WO2022219061A1

WO2022219061A1 - Sporebiotics for the prevention and/or treatment of il-17 related disorders

Info

Publication number: WO2022219061A1
Application number: PCT/EP2022/059910
Authority: WO
Inventors: Leen SLAETS; Niels HELLINGS; Raf DYBAJLO; Wannes THYS; Lucas WAUTERS; Tim VANUYTSEL
Original assignee: Universiteit Hasselt; My Research
Priority date: 2021-04-13
Filing date: 2022-04-13
Publication date: 2022-10-20

Abstract

The invention in general relates to the use of probiotics for the management of IL-17 related disorders. More specifically, the present invention relates to the use of Bacillus coagulans or Bacillus subtilis; in particular spore-forming probiotics. or a combination of both strains for the prevention or treatment of IL-17 related disorders, for example neurodegenerative disorders such as multiple sclerosis, Parkinson's disease, Alzheimer disease or Amyotrophic Lateral Sclerosis (ALS). The present invention also discloses the use of Bacillus coagulans or Bacillus subtilis or a combination of both strains as alternative therapy or adjunct therapy for IL-17 suppression treatment.

Description

SPOREBIOTICS FOR THE PREVENTION AND/OR TREATMENT OF IL-17 RELATED

DISORDERS

FIELD OF THE INVENTION The invention in general relates to the use of probiotics for the management of IL-17 related disorders; in particular spore-forming probiotics. More specifically, the present invention relates to the use of Bacillus coagulans or Bacillus subtilis or a combination of both strains for the prevention or treatment of IL-17 related disorders, for example neurodegenerative disorders such as multiple sclerosis, Parkinson’s disease, Alzheimer disease or Amyotrophic Lateral Sclerosis (ALS). The present invention also discloses the use of Bacillus coagulans or Bacillus subtilis or a combination of both strains as alternative therapy or adjunct therapy for IL-17 suppression treatment.

BACKGROUND TO THE INVENTION To date, spore-forming probiotic bacteria have been used for their protective and therapeutic potential to benefit health of the host. Spore-based probiotics are soil-based microorganisms that form spores and found in dirt and vegetation. Unlike most traditional probiotics which are similar to those naturally found in the human Gl tract such as lactobacilli and bifidobacteria , spore-based probiotics derived from Bacillus species are delivered as dormant spores. The endospores encapsulate the beneficial bacteria making them extremely stable and highly resistant to stomach acid’s low pH, resulting in the delivery of more effective probiotics to the small intestine where they then revert to active, growing bacteria. Sporebiotics have the ability to interact with the internal milieu of the host by producing a variety of antimicrobial peptides and small extracellular effector molecules.

Interestingly, gram-positive bacteria and spore-forming probiotic strains like Bacillus may outperform traditional probiotic supplements because of gastric-acid resistant endospores with improved storage conditions and survival in the small intestine. Beneficial effects of Bacillus coagulans and Bacillus subtilis strains have been shown on gut permeability and inflammation in in vitro models (Marzorati etal. 2020). In addition, a combination of oral spore-based probiotics (B. indicus, B. subtilis, B. coagulans, B. licheniformis and B. clausii) was shown to reduce postprandial endotoxemia in humans, a pathological condition which may result from increased gut permeability (McFarlin et at. 2017). Strict regulation of the inflammatory reactions is crucial to maintain immunological homeostasis. CD4-positive T cells, a type of T cell called helper T cells, regulate the function of other immune cells by producing cytokines such as Interleukin-17 (IL-17) that help activate immune cells in the microenvironment. In addition to its important role in protective immunity, IL-17 plays a critical role in the pathogenesis of various diseases. High IL-17 levels accompanied by excessive generation of Th17 cells have been found in multiple types of disorders including neurodegenerative disorders, musculoskeletal disorders, integumentary disorders, endocrine disorders, respiratory disorders, and cardiovascular disorders. Because of the importance of IL-17 in regulatory functions, IL-17 inhibitors are being investigated as possible treatments for such disorders. Based on emerging evidence from animal models, IL-17 has been suggested as a target for therapies to improve recovery post-stroke and to reduce the formation of skin cancer. In January 2015, the FDA approved the use of secukinumab (trade name Cosentyx), an IL-17 inhibiting monoclonal antibody, for the treatment of moderate to severe plaque psoriasis. In addition, Cosentyx has been approved in Japan for use in treating psoriatic arthritis.

It was unexpectedly found in the present invention, that long-term sporebiotic treatment decreased Th17-signaling with a reduction in IL-17 and in the frequency of Th17 cells resulting in a clinical efficacy of sporebiotics. The present invention describes the use of Bacillus coagulans or Bacillus subtilis (in particular spores thereof) or a combination of both strains for the prophylaxis and/or treatment of IL-17 related disorders, for example neurodegenerative disorders such as multiple sclerosis, Parkinson’s disease, Alzheimer disease or Amyotrophic Lateral Sclerosis (ALS). Additionally the present invention also discloses the use of Bacillus coagulans or Bacillus subtilis or a combination of both strains as alternative therapy or adjunct therapy for IL-17 suppression treatment. In another aspect, the present invention discloses the use of Bacillus coagulans or Bacillus subtilis spores or a combination of both strains for the prevention or treatment of other disorders selected from musculoskeletal disorders such as arthritis; integumentary disorders such as psoriasis; endocrine disorders such as diabetes type I; respiratory disorders such as asthma; cardiovascular disorders such as atherosclerosis, or a disorder characterized by increased levels of IL-17 and/or Th17 cells in comparison to a healthy subject.

SUMMARY OF THE INVENTION

In a first aspect, the present invention provides a combination comprising Bacillus coagulans or Bacillus subtilis spores for use in the prevention and/or treatment of an IL-17 disorder in a subject.

In particular, the invention is directed to a combination for use as defined herein wherein the IL- 17 related disorder is selected from the list comprising: a neurodegenerative disorder, a musculoskeletal disorder, an integumentary disorder, an endocrine disorder, a respiratory disorder, a cardiovascular disorder or a disorder characterized by increased levels of IL-17 and/or Th17 cells in comparison to a healthy subject.

In a specific embodiment, the invention is related to a combination for use as defined herein wherein the IL-17 related disorder is a neurodegenerative disorder such as multiple sclerosis, Parkinson’s disease, Alzheimer’s disease or Amyotrophic Lateral Sclerosis (ALS).

In a further embodiment, said combination for use according to the present invention comprises B. subtilis and B. coagulans spores in a 1 :1 , a 1 :2 or a 2:1 ratio.

In a further aspect, the combination for use according to the present invention comprises B. subtilis spores, derived from strain MY02 as deposited with the BCCM with accession number LMG P-31319.

In another aspect, the combination for use according to the present invention comprises B. coagulans spores, derived from strain MY01 as deposited with the BCCM with accession number LMG P-31318.

In yet a further embodiment, the present invention provides a pharmaceutical formulation for use in the prevention and/or treatment of an IL-17 related disorder in a subject, wherein said formulation comprises a combination comprising B. subtilis and B. coagulans spores according to the different embodiments as described above. Said pharmaceutical formulation may further comprise one or more pharmaceutically acceptable carriers, diluents or excipients.

In a further embodiment, the pharmaceutical formulation according to this invention is for use in the prevention and/or treatment of an IL-17 related disorder selected from the list comprising: a neurodegenerative disorder, a musculoskeletal disorder, an integumentary disorder, an endocrine disorder, a respiratory disorder, a cardiovascular disorder or a disorder characterized by increased levels of IL-17 and/or Th17 cells in comparison to a healthy subject. ln an even more preferred embodiment, the pharmaceutical formulation according to the invention is for use in the prevention and/or treatment of a neurodegenerative disorder such as multiple sclerosis, Parkinson’s disease, Alzheimer disease or Amyotrophic Lateral Sclerosis (ALS) in a subject.

In another embodiment, the pharmaceutical formulation of the present invention is a product of the dairy industry, beverage industry, food industry, or pharmaceutical industry, or it is a natural product.

In a further aspect of the invention, the combination or pharmaceutical formulation according to the different embodiments of the invention is for use in the prevention and/or treatment of an IL- 17 related disorders in a subject, wherein the subject is a human or an animal; preferably wherein the subject is a human.

In yet another embodiment of the invention, the combination or pharmaceutical formulation according to the different embodiments of the invention is provided in combination with a standard therapy for IL-17 related disorders. Said standard therapy for IL-17 related disorders may be selected from nonsteroidal anti-inflammatory drugs, corticosteroids, neuroprotective drugs, Interleukin inhibitors, antibiotics, spasmolytics, antidepressants, antipsychotics, and botanical extracts.

BRIEF DESCRIPTION OF THE DRAWINGS

With specific reference now to the figures, it is stressed that the particulars shown are by way of example and for purposes of illustrative discussion of the different embodiments of the present invention only. They are presented in the cause of providing what is believed to be the most useful and readily description of the principles and conceptual aspects of the invention. In this regard no attempt is made to show structural details of the invention in more detail than is necessary for a fundamental understanding of the invention. The description taken with the drawings making apparent to those skilled in the art how the several forms of the invention may be embodied in practice.

Fig. 1 : Reduction in IL-17A (A) and the frequency of Th17 cells (B) after 16 weeks of sporeforming probiotics in the entire cohort. DETAILED DESCRIPTION OF THE INVENTION

The present invention will now be further described. In the following passages, different aspects of the invention are defined in more detail. Each aspect so defined may be combined with any other aspect or aspects unless clearly indicated to the contrary. In particular, any feature indicated as being preferred or advantageous may be combined with any other feature or features indicated as being preferred or advantageous.

As used in the specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. By way of example, "a compound" means one compound or more than one compound.

The term "about" or "approximately" as used herein when referring to a measurable value such as a parameter, an amount, a temporal duration, and the like, is meant to encompass variations of +/- 10% or less, preferably +1-5% or less, more preferably +/- 1 % or less, and still more preferably +/-0.1 % or less of and from the specified value, insofar such variations are appropriate to perform in the disclosed invention. It is to be understood that the value to which the modifier "about" or "approximately" refers is itself also specifically, and preferably, disclosed.

The present invention describes the use of Bacillus coagulans or Bacillus subtilis (in particular spores thereof) or a combination of both strains for the prophylaxis and/or treatment of IL-17 related disorders.

Specific examples of IL-17 related disorders include: a neurodegenerative disorder, a musculoskeletal disorder, an integumentary disorder, an endocrine disorder, a respiratory disorder, a cardiovascular disorder, or a disorder characterized by increased levels of IL-17 and/or Th17 cells in comparison to a healthy subject.

In a particular embodiment, said IL-17 related disorder is not a functional gastro-intestinal disorder, in particular said IL-17 related disorder is not functional dyspepsia.

Additionally the present invention also discloses the use of Bacillus coagulans or Bacillus subtilis (in particular spore thereof) or a combination of both strains as alternative therapy or adjunct therapy for IL-17 suppression treatment.

The inventors of the present invention have unexpectedly found that long-term sporebiotic treatment with a combination of Bacillus coagulans and Bacillus subtilis spores decreased Th17- signaling with a reduction in IL-17 and the frequency of Th17 cells resulting in a clinical efficacy of these sporebiotics. The advantage of this invention is that it provides an effective monotherapy for symptoms associated with IL-17 related disorders. In addition, the present invention provides an adjuvant therapy for IL-17 related disorders that are otherwise treated with nonsteroidal antiinflammatory drugs, corticosteroids, neuroprotective drugs, Interleukin inhibitors, or antibiotics. Another advantage of sporebiotics is the high stability and long shelf life as well as that this combination of sporebiotics has the potential to reduce small intestinal bacterial overgrowth.

In a first aspect, the present invention provides a combination or pharmaceutical formulation comprising Bacillus coagulans or Bacillus subtilis spores for use in the prevention and/or treatment of an IL-17 disorder in a subject.

As defined herein, the term “combination” is in particular used in the context of a combination of Bacillus coagulans and Bacillus subtilis spores; while the term “pharmaceutical formulation” or “formulation” is used in the context of a combination of Bacillus coagulans and Bacillus subtilis spores, further comprising one or more pharmaceutically acceptable carriers, diluents or excipients.

In a specific embodiment of the present invention the combination or formulation as defined herein comprises a bacterial component, wherein said bacterial component consists of Bacillus coagulans and Bacillus subtilis spores. This embodiment is thus limited to combinations and formulations which do not contain any other bacterial species or spores thereof than the Bacillus coagulans and Bacillus subtilis spores of the invention. In particular, the inventors have found that this specific combination of Bacillus coagulans and Bacillus subtilis spores is sufficient to obtain the observed effects on IL-17 and thus no further bacterial strains or spores are required in obtaining such effects.

In the context of the present invention, the bacteria are typically in the form of bacterial spores or as vegetative bacterial cells, or a mixture of both. In a particularly preferred instance in the present invention, the bacteria are in the form of bacterial spores which can germinate in the gastrointestinal tract. In another embodiment, they may have been treated such that they cannot germinate. For example, the spores could be treated by heat and may, for instance, have been subjected to autoclaving to prevent germination. In another embodiment, the bacteria are in the form of vegetative cells. The spores or vegetative cells may, in one embodiment, be provided in isolated form. In yet another embodiment, the microbial material can be extracts of bacterial cells. In still another embodiment, the microbial material can be metabolites of bacterial cells released during cell growth in the culture medium. In another further embodiment, the microbial material can be any combination of bacterial spores, vegetative bacterial cells, extract of bacterial cells or metabolites of bacterial cells. In a preferred embodiment, the bacteria in the different embodiments of the invention are provided as viable spores which can germinate in the gastrointestinal tract. The formulation or pharmaceutical formulation according to the different embodiments of the invention requires that the bacteria B. subtilis and B. coagulans or their corresponding spores are isolated from eventual growth culture media. The skilled artisan is well aware of the techniques available for isolating the viable bacteria or spores from a growth culture medium such as using centrifugation, filtration, micro manipulation, and the like. For medium and long term storage, the isolated bacteria are preferably maintained in a dry state, such as for example achieved using freeze drying or spray drying. In one embodiment of the invention, the B. subtilis and B. coagulans used in the manufacture of the combination or pharmaceutical formulations are spray dried B. subtilis and B. coagulans, obtained by spray drying the bacteria using a saccharide protectant, in particular a trehalose as a protectant. An exemplary process suitable for spray drying the bacteria of the present invention is for instance available from Sunny- Roberts and Knorr (Int. Diary J„ 19 (2009) 209-214).

Once isolated, the B. subtilis and B. coagulans of the present invention can be prepared by any known or otherwise effective method for pharmaceutically formulating or manufacturing the selected product from. Methods for preparing the pharmaceutical formulations according to the present invention can be found in “Remington’s Pharmaceutical Sciences”, Mid. Publishing Co. Easton, Pa. USA.

In the context of the present invention, the term “IL-17” also referred to as “Interleukin 17” refers to the IL-17 family comprising IL-17A (sometimes called "IL-17"), IL-17B, IL-17C, IL-17D, IL-17E and IL-17F. The interleukin 17 family (IL-17 family) is a family of pro-inflammatory cytokines. IL- 17 is produced by a group of T helper cells known as T helper 17 cells in response to their stimulation with IL-23. IL-17 activates several signalling cascades that, in turn, lead to the induction of inflammatory mediators such as cytokines, prostaglandins and chemokines. Acting as chemoattractants, these chemokines recruit the immune cells, such as monocytes and neutrophils to the site of inflammation. IL-17 signalling is often observed in the pathogenesis of various diseases including rheumatoid arthritis, asthma, lupus, allograft rejection, anti-tumour immunity, psoriasis, and multiple sclerosis.

Each member of the IL-17 family has a distinct pattern of cellular expression. The expression of IL-17A and IL-17F appear to be restricted to a small group of activated T cells, and upregulated during inflammation. IL-17B is expressed in several peripheral tissues and immune tissues. IL- 17C is also highly upregulated in inflammatory conditions, although in resting conditions is low in abundance. IL-17D is highly expressed in the nervous system and in skeletal muscle and IL- 17E is found at low levels in various peripheral tissues. ln a particular embodiment, the invention is directed to a combination or pharmaceutical formulation for use wherein the IL-17 related disorder is selected from the list comprising: a neurodegenerative disorder, a musculoskeletal disorder, an integumentary disorder, an endocrine disorder, a respiratory disorder, a cardiovascular disorder or a disorder characterized by increased levels of IL-17 and/or Th17 cells in comparison to a healthy subject.

In a further embodiment, the invention is directed to a combination or pharmaceutical formulation for use wherein said IL-17 related disorder is selected from a neurodegenerative disorder, a musculoskeletal disorder, an integumentary disorder.

As used herein and unless otherwise specified, the term “neurodegenerative disorder” is to be understood as an umbrella term for a number of conditions affecting neurons, primarily brain neurons. These conditions are often incurable and debilitating and often result in progressive degeneration and/or death of nerve cells.

In a further embodiment, said neurodegenerative disorder is selected from the list comprising: multiple sclerosis (MS), Parkinson’s disease (PD) and PD-related disorders, Alzheimer’s disease (AD) and other dementias, Amyotrophic Lateral Sclerosis (ALS), essential tremor, multiple system atrophy, Huntington’s disease or Motor Neuron Diseases (MND); in particular MS, PD, AD and ALS.

In a specific embodiment, the invention is related to a combination or pharmaceutical formulation comprising B. subtilis and B. coagulans spores for use in the prevention and/or treatment of a neurodegenerative disorder such as multiple sclerosis, Parkinson’s disease, Alzheimer’s disease or Amyotrophic Lateral Sclerosis (ALS).

As used herein and unless otherwise specified, the term “musculoskeletal disorder” is to be understood as an umbrella term for conditions, injuries or pain in the musculoskeletal system, including the joints, ligaments, muscles, nerves, tendons, and structures that support limbs, neck and back.

In a further embodiment, said musculoskeletal disorder is selected from the list comprising: rheumatoid arthritis, psoriatic arthritis, enthesitis-related arthritis, Bechterew’s disease, axial spondyloarthritis, active ankylosing spondylitis, enthesitis, systemic sclerosis, or tendinopathy.

As used herein and unless otherwise specified, the term “integumentary disorder” is to be understood as an umbrella term for conditions, injuries or pain in the integumentary system which consists of the skin, hair, fingernails and toenails and other structures including glands. In particular, it relates to those parts that cover the body. ln a further embodiment, said integumentary disorder is selected from the list comprising: dermatitis, atopic dermatitis, atopic neurodermatitis, eczema, hidradenitis suppurativa, pityriasis rubra pilaris, plaque psoriasis, generalized pustular psoriasis, psoriatic erythroderma, psoriasis vulgaris, palmoplantar psoriasis, chronic scalp psoriasis, genital psoriasis, pyoderma gangrenosum, hidradenitis suppurativa, discoid lupus erythematosus, lupus nephritis, giant cell arteritis, papulopustular rosacea, ichthyosis, netherton syndrome, alopecia areata, lichen planus or lichen planopilaris, bullous pemphigoid.

As used herein and unless otherwise specified, the term “endocrine disorder” is to be understood as an umbrella term for conditions related to the endocrine glands of the body. In particular, it involves an imbalance in secretion (hyposecretion or hypersecretion) of hormones.

In a further embodiment, said endocrine disorder is selected from the list comprising: diabetes mellitus type I, necrobiosis lipoidica diabeticorum.

As used herein and unless otherwise specified, the term “respiratory disorder” is to be understood as an umbrella term for conditions, injury or pain affecting the organs and tissues that control gas exchange in air-breathing human and animals including the trachea, bronchi, bronchioles, alveoli, pleurae, pleural cavity, the nerves and muscles of respiration.

In a further embodiment, said respiratory disorder is selected from the list comprising: asthma, viral respiratory infections, such as coronaviral infections (e.g. COVID-19), chronic obstructive pulmonary disease (COPD), chronic bronchitis, or pneumonia.

Further IL-17 related disorders for which the combination of the present invention may suitably be used include but are not limited to: treatment-resistant depression, idiopathic subglottic stenosis, crohn’s disease, thyroid eye disease, dry eye disease, graved orbitopathy, non- infectious uveitis.

In yet a further embodiment, said combination for use according to the present invention comprises B. subtilis and B. coagulans spores in a 1 :3, a 3:1 , a 1 :4, a 4,1, a 1 :5, 5:1 ratio.

In a further aspect, the combination for use according to the present invention comprises B. subtilis spores, derived from strain MY02 as deposited with the BCCM with accession number LMG P-31319. Said strain has been deposited with the Belgian Co-ordinated Collection of Micro-Organisms (BCCM) (Universiteit Gent, K.L. Ledeganckstraat 35, 9000 Gent, Belgium) on March 14 2019 with accession number LMG P-31319 and herein further also indicated as MY02 strain.

Said strain has been deposited with the Belgian Co-ordinated Collection of Micro-Organisms (BCCM) (Universiteit Gent, K.L. Ledeganckstraat 35, 9000 Gent, Belgium) on March 14 2019 with accession number LMG P-31318 and herein further also indicated as MY01 strain.

In yet a further aspect, the combination for use according to the present invention comprises B. subtilis and B. coagulans, wherein said B. subtilis spores are derived from strain MY02 as deposited with the BCCM with accession number LMG P-31319 and wherein said B. coagulans spores are derived from strain MY01 as deposited with the BCCM with accession number LMG P-31318.

In yet a further embodiment, the present invention provides a pharmaceutical formulation for use in the prevention and/or treatment of an IL-17 related disorder in a subject, wherein said formulation comprises a combination comprising B. subtilis and B. coagulans spores according to the different embodiments as described above. Said pharmaceutical formulation further comprises one or more pharmaceutically acceptable carriers, diluents or excipients.

For example, the formulation or pharmaceutical formulation according to the different embodiments of the invention can be formulated along with common excipients, diluents or carriers, and formed into oral tablets, capsules, sprays, or oral liquids (e.g. suspensions, solutions, emulsions), powders or any other suitable dosage form.

Non-limiting examples of suitable and pharmaceutically acceptable excipients, diluents and carrier can be found in “Handbook of Pharmaceutical Excipients”, Second Edition, American Pharmaceutical Association, 1994, and may include: fillers and extenders such as starch, sugars, mannitol, and silicic derivatives; binding agents such as carboxymethyl cellulose and other cellulose derivatives, alginates, gelatin, and polyvinyl pyrolidone; moisturizing agents such as glycerol; disintegrating agents such as calcium carbonate and sodium bicarbonate; agents for retarding dissolution such as paraffin; resorption accelerators such as quaternary ammonium compounds; surface active agents such as acetyl alcohol, glycerol monostearate, adsorptive carriers such as kaolin and bentonite; carriers such as propylene glycol and ethyl alcohol, and lubricants such as talc, calcium and magnesium stearate, and solid polyethyl glycols. In another embodiment, the pharmaceutical formulation of the present invention is a product of the dairy industry, beverage industry, food industry, or pharmaceutical industry, or it is a natural product. In yet another embodiment, the pharmaceutical formulation for use according to the different embodiments of the invention can be in the form of a tablet, a capsule, a stick or any other edible form, including but not limited to functional food, juice, drinks or sweets.

The term “edible form” as used herein is intended to cover all consumable products, especially food products, and it can be solid, jellied or liquid. The term covers both ready-made products and products which are produced using the probiotic formulation as a starter alone, or in combination with conventional starters or other probiotics. The food products can for instance be products of the dairy industry or beverage industry. Alternatively, it can be a natural product. In the present invention, a “dairy product” means any liquid or semisolid milk or whey based product having a varying fat content. The dairy content can be e.g. cow milk, goat milk, sheep 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, another sour milk product, such as villi, filling of snack bars, etc. Another important group includes milk beverages, such as whey beverages, fermented milks, condensed milks, infant baby milks, ice-cream; milk-containing food such as sweets.

The products according to the different embodiments of the invention can also be concentrated and used as ingredients. Further, the products can also be dried and used in the form of powder or lyophilizate. The products are also applicable as capsules, pills or tablets. The products can also be used in the preparation of functional food products, health and wellness promoting edible products, or other corresponding products. It may also be used in an animal feed. Possible forms are capsules, pills or tablets, for example, manufactured in conventional processes used in the preparation of such a product for example in the pharmaceutical industry. This, the form of each of the food product, food material, and/or the pharmaceutical products, and the animal feed is not particularly limited.

In yet another embodiment, the present invention provides a combination or pharmaceutical formulation comprising B. subtilis and B. coagulans spores for use in combination with a standard therapy for IL-17 suppression treatment in a subject. In particular, it provides an alternative therapy or adjunct therapy for an IL-17 related disorder in a subject. In a further aspect, said standard therapy is selected from nonsteroidal anti-inflammatory drugs, corticosteroids, neuroprotective drugs, Interleukin inhibitors, antibiotics, spasmolytics, antidepressants, antipsychotics, and botanical extracts. In a further embodiment, said standard therapy is selected from the list comprising: ixekizumab, brodalumab, secukinumab; immunomodulators such as interferon-beta, glatiramer acetate, Dimethyl Fumarate, Teriflunomide, Alemtuzumab, Fingolimod, Ocrelizumab, Natalizumab; anti- tumoral treatments such as Cladribine; anti-Parkinson treatments such as Levodopa, MAO-B inhibitors, dopamine agonists, Catechol O-methyltransferase (COMT) inhibitors, Anticholinergics, or Amantadine.

The pharmaceutical formulation for use according to the different embodiments of the invention may further comprise one or more ingredients selected from the list comprising plant extracts, such as ginger, artichoke, proteins, vitamins, minerals or digestive enzymes, such as amylase, cellulase, lipase, beta-galactosidase, papain, bromelain and proteases.

In a further aspect of the invention, the combination or pharmaceutical formulation according to the different embodiments of the invention is for use in the prevention and/or treatment of IL-17 related disorders in a subject, wherein the subject is a human or an animal; preferably wherein the subject is a human.

The combination and pharmaceutical formulation according to the invention are primarily suitable for the use in human adults and infants. Though, they are also suitable for use in animals, especially pets and production animals. Examples of these include dogs, cats, rabbits, horses, cows, pigs, goats, sheep and poultry. The term “subject” as used herein therefore includes both humans and animals.

EXAMPLES

The following example illustrates the present invention. The example is not to be construed to limit the claims in any manner whatsoever. Material and methods

Study design and procedures

Study procedures were performed at baseline (1 ), after 8 weeks of treatment with spore-forming probiotics (2) and after 8 additional weeks of open-label extension (OLE) treatment with sporeforming probiotics (3). Blood and stool samples were collected at each visit.

Study subjects

Male and female patients were >18 years old, with no active psychiatric, inflammatory or metabolic conditions. Use of immunosuppressant drugs, anti- or probiotics in the last 3 months and alcohol use of >10 units per week were also exclusionary.

Study products

The probiotics treatment consisted of a 1 :1 combination of spray-dried Bacillus coagulans MY01 and Bacillus subtilis MY02 endospores (total of 2.5 x 109 CFU per capsule) in a mixture of 50mg with 300mg maltodextrin per capsule, taken twice daily. Compliance was determined by counting capsules and defined as good if >80% was used after each treatment phase.

Sample collection and processing Systemic immune activation

Fasting plasma samples were collected for determination of high-sensitivity C-reactive protein (hsCRP) (baseline, week 8) and lipopolysaccharide (LPS)-binding protein (LBP) (baseline, week 8 and 16). Also, systemic cytokines and peripheral blood mononuclear cells (PBMC) were analysed at each study visit, with subtyping of CD4+ and gut homing (CD4+ a4b7+ CCR9+) T cell subsets.

RESULTS

Study population

From June 2019 - July 2020, 68 subjects were included and randomised in the study. During the first 8 weeks (RCT-phase), drop-out occurred in 1 patient on spore-forming probiotics (adverse event)

Systemic immune activation

No within- or between-group differences were found for hsCRP or LBP in the first 8 weeks. Based on the additional clinical efficacy of open-label spore-forming probiotics, changes in systemic cytokines and CD4+ T cells were assessed after 8 and 16 weeks. No interaction effects were found for cytokines in the first 8 weeks, but IL-17A significantly decreased after 16 weeks of spore-forming probiotics (b= -.05 ± .02, p= .03) (Figure 1A). Although circulating Treg cells decreased after 8 weeks with spore-forming probiotics, effects on CD4+ T cells were mainly found after 16 weeks with spore-forming probiotics including significantly decreased Th17 cells (b= -.02 ± .01 , p= .03) (Figure 1 B). Thus, effects of spore-forming probiotics included decreased Th17-signaling.

In addition, long-term sporebiotic treatment and reduction in IL-17 and in the frequency of Th17 cells was also associated with improved clinical symptoms in patients, resulting in a clinical efficacy of sporebiotics (data not shown).

In conclusion, the presently presented data clearly evidence the role of sporebiotics of B. subtilis and B. coagulans in the reduction of IL-17 and Th17 cells rendering them particularly suitable in the prevention and/or treatment of disorder associated with altered (specifically increased) IL- 17 levels.

REFERENCES

Marzorati, M. et al. Bacillus subtilis HU58 and bacillus coagulans SC208 probiotics reduced the effects of antibiotic-induced gut microbiome dysbiosis in an M-SHIME® model. Microorganisms 8, 1-15 (2020).

McFarlin BK et al. 2017. Oral spore-based probiotic supplementation was associated with reduced incidence of post-prandial dietary endotoxin, triglycerides, and disease risk biomarkers. World J Gastrointest Pathophysiol 8, 117-126

Claims

1 . A combination comprising B. subtilis and B. coagulans spores; for use in the prevention and/or treatment of an IL-17 related disorder in a subject wherein said disorder is selected from the list comprising: a neurodegenerative disorder, a musculoskeletal disorder, an integumentary disorder, an endocrine disorder, a respiratory disorder, a cardiovascular disorder, or a disorder characterized by increased levels of IL-17 and/or Th17 cells in comparison to a healthy subject, wherein said disorder is not functional dyspepsia.

2. The combination for use according to claim 1 ; wherein said IL-17 related disorder is selected from the list comprising: a neurodegenerative disorder, a musculoskeletal disorder, an integumentary disorder, an endocrine disorder, a respiratory disorder or a cardiovascular disorder.

3. The combination for use according to claim 1, wherein the IL-17 related disorder is selected from the list comprising: a musculoskeletal disorder, an integumentary disorder, an endocrine disorder, a respiratory disorder, a cardiovascular disorder.

4. The combination for use according to claim 1 , wherein the IL-17 related disorder is selected from the list comprising: a neurodegenerative disorder, a musculoskeletal disorder, or an integumentary disorder.

5. The combination for use according to claim 1, wherein the IL-17 related disorder is a neurodegenerative disorder such as selected from the list comprising: multiple sclerosis (MS), Parkinson’s disease (PD), Alzheimer disease or Amyotrophic Lateral Sclerosis (ALS).

6. The combination for use as defined in anyone of claims 1 to 5; wherein said B. subtilis and B. coagulans spores are in a 1 :1, a 1 :2 or a 2:1 ratio.

7. The combination for use as defined in anyone of claims 1 to 6; wherein said B. subtilis spores are derived from strain MY02 as deposited with the BCCM with accession number LMG P- 31319.

8. The combination for use as defined in anyone of claims 1 to 7; wherein said B. coagulans spores are derived from strain MY01 as deposited with the BCCM with accession number LMG P-31318.

9. A pharmaceutical formulation for use in the prevention and/or treatment of an IL-17 related disorder in a subject; wherein said formulation comprises a combination as defined in anyone of claims 1 to 6, and one or more pharmaceutically acceptable carriers, diluents or excipients; wherein said IL-17 related disorder is selected from the list comprising: a neurodegenerative disorder, a musculoskeletal disorder, an integumentary disorder, an endocrine disorder, a respiratory disorder, a cardiovascular disorder or a disorder characterized by increased levels of IL-17 and/or Th17 cells in comparison to a healthy subject; wherein said disorder is not functional dyspepsia.

10. The pharmaceutical formulation for use according to claim 8 or 9 wherein the IL-17 related disorder is a neurodegenerative disorder such as selected from the list comprising: multiple sclerosis, Parkinson’s disease, Alzheimer disease or Amyotrophic Lateral Sclerosis (ALS).

11 . The pharmaceutical formulation for use as defined in anyone of claims 8 to 10; wherein said formulation further comprises one or more ingredients selected from plant extracts, proteins, vitamins, minerals and digestive enzymes.

12. The pharmaceutical formulation for use as defined in anyone of claims 8 to 11 ; wherein said formulation is in the form of a tablet, capsule, sachet, stick or any other edible form including but not limited to functional food, juice, drinks or sweets.

13. The pharmaceutical formulation for use as defined in anyone of claims 8 to 12, wherein said formulation is a product of the dairy industry, beverage industry, or pharmaceutical industry, or is a natural product.

14. The combination for use as defined in anyone of claims 1 to 7, or the pharmaceutical formulation for use as defined in anyone of claims 8 to 13, wherein said subject is a human or an animal; preferably wherein said subject is a human.

15. The combination for use as defined in anyone of claims 1 to 7 or the pharmaceutical formulation for use as defined in anyone of claims 8 to 14, further combined with a standard therapy for the treatment and/or prevention of an IL-17 related disorder.

16. The combination for use or the pharmaceutical formulation for use according to claim 15 wherein the standard therapy for the treatment and/or prevention of an IL-17 related disorder is selected from nonsteroidal anti-inflammatory drugs, corticosteroids, neuroprotective drugs, Interleukin inhibitors, antibiotics, spasmolytics, antidepressants, antipsychotics, and botanical extracts.