# US 10,864,236 B2 · Compositions comprising bacterial strains

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

- Number: US 10,864,236 B2
- Document ID: US10864236B2
- Status: Granted
- Kind: B2
- Application number: 16/248,857
- Filed: 2019-01-16
- Published: 2019-08-15
- Granted: 2020-12-15
- Earliest claimed: 2015-06-15
- Assignees: 4D Pharma Research Limited
- Applicants: 4D Pharma Research Limited
- Inventors: George Grant; Angela Margaret Patterson; Imke Mulder; Seanin McCluskey; Emma Raftis
- Web page: https://patentgrep.com/patent/US10864236B2

## CPC

- A61K 35/745: Bifidobacteria
- A61K 9/0053: {Mouth and digestive tract, i.e. intraoral and peroral administration}
- A61K 9/19: lyophilised {, i.e. freeze-dried, solutions or dispersions (lyophilised products with subsequent particle size reduction A61K9/14; granules or pellets made by lyphilisation A61K9/1682; solid oral dosage forms made by lyophilisation A61K9/2095; lyophilisation additives A61K47/00)}
- A61K 35/74: Bacteria (therapeutic use of a bacterial protein A61K38/00)
- A61P 1/00: Drugs for disorders of the alimentary tract or the digestive system
- A61P 1/02: Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
- A61P 1/04: for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
- A61P 7/00: Drugs for disorders of the blood or the extracellular fluid
- A61P 7/10: Antioedematous agents; Diuretics
- A61P 9/00: Drugs for disorders of the cardiovascular system
- A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
- A61P 11/00: Drugs for disorders of the respiratory system
- A61P 11/02: Nasal agents, e.g. decongestants
- A61P 11/06: Antiasthmatics
- A61P 11/08: Bronchodilators
- A61P 17/00: Drugs for dermatological disorders
- A61P 17/02: for treating wounds, ulcers, burns, scars, keloids, or the like
- A61P 17/04: Antipruritics
- A61P 17/06: Antipsoriatics
- A61P 19/00: Drugs for skeletal disorders
- A61P 19/02: for joint disorders, e.g. arthritis, arthrosis
- A61P 25/00: Drugs for disorders of the nervous system
- A61P 25/08: Antiepileptics; Anticonvulsants
- A61P 25/28: for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
- A61P 27/02: Ophthalmic agents
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 35/00: Antineoplastic agents
- A61P 35/04: specific for metastasis
- A61P 37/00: Drugs for immunological or allergic disorders
- 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 1/205: Bacterial isolates
- C12R 2001/01: Bacteria or Actinomycetales {; using bacteria or Actinomycetales}
- 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)
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS

## Abstract

The invention provides compositions comprising bacterial strains for treating and preventing inflammatory and autoimmune diseases.

## Claims

### Claim 1

A method of treating asthma in a subject, comprising orally administering to the subject a pharmaceutical composition that comprises: at least 1×106 CFU/g of a dried bacteria strain of the genus Bifidobacterium, wherein the Bifidobacterium bacteria strain is positive for fermentation of raffinose, as determined by an Analytical Profile Index test comprising:
a) contacting a suspension of the Bifidobacterium bacteria strain with a Rapid ID 32A strip at 37° C. for at least 4 hours;
b) contacting the suspension with a development reagent comprising FastBlue for at least 5 minutes; and
c) measuring the color of the suspension,
wherein the Bifidobacterium bacteria strain comprises a polynucleotide sequence of a 16s rRNA gene having at least 95% sequence identity to the polynucleotide sequence of SEQ ID NO: 1 as determined by the Smith-Waterman homology search algorithm using an affine gap search with a gap open penalty of 12 and a gap extension penalty of 2, and a BLOSUM matrix of 62,
wherein the administering results in a reduction of an amount of an eosinophil, neutrophil, or lymphocyte in the subject relative to a corresponding amount of the eosinophil, neutrophil, or lymphocyte prior to the administering, and
wherein the dried Bifidobacterium bacteria strain is live.

### Claim 2 (depends on 1)

The method of claim 1, wherein a bacterial cell of the Bifidobacterium bacteria strain binds to a human cell to a lesser extent than a bacterial cell of Bifidobacterium breve strain JCM 7017 binds to a corresponding cell, as determined by an in vitro assay comprising comparing a measurement of an optical density.

### Claim 3 (depends on 1)

The method of claim 1, wherein the Bifidobacterium bacteria strain is of species Bifidobacterium breve.

### Claim 4

A method of treating asthma in a subject, comprising orally administering to the subject a pharmaceutical composition that comprises: at least 1×106 CFU/g of a dried Bifidobacterium bacteria strain, wherein the Bifidobacterium bacteria strain is the bacteria strain deposited under accession number NCIMB 42380, wherein the Bifidobacterium bacteria strain is positive for fermentation of raffinose, as determined by an Analytical Profile Index test comprising:
a) contacting a suspension of the Bifidobacterium bacteria strain with a Rapid ID 32A strip at 37° C. for at least 4 hours;
b) contacting the suspension with a development reagent comprising FastBlue for at least 5 minutes; and
c) measuring a color of the suspension,
wherein the Bifidobacterium bacteria strain comprises a polynucleotide sequence of a 16s rRNA gene having at least 95% sequence identity to the polynucleotide sequence of SEQ ID NO: 1 as determined by the Smith-Waterman homology search algorithm using an affine gap search with a gap open penalty of 12 and a gap extension penalty of 2, and a BLOSUM matrix of 62,
wherein the administering results in a reduction of an amount of an eosinophil, neutrophil, or lymphocyte in the subject relative to a corresponding amount of the eosinophil, neutrophil, or lymphocyte prior to the administering, and wherein the Bifidobacterium bacteria strain is live.

### Claim 5 (depends on 1)

The method of claim 1, wherein the Bifidobacterium bacteria strain produces an amount of an exopolysaccharide on a surface of a bacterial cell of the Bifidobacterium bacteria strain that is greater than an amount of the same exopolysaccharide produced by a bacterial cell of Bifidobacterium breve strain JCM 7017 as determined by an in vitro assay comprising:
a) binding of the exopolysaccharide to Congo Red;
b) measuring a light absorbance of the Congo Red bound to the exopolysaccharide for the bacterial cell of the Bifidobacterium bacteria strain and the bacterial cell of the Bifidobacterium breve strain JCM 7017; and
c) comparing the light absorbance of the Congo Red bound to the bacterial cell of the Bifidobacterium bacteria strain and the bacterial cell of the Bifidobacterium breve strain JCM 7017.

### Claim 6 (depends on 1)

The method of claim 1, wherein the administering reduces an amount of a cytokine in a lung tissue of the subject, wherein the cytokine is selected from the group consisting of: IL-9, IL-la, IFNg, IL-17A, IL-4, IL-5, IL-1b, RANTES, MIP-1a, KC, and MIP-2.

### Claim 7 (depends on 1)

The method of claim 1, wherein the administering reduces an amount of a house dust mite (HDM) specific immunoglobulin in the subject, as determined by an ELISA assay.

### Claim 8 (depends on 7)

The method of claim 7, wherein the immunoglobulin is an IgG1 or an IgG2a.

### Claim 9 (depends on 1)

The method of claim 1, wherein the administering reduces peribronchiolar infiltration in the subject, as measured by a mean peribronchiolar infiltration score.

### Claim 10 (depends on 1)

The method of claim 1, wherein the administering reduces perivascular infiltration in the subject, as measured by a mean perivascular infiltration score.

### Claim 11 (depends on 1)

The method of claim 1, wherein the asthma is neutrophilic asthma or allergic asthma.

### Claim 12 (depends on 1)

The method of claim 1, wherein the Bifidobacterium bacteria strain ferments raffinose to an identical extent as the Bifidobacterium bacteria strain deposited under accession number NCIMB 42380, as determined by the Analytical Profile Index test.

### Claim 13 (depends on 1)

The method of claim 1, wherein the Bifidobacterium bacteria strain ferments raffinose to an identical extent as the Bifidobacterium bacteria strain deposited under accession number NCIMB 42380, as determined by the Analytical Profile Index test.

### Claim 14 (depends on 1)

The method of claim 1, wherein the Bifidobacterium bacteria strain is present in an amount of from 1×106 to 1×1011 CFU/g.

### Claim 15 (depends on 4)

The method of claim 4, wherein the Bifidobacterium bacteria strain is present in an amount of from 1×106 to 1×1011 CFU/g.