaments for example in the treatment of cancer diseases.

## Claims

### Claim 1

A method for the production of extracellular vesicles, the method comprising the steps of
(a) Culturing a tumor cell, an epithelial cell, or a fibroblast,
(b) Bringing into contact the tumor cell, the epithelial cell, or the fibroblast with at least one ligand of an innate immunity receptor in an amount and time sufficient for the tumor cell, the epithelial cell, or the fibroblast to release extracellular vesicles with an anti-infective and/or anti-tumor activity and/or immune stimulatory activity,
(c) Isolating and/or purifying the released extracellular vesicles.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the anti-infective and/or anti-tumor activity and/or immune stimulatory activity involves an activity of inducing cytokine production, and/or an activity of the extracellular vesicles to activate immune cells, such as monocytes, natural killer (NK) cells and/or T cells.

### Claim 3 (depends on 1)

The method according to claim 1, wherein the at least one ligand of an innate immunity receptor is selected from the group consisting of ligand of retinoic acid-inducible gene I (RIG-I), ligand of melanoma differentiation antigen 5 (MDA5) and ligand of cyclic GMP-AMP synthase (cGAS); and preferably is an RNA molecule, such as 3pRNA, a DNA molecule, such as Y-form DNA, or is polyinosinic:polycytidylic acid poly (I:C).

### Claim 4 (depends on 1)

The method according to claim 1, wherein the at least one innate immunity receptor is retinoic acid-inducible gene I (RIG-I), melanoma differentiation antigen 5 (MDA5) and/or is cyclic GMP-AMP synthase (cGAS).

### Claim 5 (depends on 1)

The method according to claim 1, wherein isolating and/or purifying comprises a step of determining the presence of BAG6 on the extracellular vesicles.

### Claim 6 (depends on 1)

The method according to claim 1, wherein step (b) comprises bringing into contact the tumor cell, the epithelial cell, or the fibroblast with at least two ligands of an innate immunity receptor selected from the group consisting of an RNA molecule, such as 3pRNA, a DNA molecule, such as Y-form DNA, and polyinosinic:polycytidylic acid poly (I:C); and wherein the at least two ligands of an innate immunity receptor are two ligands of two different innate immunity receptors.

### Claim 7 (depends on 1)

The method according to claim 1, wherein step (b) comprises bringing into contact the tumor cell, the epithelial cell, or the fibroblast with at least three ligands of an innate immunity receptor selected from the group consisting of an RNA molecule, such as 3pRNA, a DNA molecule, such as Y-form DNA, and polyinosinic:polycytidylic acid poly (I:C); and wherein the at least three ligands of an innate immunity receptor are three ligands of three different innate immunity receptors.

### Claim 8 (depends on 1)

A method of producing a medicament for treating a disease in a patient, the method comprising the steps of
(a) Providing a tumor cell, an epithelial cell or a fibroblast,
(b) Performing with the tumor cell, the epithelial cell or the fibroblast the method according to claim 1, to obtain released extracellular vesicles with anti-infective and/or anti-tumor activity and/or immune stimulatory activity,
(c) Formulating a medicament with the released anti-tumor extracellular vesicles for treating a disease in a patient.

### Claim 9 (depends on 8)

The method according to claim 8, wherein the tumor cell, the epithelial cell or the fibroblast is derived from a cellular sample of a patient to be treated, and wherein the formulated medicament is for treating said patient.

### Claim 10 (depends on 9)

The method according to claim 9, wherein the tumor cell, the epithelial cell or the fibroblast is not derived from the patient to be treated.

### Claim 11 (depends on 9)

The method according to claim 9, wherein the disease is cancer, and wherein the cell is a tumor cell; or wherein the disease is a viral disease, and wherein the cell is the fibroblast.