OA18792A - Compositions comprising bacterial strains. - Google Patents

Compositions comprising bacterial strains. Download PDF

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OA18792A
OA18792A OA1201800175 OA18792A OA 18792 A OA18792 A OA 18792A OA 1201800175 OA1201800175 OA 1201800175 OA 18792 A OA18792 A OA 18792A
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composition
compositions
disease
treating
preventing
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OA1201800175
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Imke Elisabeth MULDER
Amy Beth HOLT
Seanin Marie MCCLUSKEY
Grainne Clare LENNON
Suaad AHMED
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4D Pharma Research Limited
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Abstract

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

Description

COMPOSITIONS COMPRISING BACTERIAL STRAINS
TECHNICAL FIELD
This invention is in the field of compositions comprising bacterial strains isolated from the mammalian digestive tract and the use of such compositions în the treatment of disease.
BACKGROUND TO THE INVENTION
The human intestine is thought to be stérile in utero, but it is exposed to a large variety of maternai and environmental microbes îmmediately after birth. Thereafter, a dynamic period of microbial colonization and succession occurs, which is influenced by factors such as delivery mode, environment, diet and host génotype, ali of which impact upon the composition of the gut microbiota, particulariy during early life. Subsequently, the microbiota stabilizes and becomes adult-like [l]. The human gut microbiota contains more than 500-1000 different phylotypes belonging essentially to two major bacterial divisions, the Bacteroidetes and the Finnicutes [2], The successful symbîotic relationships arising from bacterial colonization of the human gut hâve yielded a wide variety of metabolic, structural, protective and other bénéficiai functions. The enhanced metabolic activities of the colonized gut ensure that otherwise indigestible dietary components are degraded with release of by-products providing an important nutrient source for the host. Similarly, the immunological importance of the gut microbiota is well-recognized and is exemplified in germfree animais which hâve an impaired immune system that is functionally reconstituted following the introduction of commensal bacteria [3-5].
Dramatic changes in microbiota composition hâve been documented in gastrointestinal disorders such as inflammatory bowel disease (IBD). For example, the levels of Clostridium cluster XlVa bacteria are reduced in IBD patients whilst numbers of E. coli are increased, suggesting a shift in the balance of symbionts and pathobionts within the gut [6-9]. interestingly, this microbial dysbiosis is also associated with imbalances in T effector cell populations.
In récognition of the potential positive effect that certain bacterial strains may hâve on the animal gut, various strains hâve been proposed for use in the treatment of various diseases (see, for example, [10-13]). Also, certain strains, including mostly Lactohacillus and Bifidohacterium strains, hâve been proposed for use in treating various inflammatory and autoimmune diseases that are not directly linked to the intestines (see [14] and [15] for reviews). However, the relationship between different diseases and different bacterial strains, and the précisé effects of particular bacterial strains on the gut and at a systemic level and on any particular types of diseases, are poorly characterised.
There îs a requirement in the art for new methods of treating inflammatory and autoimmune diseases. There is also a requirement for the potential effects of gut bacteria to be characterised so that new thérapies using gut bacteria can be developed.
SUMMARY OF THE INVENTION
The inventors hâve developed new thérapies for treating and preventing inflammatory and autoünmune diseases. In particular, the inventors hâve developed new thérapies for treating and preventing diseases and conditions medîated by IL-17 or the Thl7 pathway. In particular, the inventors hâve identified that bacterial strains of the species Enterococcus faecium can be effective for treating and preventing diseases and conditions mediated by IL-17 or the Thl 7 pathway. As described in the examples, oral administration of compositions comprising Enterococcus faecium may reduce the severity of the inflammatory response, including the Thl7 inflammatory response, in mouse models of uveitis.
Therefore, in a first embodiment, the invention provides a composition comprising a bacterial strain of the species Enterococcus faecium, for use in a method of treating or preventing a disease or condition mediated by IL-17 or the Thl7 pathway. The inventors hâve identified that treatment with bacterial strains from this species can provide clinical benefits in mouse models of inflammatory and autoimmune diseases mediated by IL-17 and the Thl 7 pathway, may reduce levels of cytokines that are part of the Thl7 pathway, including IL-17, and may alleviate the Thl7 inflammatory response.
In particular embodiments, the invention provides a composition comprising a bacterial strain of the species Enterococcus faecium, for use in a method of treating or preventing a disease or condition selected from the group consisting of: uveitis; cancer, such as breast cancer, lung cancer, liver cancer, colon cancer, or ovarian cancer; multiple sclerosis; arthritis, such as rheumatoid arthritis, osteoarthritis, psoriatic arthritis, or juvénile idiopathic arthritis; neuromyelitis optica (Devic's disease); ankylosing spondylitis; spondyloarthritis; psoriasis; systemic lupus erythematosus; inflammatory bowel disease, such as Crohn’s disease or ulcerative colitis; celiac disease; asthma, such as allergie asthma or neutrophilie asthma; chronic obstructive pulmonary disease (COPD); scleritis; vasculitis; Behcet's disease; atherosclerosis; atopie dermatitis; emphyseina; periodontitis; allergie rhinitis; and allograft rejection. The effect shown for the bacterial strains from the species Enterococcus faecium on the Thl7 inflammatory response and on diseases mediated by IL-17 and the Thl 7 pathway may provide therapeutic benefits for other diseases and conditions mediated by IL17 and the Thl7 pathway, such as those listed above.
In particularly preferred embodiments, the invention provides a composition comprising a bacterial strain of the species Enterococcus faecium, for use in a method of treating or preventing uveitis, such as posterior uveitis. The inventors hâve identified that treatment with Enterococcus faecium strains can reduce disease incidence and disease severity in a mouse model of uveitis and can prevent or reduce retînal damage. In preferred embodiments, the invention provides a composition comprising a bacterial strain of the species Enterococcus faecium, for use in the treatment of uveitis. Compositions using Enterococcus faecium may be particularly effective for treating uveitis.
In further preferred embodiments, the invention provides a composition comprising a bacterial strain of the species Enterococcus faecium, for use in a method of treating or preventing asthma, such as neutrophilie asthma or allergie asthma. Treatment with Enterococcus faecium strains may reduce recruitment of neutrophils and eosinophils into the lungs, which can help treat or prevent asthma. In certain embodiments, the composition is for use in a method of treating or preventing neutrophilie asthma or éosinophilie asthma. The compositions of the invention may be particularly effective for treating or preventing neutrophilie asthma and éosinophilie asthma. Indeed, in certain embodiments, the composition is for use in a method of reducing a neutrophilie inflammatory response in the treatment or prévention of asthma, or the composition is for use in a method of reducing an éosinophilie inflammatory response in the treatment or prévention of asthma. In preferred embodiments, the invention provides a composition comprising a bacterial strain of the species Enterococcus faecium for use în the treatment of asthma, and in particular éosinophilie or allergie asthma. Also, Enterococcus faecium may hâve a particularly pronounced effect on neutrophils in asthma models and treatment with Enterococcus faecium may be particularly effective for treating neutrophilie asthma.
In further preferred embodiments, the invention provides a composition comprising a bacterial strain of the species Enterococcus faecium, for use in a method of treating or preventing rheumatoid arthritîs. Treatment with Enterococcus faecium strains may provide clinical benefits in a mouse model of rheumatoid arthritîs and reduce joint swellîng. In preferred embodiments, the invention provides a composition comprising a bacterial strain of the species Enterococcus faecium, for use in the treatment of rheumatoid arthritîs. Compositions using Enterococcus faecium may be particularly effective for treating rheumatoid arthritîs.
In further preferred embodiments, the invention provides a composition comprising a bacterial strain of the species Enterococcus faecium, for use in a method of treating or preventing multiple sclerosis. Treatment with Enterococcus faecium strains may reduce disease incidence and disease severity in a mouse model of multiple sclerosis. In preferred embodiments, the invention provides a composition comprising a bacterial strain of the species Enterococcus faecium, for use in the treatment of multiple sclerosis. Compositions using Enterococcus faecium may be particularly effective for treating multiple sclerosis.
In further preferred embodiments, the invention provides a composition comprising a bacterial strain of the species Enterococcus faecium, for use in a method of treating or preventing cancer, such as breast, lung or liver cancer. Compositions comprising a bacterial strain of the species Enterococcus faecium may reduce tumour growth in mouse models of breast, lung and liver cancer. In certain embodiments, the composition is for use in a method of reducing tumour size or preventing tumour growth in the treatment of cancer.
In certain embodiments, the compositions of the invention are for use in a method of reducing IL-17 production or reducing Thl7 cell différentiation in the treatment or prévention of a disease or condition mediated by IL-17 or the Thl7 pathway. In particular, the compositions of the invention may be used in reducing IL-17 production or reducing Thl7 cell différentiation in the treatment or prévention of asthma. rheumatoid arthritis, multiple sclerosis, uveitis or cancer. Preferably, the invention provides compositions comprising a bacterial strain of the species Enterococcus faecium. for use in reducing IL-17 production or reducing Thl7 cell différentiation in the treatment or prévention of asthma, rheumatoid arthritis, multiple sclerosis, uveitis or cancer.
In certain embodiments, the composition is for use in a patient with elevated IL-17 levels or Thl7 cells. The effect shown for Enterococcus faecium on uveitis, which is strongly associated with the Thl 7 inflammatory response, means that Enterococcus faecium strains may be particularly bénéficiai for such patients.
In preferred embodiments of the invention, the bacterial strain in the composition is of Enterococcus faecium. Closely related strains may also be used, such as bacterial strains that hâve a 16s rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to the 16s rRNA sequence of a bacterial strain of Enterococcus faecium. Preferably, the bacterial strain has a 16s rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to SEQ ID NO: l or 2. Preferably, the sequence identity is to SEQ ID NO:2. Preferably, the bacterial strain for use in the invention has the 16s rRNA sequence represented by SEQ ID NO:2.
In certain embodiments, the composition of the invention is for oral administration. Oral administration of the strains of the invention can be effective for treating IL-17- or Thl 7 pathwaymediated diseases and conditions. Also, oral administration is convenient for patients and practitioners and allows delivery to and ! or partial or total colonisation of the intestine.
In certain embodiments, the composition of the invention comprises one or more pharmaceutically acceptable excipients or carriers.
In certain embodiments, the composition of the invention comprises a bacterial strain that has been lyophilised. Lyophilisation is an effective and convenient technique for preparing stable compositions that allow delivery of bacteria.
In certain embodiments, the invention provides a food product comprising the composition as described above.
In certain embodiments, the invention provides a vaccine composition comprising the composition as described above.
Additionally, the invention provides a method of treating or preventing a disease or condition mediated by IL-I7 or the Thl7 pathway, comprising administering a composition comprising a bacterial strain of the species Enterococcus faecium.
In developing the above invention, the inventors hâve identîfied and characterised a bacterial strain that is particularly useful for therapy. The Enterococcus faecium strain of the invention is shown to be effective for treating the diseases described herein, such as uveitis. Therefore, in another aspect, the invention provides a cell of the Enterococcus faecium strain deposited under accession number NCIMB 42487, or a dérivative thereof. The invention also provides compositions comprising such cells, or biologically pure cultures of such cells, The invention also provides a cell of the Enterococcus faecium strain deposited under accession number NCIMB 42487, or a dérivative thereof, for use in therapy, in particular for the diseases described herein.
BRIEF DESCRIPTION OF DRAWINGS
Figure 1: Mouse model of uveitis - Lymph node proliférative response to 1RBP peptide. Media background subtracted [IRBP peptide stimulated - media background] counts per minute based on 3 H- thymidine incorporation. Ail data are presented as Mean + SEM (n=3).
Figure 2: Mouse model of uveitis - TEFI Scores in the control group. Data are presented as Mean ± SEM.
Figure 3: Mouse model of uveitis - TEFI Scores on Day 21. Data are presented as Mean ± SEM.
Figure 4: Rapid ID 32 A analysis of MRxOOlO
DISCLOSURE OF THE INVENTION
Bacterial strains
The compositions of the invention comprise a bacterial strain of the species Enterococcus faecium. The examples demonstrate that bacteria of this species are useful for treating or preventing uveitis and diseases and conditions mediated by IL-17 or the Thl 7 pathway.
The invention provides a composition comprising a bacterial strain of the species Enterococcus faecium for use in therapy, fbr example, for use in treating or preventing an inflammatory and/or autoimmune disease. In certain embodiments, the compositions of the invention comprise Enterococcus faecium and do not contain any other bacterial species. In certain embodiments, the compositions of the invention comprise a single strain of Enterococcus faecium and do not contain any other bacterial strains or species.
Enterococcus faecium is a Gram-positive, alpha-hemolytic or nonhemolytic bacterium in the genus Enterococcus that often occurs in pairs (diplococci) or short chains. The type strain of Enterococcus faecium is ATCC 19434 = CCUG 542 = CIP 103014 = CFBP 4248 = DSM 20477 = HAMBI 1710 = JCM 5804 = JCM 8727 = LMG H 423 = NB RC 100486 = NBRC 100485 = NCIMB 11508 (fonnerly NCDO 942) = NCTC 7171 [16]. The GenBank accession number for the 16S rRNA gene sequence of Enterococcus faecium strain LMG 11423 is AJ3OI83O (disclosed herein as SEQ ID NO: 1 ). This exemplary Enterococcus faecium strain is described in [ 16],
Other Enterococcus faecium strains for use in the invention include: RI 3 [17], CFR 3003 [18], AL41 [19], DSM 10663 NCIMB 10415 [20], NCIMB 10415 E1707 [21], NM113 and NM213 [22], In certain embodiments, the compositions of the invention comprise one of these strains, or a dérivative or biotype thereof. A further example of an Enterococcus faecium for use in the invention is the DO strain. The genomic sequence of this bacterium consists of a chromosome and three plasmids. The sequence of the chromosome is disclosed herein as SEQ ID NO:3 and the sequence of the three plasmids is disclosed as SEQ ID NOs:4, 5 and 6. SEQ ID NOs: 3, 4, 5 and 6 are disclosed in WO 2017/085520. The genomic sequence was obtained using whole shotgun sequences and is available using GenBank accession number NC 017960.1.
The Enterococcus faecium bacterium deposited under accession number NCIMB 42487 was tested în the Examples and is also referred to herein as strain MRX010. The terms “MRX010 and “MRxOOlO” are used interchangeably herein. A 16S rRNA sequence for the MRX010 strain that was tested is provided in SEQ ID NO:2. Strain MRX010 was deposited with the international depositary authority NCIMB, Ltd. (Ferguson Building, Aberdeen, AB21 9YA, Scotland) by 4D Pharma Research Ltd. (Life Sciences Innovation Building. Aberdeen, AB25 2ZS, Scotland) on I6th November 2015 as Enterococcus faecium and was assigned accession number NCIMB 42487.
Bacterial strains closely related to the strain tested in the examples are also expected to be effective for treating or preventing uveitis and diseases and conditions mediated by IL-17 or the Th 17 pathway. In certain embodiments, the bacterial strain for use in the invention has a 16s rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to the 16s rRNA sequence of a bacterial strain of Enterococcus faecium. Preferably, the bacterial strain for use in the invention has a 16s rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to SEQ ID NO: 1 or 2. Preferably, the sequence identity is to SEQ ID NO:2. Preferably, the bacterial strain for use in the invention has the 16s rRNA sequence represented by SEQ ID NO:2.
Bacterial strains that are biotypes of the bacterium deposited under accession number 42487 are also expected to be effective for treating or preventing uveitis and diseases and conditions mediated by IL-17 or the Th 17 pathway. A biotype is a closely related strain that has the same or very siinilar physiological and biochemical characteristics.
Strains that are biotypes of the bacterium deposited under accession number NCIMB 42487 and that are suitable for use in the invention may be identified by sequencing other nucléotide sequences for the bacterium deposited under accession number NCIMB 42487. For example, substantially the whole genome may be sequenced and a biotype strain for use in the invention may hâve at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% sequence identity across at least 80% of its whole genome (e.g. across at least 85%, 90%, 95% or 99%, or across its whole genome). For example, in some embodiments, a biotype strain has at least 98% sequence identity across at least 98% of its genome or at least 99% sequence identity across 99% of its genome. Other suitable sequences for use in identifying biotype strains may include hsp60 or répétitive sequences such as BOX, ERIC, (GTG)5, or REP or [23], Biotype strains may hâve sequences with at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% sequence identity to the corresponding sequence of the bacterium deposited under accession number NCIMB 42487.1n some embodiments, a biotype strain has a sequence with at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% sequence identity to the corresponding sequence of strain MRX010 deposited as NCIMB 42487 and comprises a 16S rRNA sequence that is at least 99% identical (e.g. at least 99.5% or at least 99.9% identical) to SEQ ID NO:2. In some embodiments, a biotype strain has a sequence with at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% sequence identity to the corresponding sequence of strain MRX010 deposited as NCIMB 42487 and has the 16S rRNA sequence of SEQ ID NO:2.
Alternatively, strains that are biotypes of the bacterium deposited under accession number NCIMB 42487and that are suitable for use in the invention may be identified by using the accession number NCIMB 42487 deposit and restriction fragment analysis and/or PCR analysis, for example by using fluorescent amplified fragment length polymorphism (FAFLP) and répétitive DNA element (rep)PCR fingerprinting, or protein profîling, or partial 16S or 23s rDNA sequencing. In preferred embodiments, such techniques may be used to identify other Enterococcusfaecium strains.
In certain embodiments, strains that are biotypes of the bacterium deposited under accession number NCIMB 42487 and that are suitable for use in the invention are strains that provide the same pattern as the bacterium deposited under accession number NCIMB 42487 when analysed by amplified ribosomal DNA restriction analysis (ARDRA), for example when using Sau3AI restriction enzyme (for exemplary methods and guidance see, for examp le, [24]). Alternatively, biotype strains are identified as strains that hâve the same carbohydrate fermentation patterns as the bacterium deposited under accession number NCIMB 42487.
In some embodiments, the bacterial strain used in the invention is:
(i) Positive for at least one of (e.g. at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or ail of): arginine dîhydrolase, β-glucosidase, mannose fermentation, glutamic acid decarboxylase, arginine arylamidase, phenylalanine arylamidase, leucine arylamidase, pyroglutamic acid arylamidase, tyrosine arylamidase, glycine arylamidase, histidine arylamidase and serine arylamidase; and/or (ii) Intermediate for N-acetyl-p-glucosamînidase;
preferably as determined by an assay of carbohydrate, amino acid and nitrate metabolism, and optionally an assay of alkaline phosphatase activity, more preferably as determined by Rapid ID 32A analysis (preferably using the Rapid ID 32A system from bioMérieux).
Other Enterococcus faecitim strains that are useful in the compositions and methods of the invention, such as biotypes of the bacterium deposited under accession number NCIMB 42487, may be identified using any appropriate method or strategy, including the assays described in the examples. For instance, strains for use in the invention may be identified by culturing in anaérobie YCFA and/or administering the bacteria to the type II collagen-induced arthritis mouse model and then assessing cytokine levels. In particular, bacterial strains that hâve similar growth patterns, metabolic type and/or surface antigens to the bacterium deposited under accession number NCIMB 42487 may be useful in the invention. A useful strain will hâve comparable immune modulatory activity to the NCIMB 42487 strain. In particular, a biotype strain will elicit comparable effects on the uveitis disease models to the effects shown in the Examples, which may be identified by using the culturing and administration protocole described in the Examples.
A particularly preferred strain of the invention is the Enterococcus faecitim strain deposited under accession number NCIMB 42487. This is the exemplary MRX010 strain tested in the examples and shown to be effective for treating disease. Therefore, the invention provides a cell, such as an isolated cell, of the Enterococcus faecitim strain deposited under accession number NCIMB 42487, or a dérivative thereof. The invention also provides a composition comprising a cell of the Enterococcus faecitim strain deposited under accession number NCIMB 42487, or a dérivative thereof. The invention also provides a biologically pure culture of the Enterococcus faecitim strain deposited under accession number NCIMB 42487. The invention also provides a cell of the Enterococcus faecitim strain deposited under accession number NCIMB 42487, or a dérivative thereof, for use in therapy, in particular for the diseases described herein. A dérivative of the strain deposited under accession number NCIMB 42487 may be a daughter strain (progeny) or a strain cultured (subcloned) from the original.
A dérivative of a strain of the invention may be modified, for example at the genetic level, without ablating the biological activity, In particular, a dérivative strain of the invention is therapeutically active. A dérivative strain will hâve comparable immune modulatory activity to the original NCIMB 42487 strain. In particular, a dérivative strain will elicit comparable effects on the uveitis disease models to the effects shown in the Examples, which may be identified by using the culturing and administration protocols described in the Examples. A dérivative of the NCIMB 42487 strain will generally be a biotype of the NCIMB 42487 strain.
Référencés to cells of the Enterococcus faecium strain deposited under accession number NCIMB 42487 encompass any cells that hâve the same safety and therapeutic efficacy characteristics as the strains deposited under accession number NCIMB 42487, and such cells are encompassed by the invention. Thus, in some embodiments, référencé to cells of the Enterococcus faecium strain deposited under accession number NCIMB 42487 refers only to the MRX010 strain deposited under NCIMB 42487 and does not refer to a bacterial strain that was not deposited under NCIMB 42487. In some embodiments, référencé to cells of the Enterococcus faecium strain deposited under accession number NCIMB 42487 refers to cells that hâve the same safety and therapeutic efficacy characteristics as the strains deposited under accession number NCIMB 42487, but which are not the strain deposited under NCIMB 42487.
In certain embodiments, the bacterial strain for use in the invention has a chromosome with sequence identity to SEQ ID NO:3. In some embodiments, the bacterial strain for use in the invention has a chromosome with at least 90% sequence identity (e.g. at least 92%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity) to SEQ ID NO:3 across at least 60% (e.g. across at least 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, 99% or 100%) of SEQ ID NO:3. For example, the bacterial strain for use in the invention may hâve a chromosome with at least 90% sequence identity to SEQ ID NO:3 across 70% of SEQ ID NO:3, or at least 90% sequence identity to SEQ ID NO:3 across 80% of SEQ ID NO:3, or at least 90% sequence identity to SEQ ID NO:3 across 90% of SEQ ID NO:3, or at least 90% sequence identity to SEQ ID NO:3 across 100% of SEQ ID NO:3, or at least 95% sequence identity to SEQ ID NO:3 across 70% of SEQ ID NO:3, or at least 95% sequence identity to SEQ ID NO:3 across 80% of SEQ ID NO:3, or at least 95% sequence identity to SEQ ID NO:3 across 90% of SEQ ID NO:3, or at least 95% sequence identity to SEQ ID NO:3 across 100% of SEQ ID NO:3, or at least 98% sequence identity to SEQ ID NO:3 across 70% of SEQ ID NO:3, or at least 98% sequence identity to SEQ ID NO:3 across 80% of SEQ ID NO:3, or at least 98% sequence identity to SEQ ID NO:3 across 90% of SEQ ID NO:3, or at least 98% identity across 95% of SEQ ID NO:3, or at least 98% sequence identity to SEQ ID NO:3 across 100% of SEQ ID NO:3, or at least 99.5% sequence identity to SEQ ID NO:3 across 90% of SEQ ID NO:3, or at least 99.5% identity across 95% of SEQ ID NO:3, or at least 99.5% identity across 98% of SEQ ID NO:3, or at least 99.5% sequence identity to SEQ ID NO;3 across 100% of SEQ ID NO:3.
In certain embodiments, the bacterial strain for use in the invention has a chromosome with sequence identity to SEQ ID NO:3, for example as described above, and a 16S rRNA sequence with sequence identity to SEQ ID NO:1 or 2, for example as described above, preferably with a 16s rRNA sequence that is at least 99% identical to SEQ ID NO: 2, more preferably which comprises the 16S rRNA sequence of SEQ ID NO:2.
ΙΟ
In certain embodiments, the bacterial strain for use in the invention has a chromosome with sequence identity to SEQ ID NO:3, for example as described above, and is effective for treating or preventing diseases and conditions mediated by IL-17 or the ThI 7 pathway..
In certain embodiments, the bacterial strain for use in the invention has a chromosome with sequence identity to SEQ ID NO:3, for example as described above, and a 16S rRNA sequence with sequence identity to SEQ ID NO: l or 2, for example as described above, and is effective for treating or preventing diseases and conditions mediated by IL-17 or the Thl 7 pathway.
In certain embodiments, the bacterial strain for use in the invention has a 16s rRNA sequence that is at least 99%, 99.5% or 99.9% identical to the I6s rRNA sequence represented by SEQ ID NO: 2 (for example, which comprises the 16S rRNA sequence of SEQ ID NO:2) and a chromosome with at least 95% sequence identity to SEQ ID NO:3 across at least 90% of SEQ ID NO:3, and which is effective for treating or preventing diseases and conditions mediated by IL-17 or the Thl 7 pathway.
In certain embodiments, the bacterial strain for use in the invention is a Enterococcus faeciiim and has a 16s rRNA sequence that is at least 99%, 99.5% or 99.9% identical to the 16s rRNA sequence represented by SEQ ID NO: 2 (for example, which comprises the 16S rRNA sequence of SEQ ID NO:2) and a chromosome with at least 98% sequence identity (e.g. at least 99% or at least 99.5% sequence identity) to SEQ ID NO:3 across at least 98% (e.g. across at least 99% or at least 99.5%) of SEQ ID NO:3, and which is effective for treating or preventing diseases and conditions mediated by IL-17 or the Th 17 pathway.
In preferred embodiments, the bacterial strains in the compositions of the invention are viable and capable of partially or totally colonising the intestine.
Therapeutic uses
As demonstrated in the examples, the bacterial compositions of the invention are effective for reducing the Thl 7 inflammatory response. In particular, treatment with compositions of the invention achieves clinical improvements in animal models of conditions mediated by IL-17 and the Thl7 pathway and may achieve a réduction in IL-17A levels and other Thl7 pathway cytokines. Therefore, the compositions of the invention may be useful for treating or preventing inflammatory and autoinimune diseases, and in particular diseases or conditions mediated by IL-17. In particular, the compositions of the invention may be useful for reducing or preventing élévation of the IL-17 inflammatory response.
Thl7 cells are a subset ofT helper cells that produce, for example, IL-17A, IL17-F, IL-21 and IL-22. Thl7 cell différentiation and IL-17 expression may be driven by IL-23. These cytokines and others form important parts of the Thl7 pathway, which is a well-established inflammatory signalling pathway that contributes to and underlies a number of inflammatory and autoimmune diseases (as
H described in, for example, [25-30]). Diseases wherein the Thl7 pathway is activated are Thl7 pathway-mediated diseases. Thl7 pathway-mediated diseases can be ameliorated or alleviated by repressing the Thl7 pathway, which may be through a réduction in the différentiation of Thl7 cells or a réduction in their activity or a réduction in the level of ThI7 pathway cytokines. Diseases mediated by the Thl7 pathway may be characterised by increased levels of cytokines produced by Thl7 cells, such as IL-17A, IL-17F, IL-21, IL-22, IL-26, IL-9 (reviewed in [31]). Diseases mediated by the Thl7 pathway may be characterised by increased expression of Th-l7-related genes, such as Stat3 or IL-23R. Diseases mediated by the Thl7 pathway may be associated with increased levels of Th 17 cells.
IL-17 is a pro-inflammatory cytokine that contributes to the pathogenesis of several inflammatory and autoimmune diseases and conditions. IL-l 7 as used herein may refer to any member of the IL-l 7 family, including IL-17A, IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F. IL-l7-mediated diseases and conditions are characterised by high expression of IL-l7 and/or the accumulation or presence of IL-l7-positive cells in a tissue affected by the disease or condition. Similarly, IL-l 7-mediated diseases and conditions are diseases and conditions that are exacerbated by high IL-17 levels or an increase in IL-l7 levels, and that are alleviated by low IL-l7 levels or a réduction in IL-l7 levels. The IL-l7 inflammatory response may be local or systemic.
Examples of diseases and conditions that may be mediated by IL-l7 or the Thl7 pathway include uveitis; cancer, such as breast cancer, lung cancer, liver cancer, colon cancer, or ovarîan cancer; multiple sclerosis; arthritis, such as rheumatoid arthritis, osteoarthritis, psoriatic arthritis, or juvénile idiopathic arthritis; neuromyelitis optica (Devic's disease); ankylosing spondylitis; spondyloarthritis; psoriasis; systemic lupus erytheinatosus; inflammatory bowel disease, such as Crohn’s disease or ulcerative colitis; celiac disease; asthma, such as allergie asthma or neutrophilie asthma; chronic obstructive pulmonary disease (COPD); scleritis; vasculitis; Behcet's disease; atherosclerosis; atopie dermatitis; emphysema; periodontitis; allergie rhinitis; and allograft rejection. In preferred embodiments, the compositions of the invention are used for treating or preventing one or more of these conditions or diseases. In further preferred embodiments, these conditions or diseases are mediated by IL-l7 or the Thl 7 pathway.
In some embodiments, the pathogenesis of the disease or condition affects the intestine. In some embodiments, the pathogenesis of the disease or condition does not affect the intestine. In some embodiments, the pathogenesis of the disease or condition is not localised at the intestine. In some embodiments, the treating or preventing occurs at a site other than at the intestine. In some embodiments, the treating or preventing occurs at the intestine and also at a site other than at the intestine. In certain embodiments, the disease or condition is systemic.
In certain embodiments, the compositions of the invention are for use in a method of reducing IL-17 production or reducing Thl 7 cell différentiation in the treatment or prévention of a disease or condition mediated by IL-17 or the Thl 7 pathway. In certain embodiments, the compositions of the invention are for use in treating or preventing an inflammatory or autoimmune disease, wherein said treatment or prévention is achieved by reducing or preventing élévation of the Thl7 inflammatory response. in certain embodiments, the compositions of the invention are for use in treating a patient with an inflammatory or autoimmune disease, wherein the patient has elevated IL-17 levels or elevated Thl7 cells or is exhibiting a Thl7 inflammatory response. In certain embodiments, the patient may hâve been dîagnosed with a chronic inflammatory or autoimmune disease or condition, or the composition of the invention may be for use in preventing an inflammatory or autoimmune disease or condition developing into a chronic inflammatory or autoimmune disease or condition. In certain embodiments, the disease or condition may not be responsive to treatment with TNF-a inhibitors. These uses of the invention may be applied to any of the spécifie disease or conditions listed in the preceding paragraph.
IL-17 and the Thl7 pathway are often associated with chronic inflammatory and autoimmune diseases, so the compositions of the invention may be particularly useful for treating or preventing chronic diseases or conditions as listed above. In certain embodiments, the compositions are for use in patients with clnonic disease. In certain embodiments, the compositions are for use in preventing the development of chronic disease.
The compositions of the invention may be useful for treating diseases and conditions mediated by IL17 or the Thl 7 pathway and for addressing the Thl 7 inflammatory response, so the compositions of the invention may be particularly useful for treating or preventing chronic disease, treating or preventing disease in patients that hâve not responded to other thérapies (such as treatment with TNF-α inhibitors), and/or treating or preventing the tissue damage and symptoms associated with IL17 and Th 17 cells. For example, IL-17 is known to activate matrix destruction in cartilage and bone tissue and IL-17 has an inhibitory effect on matrix production in chondrocytes and osteoblasts, so the compositions of the invention may be useful for treating or preventing bone érosion or cartilage damage.
In certain embodiments, treatment with compositions of the invention provides a réduction or prevents an élévation in IL-17 levels, in particular IL-17A levels. In certain embodiments, treatment with compositions of the invention provides a réduction or prevents an élévation in TNFa, IFN-γ or IL-6 levels. Such réduction or prévention of elevated levels of these cytokines may be useful for treating or preventing inflammatory and autoimmune diseases and conditions, in particular those mediated by IL-17 or the Th 17 pathway.
Uveitis
In preferred embodiments, the compositions of the invention are for use in treating or preventing uveitis. The examples demonstrate that the compositions of the invention achieve a réduction in disease incidence and disease severity in an animal model of uveitis and so they may be useful in the treatment or prévention of uveitis. Uveitis is inflammation of the uvea and can resuit in retinal tissue destruction. It can present in different anatomical forms (anterior, intermediate, posterior or diffuse) and resuit from différent, but related, causes, including systemic autoimmune disorders. IL-17 and the Thl7 pathway are centrally involved in uveitis, so the efficacy of the compositions of the invention for treating uveitis indicates that the compositions of the invention may be particularly effective for treating and preventing diseases and conditions mediated by IL-17 or the Th 17 pathway. Références [32-39] describe elevated sérum levels of interleukin-l 7A in uveitis patients, spécifie association of IL17A genetic variants with panuveitis, the rôle of Thl7-associated cytokines in the pathogenesis of experimental autoimmune uveitis, the imbalance between Thl7 Cells and regulatory T Cells during monophasic experimental autoimmune uveitis, the up-regulation of IL-17A in patients with uveitis and active Adamantiades-Behçet and Vogt-Koyanagi-Harada (VKH) diseases, the treatment of non-infectious uveitis with seeukinumab (anti-IL-l7A antibody), and Thl7 in uveitic eyes.
In certain embodiments, the uveitis is posterior uveitis. Posterior uveitis présents primarily with inflammation of the retina and choroid and the examples demonstrate that the compositions of the invention are effective for reducing retinal inflammation and damage.
In certain embodiments, treatment with the compositions of the invention results in a réduction in retinal damage. In certain embodiments, the compositions of the invention are for use in reducing or preventing retinal damage in the treatment of uveitis. In certain embodiments, the compositions are for use in treating patients with severe uveitis that are at risk of retinal damage. In certain embodiments, treatment with the compositions of the invention results in a réduction in optic dise inflammation. In certain embodiments, the compositions of the invention are for use în reducing or preventing optic dise inflammation. In certain embodiments, treatment with the compositions of the invention results in a réduction in retinal tissue infiltration by inflammatory cells. In certain embodiments, the compositions of the invention are for use in reducing retinal tissue infiltration by inflammatory cells. In certain embodiments, treatment with the compositions of the invention results in vision being maintained or improved. In certain embodiments, the compositions of the invention are for use in maintaining or improving vision.
In certain embodiments, the compositions are for use in treating or preventing uveitis associated with a non-infectious or autoimmune disease, such as Behçet disease, Crohn's disease, Fuchs hétérochromie iridocyclitis, granulomatosis with polyangiîtis, HLA-B27 related uveitis, juvénile
I4 idiopathic arthritis, sarcoidosis, spondyloarthritis, sympathetic ophthalmia, tubulointerstitial nephritis and uveitis syndrome or Vogt-Koyanagi-Harada syndrome. IL-17A has been shown to be involved in, for example, Behçet and Vogt-Koyanagi-Harada diseases.
Treatment or prévention of uveitis may refer to, for example, an alleviation of the severity of symptoms or a prévention of relapse.
Cancer
In preferred embodiments, the compositions of the invention are for use in treating or preventing cancer. IL-17 and the Thl7 pathway hâve central rôles in cancer development and progression, and so the compositions of the invention may be useful for treating or preventing cancer.
Although the rôles of IL-17 and Thl7 cells in cancer are not fully understood, numerous pro-tumour effects of IL-17 and Thl7 cells are known. For example, Thl7 cells and IL-17 can promote angiogenesîs, increase prolifération and survival of tumor cells and activate tumour-promoting transcription factors [40-42],
In certain embodiments, treatment with the compositions of the invention results in a réduction in tumour sîze or a réduction in tumour growth. In certain embodiments, the compositions of the invention are for use in reducing tumour size or reducing tumour growth. The compositions of the invention may be effective for reducing tumour size or growth. In certain embodiments, the compositions of the invention are for use in patients with solid tumeurs. In certain embodiments, the compositions of the invention are for use in reducing or preventing angiogenesîs in the treatment of cancer. IL-17 and Thl7 cells hâve central rôles in angiogenesîs. In certain embodiments, the compositions of the invention are for use in preventing metastasis.
In certain embodiments, the compositions of the invention are for use in treating or preventing breast cancer. The compositions of the invention may be effective for treating breast cancer, and IL-17 and Thl7 cells hâve important rôles in breast cancer [43], In certain embodiments, the compositions of the invention are for use in reducing tumour size, reducing tumour growth, or reducing angiogenesîs în the treatment of breast cancer. In preferred embodiments the cancer is mammary carcinoma. In preferred embodiments the cancer is stage IV breast cancer.
In certain embodiments, the compositions of the invention are for use in treating or preventing lung cancer. The compositions of the invention may be effective for treating lung cancer, and IL-17 and Thl7 cells hâve important roles in lung cancer [44], In certain embodiments, the compositions of the invention are for use in reducing tumour size, reducing tumour growth, or reducing angiogenesîs in the treatment of lung cancer. In preferred embodiments the cancer is lung carcinoma.
In certain embodiments, the compositions of the invention are for use in treating or preventing liver cancer. The compositions of the invention may be effective for treating liver cancer, and lL-17 and Thl7 cells hâve important roles in liver cancer [45], In certain embodiments, the compositions of the invention are for use in reducing tumour size, reducing tumour growth, or reducing angiogenesis in 5 the treatment of liver cancer. In preferred embodiments the cancer is hepatoma (hepatocellular carcinoma).
In certain embodiments, the compositions of the invention are for use in treating or preventing carcinoma. The compositions of the invention may be particularly effective for treating carcinoma. In certain embodiments, the compositions of the invention are for use in treating or preventing nonI0 immunogenic cancer. The compositions of the invention may be effective for treating nonimmunogenic cancers.
In further embodiments, the compositions of the invention are for use in treating or preventing acute lymphoblastic leukemia (ALL), acute myeloid leukemia, adrenocortical carcinoma, basal-cell carcinoma, bile duct cancer, bladder cancer, bone tumor, osteosarcoma/malignant fibrous 15 histiocytoma, brainstem glioma, brain tumor, cerebellar astrocytoma, cerebral astrocytoma/malignant glioma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumors, breast cancer, bronchial adenomas/carcinoids, Burkitt's lymphoma, carcînoid tumor, cervical cancer, chronic lymphocytic leukemia, chronic myelogenous leukemia, chronic myeloproliferative disorders, colon cancer, cutaneous T-cell lymphoma. endométrial cancer, ependymoma, esophageal cancer, 20 Ewing's sarcoma, intraocular melanoma, retinoblastoma, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, glioma, childhood visual pathway and hypothalamic, Hodgkin lymphoma, melanoma, islet cell carcinoma, Kaposi sarcoma, rénal cell cancer, laryngeal cancer, leukaemias, lymphomas, mesothelioma, neuroblastoma, non-Hodgkin lymphoma, oropharyngeal cancer, osteosarcoma, ovarian cancer, 25 pancreatic cancer, parathyroid cancer, pharyngeal cancer, pituitary adenoma, plasma cell neoplasia, prostate cancer, rénal cell carcinoma, retinoblastoma, sarcoma, testicular cancer, thyroid cancer, or uterine cancer.
The compositions of the invention may be particularly effective when used in combination with further therapeutic agents. The immune-modulatory effects of the compositions of the invention may 30 be effective when combined with more direct anti-cancer agents. Therefore, in certain embodiments, the invention provides a composition comprising a bacteriai strain of the species Enlerococcus faecium and an anticancer agent. In preferred embodiments the anticancer agent îs an immune checkpoint inhibitor, a targeted antibody immunotherapy, a CAR-T cell therapy, an oncolytic virus, or a cytostatic drug. In preferred embodiments, the composition comprises an anti-cancer agent 35 selected from the group consisting of: Yervoy (ipilimumab, BMS); Keytruda (pembrolizumab,
Merck); Opdivo (nivolumab, BMS); MEDI4736 (AZ/Medlmmune); MPDL3280A (Roche/Genentech); Tremelimumab (AZ/Medlmmune); CT-Oll (pidilizumab, CureTech); BMS986015 (lirilumab, BMS); MEDI0680 (AZ/Medlmmune); MSB-00I0718C (Merck); PF-05082566 (Pfizer); MEDI6469 (AZ/Medlmmune); BMS-986016 (BMS); BMS-663513 (urelumab, BMS); IMP321 (Prima Bîomed); LAG525 (Novartis); ARGX-110 (arGEN-X); PF-05082466 (Pfizer); CDX-1127 (varlilumab; CellDex Therapeutics); TRX-518 (G1TR Inc.); MK-4166 (Merck); JTX2011 (Jounce Therapeutics); ARGX-115 (arGEN-X); NLG-9189 (indoximod, NewLink Genetics); INCB024360 (Incyte); IPH2201 (Innate lmmotherapeutics/ΑΖ); NLG-919 (NewLink Genetics); antiVISTA (JnJ); Epacadostat (INCB24360, Incyte); F001287 (Flexus/BMS); CP 870893 (University of Pennsylvania); MGA271 (Macrogenix); Emactuzumab (Roche/Genentech); Galunisertib (Eli Lilly); Ulocuplumab (BMS); BKT140/BL8040 (Biokine Therapeutics); Bavituximab (Peregrine Pharmaceuticals); CC 90002 (Celgene); 852A (Pfizer); VTX-2337 (VentiRx Pharmaceuticals); IMO2055 (Hybridon, Idera Pharmaceuticals); LY2157299 (Eli Lilly); EW-7197 (Ewha Women's University, Korea); Vemurafenib (Plexxikon); Dabrafenib (Genentech/GSK); BMS-777607 (BMS); BLZ945 (Memorial Sloan-Kettering Cancer Centre); Unituxin (dinutuximab, United Therapeutics Corporation); Blincyto (blinatumomab, Amgen); Cyramza (ramucirumab, Eli Lilly); Gazyva (obinutuzumab, Roche/Biogen); Kadcyla (ado-trastuzumab emtansine, Roche/Genentech); Perjeta (pertuzumab, Roche/Genentech); Adcetris (brentuximab vedotin, Takeda/Millennium); Arzerra (ofatumumab, GSK); Vectibix (panitumumab, Amgen); Avastin (bevacizumab, Roche/Genentech); Erbitux (cetuximab, BMS/Merck); Bexxar (tositumomab-1131, GSK); Zevalîn (ibritumomab tiuxetan, Biogen); Campath (alemtuzumab, Bayer); Mylotarg (gemtuzumab ozogamicin, Pfizer); Herceptin (trastuzumab, Roche/Genentech); Rituxan (rituximab, Genentech/Biogen); volociximab (Abbvie); Enavatuzumab (Abbvie); ABT-414 (Abbvie); Elotuzumab (Abbvie/BMS); ALX-0141 (Ablynx); Ozaralizumab (Ablynx); Actimab-C (Actinium); Actimab-P (Actinium); Milatuzumab-dox (Actinium); Emab-SN-38 (Actinium); Naptumonmab estafenatox (Active Biotech); AFM 13 (Affimed); AFM11 (Affimed); AGS-16C3F (Agensys); AGS-16M8F (Agensys); AGS-22ME (Agensys); AGS-15ME (Agensys); GS-67E (Agensys); ALXN6000 (samalizumab, Alexion); ALT836 (Altor Bioscience); ALT-801 (Altor Bioscience); ALT-803 (Altor Bioscience); AMG780 (Amgen); AMG 228 (Amgen); AMG820 (Amgen); AMG172 (Amgen); AMG595 (Amgen); AMG110 (Amgen); AMG232 (adecatumumab, Amgen); AMG211 (Amgen/MedImmune); BAY2010112 (Amgen/Bayer); Rilotumumab (Amgen); Denosumab (Amgen); AMP-514 (Amgen); MEDI575 (AZ/Medlmmune); MEDI3617 (AZ/Medlmmune); MEDI6383 (AZ/Medlmmune); MEDI551 (AZ/Medlmmune); Moxetumomab pasudotox (AZ/Medlmmune); MEDI565 (AZ/Medlmmune); MED10639 (AZ/Medlmmune); MEDI0680 (AZ/Medlmmune); MEDI562 (AZ/Medlmmune); AV-380 (AVEO); AV203 (AVEO); AV299 (AVEO); BAY79-4620 (Bayer); Anetumab ravtansine (Bayer); vantictumab (Bayer); BAY94-9343 (Bayer); Sibrotuzumab (Boehringer Ingleheim); Bl-836845 (Boehringer Ingleheim); B-701 (BioClin); B1IB015 (Biogen);
Obinutuzumab (Biogen/Genentech); BI-505 (Bioinvent); BI-1206 (Bioinvent); TB-403 (Bioinvent); BT-062 (Biotest) BIL-OlOt (Biosceptre); MDX-1203 (BMS); MDX-1204 (BMS); Necitumumab (BMS); CAN-4 (Cantargia AB); CDX-Ol l (Celldex); CDX1401 (Celldex); CDX301 (Celldex); U31565 (Daiichi Sankyo); patritumab (Daiichi Sankyo); tigatuzumab (Daiichi Sankyo); nimotuzumab (Daiichi Sankyo); DS-8895 (Daiichi Sankyo); DS-8873 (Daiichi Sankyo); DS-5573 (Daiichi Sankyo); MORab-004 (Eisai); MORab-009 (Eisai); MORab-003 (Eisai); MORab-066 (Eisai); LY3012207 (Eli Lilly); LY2875358 (Eli Lilly); LY2812176 (Eli Lilly); LY3012217(Eli Lilly); LY2495655 (Eli Lilly); LY3012212 (Eli Lilly); LY30122H (Eli Lilly); LY3009806 (Eli Lilly); cixutumumab (Eli Lilly); Flanvotumab (Eli Lilly); IMC-TRl (Eli Lilly); Ramucirumab (Eli Lilly); Tabalumab (Eli Lilly); Zanolimumab (Emergent Biosolution); FG-3019 (FibroGen); FPA008 (Five Prime Therapeutics); FP-1039 (Five Prime Therapeutics); FPA144 (Five Prime Therapeutics); catumaxomab (Fresenius Biotech); 1MAB362 (Ganymed); IMAB027 (Ganymed); HuMax-CD74 (Genmab); HuMax-TFADC (Genmab); GS-5745 (Gilead); GS-6624 (Gilead); OMP-21M18 (demcizumab, GSK); mapatumumab (GSK); IMGN289 (ImmunoGen); IMGN901 (ImmunoGen); 1MGN853 (ImmunoGen); IMGN529 (ImmunoGen); IMMU-130 (Immunomedics); milatuzumabdox (Immunomedics); IMMU-115 (Immunomedics); IMMU-132 (Immunomedics); IMMU-106 (Immunomedics); IMMU-102 (Immunomedics); Epratuzumab (Immunomedics); Clivatuzumab (Immunomedics); IPH41 (Innate Immunotherapeutics); Daratumumab (Janssen/Genmab); CNTO-95 (Intetumumab, Janssen); CNTO-328 (siltuximab, Janssen); KB004 (KaloBios); mogamulizumab (Kyowa Hakko Kirrin); KW-2871 (ecromeximab, Life Science); Sonepcizumab (Lpath); Margetuximab (Macrogenics); Enoblituzumab (Macrogenics); MGD006 (Macrogenics); MGF007 (Macrogenics); MK-0646 (dalotuzumab, Merck); MK-3475 (Merck); Sym004 (Symphogen/Merck Serono); D117E6 (Merck Serono); MOR208 (Morphosys); MOR202 (Morphosys); Xmab5574 (Morphosys); BPC-1C (ensituximab, Précision Biologics); TAS266 (Novartis); LFA 102 (Novartis); BHQ880 (Novartis/Morphosys); QGE031 (Novartis); HCD122 (lucatumumab, Novartis); LJM716 (Novartis); AT355 (Novartis); OMP-21M18 (Demcizumab, OncoMed); OMP52M51 (Oncomed/GSK); OMP-59R5 (Oncomed/GSK); vantictumab (Oncomed/Bayer); CMC-544 (inotuzumab ozogamicin, Pfizer); PF-03446962 (Pfizer); PF-04856884 (Pfizer); PSMA-ADC (Progenics); REGN1400 (Regeneron); REGN910 (nesvacumab, Regeneron/Sanofi); REGN421 (enoticumab, Regeneron/Sanofi); RG722I, RG7356, RG7155, RG7444. RG7116, RG7458, RG7598, RG7599, RG7600, RG7636, RG7450, RG7593, RG7596, DCDS3410A, RG7414 (parsatuzumab), RG7160 (imgatuzumab), RG7159 (obintuzumab), RG7686, RG3638 (onartuzumab), RG7597 (Roche/Genentech); SAR307746 (Sanofi); SAR566658 (Sanofi); SAR650984 (Sanofi); SARI53192 (Sanofi); SAR3419 (Sanofi); SAR256212 (Sanofi), SGN-L1V1A (lintuzumab, Seattle Genetics); SGN-CD33A (Seattle Genetics); SGN-75 (vorsetuzumab mafodotin, Seattle Genetics); SGN-I9A (Seattle Genetics) SGN-CD70A (Seattle Genetics); SEA-CD40 (Seattle Genetics); ibritumomab tiuxetan (Spectrum); MLN0264 (Takeda); ganitumab (Takeda/Amgen); CEP-37250 (Teva); TB-403
I8 (Thrombogenic); VB4-845 (Viventia); Xmab25l2 (Xencor); Xmab5574 (Xencor); nimotuzumab (YM Biosciences); Carlumab (Janssen); NY-ESO TCR (Adaptimmune); MAGE-A-IO TCR (Adaptimmune); CTL019 (Novartis); JCAR015 (Juno Therapeutics); KTE-C19 CAR (Rite Pharma); UCART19 (Cellectis); BPX-401 (Bellicum Pharmaceuticals); BPX-601 (Bellicum Pharmaceuticals); ATTCK20 (Unum Therapeutics); CAR-NKG2D (Celyad); Onyx-015 (Onyx Pharmaceuticals); HIOI (Shanghai Sunwaybio); DNX-2401 (DNAtrix); VCN-Ol (VCN Biosciences); Colo-Adl (PsiOxus Therapeutics); ProstAtak (Advantagene); Oncos-102 (Oncos Therapeutics); CG0070 (Cold Genesys); Pexa-vac (JX-594, Jennerex Biotherapeutics); GL-ONCl (Genelux); T-VEC (Amgen); G207 (Medigene); HFIO (Takara Bio); SEPREHVIR (HSV1716, Virttu Biologics); OrienXOlO (OrienGene Biotechnology); Reolysin (Oncolytics Biotech); SW-001 (Neotropix); Cacatak (CVA2I, Viralytics); Alimta (Eli Lilly), cisplatin, oxaliplatin. irinotecan, folinic acid, methotrexate, cyclophosphamide, 5-fluorouracil, Zykadia (Novartis), Tafinlar (GSK), Xalkori (Pfizer), Iressa (AZ), Gilotrif (Boehringer Ingelheim), Tarceva (Astellas Pharma), Halaven (Eisai Pharma), Veliparib (Abbvie), AZD9291 (AZ), Alectinib (Chugai), LDK.378 (Novartis), Genetespib (Synta Pharma), Tergenpumatucel-L (NewLink Genetics), GVlOOl (Kael-GemVax), Tivantinib (ArQuIe); Cytoxan (BMS); Oncovin (Eli Lilly); Adriamycin (Pfizer); Gemzar (Eli Lilly); Xeloda (Roche); Ixempra (BMS); Abraxane (Celgene); Trelstar (Debiophami); Taxotere (Sanofi); Nexavar (Bayer); IMMU-132 (Immunomedics); E7449 (Eisai); Thermodox (Celsion); Cometriq (Exellxis); Lonsuri’ (Taiho Pharmaceuticals); Camptosar (Pfizer); UFT (Taiho Pharmaceuticals); and TS-l (Taiho Pharmaceuticals).
In some embodiments, the one or more Enterococcus faecium bacterial strains is/are the only therapeutically active agent(s) in a composition of the invention. In some embodiments, the bacterial strain(s) in the composition is/are the only therapeutically active agent(s) in a composition of the invention.
Asthma
In preferred embodiments, the compositions of the invention are for use in treating or preventing asthma. The compositions ofthe invention may achieve a réduction in the recruitment of neutrophils and/or eosinophils into the airways following sensitisation and challenge with house dust mite extract and so they may be useful in the treatment or prévention of asthma. Asthma is a chronic disease characterised by inflammation and restriction of the airways. The inflammation in asthma may be mediated by IL-17 and/or Thl7 cells, and so the compositions of the invention may be particulariy effective for preventing or treating asthma. The inflammation in asthma may be mediated by eosinophils and/or neutrophils.
In certain embodiments, the asthma is éosinophilie or allergie asthma. Eosinophilie and allergie asthma are characterised by increased numbers of eosinophils in peripheral blood and in airway sécrétions and is associated pathologically with thickening of the basement membrane zone and pharmacologically by corticosteroid responsiveness [46]. Compositions that reduce or inhibit eosinophil recruitment or activation may be useful for treating or preventing éosinophilie and allergie asthma.
In additional embodiments, the compositions of the invention are for use in treating or preventing neutrophilie asthma (or non-eosinophilic asthma). High neutrophil numbers are associated with severe asthma that may be insensitive to corticosteroid treatment. Compositions that reduce or inhibit neutrophil recruitment or activation may be useful for treating or preventing neutrophilie asthma.
Eosinophilie and neutrophilie asthma are not mutually exclusive conditions and treatments that help 10 address either the eosinophil and neutrophil responses may be useful for treating asthma în general.
Increased IL-l 7 levels and activation of the Thl7 pathway are associated with severe asthma, so the compositions of the invention may be useful for preventing the development of severe asthma or for treating severe asthma.
In certain embodiments, the compositions of the invention are for use in methods reducing an 15 éosinophilie inflammatory response in the treatment or prévention of asthma, or for use in methods of reducing a neutrophilie inflammatory response in the treatment or prévention of asthma. As noted above, high levels of eosinophils in asthma is associated pathologically with thickening of the basement membrane zone, so reducing éosinophilie inflammatory response in the treatment or prévention of asthma may be able to specifically address this feature of the disease. Also, elevated 20 neutrophils, either în combination with elevated eosinophils or in their absence, is associated with severe asthma and chronic airway narrowing. Therefore, reducing the neutrophilie inflammatory response may be particularly useful for addressing severe asthma.
In certain embodiments, the compositions reduce peribronchiolar infiltration in allergie asthma, or are for use in reducing peribronchiolar infiltration in the treatment of allergie asthma. In certain 25 embodiments, the compositions reduce peribronchiolar and/or perivascular infiltration in neutrophilie asthma, or are for use in reducing peribronchiolar and/or perivascular infiltration in the treatment of allergie neutrophilie asthma.
In certain embodiments, treatment with compositions of the invention provides a réduction or prevents an élévation in TNFa levels.
In certain embodiments, the compositions of the invention are for use in a method of treating asthma that results in a réduction of the éosinophilie and/or neutrophilie inflammatory response. In certain embodiments, the patient to be treated has, or has previously been identified as having, elevated neutrophil or eosinophil levels, for example as identified through blood sampling or sputum analysis.
The compositions of the invention may be useful for preventing the development of asthma in a newbom when administered to the new-bom, or to a prégnant woman. The compositions may be useful for preventing the development of asthma in children. The compositions of the invention may be useful for treating or preventing adult-onset asthma. The compositions of the invention may be useful for managing or alleviating asthma. The compositions of the invention may be particularly useful for reducing symptoms associated with asthma that is aggravated by allergens, such as house dust mites.
Treatment or prévention of asthma may refer to, for example, an alleviation of the severity of symptoms or a réduction in the frequency of exacerbations or the range of triggers that are a problem for the patient.
Arthritis
In preferred embodiments, the compositions of the invention are for use in treating or preventing rheumatoid arthritis (RA). The compositions of the invention may achieve a réduction in the clinîcal signs of RA in a mouse model, reduce cartilage and bone damage, and reduce the IL-17 inflammatory response, and so they may be useful in the treatment or prévention of RA. RA is a systemic inflammatory disorder that primarily affects joints. RA is associated with an Inflammatory response that results in swellîng of joints, synovial hyperplasia, and destruction of cartilage and bone. IL-17 and Thl7 cells may hâve a key rôle in RA, for example because IL-17 inhibits matrix production in chondrocytes and osteoblasts and activâtes the production and function of matrix metalloproteinases and because RA disease activity is correlated to IL-17 levels and Th-l7 cell numbers [47,48], so the compositions of the invention may be particularly effective for preventing or treating RA.
In certain embodiments, the compositions of the invention are for use in lowerîng IL-17 levels or preventing élévation of IL-17 levels in the treatment or prévention of RA. In certain embodiments, treatment with compositions of the invention provides a réduction or prevents an élévation in IL-I7 levels, in particular IL-17A levels. In certain embodiments, treatment with compositions of the invention provides a réduction or prevents an élévation in IFN-y or IL-6 levels.
In certain embodiments, treatment with the compositions of the invention results in a réduction in the swelling of joints. In certain embodiments, the compositions of the invention are for use in patients with swollen joints or patients identified as at rîsk of having swollen joints. In certain embodiments, the compositions of the invention are for use in a method of reducing joint swelling in RA.
In certain embodiments, treatment with the compositions of the invention results in a réduction in cartilage damage or bone damage. In certain embodiments, the compositions of the invention are for use in reducing or preventing cartilage or bone damage in the treatment of RA. In certain embodiments, the compositions are for use in treating patient with severe RA that are at risk of cartilage or bone damage.
Increased IL-17 levels and Thl7 cell numbers are associated with cartilage and bone destruction in RA [47,48]. IL-17 is known to activate matrix destruction in cartilage and bone tissue and IL-17 has an inhibitory effect on matrix production in chondrocytes and osteoblasts. Therefore, in certain embodiments. the compositions of the invention are for use in preventing bone érosion or cartilage damage in the treatment of RA. In certain embodiments, the compositions are for use in treating patients that exhibit bone érosion or cartilage damage or patients identified as at risk of bone érosion or cartilage damage.
TNF-α is also associated with RA, but TNF-α is not involved in the pathogenesis of the later stages of the disease. In contrast, IL-17 has a rôle throughout ail stages of chronic disease [49]. Therefore, in certain embodiments the compositions of the invention are for use in treating chronic RA or latestage RA, such as disease that includes joint destruction and loss of cartilage. In certain embodiments, the compositions of the invention are for treating patients that hâve previously received anti-TNF-α therapy. In certain embodiments, the patients to be treated do not respond or no longer respond to anti-TNF-α therapy.
The compositions of the invention may be useful for modulating a patient's immune system, so in certain embodiments the compositions of the invention are for use in preventing RA in a patient that has been identified as at risk of RA, or that has been diagnosed with early-stage RA. The compositions of the invention may be useful for preventing the development of RA.
The compositions of the invention may be useful for managing or alleviating RA. The compositions of the invention may be particularly useful for reducing symptoms associated with joint swelling or bone destruction. Treatment or prévention of RA may refer to, for example, an alleviation of the severity of symptoms or a réduction in the frequency of exacerbations or the range of triggers that are a problem for the patient.
Multiple sclerosis
In preferred embodiments, the compositions of the invention are for use in treating or preventing multiple sclerosis. The compositions of the invention may achieve a réduction in the disease incidence and disease severity in a mouse model of multiple sclerosis (the EAE model), and so they may be usefiil in the treatment or prévention of multiple sclerosis. Multiple sclerosis is an inflammatory disorder associated with damage to the myelin sheaths of neurons, particularly in the brain and spinal column. Multiple sclerosis is a chronic disease, which is progressively incapacitating and which evolves in épisodes. IL-17 and Thl7 cells may hâve a key rôle in multiple sclerosis, for example because IL-17 levels may correlate with multiple sclerosis lésions, IL-17 can disrupt blood brain barrier endothélial cell tight junctions, and Thl7 cells can migrate into the central nervous system and cause neuronal loss [50,51]. Therefore, the compositions of the invention may be particularly effective for preventing or treating multiple sclerosis.
In certain embodiments, treatment with the compositions of the invention results in a réduction in disease incidence or disease severity. In certain embodiments, the compositions of the invention are for use in reducing disease incidence or disease severity. In certain embodiments, treatment with the compositions of the invention prevents a décliné in motor function or results in improved motor function. In certain embodiments, the compositions of the invention are for use in preventing a décliné in motor function or for use in improving motor function. In certain embodiments, treatment with the compositions of the invention prevents the development of paralysis. In certain embodiments, the compositions of the invention are for use in preventing paralysis in the treatment of multiple sclerosis.
The compositions of the invention may be useful for modulating a patient's immune system, so in certain embodiments the compositions of the invention are for use in preventing multiple sclerosis in a patient that has been identified as at risk of multiple sclerosis, or that has been diagnosed with early-stage multiple sclerosis or “relapsing-remitting” multiple sclerosis. The compositions of the invention may be useful for preventing the development of sclerosis.
The compositions of the invention may be useful for managing or alleviating multiple sclerosis. The compositions of the invention may be particularly useful for reducing symptoms associated with multiple sclerosis. Treatment or prévention of multiple sclerosis may refer to, for example, an alleviation of the severity of symptoms or a réduction in the frequency of exacerbations or the range of triggers that are a problem for the patient.
Modes of administration
Preferably, the compositions of the invention are to be administered to the gastrointestinal tract in order to enable delivery to and / or partial or total colonisation of the intestine with the bacterial strain of the invention. Generally, the compositions of the invention are administered orally, but they may be administered rectally, intranasally, or via buccal or sublingual routes.
In certain embodiments, the compositions of the invention may be administered as a foam, as a spray or a gel.
In certain embodiments, the compositions of the invention may be administered as a suppository, such as a rectal suppository, for example in the form of a theobroma oil (cocoa butter), synthetic hard fat (e.g. suppocire, witepsol), glycero-gelatîn, polyethylene glycol, or soap glycerin composition.
In certain embodiments, the composition of the invention is administered to the gastrointestinal tract via a tube, such as a nasogastric tube, orogastric tube, gastric tube, jejunostomy tube (J tube), percutaneous endoscopie gastrostomy (PEG), or a port, such as a chest wall port that provides access to the stomach, jéjunum and other suitable access ports.
The compositions of the invention may be administered once, or they may be administered sequentîally as part of a treatment regimen. In certain embodiments, the compositions of the invention are to be administered daily.
In certain embodiments of the invention, treatment according to the invention is accompanied by assessment of the patient’s gut microbiota. Treatment may be repeated if delivery of and / or partial or total colonisation with the strain of the invention is not achieved such that efficacy is not observed, or treatment may be ceased if delivery and 1 or partial or total colonisation is successfiil and efficacy is observed.
In certain embodiments, the composition of the invention may be administered to a prégnant animal, for example a mammal such as a human in order to prevent an inflammatory or autoimmune disease developing in her child in utero and / or after it is bom.
The compositions of the invention may be administered to a patient that has been diagnosed with a disease or condition mediated by IL-17 or the Thl7 pathway, or that has been identified as being at risk of a disease or condition mediated by IL-17 or the Thl 7 pathway. The compositions may also be administered as a prophylactic measure to prevent the development of diseases or conditions mediated by IL-17 or the Thl 7 pathway in a healthy patient.
The compositions of the invention may be administered to a patient that has been identified as having an abnormal gut microbiota. For example, the patient may hâve reduced or absent colonisation by En terococcus faecitim.
The compositions of the invention may be administered as a food product, such as a nutritional supplément.
Generally, the compositions of the invention are for the treatment of humans, although they may be used to treat animais including monogastric mammals such as poultry, pigs, cats, dogs, horses or rabbits. The compositions of the invention may be useful for enhancing the growth and performance of animais. If administered to animais, oral gavage may be used.
Compositions
Generally, the composition of the invention comprises bacteria. In preferred embodiments of the invention, the composition is formulated in freeze-dried form. For example, the composition of the invention may comprise granules or gelatin capsules, for example hard gelatin capsules, comprising a bacterial strain of the invention.
Preferably, the composition of the invention comprises lyophilised bacteria. Lyophilisation of bacteria is a well-established procedure and relevant guidance is available in, for example, référencés [52-54],
Alternatively, the composition of the invention may comprise a live, active bacterial culture.
In some embodiments, the bacterial strain in the composition of the invention has not been inactivated, for example, has not been heat-inactivated. In some embodiments, the bacterial strain in the composition of the invention has not been killed, for example, has not been heat-killed. In some embodiments, the bacterial strain in the composition of the invention has not been attenuated, for example, has not been heat-attenuated. For example, in some embodiments, the bacterial strain in the composition of the invention has not been killed, inactivated and/or attenuated. For example, in some embodiments, the bacterial strain in the composition of the invention is live. For example, in some embodiments, the bacterial strain in the composition of the invention is viable. For example, in some embodiments, the bacterial strain in the composition of the invention is capable of partially or totally colonising the intestine. For example, in some embodiments, the bacterial strain in the composition of the invention is viable and capable of partially or totally colonising the intestine.
In some embodiments, the composition comprises a mixture of live bacterial strains and bacterial strains that hâve been killed.
In preferred embodiments, the composition of the invention is encapsulated to enable delivery of the bacterial strain to the intestine. Encapsulation protects the composition from dégradation until delivery at the target location through, for example, rupturing with Chemical or physical stimuli such as pressure, enzymatic activity, or physical disintegration, which may be triggered by changes in pH. Any appropriate encapsulation method may be used. Exemplary encapsulation techniques include entrapment within a porous matrix, attachment or adsorption on solid carrier surfaces, selfaggregation by flocculation or with cross-linking agents, and mechanical containment behind a microporous membrane or a microcapsule. Guidance on encapsulation that may be useful for preparing compositions of the invention is available in, for example, référencés [55] and [56].
The composition may be administered orally and may be in the form of a tablet, capsule or powder. Encapsulated products are preferred because Enterococcus faeciutn are anaerobes. Other ingrédients (such as vitamin C, for example), may be included as oxygen scavengers and prebiotic substrates to improve the delivery and / or partial or total colonisation and survival m vivo. Alternatively, the probiotic composition of the invention may be administered orally as a food or nutritional product. such as milk or whey based fermented dairy product, or as a pharmaceutical product.
The composition may be formulated as a probiotic.
A composition of the invention includes a therapeutically effective amount of a bacterial strain of the invention. A therapeutically effective amount of a bacterial strain is sufficient to exert a bénéficiai effect upon a patient. A therapeutically effective amount of a bacterial strain may be sufficient to resuit in delivery to and / or partial or total colonisation of the patient’s intestine.
A suitable daily dose of the bacteria, for example for an adult human, may be from about l x IO3 to about l x IO11 colony forming units (CFU); for example, from about l x IO7 to about l x 1O10 CFU; in another example from about 1 x 106 to about 1 x IO10 CFU.
In certain embodiments, the composition contains the bacterial strain in an amount of from about 1 x 106to about 1 x 10 CFU/g, respect to the weight of the composition; for example, from about 1 x 10s to about 1 x 1 Oto CFU/g. The dose may be, for example, 1 g, 3g, 5g, and 10g.
Typically, a probiotic, such as the composition of the invention, is optionally combined with at least one suitable prebiotic compound. A prebiotic compound is usually a non-digestible carbohydrate such as an oligo- or polysaccharide, or a sugar alcohol, which is not degraded or absorbed în the upper digestive tract. Known prebiotics include commercial products such as inulin and transgalactooligosaccharides.
In certain embodiments, the probiotic composition of the présent invention includes a prebiotic compound in an amount of from about 1 to about 30% by weight, respect to the total weight composition, (e.g. from 5 to 20% by weight). Carbohydrates may be selected from the group consisting of: fructo- oligosaccharides (or FOS), short-chain fructo-oligosaccharides, inulin, isomaltoligosaccharides, pectins, xylo-oligosaccharides (or XOS), chitosan-oligosaccharîdes (or COS), betaglucans, arable gum modified and résistant starches, polydextrose, D-tagatose, acacia fibers, carob, oats, and citrus fibers. In one aspect, the prebiotics are the short-chain fructo-oligosaccharides (for simplicity shown herein below as FOSs-c.c); said FOSs-c.c. are not digestible carbohydrates, generally obtained by the conversion of the beet sugar and including a saccharose molécule to which three glucose molécules are bonded.
The compositions of the invention may comprise pharmaceutically acceptable excipients or carriers. Examples of such suitable excipients may be found in the reference [57]. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical art and are described, for example, in reference [58], Examples of suitable carriers include lactose, starch, glucose, methyl cellulose, magnésium stéarate, mannitol, sorbîtol and the like. Examples of suitable diluents include éthanol, glycerol and water. The choice of pharmaceutical carrier, excipient or diluent can be selected with regard to the intended route of administration and standard pharmaceutical practice. The pharmaceutical compositions may comprise as, or in addition to, the carrier, excipient or diluent any suitable binder(s), lubricant(s), suspending agent(s), coating agent(s), solubilising agent(s). Examples of suitable binders include starch, gelatin, natural sugars such as glucose, anhydrous lactose, free flow lactose, beta-lactose, com sweeteners, natural and synthetic gums, such as acacia, tragacanth or sodium alginate, carboxymethyl cellulose and polyethylene glycol. Examples of suitable lubricants include sodium oleate, sodium stéarate, magnésium stéarate, sodium benzoate, sodium acetate, sodium chloride and the like. Preservatives, stabilizers, dyes and even flavouring agents may be provided in the pharmaceutical composition. Examples of preservatives include sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid. Antioxidants and suspending agents may be also used.
The compositions of the invention may be formulated as a food product. For example, a food product may provide nutritional benefît in addition to the therapeutic effect of the invention, such as in a nutritional supplément. Similarly, a food product may be formulated to enhance the taste of the composition of the invention or to make the composition more attractive to consume by being more similar to a common food item, rather than to a pharmaceutical composition. In certain embodiments, the composition of the invention is formulated as a milk-based product. The term milk-based product means any liquid or semi-solid milk- or whey- based product having a varying fat content. The milk-based product can be, e.g., cow's milk, goat's milk, sheep's milk, skimmed milk, whole milk, milk recombined from powdered milk and whey without any processing, or a processed product, such as yoghurt, curdled milk, curd, sour milk, sour whole milk, butter milk and other sour milk products. Another important group includes milk beverages, such as whey beverages, fermented milks, condensed milks, infant or baby milks; flavoured milks, ice cream; milk-containing food such as sweets.
In some embodiments, the compositions of the invention comprise one or more bacterial strains of the species Enterococcus faecium and do not contain bacteria from any other genus, or which comprise only de mininiis or biologically irrelevant amounts of bacteria from another genus. Thus, in some embodiments, the invention provides a composition comprising one or more bacterial strains of the species Enterococcus faecium, which does not contain bacteria from any other genus or which comprises only de mininiis or biologically irrelevant amounts of bacteria from another genus, for use in therapy.
In some embodiments, the compositions of the invention comprise one or more bacterial strains of the species Enterococcus faecium and do not contain bacteria from any other Enterococcus species, or which comprise only de mininiis or biologically irrelevant amounts of bacteria from another Enterococcus species. Thus, in some embodiments, the invention provides a composition comprising one or more bacterial strains of the species Enterococcus faecium., which does not contain bacteria from any other Enterococcus species or which comprises only de mininiis or biologically irrelevant amounts of bacteria from another Enterococcus species, for use in therapy.
In certain embodiments, the compositions of the invention contain a single bacterial strain or species and do not contain any other bacterial strains or species. Such compositions may comprise only de minimis or biologically irrelevant amounts of other bacterial strains or species. Such compositions may be a culture that is substantially free from other species of organism.
In some embodiments, the invention provides a composition comprising a single bacterial strain of the species Enterococcus faecium, which does not contain bacteria from any other strains or which comprises only de minimis or biologically îrrelevant amounts of bacteria from another strain for use in therapy.
In some embodiments, the compositions of the invention comprise more than one bacterial strain. For example, in some embodiments, the compositions of the invention comprise more than one strain from within the same species (e.g. more than l, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40 or 45 strains), and, optionally, do not contain bacteria from any other species. In some embodiments, the compositions of the invention comprise less than 50 strains from within the same species (e.g. less than 45, 40, 35, 30, 25, 20, 15, 12, 10, 9, 8, 7, 6, 5, 4 or 3 strains), and, optionally, do not contain bacteria from any other species. In some embodiments, the compositions of the invention comprise
1- 40, l-30, l-20, 1-19, 1-18, 1-15, l-ΊΟ, l-9, 1-8, l-7, l-6, l-5, l-4, l-3, 1-2,2-50, 2-40, 2-30, 2-20,
2- 15, 2-10, 2-5, 6-30, 6-15, 16-25, or 31-50 strains from within the same species and, optionally, do not contain bacteria from any other species. The invention comprises any combination of the foregoing.
In some embodiments, the composition comprises a microbial consortium. For example, in some embodiments, the composition comprises the Enterococcus faecium bacterial strain as part of a microbial consortium. For example, in some embodiments, the Enterococcus faecium bacterial strain is present in combination with one or more (e.g. at least 2, 3, 4, 5, 10, 15 or 20) other bacterial strains from other généra with which it can live symbiotically in vivo in the intestine. For example, in some embodiments, the composition comprises a bacterial strain of Enterococcus faecium in combination with a bacterial strain from a different genus. In some embodiments, the microbial consortium comprises two or more bacterial strains obtained from a faeces sample of a single organism, e.g. a human. In some embodiments, the microbial consortium is not found together in nature. For example, in some embodiments, the microbial consortium comprises bacterial strains obtained from faeces samples of at least two different organisms. In some embodiments, the two different organisms are from the same species, e.g. two different humans. In some embodiments, the two different organisms are an infant human and an adult human. In some embodiments, the two different organisms are a human and a non-human mammal.
In some embodiments, the composition of the invention additionally comprises a bacterial strain that has the same safety and therapeutic efficacy characteristics as strain MRX010, but which is not MRX010 deposited as NCIMB 42487, or which is not a Enterococcus faecium.
In some embodiments, the composition of the invention does not additionally comprise Bacilhis subtilis and/or Bacilhis coagidans.
In some embodiments in which the composition of the invention comprises more than one bacterial strain, species or genus, the individual bacterial strains, species or généra may be for separate, simultaneous or sequential administration. For example, the composition may comprise ail of the more than one bacterial strain, species or généra, or the bacterial strains, species or généra may be stored separately and be adminîstered separately, simultaneously or sequentially. In some embodiments, the more than one bacterial strains, species or généra are stored separately but are mixed together prior to use.
In some embodiments, the bacterial strain for use in the invention is obtained from human adult faeces. In some embodiments in which the composition of the invention comprises more than one bacterial strain, ail of the bacterial strains are obtained from human adult faeces or if other bacterial strains are present they are present only in de minimis amounts. The bacteria may hâve been cultured subséquent to being obtained from the human adult faeces and being used în a composition of the invention.
As mentioned above, in some embodiments, the one or more Enterococcus faecium bacterial strains is/are the only therapeutically active agent(s) in a composition of the invention. In some embodiments, the bacterial strain(s) in the composition is/are the only therapeutically active agent(s) in a composition of the invention.
The compositions for use in accordance with the invention may or may not require marketing approval.
In certain embodiments, the invention provides the above pharmaceutical composition, wherein said bacterial strain is lyophilised. In certain embodiments, the invention provides the above pharmaceutical composition, wherein said bacterial strain is spray dried. In certain embodiments, the invention provides the above pharmaceutical composition, wherein the bacterial strain is lyophilised or spray dried and wherein it is live. In certain embodiments, the invention provides the above pharmaceutical composition, wherein the bacterial strain is lyophilised or spray dried and wherein it is viable. In certain embodiments, the invention provides the above pharmaceutical composition, wherein the bacterial strain is lyophilised or spray dried and wherein it is capable of partially or totally colonising the intestine. In certain embodiments, the invention provides the above pharmaceutical composition, wherein the bacterial strain is lyophilised or spray dried and wherein it is viable and capable of partially or totally colonising the intestine.
In some cases, the lyophilised or spray dried bacterial strain is reconstituted prior to administration. In some cases, the reconstitution is by use of a diluent described herein.
The compositions of the invention can comprise pharmaceutically acceptable excipients, diluents or carriers.
In certain embodiments, the invention provides a pharmaceutical composition comprising: a bacterial strain as used in the invention; and a pharmaceutically acceptable excipient, carrier or diluent; wherein the bacterial strain is in an amount sufficient to treat a disorder when administered to a subject in need thereof; and wherein the disorder is selected from the group consisting of uveitis; cancer, such as breast cancer, lung cancer, liver cancer, colon cancer, or ovarian cancer; multiple sclerosis; arthritis, such as rheumatoid arthritis, osteoarthritis, psoriatic arthritis, or juvénile idiopathîc arthritis; neuromyelitis optica (Devic's disease); ankylosing spondylilis; spondyloarthritis; psoriasis; systemic lupus erythematosus; inflammatory bowel disease, such as Crohn’s disease or ulcerative colitis; celiac disease; asthma, such as allergie asthma or neutrophilie asthma; chronic obstructive pulmonary disease (COPD); scleritis; vasculitis; Behcet's disease; atherosclerosis; atopie dermatitis; emphysema; periodontîtîs; allergie rhinitis; and allograft rejection.
In certain embodiments, the invention provides pharmaceutical composition comprising: a bacterial strain as used in the invention; and a pharmaceutically acceptable excipient, carrier or diluent; wherein the bacterial strain is in an amount sufficient to treat or prevent a disease or condition mediated by IL-17 or the Thl7 pathway. In preferred embodiments, said disease or condition is selected from the group consisting of uveitis; cancer, such as breast cancer, lung cancer, liver cancer, colon cancer, or ovarian cancer; multiple sclerosis; arthritis, such as rheumatoid arthritis, osteoarthritis, psoriatic arthritis, or juvénile idiopathîc arthritis; neuromyelitis optica (Devic's disease); ankylosing spondylitis; spondyloarthritis; psoriasis; systemic lupus erythematosus; inflammatory bowel disease, such as Crohn’s disease or ulcerative colitis; celiac disease; asthma, such as allergie asthma or neutrophilie asthma; chronic obstructive pulmonary disease (COPD); scleritis; vasculitis; Behcet's disease; atherosclerosis; atopie dermatitis; emphysema; periodontîtîs; allergie rhinitis; and allograft rejection.
In certain embodiments, the invention provides the above pharmaceutical composition, wherein the amount of the bacterial strain is from about l χ IO3 to about l * IO11 colony forming units per grain with respect to a weight of the composition.
In certain embodiments. the invention provides the above pharmaceutical composition, wherein the composition is administered at a dose of l g, 3 g, 5 g or 10 g.
In certain embodiments, the invention provides the above pharmaceutical composition, wherein the composition is administered by a method selected from the group consisting of oral, rectal, subcutaneous, nasal, buccal, and sublingual.
In certain embodiments, the invention provides the above pharmaceutical composition, comprising a carrier selected from the group consisting of lactose, starch, glucose, methyl cellulose, magnésium stéarate, mannitol and sorbitol.
In certain embodiments, the invention provides the above pharmaceutical composition, comprising a diluent selected from the group consisting of éthanol, glycerol and water.
In certain embodiments, the invention provides the above pharmaceutical composition, comprising an excipient selected from the group consisting of starch, gelatin, glucose, anhydrous lactose, freeflow lactose, beta-lactose, corn sweetener, acacia, tragacanth, sodium alginate, carboxymethyl cellulose, polyethylene glycol, sodium oleate, sodium stéarate, magnésium stéarate, sodium benzoate, sodium acetate and sodium chloride.
In certain embodiments, the invention provides the above pharmaceutical composition, further comprising at least one of a preservative, an antioxidant and a stabilizer.
In certain embodiments, the invention provides the above pharmaceutical composition, comprising a preservative selected from the group consisting of sodium benzoate, sorbic acid and esters of phydroxybenzoic acid.
In certain embodiments, the invention provides the above pharmaceutical composition, wherein when the composition is stored in a sealed container at about 4°C or about 25°C and the container is placed in an atmosphère having 50% relative humidity, at least 80% of the bacterial strain as measured in colony forming units, remains after a period of at least about: l month. 3 months, 6 months, l year, l .5 years, 2 years, 2.5 years or 3 years.
In some embodiments, the composition of the invention is provided in a sealed container comprising a composition as described herein. In some embodiments, the sealed container is a sachet or bottle. In some embodiments, the composition of the invention is provided in a syringe comprising a composition as described herein.
The composition of the present invention may, in some embodiments, be provided as a pharmaceutical formulation. For example, the composition may be provided as a tablet or capsule. In some embodiments, the capsule is a gélatine capsule (“gel-cap”).
In some embodiments, the compositions of the invention are administered orally. Oral administration may involve swallowing, so that the compound enters the gastrointestinal tract, and/or buccal, lingual, or sublingual administration by which the compound enters the blood stream directly from the mouth.
Pharmaceutical formulations suitable for oral administration include solid plugs, solid microparticulates, semi-solid and liquid (including multiple phases or dispersed Systems) such as tablets; soft or hard capsules containing multi- or nano-particulates, liquids (e.g. aqueous solutions), émulsions or powders; lozenges (including liquid-filled); chews; gels; fast dispersing dosage forms; films; ovules; sprays; and buccal/mucoadhesive patches.
In some embodiments the pharmaceutical formulation is an enteric formulation, i.e. a gastro-résistant formulation (for example, résistant to gastric pH) that is suitable for delivery of the composition of the invention to the intestine by oral administration. Enteric formulations may be particularly useful when the bacteria or another component of the composition is acid-sensitive, e.g. prône to dégradation under gastric conditions.
In some embodiments, the enteric formulation comprises an enteric coating. In some embodiments, the formulation is an enteric-coated dosage form. For example, the formulation may be an entericcoated tablet or an enteric-coated capsule, or the like. The enteric coating may be a conventional enteric coating, for example, a conventional coating for a tablet, capsule, or the like for oral delivery. The formulation may comprise a film coating, for example, a thin film layer of an enteric polymer, e.g. an acid-insoluble polymer.
In some embodiments, the enteric formulation is intrînsically enteric, for example, gastro-résistant without the need for an enteric coating. Thus, in some embodiments, the formulation is an enteric formulation that does not comprise an enteric coating. In some embodiments, the formulation is a capsule made from a thermogelling material. In some embodiments, the thermogelling material is a cellulosic material, such as methylcellulose, hydroxymethylcellulose or hydroxypropylmethylcellulose (HPMC). In some embodiments, the capsule comprises a shell that does not contain any film forming polymer. In some embodiments, the capsule comprises a shell and the shell comprises hydroxypropylmethylcellulose and does not comprise any film forming polymer (e.g. see [59 ]). In some embodiments, the formulation is an intrînsically enteric capsule (for example, Vcaps® from Capsugel).
In some embodiments, the formulation is a soft capsule. Soft capsules are capsules which may, owing to additions of softeners, such as, for example, glycerol, sorbitol, maltitol and polyethylene glycols, present in the capsule shell, hâve a certain elasticity and softness. Soft capsules can be produced. for example, on the basis of gélatine or starch. Gelatine-based soft capsules are commercially available from various suppliers. Depending on the method of administration, such as, for example, orally or rectally, soft capsules can hâve various shapes, they can be, for example, round, oval, oblong or torpédo-shaped. Soft capsules can be produced by conventional processes, such as, for example, by the Scherer process, the Accogel process or the droplet or blowing process.
Culturing methods
The bacterial strains for use in the présent invention can be cultured using standard microbiology techniques as detailed in, for example, references [60-62].
The solid or liquid medium used for culture may be YCFA agar or YCFA medium. YCFA medium may include (per lOOml, approximate values): Casitone (l.O g), yeast extract (0.25 g), NaHCOj (0.4 g), cysteine (0.1 g), K2HPO4 (0.045 g), KH2PO4 (0.045 g), NaCl (0.09 g), (NH4)2SO4 (0.09 g), MgSO4 · 7H2O (0.009 g), CaCl2 (0.009 g), resazurin (0.1 mg), hemin (l mg), biotin (l pg), cobalamin (I pg).p-aminobenzoic acid (3 pg), folie acid (5 pg), and pyridoxamine (15 pg).
Bacterial strains for use in vaccine compositions
The inventors hâve identified that the bacterial strains of the invention are useful for treating or preventing diseases or conditions mediated by IL-l 7 or the Thl7 pathway. This is likely to be a resuit of the effect that the bacterial strains of the invention hâve on the host immune system. Therefore, the compositions of the invention may also be useful for preventing diseases or conditions mediated by IL-17 or the Thl7 pathway, when administered as vaccine compositions. In certain such embodiments, the bacterial strains of the invention are viable. In certain such embodiments, the bacterial strains of the invention are capable of partially or totally colonising the intestine. In certain such embodiments, the bacterial strains of the invention are viable and capable of partially or totally colonising the intestine. In other certain such embodiments, the bacterial strains of the invention may be killed, inactivated or attenuated. In certain such embodiments, the compositions may comprise a vaccine adjuvant. In certain embodiments, the compositions are for administration via injection, such as via subeutaneous injection.
General
The practice of the présent invention will employ, unless otherwise indicated, conventional methods of chemistry, biochemistry, molecular biology, immunology and pharmacology, within the skill of the art. Such techniques are explained fully in the literature. See, e.g., references [63] and [64-70], etc.
The term “comprising” encompasses “including as well as consisting e.g. a composition comprising X may consist exclusively of X or may include something additional e.g. X + Y.
The terni “about” in relation to a numerical value x is optional and means, for example, ,v+!0%.
The word “substantially” does not exclude “completely” e.g. a composition which is “substantially free” from Y may be completely free from Y. Where necessary, the word “substantially” may be omitted from the définition of the invention.
Référencés to a percentage sequence identity between two nucléotide sequences means that, when aligned, that percentage of nucléotides are the same in comparing the two sequences. This alignment and the percent homology or sequence identity can be determined using software programs known in the art, for example those described in section 7.7.18 of ref. [71], A preferred alignment is 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, BLOSUM matrix of 62. The SmithWaterman homology search algorithm is disclosed in ref [72].
Unless specifically stated, a process or method comprising numerous steps may comprise additional steps at the beginning or end of the method, or may comprise additional intervening steps. Also, steps may be combined, omitted or performed in an alternative order, if appropriate.
Various embodiments of the invention are described herein. It will be appreciated that the features specified in each embodiment may be combined with other specified features, to provide further embodiments. In particular, embodiments highlighted herein as being suitable, typical or preferred may be combined with each other (except when they are mutually exclusive).
MODES FOR CARRYING OUT THE INVENTION
Example 1 - Efficacy of bacterial inocula in a mouse model of uveitis
Sitmmary
This study used a mouse model of interphotoreceptor retinoid-binding protein (IRBP)-induced uveitis to test the effects of bacterial administration on uveitis. Uveitis is a sight-threatening condition resulting from intraocular inflammation and retînal tissue destruction. This disease can be studied in rodents in a model of experimental autoimmune uveoretinitis (EAU) [73]. EAU is an organ-specific disorder where Thl/Thl7 cells are directed toward retinal antigens and produce cytokines that actîvate résident and infiltrating mononuclear cells leading to tissue destruction. EAU can be induced in mice by challenge with retinal antigens including interphotoreceptor retinoid btnding protein peptide (IRBPp). Disease onset normally occurs from day 8-9 and peaks after days 14-15. Signs of clinical disease can be monitored using topical endoscopie fiindal imaging (TEF1).
Strain
MRX010: Enterococcus faecium, bacteria deposited under accession number NCIMB 42487.
Biotherapeutic was provided in glycerol stock. Microbiological growth media (YCFA) was used for the culture of the agent.
Mice
The mice were strain C57BL/6 and were over 6 weeks old at the beginning of the study. 72 mice were used (+ 36 Satellite animais). Unhealthy animais were excluded from the study. Animais were housed in spécifie pathogen free (spf) conditions, in a thermostatically monitored holding room (22 ± 4°C). Animais were allowed to acclimatise under standard animal house conditions for a minimum of one week prior to use. The health status of the animais was monitored throughout this period and the suitability of each animal for experimental use was assessed prior to study start. Mice were housed in groups of up to 10 animais per cage for the duration of the study. Irradiated pellet diet (Lab diet, EU Rodent diet 22%, 5LF5) and water were available ad libitum throughout the acclimatisation and study periods. It is unlikely that any constituent of the diet or water interfered with the study.
Experimental Outline
Adult female C57BL/6 mice were randomly allocated to experimental groups and allowed to acclimatise for one week. Treatments were administered according to the schedule below. On Day 0, animais were administered with an émulsion containing 200 pg of interphotoreceptor retinoid binding protein peptide l-20 (IRBP pl-20) in complété Freund’s adjuvant (CFA) supplemented with
2.5 mg/ml Mycobacterium Tuberculosis H37 Ra by subeutaneous injection. Also on Day 0, animais were administered with l .5 pg Bordetella Pertussis toxin by intra-peritoneal injection. From Day 14, animais are weighed three times per week, From Day -1 until the end of the experiment on Day 42, animais are monitored twice per week for clinical signs of uveitis using topical endoscopie fundal imaging (TEFI).
Administration schedule
Ail Groups are n=12
Vehicle for oral administration is YCFA medium.
Administration volume for twice daily oral administration is 5 ml/kg.
Group Treatment Dose Route Frequency Disease Induction
I Tehicle 5 ml/kg PO BID Day -14-End Day 0: IRBP/CFA, SC Day 0: PTx, IP
2 MRX010 5 ml/kg
PO: oral administration, BID: twice daily, SC: subeutaneous injection, IP: intra-peritoneal injection. IRBP: interphotoreceptor binding protein. CFA: complété Freund's adjuvant, PTx: Pertussis toxin
A positive control group was also tested using treatment with the drug cyclosporin A.
Readouts
Bodyweights. From Day -14, animais are weighed three times a week. Animais with a bodyweight loss equal to or greater than 15% of their initial (Day 0) bodyweight on two consecutive occasions are culled.
Non-specific clinical observations. From Day -14 until the end of the experiment, animais are checked daily for non-specific clinical signs to include abnormal posture (hunched), abnormal coat 5 condition (piloerection) and abnormal activîty levels (reduced or increased activity).
Clinical Scores: Retinal imaging by topical endoscopie fundal imaging (TEFI). From Day -l until the end of the experiment, animais are scored twice per week for clinical signs of uveitis. Retinal images are captured using TEFI in non-anaesthetised but restrained animais following pupil dilatation using Tropicamide I% then Phenylephrine hydrochloride 2.5%. Retinal images are scores I0 using the following system. The maximum cumulative score is 20.
Score Optic dise Inflammation Retinal vessels Retinal tissue Infiltration Structural damage
1 Minimal 1-4 mild euffings 1-4 small lésions or 1 linear lésion Retinal lésions or atrophy involving 14 to % of retinal area
2 Mild >4 mild euffings or 1 -3 moderate euffings 5-10 small lésions or 2-3 linear lésions Panretinal atrophy with multiple small lésions (scars) or <3 linear lésions (scars)
3 Moderate >3 moderate euffings >10 small lésions or >3 linear lésions Panretinal atrophy with >3 linear lésions or confluent lésions (scars)
4 Severe >1 severe euffings Linear lésion confluent Retinal detachment with fblding
5 Not visible (white-out or severe detachment)
Results
The results of the study are shown in Figures l -3.
Cell Prolifération (satellite animais, day 21). Draining lymph nodes (DLN) were removed and cells were isolated. After counting, cells were cultured for 72h in the presence or absence of IRBP 15 peptide. Supematants for cytokine analysis were removed prior to pulsing with 3H-Thymidine.
Cells were then cultured for a further 18h prior to harvesting and détermination of prolifération by incorporation of 3 H-thymidine using a beta-counter.
The groups showed cell prolifération to IRBP stimulus above that seen in the media control wells (these background values were subtracted from the IRBP resuit to provide data as Acpm). Both groups gave strong proliférative responses to a positive control stimulus (anti-CD3/CD28) showing cells in culture were viable and able to proliferate (data not shown).
Proliférative responses to IRBP peptide were analysed by one-way ANOVA followed by Dunnett’s post-test for comparison of stimulated responses in the experimental group with the control group.
Proliférative responses to IRBP peptide in disease control animais were of the magnitude expected for this model. The positive control drug, Cyclosporin A, reduced prolifération, although this réduction did not achieve statîstical significance (Figure l).
The treatment groups (including vehicle alone) showed non-significantly increased prolifération above that seen in control animais.
Proliférative responses to IRBP peptide were analysed by one-way ANOVA followed by Dunnett’s post-test for comparison of stimulated responses in treatment groups with the vehicle group. No statîstical différences were seen.
Clinicai Scores: Retinal imagîng by topical endoscopie fundal imaging (TEFI). TEFI scores data measured in the Control group from Day 0 until Day 2I were analysed by Kruskal-Wallis test for non-parametric data followed by Dunn’s post-test for multiple comparisons between experimental days.
IRBP administration induced a significant increase in the TEFI scores measured from Day 14 (p < 0.01 ) and on Day 21 (p < 0.0001 ) when compared to Day 0 in the Control group (Figure 2).
TEFI scores data measured in ail experimental groups on Day 21 were analysed by Kruskal-Wallis test for non-parametric data followed by Dunn’s post-test for multiple comparisons between experimental groups.
At this stage in the experiment, there was no significant différence between experimental groups, but TEFI scores were lower in the MRX010 treated group than in the négative control group. Indeed, TEFI scores in the MRX010 treated group were comparable to the positive control. cyclosporin A treated group.
Conclusions. Proliférative responses to IRBP peptide were seen in lymph node cultures from ail experimental groups, excluding naïve animais, indicating successful disease induction. Clinicai scores determined by TEFI increased from Day 14, as expected in this model of IRBP-induced uveitis. By Day 21, significant différences between experimental groups are not yet visible, but a striking réduction in disease incidence and disease severity was observed in the MRX010 treated group, which was a comparable réduction to that seen for the positive control group. In particular, these data indicate that treatment with the strain MRX010 reduced retinal damage, optic dise inflammation and/or retinal tissue infiltration by inflammatory cells (see TEFI retinal image scoring system above). These data indicate the strain MRX010 may be useful for treating or preventing uveitis.
Example 2 - Stability testing
A composition described herein containing at least one bacteriai strain described herein is stored in a sealed container at 25 C or 4°C and the container is placed in an atmosphère having 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90% or 95% relative humidity. After l month, 2 months, 3 months, 6 months, l year, 1.5 years, 2 years, 2.5 years or 3 years, at least 50%, 60%, 70%, 80% or 90% of the bacteriai strain shall remain as measured in colony forming units determined by standard protocols.
Example 3 - Characterisation of enzymatic activity
Summaiy
The Analytical Profile Index (API®) test System consists of strips that contain miniaturised biochemical tests that assay for enzymatic activity in bacteriai species. MRX010 (the bacterium deposited under accession number NCIMB 42487) was characterised using Rapid ID 32A - This system is designed specifically for anaérobie species and encompasses tests for carbohydrate, amîno acid and nitrate nietabolism as well as alkaline phosphatase activity.
Rapid ID 32A testing was carried out on bacteriai colonies as per manufacturées instructions. Briefly, bacteria were cultured on YCFA agar for 24 hours at 37 °C in an anaérobie workstation. Colonies were removed from plates using a stérile 5 pl inoculating loop and resuspended in a 2 ml ampoule of API® Suspension Medium until a density roughly équivalent to that of McFarland standard No. 4 was achieved. Fifty-five microlitres of bacteriai suspension was added to each cupule on a Rapid ID 32A strip, and the urease test was overlayed with two drops of minerai oil. Strips were covered with a plastic lid and incubated aerobically at 37 °C for 4 hours, following which the bottom row of cupules were developed using the following reagents: NIT: 1 drop each of NIT1 and NIT2; IND: 1 drop of James reagent; ail remaining cupules: 1 drop of FastBlue reagent. Strips were incubated at room température for 5 minutes, following which the colour of each cupule was recorded and assigned a value of négative, intermediate positive or positive.
Resulls
The results of the Rapid ID 32A analysis are shown in Figure 4. MRX010 tested positive for arginine dihydrolase, β-glucosidase, mannose fermentation, glutamic acid decarboxylase, arginine arylamidase, phenylalanine arylamidase, leucine arylamidase, pyroglutamic acid arylamidase, tyrosine arylamidase, glycine arylamidase, histidine arylamidase and serine arylamidase. MRX010 showed an inteimediate reaction for N-acetyl-P-glucosaminidase.
Sequences
SEQ ID NO: l (Enterococcusfaecium 16S rRNA gene, strain LMG 11423 - AJ301830)
1 agagtttgat cctggctcag gacgaacgct ggcggcgtgc ctatacatgc aagtcgaacg
61 cttctttttc caccggagct tgctccaccg gaaaaagagg agtggcgaac gggtgagtaa
121 cacgtgggta acctgcccat cagaaaggga taacacttgg aaacaggtgc taataccgta
181 taacaaatca aaaccgcatg gtt ttgatt t gaaaggcgct ttcgggtgtc gctgatggat
241 ggacccgcgg tgcattagct agttggtgag gùaacggctc accaaggcca cgatgcatag
301 ccgcacctga gagggtgatc ggccacattg ggactgagac acggcccaaa ctctacggga
361 ggcagcagta gggaatcttc ggcaatggac gaaagtctga ccgagcaacg ccgcgtgagt
421 gaagaaggt t t tcggatcgt aaaactctgt tgttagagaa gaacaaggat gagagtaact
481 gttcatccct tgacggtatc taaccagaaa gccacggcta actacgtgcc agcagccgcg
541 gtaatacgta ggtggcaagc gttgtccgga ttcactgggc gtaaagcgag cgcaggcggt
601 tcttaagtct gatgtgaaag cccccggctc aaccggggag ggtcattgga aactgggaga
661 cttgagtgca gaagaggaga gtggaattcc atgtgtagcg gtgaaatgcg tagatatatg
721 gaggaacacc agtggcgaag gcggctctct ggtctgtaac tgacgctgag gctcgaaagc
781 gtggggagca aacaggatta gataccctgg tagtccacgc cgtaaacgat gagtgctaag
841 tgttggaggg t ttccgccct tcagtgctgc agctaacgca t taagcactc cgcctgggga
901 gtacgaccgc aaggt tgaaa ctcaaaggaa ttgacggggg cccgcacaag cggtggagca
961 tgtggtt taa ttcgaagcaa cacgaagaac cttaccaggt cttgacatcc tttgaccact
1021 ctagagatag agcttcccct tcgggggcaa agtgacaggt ggtgcarggt tgtcgtcagc
1081 tcgtgtcgtg agatgttggg ttaagtcccg caacgagcgc aaccct tact gttagttgcc
1141 atcatccagt tgggcactct agcaagactg ccggtgacaa accggaggaa ggtggggatg
1201 acgtcaaatc atcatgcccc ttatgacctg ggctacacac gtgctacaat gggaagtaca
1261 acgagttgcg aagtcgcgag gctaagctaa tctcttaaag ct tctctcag t tcggat tgc
1321 aggctgcaac tcgcctgcat gaagccggaa tcgctagtaa tcgcggatca gcacgccgcg
1381 tgaatacgtt cccgggcctC gtacacaccg cccgtcacac cacgagagt t tgtaacaccc
1441 gaagtcggtg aggtaacctt ttggagccag ccgcctaagg tgggatagat gattggggtg
1501 aagtcgtaac aaggtagccg tatctgaagg tgcggctgga tcacctcctt tctaaggaat
1561 attacggata ctacacactt tttttttact tttttcattt tgaatttact ctcaaacact
1621 cctgttcatt gacactgcat atctgaagta t
SEQ ID NO:2 (consensus 16S rRNA sequence for Enterococcus faecium strain MRX010)
TTAGGCGGCTGGCTCCAAAAGGTTACCTCACCGACTTCGGGTGTTACAAACTCTCGTGGTGTGACGGGCGGTGTGTA
CAAGGCCCGGGAACGTATTCACCGCGGCGTGCTGATCCGCGATTACTAGCGATTCCGGCTTCATGCAGGCGAGTTGC
AGCCTGCAATCCGAACTGAGAGAAGCTTTAAGAGATTAGCTTAGCCTCGCGACTTCGCAACTCGTTGTACTTCCCAT
TGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCG
GCAGTCTTGCTAGAGTGCCCAACTGAATGATGGCAACTAACAATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAAC
ATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTCACTTTGCCCCCGAAGGGGAAGCTCTATCTCTAGAGT
GGTCAAAGGATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGG
CCCCCGTCAATTCCTTTGAGTTTCAACCTTGCGGTCGTACTCCCCAGGCGGAGTGCTTAATGCGTTAGCTGCAGCAC
TGAAGGGCGGAAACCCTCCAACACTTAGCACTCATCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTTGCT
CCCCACGCTTTCGAGCCTCAGCGTCAGTTACAGACCAGAGAGCCGCCTTCGCCACTGGTGTTCCTCCATATATCTAC
GCATTTCACCGCTACACATGGAATTCCACTCTCCTCTTCTGCACTCAAGTCTCCCAGTTTCCAATGACCCTCCCCGG
TTGAGCCGGGGGCTTTCACATCAGACTTAAGAAACCGCCTGCGCTCGCTTTACGCCCAATAAATCCGGACAACGCTT
GCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGATACCGTCAAGGGATGAACA
GTTACTCTCATCCTTGTTCTTCTCTAACAACAGAGTTTTACGATCCGAAAACCTTCTTCACTCACGCGGCGTTGCTC
GGTCAGACTTTCGTCCATTGCCGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTTTGGGCCGTGTCTCAGTCCCAAT
GTGGCCGATCACCCTCTCAGGTCGGCTATGCATCGTGGCCTTGGTGAGCCGTTACCTCACCAACTAGCTAATGCACC
GCGGGTCCATCCATCAGCGACACCCGAAAGCGCCTTTCAAATCAAAACCATGCGGTTTTGATTGTTATACGGTATTA
GCACCTGTTTCCAAGTGTTATCCCCTTCTGATGGGCAGGTTACCCACGTGTTACTCACCCGTTCGCCACTCCTCTTT
TTCCGGTGGAGCAAGCTCCGGTGGAAAAAGAAGCGTTCGACTGCA
SEQ ID NO:3 (chromosome sequence of Enterococcusfaecium strain DO) - see electronic sequence listing.
SEQ ID NO:4 (plasmid l sequence of Enterococcusfaecium strain DO) - see electronic sequence listing.
SEQ ID NO:5 (plasmid 2 sequence of Enterococcusfaecium strain DO) - see electronic sequence listing.
SEQ ID NO:6 (plasmid 3 sequence of Enterococcus faecium strain DO) - see electronic sequence listing.
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Claims (25)

1. A composition comprising a bacterial strain of the species Enterococcus faeciiim, for use in a method of treating or preventing an inflammatory or autoimmune disease, wherein the composition does not comprise a bacterial strain of the species Bacillus subiilis.
2. The composition of claim l, wherein the composition is for use in a method of treating or preventing an inflammatory or autoimmune disease selected from the group consisting of: uveitis; multiple sclerosis; arthrîtis, including rheumatoid arthritis, osteoarthritis, psoriatic arthritis, or juvénile idiopathic arthritis; neuromyelitis optica (Devic's disease); ankylosing spondylitis; spondyloarthritis; psoriasis; systemic lupus erythematosus; inflammatory bowel disease, including Crohn’s disease or ulcerative colitis; celiac disease; asthma, including allergie asthma or neutrophilie asthma; chronic obstructive pulmonary disease (COPD); scleritis; vasculitis; Behcet's disease; atherosclerosis; atopie dermatitis; emphysema; periodontitis; allergie rhinitis; and allograft rejection.
3. The composition of claim 2, wherein the composition is for use in a method of treating or preventing uveitis.
4. The composition of claim 3, wherein the composition is for use in a method of reducing or preventing retinal damage in uveitis.
5. The composition of claim 2, wherein the composition is for use in in a method of treating or preventing asthma, including neutrophilie asthma or allergie asthma.
6. The composition of claim 5, wherein the composition is for use in a method of reducing neutrophilia or eosinophilia in the treatment of asthma.
7. The composition of claim 2, wherein the composition is for use in in a method of treating or preventing rheumatoid arthritis.
8. The composition of claim 7, wherein the composition is ibr use in a method of reducing joint swelling in rheumatoid arthritis.
9. The composition of claim 2, wherein the composition is for use in in a method of treating or preventing multiple sclerosis.
10. The composition of claim 9, wherein the composition is for use in a method of reducing disease incidence or disease severity.
11. The composition of any preceding claim, wherein the composition is for use in a method of reducing IL-17 production or reducing Thl7 cell différentiation in the treatment or prévention of the inflammatory or autoimmune disease mediated by IL-17 or the Thl7 pathway.
12. The composition of any preceding claim, wherein the composition is for use in a patient with elevated IL-17 levels or Thl 7 cells.
13. The composition of any preceding claim, wherein the bacterial strain has a 16s rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to SEQ ID NO:2, or wherein the bacterial strain has the 16s rRNA sequence represented by SEQ ID NO:2.
14. The composition of any preceding claim, wherein the bacterial strain is lyophilised.
15. The composition of any preceding claim, wherein the bacterial strain is viable and capable of partially or totally colonising the intestine.
16. The composition of any preceding claim, wherein the composition comprises a single strain of Enterococcus faecium and does not contain any other bacterial strains or species or comprises only de minimis or bîologically irrelevant amounts of other bacterial strains or species.
17. The composition of any preceding claim, wherein the composition does not comprise a bacterial strain ofthe species Bacillus coagulons.
18. A food product or vaccine comprising the composition of any preceding claim, for the use of any preceding claim.
19. A cell of the Enterococcus faecium strain deposited under accession number NCIMB 42487, or a dérivative thereof.
20. A composition comprising the cell of claim 19.
21. The composition of claim 20, comprising a pharmaceutically acceptable carrier or excipient.
22. A bîologically pure culture of the Enterococcus faecium strain deposited under accession number NCIMB 42487, or a dérivative thereof.
23. A cell of the Enterococcus faecium strain deposited under accession number NCIMB 42487, or a dérivative thereof, for use in therapy.
24. The cell of claim 23, wherein the cell is for use in a method defined in any of daims 1-12.
25. A composition comprising one or more bacterial strains of the species Enterococcus faecium, which does not contain bacteria from any other species or which comprises only de minimis or bîologically irrelevant amounts of bacteria from another species, for use in therapy.
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