# US 10,869,898 B2 · Methods and compositions for immunomodulation

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- Number: US 10,869,898 B2
- Document ID: US10869898B2
- Status: Granted
- Kind: B2
- Application number: 15/301,046
- Filed: 2015-03-13
- Published: 2017-01-26
- Granted: 2020-12-22
- Earliest claimed: 2014-04-01
- Assignees: Rubius Therapeutics, Inc.
- Applicants: Rubius Therapeutics, Inc.
- Inventors: Jordi Mata-Fink; John Round; Noubar B. Afeyan; Avak Kahvejian
- Web page: https://patentgrep.com/patent/US10869898B2

## CPC

- A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
- A61P 1/04: for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
- A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- A61P 7/02: Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
- A61P 7/04: Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
- A61P 13/12: of the kidneys
- A61P 19/02: for joint disorders, e.g. arthritis, arthrosis
- A61P 21/02: Muscle relaxants, e.g. for tetanus or cramps
- A61P 21/04: for myasthenia gravis
- A61P 25/00: Drugs for disorders of the nervous system
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 37/02: Immunomodulators
- A61P 37/06: Immunosuppressants, e.g. drugs for graft rejection
- A61P 37/08: Antiallergic agents (antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14)
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C12N 5/06: {Animal cells or tissues; Human cells or tissues (preservation of excised living parts A01N1/10)}
- C12N 5/0641: {Erythrocytes}
- C12N 5/0644: {Platelets; Megakaryocytes}
- A61K 2035/122: {for inducing tolerance or supression of immune responses}
- A61K 2035/124: {the cells being hematopoietic, bone marrow derived or blood 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
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Provided are cells containing exogenous antigen and uses thereof.

## Claims

### Claim 1

A method of inducing immune tolerance in a subject having immune activation in response to an antigen that mediates an autoimmune disease, an inflammatory disease, or an allergic disease, the method comprising:
administering to the subject a pharmaceutical composition comprising a plurality of enucleated erythroid cells, the plurality of enucleated erythroid cells comprising a fusion polypeptide comprising the antigen, or an antigenic fragment thereof,
wherein the antigen, or the antigenic fragment thereof, of the fusion polypeptide is localized on the surface of the enucleated erythroid cells; the fusion polypeptide does not comprise a ligand binding domain; the fusion polypeptide was expressed from a transgene introduced into an erythroid cell precursor used to generate the plurality of enucleated erythroid cells, and
the pharmaceutical composition is administered in an amount and frequency sufficient to mediate the induction of the immune tolerance in the subject.

### Claim 2 (depends on 1)

The method of claim 1, wherein the subject has the autoimmune disease.

### Claim 3 (depends on 1)

The method of claim 1, wherein the subject has multiple sclerosis.

### Claim 4 (depends on 3)

The method of claim 3, wherein the antigen is a myelin basic protein, a proteolipid protein, or a myelin oligodendrocyte glycoprotein.

### Claim 5 (depends on 1)

The method of claim 1, wherein the subject has type 1 diabetes.

### Claim 6 (depends on 5)

The method of claim 5, wherein the antigen is a preproinsulin, a proinsulin, or an insulin.

### Claim 7 (depends on 1)

The method of claim 1, wherein the subject has membranous nephropathy.

### Claim 8 (depends on 7)

The method of claim 7, wherein the antigen is a phospholipase A2 receptor.

### Claim 9 (depends on 1)

The method of claim 1, wherein the subject has antiphospholipid syndrome (APS) or catastrophic APS.

### Claim 10 (depends on 9)

The method of claim 9, wherein the antigen is a beta-2 glycoprotein 1.

### Claim 11 (depends on 1)

The method of claim 1, wherein the plurality of enucleated erythroid cells are not hypotonically loaded.

### Claim 12 (depends on 1)

The method of claim 1, wherein a cell of the plurality of enucleated erythroid cells exhibits substantially the same osmotic membrane fragility as an isolated, uncultured erythroid cell not comprising the fusion polypeptide.

### Claim 13 (depends on 1)

The method of claim 1, wherein the plurality of enucleated erythroid cells comprises erythrocytes.

### Claim 14 (depends on 1)

The method of claim 1, wherein the plurality of enucleated erythroid cells are human cells.

### Claim 15 (depends on 1)

The method of claim 1, wherein the fusion polypeptide comprises an A domain and an S domain, wherein the surface domain comprises the antigen, or the antigenic fragment thereof.

### Claim 16 (depends on 1)

The method of claim 1, wherein the fusion polypeptide comprises at least one of an S domain, an A domain or a U domain, wherein the S domain is a surface domain on the enucleated erythroid cell, wherein the A domain is an anchor in or on the cell membrane, wherein the U domain faces the intracellular, unexposed side of the enucleated erythroid cell, and wherein at least one of the S domain, the A domain, and the U domain are of different polypeptide origin.

### Claim 17 (depends on 16)

The method of claim 16, wherein at least one of the S domain and the A domain comprises at least 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, or at least 500 amino acids.

### Claim 18 (depends on 1)

The method of claim 1, wherein the fusion polypeptide comprises a transmembrane domain.

### Claim 19 (depends on 2)

The method of claim 2, wherein the method comprises administering the pharmaceutical composition at least twice over a treatment period to the subject.

### Claim 20 (depends on 1)

The method of claim 1, comprising administering the pharmaceutical composition a sufficient number of times over a treatment period such that the percentage of antigen-specific immune cells is substantially decreased during the treatment period.

### Claim 21 (depends on 1)

The method of claim 1, wherein the pharmaceutical composition is administered intravenously.

### Claim 22 (depends on 1)

The method of claim 1, wherein at least 90% of enucleated erythroid cells in the pharmaceutical composition comprise the fusion polypeptide comprising the antigen, or the antigenic fragment thereof.

### Claim 23 (depends on 1)

The method of claim 1, wherein the plurality of enucleated erythroid cells comprises reticulocytes.

### Claim 24 (depends on 1)

The method of claim 1, wherein the antigen is a myelin basic protein, a proteolipid protein, a myelin oligodendrocyte glycoprotein, a preproinsulin, a proinsulin, an insulin, a phospholipase A2 receptor, or a beta-2 glycoprotein 1.

### Claim 25 (depends on 1)

The method of claim 1, wherein the subject is a human subject.