IP Library Granted Patent US 10,744,167
Granted Patent B2
US 10,744,167 · App. 15/700,007 · Granted Aug 18, 2020

Compositions comprising bacterial strains

Inventors: George Grant (Aberdeen, GB); Angela Margaret Patterson (Norwich, GB); Imke Mulder (Aberdeen, GB); Seanin McCluskey (Leeds, GB); Emma Raftis (Leeds, GB)
Assignee: 4D Pharma Research Limited
A61K35/745A61K9/0053A61K9/19A61K35/74C12R1/01
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Quick Facts
Patent No.
US 10,744,167
App. No.
15/700,007
Granted
Aug 18, 2020
Kind
B2
Abstract

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

Claims (47)

1. A method of reducing inflammation caused by Th17 differentiation in a subject, comprising orally administering to the subject a pharmaceutical composition that comprises

a therapeutically effective amount of a bacteria strain of genus Bifidobacterium;

wherein the Bifidobacterium bacteria strain is lyophilized;

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 therapeutically effective amount comprises from about 1×10 6 to about 1×10 11 CFU/g of the Bifidobacterium bacteria strain with respect to a total weight of the pharmaceutical composition; and

wherein the administering of the therapeutically effective amount of the Bifidobacterium bacteria strain results in a reduction of inflammation caused by Th17 differentiation.

2. 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 human cell, as determined by an in vitro assay comprising comparing a measurement of an optical density.

3. The method of claim 1 , wherein the administering of the therapeutically effective amount of the pharmaceutical composition to the subject decreases an amount of IL-17A, IL-17B, IL-17C, IL-17D, IL-17E, IL-17F, IFN-y, IL-10, IL-6, or TNFa; relative to an amount prior to the administering.

4. A method of reducing inflammation caused by Th17 differentiation in a subject, comprising orally administering to the subject a pharmaceutical composition that comprises

a therapeutically effective amount of a bacteria strain of species Bifidobacterium breve , wherein the Bifidobacterium breve bacteria strain is lyophilized;

wherein the Bifidobacterium breve bacteria strain is positive for fermentation of raffinose as determined by an Analytical Profile Index test comprising:

(a) contacting a suspension of the Bifidobacterium breve 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 therapeutically effective amount comprises from about 1×10 6 to about 1×10 11 CFU/g of the Bifidobacterium breve bacteria strain with respect to a total weight of the pharmaceutical composition; and wherein the administering of the therapeutically effective amount of the Bifidobacterium breve bacteria strain results in a reduction of inflammation caused by Th17 differentiation.

5. The method of claim 1 , wherein the Bifidobacterium bacteria strain is a Bifidobacterium bacteria strain deposited under accession number NCIMB 42380.

6. The method of claim 1 , wherein the pharmaceutical composition further comprises a pharmaceutically acceptable excipient, diluent, or carrier.

7. 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; and

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 bacteria bacterial cell of the Bifidobacterium breve strain JCM 7017.

8. The method of claim 1 , wherein the subject has a condition that comprises uveitis, multiple sclerosis, an arthritis, rheumatoid arthritis, osteoarthritis, psoriatic arthritis, spondyloarthriti s, ankylosing spondyliti s, juvenile idiopathic arthritis, neuromyelitis optica, psoriasis, systemic lupus erythematosus, an inflammatory bowel disease, celiac disease, an asthma, allergic asthma, neutrophilic asthma, chronic obstructive pulmonary disease (COPD), scleritis, vasculitis, Behcet's disease, atherosclerosis, atopic dermatitis, emphysema, periodontiti s, allergic rhiniti s, or allograft rejection.

9. The method of claim 8 , wherein the condition is arthritis.

10. The method of claim 9 , wherein the arthritis is rheumatoid arthritis, osteoarthritis, psoriatic arthritis, spondyloarthritis, ankylosing spondylitis, or juvenile idiopathic arthritis.

11. A pharmaceutical composition, in the form of a tablet, capsule, or microcapsule, that comprises:

a therapeutically effective amount of a bacteria strain of genus Bifidobacterium ; and

a pharmaceutically acceptable excipient, diluent, or carrier;

wherein the Bifidobacterium bacteria strain is lyophilized;

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; and

wherein the therapeutically effective amount comprises from about 1×10 6 to about 1×10 11 CFU/g of the Bifidobacterium bacteria strain with respect to a total weight of the pharmaceutical composition and is sufficient to reduce inflammation caused by Th17 differentiation when administered to a subject.

12. The pharmaceutical composition of claim 11 , wherein the therapeutically effective amount of the Bifidobacterium bacteria strain is an amount sufficient to treat a condition mediated by an increase in at least one cytokine in a subject.

13. The pharmaceutical composition of claim 11 , wherein the Bifidobacterium bacteria strain comprises a 16s rRNA gene sequence with at least 95% sequence identity SEQ ID NO:1 as determined by a 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.

14. The pharmaceutical composition of claim 11 , wherein the Bifidobacterium bacteria strain comprises a 16s rRNA gene sequence of SEQ ID NO:1.

15. The pharmaceutical composition of claim 11 , wherein the Bifidobacterium bacteria strain totally or partially colonises an intestine when administered to a subject.

16. The pharmaceutical composition of claim 11 , wherein a bacterial cell of the Bifidobacterium bacteria strain produces an amount of exopolysaccharide 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:

(i) binding of the exopolysaccharide to Congo Red; and

(ii) 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

(iii) comparing the light absorbance of the Congo Red bound to the bacterial cell of the Bifidobacterium bacteria strain and the bacteria bacterial cell of the Bifidobacterium breve strain JCM 7017.

17. The pharmaceutical composition of claim 11 , wherein the pharmaceutical composition further comprises an adjuvant.

18. The method of claim 1 , wherein an Analytical Profile Index test profile for the Bifidobacterium bacteria strain for a-galactosidase, β-galactosidase, α-glucosidase, β-glucosidase, α-arabinose, and mannose is the same as an Analytical Profile Index test profile for a Bifidobacterium bacteria strain deposited under accession number NCIMB 42380 for α-galactosidase, β-galactosidase, α-glucosidase, β-glucosidase, α-arabinose, and mannose.

19. The pharmaceutical composition of claim 11 , wherein an Analytical Profile Index test profile for the Bifidobacterium bacteria strain for α-galactosidase, β-galactosidase, α-glucosidase, β-glucosidase, α-arabinose, and mannose is the same as an Analytical Profile Index test profile for a Bifidobacterium bacteria strain deposited under accession number NCIMB 42380 for α-galactosidase, β-galactosidase, α-glucosidase, β-glucosidase, α-arabinose, and mannose.

Assignments (4)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 14, 2023
From: 4D PHARMA PLC; 4D PHARMA RESEARCH LIMITED
To: CJ BIOSCIENCE, INC.
Reel/Frame 063992/0427 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 31, 2022
From: OXFORD FINANCE LUXEMBOURG S.A R.L.
To: ARMISTICE CAPITAL MASTER FUND LTD.
Reel/Frame 061806/0371 →
INTELECTUAL PROPERTY SECURITY AGREEMENT Recorded Jul 30, 2021
From: 4D PHARMA PLC; 4D PHARMA RESEARCH LIMITED; 4D PHARMA CORK LIMITED; 4D PHARMA DELAWARE INC.
To: OXFORD FINANCE LUXEMBOURG S.A R.L.
Reel/Frame 057042/0715 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 24, 2018
From: GRANT, GEORGE; MULDER, IMKE; RAFTIS, EMMA; PATTERSON, ANGELA MARGARET; MCCLUSKEY, SEANIN
To: 4D PHARMA RESEARCH LIMITED
Reel/Frame 044709/0652 →
Priority Claims (2)
GB 1510467.2 · Jun 15, 2015 · national
GB 1520501.6 · Nov 20, 2015 · national
Continuity (2)
Continuation PCTGB2016051776 · Jun 15, 2016
Related Publication 20170368110A1 · Dec 28, 2017