Composition for the prevention and treatment of intestinal diseases characterised by pain and inflammation

Abstract

The present invention relates to the field of therapeutic compositions for gastrointestinal disorders. More particularly, the invention refers to compositions comprising resveratrol, saffron extract and vitamin D, and to the use thereof for the treatment of functional gastrointestinal disorders, and in particular for the treatment of irritable bowel syndrome (IBS).

Description

[0001] COMPOSITION FOR THE PREVENTION AND TREATMENT OF INTESTINAL DISEASES

[0002] CHARACTERISED BY PAIN AND INFLAMMATION

[0003] FIELD OF THE INVENTION

[0004] The present invention relates to the field of therapeutic compositions for gastrointestinal disorders. More particularly, the invention refers to compositions comprising resveratrol, saffron extract and vitamin D, and to the use thereof for the treatment of functional gastrointestinal disorders, and in particular for the treatment of irritable bowel syndrome (IBS).

[0005] BACKGROUND

[0006] Functional gastrointestinal disorders (FGIDs) represent a class of disorders of the gastrointestinal tract characterised by a lack of identifiable structural anomalies. These disorders present symptoms both of a gastrointestinal nature, such as abdominal pain, bowel irregularities, diarrhoea, constipation, bloating, feeling of fullness, nausea and vomiting, and of a psychological nature, such as anxiety, depression, persistent fatigue, sleep disorders and impairment of work and social activities. Consequently, the quality of life of patients suffering from FGIDs is compromised.

[0007] Irritable bowel syndrome (IBS), dyspepsia and functional constipation are common examples of FGIDs. However, other manifestations are often underestimated due to lack of medical consultation or diagnoses not made.

[0008] Irritable bowel syndrome (IBS) is a common functional gastrointestinal disorder characterised by abdominal pain, bloating and changes in bowel habits, including diarrhoea, constipation or both. Notwithstanding the high prevalence of IBS, its exact aetiology remains unclear, and the current treatment options often provide only partial relief of the symptoms or are associated with undesirable side effects.

[0009] Documents that address the problem of IBS in various ways are known in the prior art. For example, CN 106389395 discloses the use of compositions comprising resveratrol for the treatment of IBS.

[0010] Tsai et al., Phytomedicine, Elsevier, Amsterdam, NL, 1 February 2018 (2018-02-01), pages 55-59, discuss fundic relaxation and how it may be influenced by the use of resveratrol.

[0011] Huang Hangkai et al., Nutrition Journal, vol. 21, no. 1, 5 May 2022 (2022-05-05), describe the use of vitamin D in individuals suffering from IBS. The article sets forth some preliminary considerations while stressing, however, that additional studies are necessary in order to draw accurate conclusions.

[0012] There is thus a need to find novel compositions for the treatment of functional gastrointestinal disorders, and in particular for the treatment of IBS, which can provide effective relief of the symptoms with reduced side effects.

[0013] The present invention addresses this need by providing novel compositions comprising resveratrol, saffron extract and vitamin D for the treatment of IBS and functional gastrointestinal disorders.

[0014] SUMMARY OF THE INVENTION

[0015] In one aspect, the present invention provides a composition comprising resveratrol, saffron extract, and vitamin D - as claimed.

[0016] In another aspect, the present invention relates to uses and methods in which the composition described herein is used for the treatment of functional gastrointestinal disorders (FGIDs), in particular for the treatment of IBS. The present description also relates to the process for preparing the compositions described herein.

[0017] BRIEF DESCRIPTION OF THE FIGURES Figure 1 represents the cell viability of the experimental model following the induction of damage with LPS, which mimics the condition of IBS, and treatment with the single components and the combination thereof. Viability, tested by means of the MTT assay, was observed for 6 hours. The control line (0) represents the model under baseline conditions.

[0018] Figure 2 represents oxidative stress calculated as the production of reactive oxygen species (ROS) triggered by LPS-induced damage, and following treatment with the single components and the combination thereof. Oxidative stress was observed for 6 hours, the control line (0) represents the model under baseline conditions.

[0019] Figure 3 shows the trans-epithelial electrical resistance (TEER) of the intestinal barrier, an indicator of barrier integrity. The TEER value was recorded in the healthy control (dotted line) and following LPS-induced damage and the treatment with single components and the combination thereof. The membrane integrity was observed for 6 hours.

[0020] Figure 4 represents the expression of the proteins making up the tight junctions at the level of the intestinal barrier, namely claudin, occludin and zonula occludens-1. The expression of these three proteins was tested in healthy cells and following LPS-induced damage and treatment with the single components and the combination thereof.

[0021] Figure 5 represents the rate of absorption of the single components and the combination thereof through the intestinal membrane following LPS-induced damage. The absorption was recorded for 6 hours.

[0022] Figure 6 represents the state of inflammation of the experimental model following treatment with LPS. The expression of TNF-a, pro-inflammatory cytokine, is measured as an inflammation marker. The expression of this cytokine was compared against the baseline condition (line 0) and following treatment with the single components and the combination thereof.

[0023] Figure 7 represents the state of mitochondrial dysfunction arising in the cellular model following LPS-induced damage. The expression of NLRP3 as a marker of mitochondrial dysfunction was tested 6 hours after the induction of damage with LPS and following treatment with the single components and the combination thereof.

[0024] Figure 8 shows the cell viability of the experimental model under a condition of damage caused by glutamate and LPS. Viability, measured by means of the MTT assay, was assessed following treatment with the single components and with the combination thereof. The control line (0) represents the cell viability of the experimental model under healthy conditions.

[0025] Figure 9a represents the oxidative stress calculated as the production of reactive oxygen species (ROS) triggered by the glutamate- and LPS-induced damage, and following treatment with the single components and the combination thereof. The control line (0) represents the model under baseline conditions.

[0026] Figure 9b represents the mitochondrial membrane potential following the glutamate- and LPS-induced damage and following treatment with the single components and the combination thereof. The control line (0) represents the model under baseline conditions.

[0027] Figure 10 represents the state of inflammation as measured through the expression of COX-2, a pro-inflammatory enzyme induced by glutamate- and LPS-induced damage. The analysis was performed on the single components and on the combination thereof. The control line (0) represents the model under baseline conditions.

[0028] Figure 11 represents the expression of neurotransmitters altered by the glutamate- and LPS-induced damage. The neurotransmitters taken into consideration are tryptophan, serotonin and GABA and they were analysed following treatment with the single components and the combination thereof. The control line (0) represents the model under baseline conditions.

[0029] Figure 12 represents the expression of M2 and M5 muscarinic receptors following the glutamate- and LPS-induced damage. The expression was measured following treatment with the single components and the combination thereof. The control line (0) represents the model under baseline conditions.

[0030] DETAILED DESCRIPTION OF THE INVENTION

[0031] The compositions of the present invention (hereinafter also called "the composition(s)") are compositions comprising resveratrol, saffron extract and vitamin D - as claimed. The compositions described herein may also contain additional active components, as well as acceptable pharmaceutical or food grade additives and / or excipients.

[0032] Resveratrol

[0033] The term "resveratrol" (C14H12O3) as used herein refers to a chemical compound also known as trans-resveratrol, or 3,5,4'-trihydroxy-trans-stilbene. Resveratrol is a stilbenol that is stilbene in which the phenyl groups are substituted at positions 3, 5, and 4' by hydroxy groups. It is produced in plants with the help of the enzyme stilbene synthase. It exists in the form of cis-(Z) and trans-(E) isomers. The trans form can undergo isomerisation to the cis form when heated or exposed to ultraviolet irradiation. However, recent studies have demonstrated that trans-resveratrol undergoes a negligible amount of oxidation in the normal atmosphere at room temperature. Resveratrol is present in high concentrations in red grapes, in some berry varieties, peanuts and several herbs such as Fallopia japonica (also known as Polygonum cuspidatum). It is known for its potential antioxidant, anti-inflammatory, and cardioprotective properties.

[0034] The resveratrol used in the compositions described herein is food and / or pharmaceutical grade and may be purchased commercially. The resveratrol comprised in the compositions of the present invention is preferably trans-resveratrol. This can be obtained as a 100% pure synthetic product.

[0035] In an alternative preferred embodiment, the resveratrol used is at least 98% transresveratrol by weight out of the total weight of the extract. Preferably, said resveratrol is obtained as an extract of Polygonum cuspidatum. Preferably, said resveratrol is in powder form.

[0036] Saffron extract

[0037] The term "saffron extract" as used herein refers to any commercially purchasable food and / or pharmaceutical grade saffron extract. For example, in the compositions described herein, use can be made of saffron extract complying with European Pharmacopoeia (PhEur) standards for use in pharmaceutical products.

[0038] The saffron extract used in the compositions described herein can be an extract obtained from the flower pistil (stigmas) (with or without the flower) of the plant Crocus sativus L., also known as saffron. Saffron is a natural substance that has been used for centuries for its properties which are beneficial for the body. This health-promoting property is recognised by the Italian Ministry of Health, as stated in Annex 1 to the Ministerial Decree of 10 August 2018 and subsequent amendments and additions, and is further confirmed by the scientific literature. In some recent studies, the effectiveness of saffron in improving mood was compared to that of an antidepressant drug belonging to the category of serotonin reuptake inhibitors (SSRIs).

[0039] The saffron extract comprised in the compositions described herein can be extracted using an alcohol-based extraction process. Ethanol is preferably used as the extraction solvent. In the process, the pistils / stigmas of the flower are collected and subsequently subjected to extraction. The extraction is performed using techniques known in the art, in such a way as to preserve the desired compounds, including crocin, safranal and picrocrocin. Preferably, the extract used in the compositions described herein is in powder form. However, it is possible to use tinctures and oils, or any other form that preserves its active substances.

[0040] The composition of the saffron can vary slightly according to the place and conditions of growth and the plant variety. In any case, the scientific literature on saffron extract is vast and comprises extracts of varying nature and content, providing evidence of effectiveness independent of these two factors. In the present invention use is made of extracts that are suitable for food and / or pharmaceutical use and comprise: crocin, picrocrocin and safranal.

[0041] The term "crocin", as used herein, refers to a chemical compound which is a carotenoid responsible for the colour of saffron. It is a natural pigment that is also found in carrots and other orange vegetables.

[0042] The term "picrocrocin", as used herein, refers to a chemical compound responsible for the bitter taste of saffron.

[0043] The term "safranal", as used herein, refers to a volatile oil which contributes to the characteristic aroma of saffron.

[0044] In addition to these main components, the saffron used in the present invention can contain other components in smaller amounts, namely zafferina, lycopene and anthocyanins. These compounds can further contribute to the beneficial properties of saffron.

[0045] In one embodiment, the saffron extract used in the compositions described herein comprises: crocins, and safranal. It preferably comprises picrocrocin, crocins, and safranal.

[0046] In one embodiment, the saffron extract used in the compositions described herein comprises:

[0047] Picrocrocin in an amount of from 0.3% to 5% by weight out of the total weight of the saffron extract, preferably from 0.4% to 3% by weight out of the total weight of the saffron extract, even more preferably from 0.5% to 3.5% by weight with respect to the total weight of the saffron extract. The amount of picrocrocin can be determined by means of HLPC (high-performance liquid chromatography) measurements.

[0048] Crocins in an amount of from 0.3% to 10% by weight out of the total weight of the saffron extract, preferably from 2% to 9% by weight out of the total weight of the saffron extract, even more preferably from 4% to 8% by weight with respect to the total weight of the saffron extract. The amount of crocins can be determined by means of HLPC (high-performance liquid chromatography) measurements.

[0049] Safranal in an amount of from 0.2% to 2% by weight out of the total weight of the saffron extract, preferably from 0.4% to 1.5% by weight out of the total weight of the saffron extract, even more preferably from 0.5% to 1% by weight with respect to the total weight of the saffron extract. The amount of safranal can be determined by means of HLPC (high-performance liquid chromatography) measurements.

[0050] In a preferred embodiment, the saffron extract used in the compositions described herein derives from plants cultivated in Spain, France and / or the Middle East (Iran).

[0051] In the compositions of the present invention, an extract having the following composition can be used:

[0052] Picrocrocin: 7-10 mg per gram of extract (0.7%-1.0% by weight out of the total weight of the extract)

[0053] Crocins: 9-13 mg per gram of extract (0.9%-1.3% by weight out of the total weight of the extract)

[0054] Safranal:4-7 mg per gram of extract (0.4%-0.7% by weight out of the total weight of the extract)

[0055] Kaempferol: 0.1-0.6 mg per gram of extract (0.01%-0.06% by weight out of the total weight of the extract).

[0056] The term "kaempferol" as used herein refers to a flavanol, a type of flavonoid, present in various plants, including saffron extract. It is a plant pigment that imparts colours such as yellow and red to many plants. It has demonstrated various biological properties, including potential antioxidant and anti-inflammatory effects.

[0057] In a preferred embodiment, the saffron extract used in the compositions described herein has a minimum content of 0.34% safranal, 0.43% crocin and 0.37% picrocrocin.

[0058] The saffron extract can be an extract of Crocus Sativus L. in the form of a commercially purchasable powder (beige / yellow / orange). This extract can have a content of 0.34% safranal, 0.43% crocin and 0.37% picrocrocin as measured by HPLC, and is food / pharmaceutical grade.

[0059] More preferably, the saffron extract used in the compositions described herein has a minimum content of 0.34% safranal and has a minimum content of 0.43% crocin by weight out of the weight of the extract.

[0060] The saffron extract can be a dry extract of Crocus Sativus L. in the form of a commercially purchasable powder (pale pink / orange). This extract has a content of safranal greater than or equal to 2% and of crocin greater than or equal to 3%, as measured by means of UV methods, and is food / pharmaceutical grade.

[0061] As understood in the present context, having a "minimum content" means that the saffron extract must contain at least a certain percentage of a specific component, such as safranal or crocin. In the example given above, the saffron extract must have a minimum concentration of 0.3% safranal and a minimum concentration of 0.4% crocin.

[0062] This means that the amount of safranal present in the saffron extract should be at least 0.3% of the total weight of the extract, while the amount of crocin should be at least 0.4% of the total weight of the extract. These minimum percentages assure that the extract is particularly effective.

[0063] Vitamin D

[0064] The term 'vitamin D' as used in the present context refers to a group of liposoluble vitamins. The two main forms of vitamin D are vitamin D2 (or ergocalciferol) and vitamin D3 (or cholecalciferol). Vitamin D is a liposoluble vitamin that is essential for human health, known mainly for its role in regulating the metabolism of calcium and phosphorous and in bone health. In the context of this description, the term 'vitamin D' must be interpreted as including all the vitamers and precursors related thereto. The term includes all the biologically active forms of vitamin D (for example calcitriol), as well as analogues, derivatives and precursors related thereto. The vitamin D present in the compositions described herein can be in the form of a salt, for example a pharmaceutically acceptable salt.

[0065] The vitamin D comprised in the compositions described herein is preferably vitamin D3. The vitamin D used in the compositions described herein can be purchased commercially and is food and / or pharmaceutical grade.

[0066] For example, use can be made of commercially available vitamin D3 purchasable from DSM as "dry Vitamin D3" comprising 90000-110000 I U / g of vitamin D3 (cholecalciferol) in medium-chain triglycerides finely dispersed in a corn starch-coated matrix of acacia gum and sucrose.

[0067] The present invention provides a composition comprising or, alternatively, consisting of resveratrol, saffron, and vitamin D (as defined in the present description) and optionally at least one acceptable pharmaceutical or food grade additive and / or excipient. The compositions of the invention are advantageously formulated for oral (or sublingual) administration. The dosage form of the composition can be a solid form, such as a tablet, chewable tablet, effervescent tablet, capsule, soft capsule, lozenge, granules or powder (granules or powder to be dissolved in water or orodispersible), or a semisolid form, such as soft gel, or a liquid form, such as a solution, suspension, dispersion, emulsion or syrup; the composition is preferably in solid form for oral use, more preferably in powder form.

[0068] The composition comprises resveratrol in an amount of from 75% to 89% by weight with respect to the total weight of resveratrol, vitamin D and saffron extract. In a particular embodiment, the composition comprises resveratrol in an amount of from 80 to 85% by weight with respect to the total weight of resveratrol, vitamin D and saffron extract, for example 83%.

[0069] In one embodiment, the composition comprises saffron extract in an amount of from 10% to 20% by weight with respect to the total wei

[0070] vitamin D and saffron extract are calculated with respect to the total weight of the components resveratrol, vitamin D and saffron extract in the mixture of these ingredients, and not with respect to the total weight of the final composition comprising any other components, excipients or vehicles.

[0071] The composition comprising resveratrol, vitamin D and saffron extract, can also be called mixture (MIX). Said mixture is then introduced into the composition comprising any other components, excipients or vehicles.

[0072] In one embodiment, the present invention relates to a mixture consisting of resveratrol, vitamin D and saffron extract.

[0073] Unless otherwise specified, the expression the composition comprises a component in an amount "in a range from x to y" means that said component can be present in the composition in all the amounts present in said interval, even if not explicitly stated, including the endpoints of the range.

[0074] In one embodiment, the composition comprises resveratrol in an amount of from 50 mg to 500 mg (for example, 100 mg, 150 mg, 200 mg, 300 mg, 500 mg). Preferably in an amount of from 100 mg to 200 mg (for example, 150 mg).

[0075] In one embodiment, the composition comprises saffron extract in an amount of from 5 mg to 100 mg (for example, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg). Preferably in an amount of from 15 mg to 35 mg (for example 30 mg). Said saffron extract having a content of safranal between 0.3% and 10%.

[0076] In one embodiment, the composition comprises vitamin D, preferably vitamin D3, in an amount of from 5 pg to 45 pg (for example, 10 pg, 20 pg, 30 pg, 40 pg). Preferably in an amount of from 15 pg to 35 pg (for example 20 pg).

[0077] In a preferred embodiment, the composition comprises resveratrol in an amount of from 50 mg to 300 mg, saffron extract in an amount of from 5 mg to 100 mg, vitamin D, preferably vitamin D3, in an amount of from 5 pg to 45 pg.

[0078] In a preferred embodiment, the composition comprises resveratrol in an amount of from 100 mg to 200 mg, saffron extract in an amount of from 15 mg to 35 mg, vitamin D, preferably vitamin D3, in an amount of from 15 pg to 35 pg.

[0079] In one embodiment, the composition comprises resveratrol, vitamin D (preferably D3), crocin, safranal, and picrocrocin. Preferably said crocin is in an amount of from 0.1 mg to 4 mg, said safranal is in an amount of from 0.1 mg to 3 mg, and said picrocrocin is preferably in an amount of from 0.1 mg to 3.0 mg.

[0080] In addition, the compositions described herein can comprise additional vitamins and minerals, selected as supplements to address dietary deficiencies of individuals suffering from functional gastrointestinal disorders.

[0081] The compositions described herein can further comprise at least one vitamin of the B group, preferably a vitamin B selected from the group consisting of vitamin Bl, B2, B6, B9, B12 and combinations thereof. Preferably, said at least one vitamin of the B group is included in amounts corresponding to 100% of the Nutrient Reference Value (NRV).

[0082] For example, the composition can comprise a vitamin B selected from the group consisting of vitamin Bl, B2, B6 and combinations thereof, in an amount of from 0.5 to 5 mg. The composition can further comprise a vitamin B9, in an amount of from 50 to 500 pg. The composition can further comprise a vitamin B12, in an amount of from 1 to 10 Pg-

[0083] The compositions described herein can further comprise calcium, zinc or combinations thereof. Preferably, the calcium is in an amount of from 100 to 1000 mg, (for example 400 mg). Preferably, the zinc is in an amount of from 1 to 50 mg (for example, 10 mg of zinc). In a preferred embodiment, the composition comprises resveratrol, saffron extract, vitamin D3, calcium, zinc and at least one vitamin of the B group.

[0084] The compositions described herein can further comprise a compound selected from the group consisting of: tryptophan, green tea extract (Camellia sinensis) containing epigallocatechin-3-gallate, butyrate and combinations thereof.

[0085] The term "tryptophan" (IUPAC name L-tryptophan) as used in the present context refers to an essential amino acid that is not produced by the human body but must rather be introduced through the diet and supplementation. In the compositions described herein, if used in addition to saffron extract, it will enhance the beneficial effects thereof, in particular in the treatment of functional gastrointestinal disorders (FGIDs).

[0086] In one embodiment, tryptophan can be used in the present invention as an alternative to saffron extract, all the other components and amounts remaining unchanged.

[0087] Preferably, the tryptophan, included in addition to the saffron, is used in an amount of from 5 mg to 500 mg of tryptophan, preferably from 50 mg to 400 mg (for example 300 mg).

[0088] The term "green tea extract (Camellia sinensis) containing epigallocatechin-3-gallate" refers to a green tea extract comprising polyphenols and epigallocatechin-3-gallate (EGCG) (most active catechin). In the compositions described herein, if used in addition to resveratrol, it will enhance the beneficial effects thereof, in particular in the treatment of functional gastrointestinal disorders (FGIDs).

[0089] In one embodiment, the green tea extract can be used as an alternative to resveratrol, all the other components and amounts remaining unchanged.

[0090] Preferably, the green tea extract, included in addition to the resveratrol, is used in an amount of from 10 mg to 1000 mg, preferably from 50 mg to 500 mg (for example 400 mg), said extract having an EGCG content of 40% and being decaffeinated.

[0091] The term "butyrate" as used herein, refers to a short-chain fatty acid (SCFA) approved as a molecule for dietary use. Often also defined as postbiotics, SFCAs are the product of metabolism of the gut microbiota. In the compositions described herein, if used in addition to resveratrol, saffron extract and vitamin D, it will enhance the beneficial effects thereof, in particular in the treatment of functional gastrointestinal disorders (FGIDs).

[0092] Preferably, the butyrate is used in an amount of from 10 mg to 1000 mg, preferably from 50 mg to 500 mg (for example 600 mg).

[0093] The compositions described herein can further comprise prebiotic substances.

[0094] The term "prebiotics" as used herein refers to "selectively fermented ingredients that result in specific changes in the composition and / or activity of the gastrointestinal microbiota, thus conferring benefit(s) upon host health" (definition taken from the "International Scientific Association of Probiotics and Prebiotics (ISAPP)"). The prebiotics optionally used in the compositions described herein are resistant to the pH of the stomach, cannot be hydrolysed and digested by human enzymes, nor absorbed by the gastrointestinal tract. These technical characteristics mean that prebiotics arrive intact in the gut, where they are fermented by the microbiota, thus promoting the activity and metabolism thereof.

[0095] In the compositions described herein, the following can be used as prebiotics: indigestible sugar fibres, such as inulin, fructooligosaccharides (FOS) and galactooligosaccharides (GOS). Since they are indigestible, the intake thereof increases the faecal mass, and as they draw water at the gut level, they improve the condition of constipation, thus reducing abdominal discomfort. Furthermore, by acting on the metabolism of the commensal microbiota, they induce the production of short-chain fatty acids, thus acting indirectly on inflammation, balance and absorption.

[0096] In one embodiment, inulin can be comprised in the compositions described herein, preferably in an amount of from 10 mg to 10 g, more preferably from 50 mg -1000 mg.

[0097] The compositions described herein can further comprise "orange polyphenols". The term "orange polyphenols" as used herein refers to a group of chemical compounds present in many plant foods, including oranges. Chemically speaking, they are organic compounds characterised by the presence of several phenolic groups. These compounds are known for their antioxidant properties, which can contribute to human health by protecting cells from damage caused by free radicals. In recent years, scientific studies have shown that there are also other classes of molecules which are not carbohydrates but could fall within the classification of a prebiotic. An example of these is polyphenols. It is well known that polyphenols are poorly absorbable, so much so that 90-95% of them reach the colon intact, where they are metabolised by the microbiota. Consequently, the intake of polyphenols is capable of modulating the composition of the microbiota itself. In the present context, orange polyphenols can be used as prebiotic substances. In the compositions described herein, if used in addition to resveratrol, they will enhance the beneficial effects thereof, in particular in the treatment of functional gastrointestinal disorders (FGIDs).

[0098] In one embodiment, orange polyphenols can be used as an alternative to resveratrol, all the other components and amounts remaining unchanged.

[0099] Preferably, the orange polyphenols included in addition to resveratrol are used in an amount of from 10 mg to 1000 mg, preferably from 50 mg to 500 mg (for example 350 mg).

[0100] Probiotics The composition further comprises at least one bacterial strain selected from the group consisting of:

[0101] • Lactobacillus acidophilus;

[0102] • Lactobacillus paracasei;

[0103] • Bifidobacterium lactis; and combinations thereof.

[0104] The composition further comprises at least one bacterial strain selected from the group consisting of:

[0105] • Lactobacillus acidophilus NCFM® = 2.5 billion CFU, strain number ATCC 700396;

[0106] • Lactobacillus paracasei Lpc-37® = 2.5 billion CFU, strain number ATCC SD5275;

[0107] • Bifidobacterium lactis Bi-07™ = 2.5 billion CFU, strain number ATCC SD5220;

[0108] • Bifidobacterium lactis BI-04® = 2.5 billion CFU, strain number ATCC SD5219;

[0109] • Bifidobacterium lactis HN019™ = 2 billion CFU, strain number AGAL NM97 / 09513; and

[0110] • combinations thereof.

[0111] The mixture of probiotics can be purchased commercially (trade name HowArU Gl Complete, property of IFF).

[0112] The composition described herein can be administered simultaneously with a composition of probiotics comprising at least one bacterial strain selected from the group consisting of:

[0113] • Lactobacillus acidophilus NCFM® = 2.5 billion CFU, strain number ATCC 700396;

[0114] • Lactobacillus paracasei Lpc-37® = 2.5 billion CFU, strain number ATCC SD5275;

[0115] • Bifidobacterium lactis Bi-07™ = 2.5 billion CFU, strain number ATCC SD5220;

[0116] • Bifidobacterium lactis BI-04® = 2.5 billion CFU, strain number ATCC SD5219;

[0117] • Bifidobacterium lactis HN019™ = 2 billion CFU, strain number AGAL NM97 / 09513; and combinations thereof. Said composition of probiotics can be administered together with the composition of the present invention to support the beneficial effects thereof in the treatment of functional gastrointestinal disorders (FGIDs), in particular IBS.

[0118] In the present context, the term "administered together" or "administered simultaneously" means administered at the same time, at most 1 hour apart or on the same day.

[0119] The probiotics described above are not contained in the same composition comprising resveratrol, saffron extract and vitamin D but in a different composition.

[0120] The present invention also provides a kit comprising two separate compositions: the composition comprising resveratrol, saffron extract and vitamin D, in every form described herein, and a composition comprising probiotics.

[0121] In a preferred embodiment, the present invention provides a kit comprising: the composition according to the present invention; and a composition comprising at least one bacterial strain selected from the group consisting of:

[0122] • Lactobacillus acidophilus, preferably wherein Lactobacillus acidophilus is Lactobacillus acidophilus NCFM® = 2.5 billion CFU, strain number ATCC 700396;

[0123] • Lactobacillus paracasei, preferably wherein Lactobacillus paracasei is Lactobacillus paracasei Lpc-37® = 2.5 billion CFU, strain number ATCC SD5275;

[0124] • Bifidobacterium lactis, preferably wherein Bifidobacterium lactis is Bifidobacterium lactis Bi-07™ = 2.5 billion CFU, strain number ATCC SD5220;

[0125] • Bifidobacterium lactis, preferably wherein Bifidobacterium lactis is Bifidobacterium lactis BI-04® = 2.5 billion CFU, strain number ATCC SD5219;

[0126] • Bifidobacterium lactis, preferably wherein Bifidobacterium lactis is Bifidobacterium lactis HN019™ = 2 billion CFU, strain number AGAL NM97 / 09513; and

[0127] • combinations thereof. In a preferred embodiment the present invention provides a kit comprising:

[0128] The composition according to the present invention;

[0129] A composition comprising at least one bacterial strain selected from the group consisting of:

[0130] • Lactobacillus acidophilus NCFM® = 2.5 billion CFU, strain number ATCC 700396;

[0131] • Lactobacillus paracasei Lpc-37® = 2.5 billion CFU, strain number ATCC SD5275;

[0132] • Bifidobacterium lactis Bi-07™ = 2.5 billion CFU, strain number ATCC SD5220;

[0133] • Bifidobacterium lactis BI-04® = 2.5 billion CFU, strain number ATCC SD5219;

[0134] • Bifidobacterium lactis HN019™ = 2 billion CFU, strain number AGAL NM97 / 09513; and

[0135] • combinations thereof.

[0136] More preferably, the composition of probiotics in the kit comprises:

[0137] • Lactobacillus acidophilus NCFM® = 2.5 billion CFU, strain number ATCC 700396;

[0138] • Lactobacillus paracasei Lpc-37® = 2.5 billion CFU, strain number ATCC SD5275;

[0139] • Bifidobacterium lactis Bi-07™ = 2.5 billion CFU, strain number ATCC SD5220;

[0140] • Bifidobacterium lactis BI-04® = 2.5 billion CFU, strain number ATCC SD5219;

[0141] • Bifidobacterium lactis HN019™ = 2 billion CFU, strain number AGAL NM97 / 09513; and

[0142] • combinations thereof.

[0143] The probiotics described herein can be purchased commercially.

[0144] The term "kit" in the present context means a combination of two separate compositions. Said compositions are administered together to support the beneficial effects of the composition according to the present invention.

[0145] The composition according to the present invention, and the composition of probiotics described herein can further contain at least one pharmaceutical or food grade additive and / or excipient. As used in the present context, "pharmaceutical or food grade additive and / or excipient" refers to a substance devoid of therapeutic activity, suitable for pharmaceutical or food use, and selected from the auxiliary substances known to the person skilled in the art, e.g. diluents, solvents (including water, glycerine and ethyl alcohol), solubilisers, thickeners, sweeteners, flavourings, colourings, lubricants, surfactants, antimicrobials, antioxidants, preservatives, pH stabilising buffers and mixtures thereof. Non-limiting examples of such substances are phosphate buffers (for example, dicalcium phosphate), a stearate of an alkali or alkaline earth metal (for example magnesium), silicon dioxide, mono- and diglycerides of fatty acids, microcrystalline cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, starch or corn starch, and natural or artificial flavourings (for example, iron oxides).

[0146] The composition of the invention can be a pharmaceutical composition, a composition for medical devices (Medical Device Regulation (EU) 2017 / 745 (MDR)), a dietary supplement and / or a food for special medical purposes (FSMP).

[0147] Examples of compositions

[0148] Composition 1

[0149] Functional Daily amount ingredient Resveratrol derived 100-300 mg either from

[0150] Polygonum cospidatum d.e. cone. 98%, or in the form of Transresveratrol

[0151] Saffron d.e. 10-50 mg

[0152] Vitamin D 10-30 pg Composition 2 >

[0153] Functional Daily amount ingredient Resveratrol derived 100-300 mg either from Polygonum cospidatum d.e. cone. 98%, or in the form of Transresveratrol

[0154] Saffron d.e. 10-50 mg

[0155] Vitamin Bl 0.5-1.5 mg

[0156] (thiamine)

[0157] Vitamin B2

[0158] (riboflavin)

[0159] Vitamin B6 0.5-2 mg

[0160] (pyridoxine)

[0161] Vitamin B9 100-300 pg

[0162] (folic acid)

[0163] Vitamin B12 1-2.5 pg

[0164] (cobalamin)

[0165] Calcium 200-500 mg

[0166] Zinc 5-20 mg

[0167] Probiotic12 billion CFU

[0168] Composition 3

[0169] Functional Daily amount ingredient Resveratrol derived 150 mg either from Polygonum cospidatum d.e. cone. 98%, or in the form of Transresveratrol

[0170] Saffron d.e. 30 mg

[0171] Vitamin D 20 pg

[0172] Vitamin Bl 1.1 mg

[0173] (thiamine)

[0174] Vitamin B2 1.4 mg

[0175] (riboflavin)

[0176] Vitamin B9 200 pg

[0177] (folic acid)

[0178] Calcium 400 mg

[0179] Zinc 10 mg

[0180] Probiotic 12 billion CFU

[0181] Preparation process

[0182] The compositions described herein can be obtained through means and methods commonly known in the prior art. For example, the compositions can be prepared by mixing the active ingredients with active excipients suitable for food and / or pharmacological use in the necessary amounts.

[0183] Properties and applications One aspect of the present invention refers to the composition comprising resveratrol, saffron extract, and vitamin D, according to any of the embodiments or aspects described for use as a medicament.

[0184] The compositions described herein show a surprising effectiveness in the treatment of functional gastrointestinal disorders, in particular in the treatment of IBS.

[0185] The composition according to the present invention (or the kit) can be used in a method of treatment (preventive or curative) of functional gastrointestinal disorders.

[0186] The composition or the kit according to the present invention can be used in a method of curative or preventive treatment of functional gastrointestinal disorders through the administration thereof to an individual in need in a therapeutically effective amount.

[0187] Preferably, the composition (or the kit) according to the present invention is for use in the treatment (preventive or curative) of irritable bowel syndrome (IBS), dyspepsia or functional constipation in an individual in need through the administration of a therapeutically effective amount of the composition of the present invention to said individual.

[0188] The term "functional gastrointestinal disorders", in the present context, refers to a class of disorders involving the gastrointestinal tract for which no structural anomaly is identifiable. Said disorders are as indicated in the diagnostic criteria, called the Rome Criteria (edition in force: 4th Edition of 2016).

[0189] The term "irritable bowel syndrome" (IBS), in the present context, may refer to a functional gastrointestinal disorder classified by the following symptoms according to the Rome Criteria: recurrent abdominal pain, on average for at least 1 day a week in the past 3 months, associated with a factor related to defecation (i.e. a change in the frequency or form of stools); the criteria must be fulfilled in the last 3 months, with symptom onset at least 6 months prior to diagnosis. This definition is as given in the publicly consultable databases of the Rome Criteria in force at the time of filing of the application. The definition may be updated, but this does not vary the identification of the treatment claimed in the present application.

[0190] IBS patients are distinguished, according to the Bristol Stool Form Scale, into IBS with diarrhoea (IBS-D), IBS with constipation (IBS-C), IBS with a mixed stool pattern (IBS-M) and unclassified IBS, based on the predominant frequency of stool consistency.

[0191] The compositions according to the present invention are for use in a method of treatment of IBS, in particular for the treatment of mixed IBS (i.e. with constipation and diarrhoea).

[0192] Symptoms and / or disorders connected to said functional gastrointestinal disorders are understood, for example, as: abdominal pain, bowel irregularity, diarrhoea, constipation, bloating sensation, feeling of fullness even after a small meal, nausea and vomiting, but also symptoms of a psychological nature, such as anxiety, depression, persistent feeling of tiredness, restless sleep, and others known to the person skilled in the art.

[0193] The subject matter of the present invention relates to use for the treatment, for nonmedical (non-therapeutic) purposes, of conditions or sensations in an individual connected to problems tied to functional gastrointestinal disorders, through the administration of an adequate amount of the composition of the present invention to said individual.

[0194] Surprisingly, the compositions and kit described herein have demonstrated a synergistic capacity of the ingredients in combating intestinal pain and discomfort by acting on inflammation and oxidative stress as mechanisms at the basis of functional gastrointestinal disorders. The compositions (or the kit) have shown a surprising capacity to restore the integrity of the intestinal barrier and bring intestinal motility and the gut microbiota back to a situation of balance.

[0195] The action of resveratrol allows for improving the body's depurative functions, as well as acting as a tonic against physical and mental fatigue. The action of saffron extract improves the symptoms tied to depression and anxiety, as well as the inflammatory states. Finally, vitamin D contributes to the normal absorption / use of calcium and phosphorous, thus contributing to maintaining normal muscle function. Vitamin D also contributes to the normal functioning of the immune system. The synergistic action of the three components enables these properties to be amplified in a significant and surprising manner.

[0196] The FGIDs treated with the compositions of the present invention are not chronic inflammatory intestinal diseases, i.e. diseases that fall under the acronym IBD and are mainly represented by Chron's disease and ulcerative colitis. These diseases have a strong autoimmune component, where the activation of the immune system causes not only inflammation, as in the case of IBS, but also actual damage to intestinal tissue, with necrosis and haemorrhaging. Consequently, these diseases lead to dysbiosis, pain, symptoms of impaired motility and hypersensitivity. The laxity of the gastrointestinal membrane is such as to cause malabsorption and poor digestion of some micro and macro nutrients.

[0197] Although the compositions described herein are intended for the treatment of FGIDs, they may also be administered as an adjuvant of at least one other therapy or composition capable of treating chronic inflammatory intestinal diseases such as IBD, Chron's disease and ulcerative colitis.

[0198] In the context of the present invention, the term "individual(s)" indicates mammals (animals and humans), preferably human beings.

[0199] The term "therapeutically effective amount" refers to the amount of composition or compound or formulation that elicits the biological or medicinal response in a tissue, system or individual that is sought and defined by the person skilled in the art.

[0200] EXAMPLES

[0201] A composition comprising resveratrol (A), saffron (B) and vitamin D (C) is tested in vitro to demonstrate the synergism of the components.

[0202] All the experiments described below are carried out on the following experimental groups:

[0203] - negative control = condition simulating the parameters of a healthy intestine at the baseline value.

[0204] - positive control = condition simulating the parameters of a damaged intestine at the baseline value (i.e. prior to performing a treatment)

[0205] - treatment with the 3 single active components (A; B; C)

[0206] - treatment with all 2-component mixture combinations (A+B, B+C, A+C)

[0207] - treatment with the mixture A + B + C (composition according to the present invention) The experimental studies are conducted by dividing the project into 3 phases; the first simulates gastric passage, the second intestinal passage and, lastly, the third takes into consideration the gut-brain axis.

[0208] Gastric passage:

[0209] Use is made of a 3D model of the stomach amply described in the literature which enables a culture of gastric epithelial cells to be associated with stomach fibroblasts in a hydrogel matrix. This technology provides an in vitro representation of the gastric mucosa and in this way it is possible to study the permeability of the single active components and the mixtures.

[0210] Furthermore, digestion is mimicked by introducing gastric enzymes. The product of gastric digestion is used for the subsequent phase. Intestinal passage:

[0211] In this case as well, use is made of a 3D model of the intestinal barrier validated in the literature. It is possible to study the effect of the single components and the compositions thereof on the intestinal mucosa both under healthy conditions (thus to verify the safety of the composition), and under pathological conditions (in this case to verify the effectiveness of the composition). In particular, an assessment is made of the state of inflammation and the changes in the expression of tight junction proteins, as markers of the integrity of the intestinal barrier.

[0212] Gut-brain axis:

[0213] Lastly, samples metabolised by intestinal cells (according to the previously used models) are used to stimulate the cerebral epithelium placed in the basolateral compartment. For the creation of the gut-brain axis, a co-culture model is set up, as described in the literature, in which use is made of SH-SY5Y neuronal cells cultured in the presence of glutamate, introduced because of its central role in mood regulation, and the single active components and the composition of the invention; then the influence of the latter on the mechanisms intracellular involved in mood regulation is tested.

[0214] The experiments were divided into two phases. In the first phase, a co-culture model of CaCo-2 (intestinal cells) + THP-1 (monocytes) pretreated with LPS (lipopolysaccharide) 100 ng / mL for 48h was used to mimic a condition of IBS. This 3D cellular model has been approved by both the FDA and EMA for analysing absorption and the mechanisms of transport through the intestinal barrier.

[0215] The second phase, on the other hand, mimicked the connection between the gut and brain in the case of IBS; therefore, the cellular model was built using an intestinal coculture (CaCo-2 / THP-l pretreated with LPS lOOng / mL for 48h) in vitro model of IBS, to which a Transwell® insert was added, and SH-SY5Y neuronal cells, in which a condition of alteration was induced with glutamate, were seeded in the basolateral compartment. In neuronal cells glutamate induces a condition that mimics central depression in a human being, with a state of reduced ability to synthesise the neurotransmitters involved in mood management.

[0216] As regards the data analysis, unless otherwise indicated, a p value < 0.05 is considered statistically significant. The synergism was calculated considering the sum of the biological activities of the single components compared to the composition of the invention (MIX), following universally recognised standards for the evaluation of synergism. The experiments reported here serve as proof of concept to demonstrate the synergism of the active components.

[0217] The results obtained in phase 1 (condition of IBS) are reported below. Cellular damage was induced by treating the cells with LPS to mimic the condition of IBS. Based on the viability study, one may note that all the active components improve viability in the 6 hours, but only partially counteract the damage induced with LPS (see Figure 1). In contrast, the composition of the present invention is capable of restoring the viability values, even surpassing the physiological levels. These data thus show a clear synergistic effect of the composition compared with the single components.

[0218] Subsequently, oxidative stress was analysed to evaluate how the single active ingredients and the composition of the invention influence this condition, inducing an improvement.

[0219] As may be seen from Figure 2, the damage induced with LPS gave rise to a condition of oxidative stress, which the active components partially reduced. The combination of active components assures better effectiveness, and the composition according to the present invention shows a significantly greater effect and proves to be synergistic compared to the single active components.

[0220] The membrane integrity (TEER) was also analysed. The damage caused by LPS results in a laxity of the membrane and indeed the resistance value is greatly reduced compared to the healthy control. The single substances partly contain the damage, bringing this value similar to that of the control, but only the composition of the present invention is capable of significantly increasing this parameter. When the TEER values are normalised relative to the control, one notes that the composition of the invention is capable of exceeding the line of the healthy control, thus showing its synergistic effectiveness as compared with the single raw materials (Figure 3).

[0221] The expressions of the proteins making up the tight junctions, namely Claudin, Occludin and Zonula occludens, were analysed to confirm the protective action of the composition of the invention as regards membrane integrity (Figure 4). In all three cases, the damage caused by LPS significantly reduces the expression of these proteins. Treatment with the active components in combination according to the present invention results in an effectiveness such as to exceed the values of the control. In this case as well, the composition made up of saffron, resveratrol and vitamin D3 reaches statistical significance and show a synergistic action compared with the single components.

[0222] As regards absorption, in accordance with what was previously observed in relation to the values of membrane integrity, the damage caused by LPS, which gives rise to a laxity of the barriers, results in an excessive absorption. Treatment with the single active components is only partly able to limit the damage, bringing the absorption rate to between 70 and 80%. The J max of the composition of the present invention, by contrast, is wholly comparable to that of the non-pathological condition and significant when compared with the single agents (Figure 5).

[0223] The evaluation of the inflammatory profile offers results that are equally promising for the present invention. Treatment with LPS increases the production of pro- inflammatory cytokines such as, for example, TNF-a. The active components administered individually have a slight action of countering inflammation, whereas the composition of the present invention shows a statistically superior effectiveness and is also synergistic when compared with the activity of the single components. Forthe sake of completeness, we note that the synergism was calculated by subtracting the activity of the single active components from the LPS value. The value of the sum of the single active components was then compared with the value of the composition of the present invention to establish whether the latter could be considered synergistic. In all cases, one sees that the final value is much greater than the sum of the single active components (Figure 6).

[0224] As regards mitochondrial dysfunction, treatment with LPS, by increasing the production of pro-inflammatory cytokines, also triggers inflammasome activation and an increase in the markers typical of mitochondrial dysfunction, such as NLRP3. The active components administered individually have only a partial action of countering inflammasomes, whereas the composition of the invention shows a statistically superior effectiveness and is also synergistic when compared with the activity of the single active components (Figure 7).

[0225] In the second part of the experimentation (phase 2), the effects on a model of gut-brain axis correlation in the case of IBS were evaluated.

[0226] This experimental model envisages the presence of two pathogenic agents: LPS at the intestinal level, to mimic a condition of IBS, and glutamate at the neuronal level, to replicate the effects of a depression at the central level, which is an index of the crosstalk along the gut-brain axis. The data show the effects of LPS-induced damage, which causes a reduction in cell viability. Treatment with the composition of the invention effectively counters mortality, bringing viability back to levels comparable to those of the control. In greater detail, the composition of the invention is capable of improving viability and bringing it back to positive values, thus showing its synergistic effectiveness (Figure 8).

[0227] The damage also influenced oxidative stress, promoting ROS production. The single active components only partly counteract the damage, whereas the composition according to the present invention restores production to baseline levels. In fact, the latter shows to be statistically superior and synergistic when compared with the single active components. The mitochondrial membrane potential also undergoes considerable damage following the administration of LPS. In this case as well, the data show a greater effectiveness of the composition of the invention versus the active components, thus demonstrating the synergistic effect (Figure 9).

[0228] The evaluation of the inflammatory profile is consistent with the results obtained; treatment with LPS increases the production of inflammation factors such as, for example, COX-2. The active components administered individually have an only partial action of counteracting inflammation, whereas the composition of the invention shows a statistically superior effectiveness, and shows a marked synergism when compared with the activity of the single components (Figure 10).

[0229] Symptoms related to mood disorders also play a fundamental role in the physiopathology of IBS. In this experimental protocol, this aspect was mimicked by pretreatment with glutamate. This treatment causes a reduced production of the essential amino acid tryptophan, precursor of serotonin, and a reduced production of neurotransmitters such as serotonin and GABA. Treatment with the composition of the present invention improves the production of these fundamental mood regulating factors. In all cases the composition shows to be synergistic when compared with the single components (Figure 11).

[0230] Finally, the expression of the M2 and M5 muscarinic acetylcholine receptors also plays a fundamental role in the treatment of symptoms of depression.

[0231] In this case as well, the composition of the present invention is able in both cases to push the expression towards positive values, thus showing its synergistic effectiveness (Figure 12).

[0232] In conclusion, the composition according to the present invention has been demonstrated to be synergistic as compared with the single active components making it up for every tested factor regarding the physiopathology of irritable bowel syndrome.

[0233] The composition according to the present invention results in an unexpected synergistic effect in the treatment of functional gastrointestinal disorders, and in particular for the treatment of irritable bowel syndrome (IBS).

[0234] The materials, the methods and the examples provided are purely illustrative and not intended to be limiting.

[0235] Unless otherwise defined, all the technical and scientific terms used in the present document have the same meaning as commonly understood by a person of ordinary skill in the art to which the present invention belongs.

[0236] It is clear that, although the disclosure has been set forth in combination with the detailed description, the foregoing description hasthe aim of illustrating and not limiting the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages and modifications fall within the scope of the claims.

[0237] EMBODIMENTS

[0238] 1. A composition comprising: resveratrol; saffron extract; and vitamin D.

[0239] 2. The composition according to embodiment 1, wherein the composition comprises resveratrol and saffron extract in a ratio (resveratrol / saffron extract) of 5:1 to 1:1.

[0240] 3. The composition according to any of the preceding embodiments, wherein the composition comprises resveratrol in an amount of from 75% to 89% by weight, with respect to the total weight of resveratrol, vitamin D and saffron extract.

[0241] 4. The composition according to any of the preceding embodiments, wherein the composition comprises saffron extract in an amount of from 10% to 20% by weight, with respect to the total weight of resveratrol, vitamin D and saffron extract.

[0242] 5. The composition according to any of the preceding embodiments, wherein the composition comprises vitamin D, preferably vitamin D3 in an amount of from 0.004% to 0.02% by weight, with respect to the total weight of resveratrol, vitamin D and saffron extract.

[0243] 6. The composition according to any of the preceding embodiments, wherein the composition further comprises at least one vitamin of the B group selected from the group consisting of Bl, B2, B6, B9, B12 and combinations thereof.

[0244] 7. The composition according to any of the preceding embodiments, wherein the composition further comprises calcium, zinc and combinations thereof.

[0245] 8. A kit comprising:

[0246] The composition according to any of the preceding embodiments; and

[0247] A composition comprising at least one bacterial strain selected from the group consisting of:

[0248] • Lactobacillus acidophilus NCFM® = 2.5 billion CFU, strain number ATCC 700396;

[0249] • Lactobacillus paracasei Lpc-37® = 2.5 billion CFU, strain number ATCC SD5275;

[0250] • Bifidobacterium lactis Bi-07™ = 2.5 billion CFU, strain number ATCC SD5220;

[0251] • Bifidobacterium lactis BI-04® = 2.5 billion CFU, strain number ATCC SD5219;

[0252] • Bifidobacterium lactis HN019™ = 2 billion CFU, strain number AGAL NM97 / 09513; and

[0253] • combinations thereof.

[0254] 9. The composition or the kit according to any of the preceding embodiments for use in a method for the treatment of functional gastrointestinal disorders (FGIDs).

[0255] 10. The composition or kit for use according to embodiment 9, for use in the treatment of the irritable bowel syndrome (IBS), dyspepsia or functional constipation.

Claims

CLAIMS1. A composition comprising: resveratrol; saffron extract; and vitamin D wherein the composition comprises resveratrol in an amount of from 75% to 89% by weight with respect to the total weight of resveratrol, vitamin D and saffron extract.

2. The composition according to claim 1, wherein the composition comprises resveratrol and saffron extract in a ratio (resveratrol / saffron extract) of 5:1 to 1:1.

3. The composition according to any of the preceding claims, wherein the composition comprises saffron extract in an amount of from 10% to 20% by weight with respect to the total weight of resveratrol, vitamin D and saffron extract.

4. The composition according to any of the preceding claims, wherein the composition comprises vitamin D, preferably Vitamin D3 in an amount of from 0.004% to 0.02% by weight with respect to the total weight of resveratrol, vitamin D and saffron extract.

5. The composition according to any of the preceding claims, wherein the composition further comprises at least one vitamin of the B group selected from vitamin Bl, B2, B6, B9, B12 and combinations thereof.

6. The composition according to any of the preceding claims, wherein the composition further comprises calcium, zinc or combinations thereof.

7. A kit comprising:The composition according to any of the preceding claims;A composition comprising at least a bacteria strain selected from the group consisting of:• Lactobacillus acidophilus, preferably wherein Lactobacillus acidophilus is Lactobacillus acidophilus NCFM® = 2.5 billion CFU, strain number ATCC 700396;• Lactobacillus paracasei, preferably wherein Lactobacillus paracasei is Lactobacillus paracasei Lpc-37® = 2.5 billion CFU, strain number ATCC SD5275;• Bifidobacterium lactis, preferably wherein Bifidobacterium lactis is Bifidobacterium lactis Bi-07™ = 2.5 billion CFU, strain number ATCC SD5220;• Bifidobacterium lactis, preferably wherein Bifidobacterium lactis is Bifidobacterium lactis BI-04® = 2.5 billion CFU, strain number ATCC SD5219;• Bifidobacterium lactis, preferably wherein Bifidobacterium lactis is Bifidobacterium lactis HN019™ = 2 billion CFU, strain number AGAL NM97 / 09513; and• combinations thereof.

8. The composition or the kit according to any of the preceding claims for use in the treatment of functional gastrointestinal disorders (FGIDs).

9. The composition or the kit for use according to claim 8, wherein the composition is for use in the treatment of irritable bowel syndrome (IBS), dyspepsia or functional constipation.

Images