Prebiotic compositions

Description of the invention

The invention relates to a nutritional or pharmaceutical composition containing fructo-oligosaccharides (FOS) and galacto-oligosaccharides (GOS). Furthermore, the invention relates to uses of said composition. The invention finds utility in the treatment and prevention of gastrointestinal diseases such as inflammatory bowel disease (IBD), diarrhea and constipation.

Prebiotics are non-digestible food ingredients that have a beneficial effect on the health of the recipient. In order to be classified as a prebiotic, a food ingredient must meet the following criteria: i) not be hydrolyzed or absorbed in the upper gastrointestinal tract, ii) selectively fermented by one or a limited number of potentially beneficial commensal bacteria species in the large intestine, such as Lactobacillus and bifidobacteria, stimulating their growth and / or metabolic activation, iii) be able to change the composition of the colon microflora to a healthier one, for example by increasing the number of saccharolytic species and reducing putrefactive microorganisms. Bifidobacteria are obligate anaerobes that metabolize carbohydrates, mainly into acetic and lactic acids, which are further metabolized systemically. They are believed to play an important role in gut homeostasis, inhibiting the growth of putrefactive and pathogenic bacteria, the production of certain vitamins, activating intestinal functions, supporting digestion and absorption, and stimulating the immune response. Since bifidobacteria are sensitive to oxygen and high temperature, their use in food is limited. Hence, there has been increasing interest in prebiotics that are effective, withstand standard food processing and selectively targeting the beneficial gastrointestinal bacteria.

One of the desirable features of a prebiotic is the ability to reach further regions of the large intestine. This part of the gut is home to several chronic conditions, including colorectal cancer and ulcerative colitis. It is believed that the microflora in this part of the intestine may play an important role in the development or occurrence of such diseases. Dietary carbohydrate is the main substrate that is fermented in the proximal part of the large intestine, and because it is degraded by bacterial fermentation, protein becomes the main substrate in the more distal parts of the intestine. The products of bacterial metabolism include toxic and potentially carcinogenic compounds such as amines, ammonia and phenol compounds.

Constipation is a problem that often occurs in hospital patients as a result of a lack of dietary fiber, which increases transit time and reduces faecal mass. Illness, medications, immobilization, and reduced fluid intake also contribute to this problem . Studies have shown an increase in stool weight and an increase in the frequency of bowel movements by providing patients with increased amounts of dietary fiber in the hospital diet and / or the use of oral supplements or enteral feeding.

Inflammatory bowel disease (IBD) is a group of diseases that cause inflammation or ulceration in the small and large intestines. Most often, IBD is classified as ulcerative colitis or Crohn's disease. IBD may be a predisposition to colorectal cancer. The etiology of IBD is unknown and current drug treatments are not effective. It is suspected that various factors are involved in the aetiology of IBD, in particular genetic, environmental, immune and bacterial factors.

Ulcerative colitis is an inflammatory reaction that usually occurs in the rectum and the lower parts of the large intestine, or affects the entire large intestine. Symptoms include haemorrhage, bloody diarrhea, rectal pain and bleeding, raised temperature, weight loss, and can cause complications such as colon perforation, conjunctivitis, abscess, mouth ulcers, and skin lesions. Some studies suggest a role for sulfate-reducing bacteria such as the genus Desulfovibrio in ulcerative colitis. These bacteria, which reduce sulfate to sulfide, are found in 50% of the healthy population but are common in people suffering from ulcerative colitis (97 to 100%); they increase sulphide production in IBD patients and cause disorders such as impaired butyrate oxidation, damage to the epithelial cell barrier, induction of translocation of bacteria and antigens from food, and inflammation. Other bacteria such as E. coli or Helicobacter spp., although their role seems to be less certain. Ulcerative colitis cannot be cured with current drug therapies, but can be controlled with anti-inflammatory drugs. Drugs that can be used to treat patients with mild to moderate disease, such as sulfasalazine, can have adverse side effects such as nausea, vomiting, diarrhea and headache. Corticosteroids can be used in patients with more severe forms of the disease, but

They may in particular increase the risk of infection. In serious cases, surgery is required to remove the diseased intestine.

Crohn's disease is different from ulcerative colitis because it causes inflammation in the deeper layers of the intestinal wall and can occur anywhere in the gastrointestinal tract, with the small intestine most commonly affecting it. In addition, it is a chronic condition and may appear at different times in life.

Irritable bowel syndrome (IBS) is characterized by a combination of persistent and recurring abdominal pain and irregular bowel movements, including diarrhea, constipation, or both. IBS cannot be cured with any existing drug therapy.

In recent years, consumer demand for food has increased, which in addition to its nutritional value may also have a beneficial effect on health. In particular, there has been increased interest in the development of functional prebiotics containing food products with a prolonged fermentation time which are able to reach further parts of the intestine and there to increase the number of bifidobacteria.

In vivo studies in humans have shown that the addition of fructo-oligosaccharides (FOS) to the diet leads to an increase in fecal bifidobacteria and is an effective prebiotic. However, high levels of FOS can lead to overproduction of gases in human volunteers, so the lowest effective amounts of FOS should be used in the production of prebiotic foods. There is a need to develop new prebiotic compositions.

Patent document PL 193872 B1 (WO00 / 08948) discloses a composition consisting of a mixture of components A and B, where component A may be an oligosaccharide consisting of 2-6 monosaccharides and component B is a polysaccharide containing from 7 monosaccharides. This document also discloses that component A may be a mixture of β-galactooligosaccharides and fructooligosaccharides. However, PL 193872 B1 does not disclose specific examples of compositions where component A was such a mixture, or the beneficial technical effects that could be obtained by using such a combination.

The present inventors have surprisingly found that the prebiotic properties of FOS are significantly improved in the presence of galacto-oligosaccharides (GOS) and that the effect of FOS and GOS is more potent than additive, i.e. their synergistic effect on promoting the growth of beneficial bacteria such as bifidobacteria and lactobacilli has been observed. .

As a result of this synergistic effect, it is possible to obtain an equal or increased FOS effect at lower doses. This has the advantage that a strong prebiotic effect can be obtained in vivo, avoiding the need to consume any single probiotic in doses that may cause side effects. Moreover, the maximum beneficial prebiotic effect that can be obtained is greater than the effect produced by these prebiotics alone.

The subject of the invention is a nutritional or pharmaceutical composition containing fructooligosaccharides (FOS) and galactooligosaccharides (GOS), characterized in that FOS contains up to 95% by weight of oligofructose containing chains with a degree of polymerization ranging from 2 to 6, and GOS contains up to 95% by weight of oligogalactose containing chains with a degree of polymerization ranging from 2 to 7, wherein the amount of the mixture of FOS and GOS in the composition is from 0.1 to 60% by weight, and the weight ratio of FOS: GOS is from 0.2 to 5.

Preferably, the amount of the mixture of FOS and GOS is from 2 to 55% by weight, based on the total weight of the composition.

Preferably, the composition is a high caloric composition greater than 400 kcal.

Preferably, the composition comprises at least 1% protein, based on the total caloric value of the powdered composition.

Preferably, the composition additionally comprises glutamine.

Preferably, the composition is a complete dietary preparation.

Preferably, in a 100 ml formulation, the composition comprises: 1 to 3 g FOS, 1 to 3 g GOS, 2 to 20 g protein, 1 to 30 g carbohydrate, 0.5 to 20 g fat, as well as minerals and vitamins.

Preferably, in a 100 ml formulation, the composition comprises 1.25 g FOS, 1.25 g GOS, 9 g protein, 22 g carbohydrate and 9 g fat.

Preferably, the composition is a ready-to-eat composition.

The invention also relates to the use of a composition according to the invention for the manufacture of a medicament or nutritional composition for the treatment or prevention of gastrointestinal disorders such as diarrhea or constipation.

PL 213 665 B1

The invention also relates to the use of a composition according to the invention for the preparation of a medicament or nutritional composition for the maintenance and / or restoration of the intestinal flora.

The invention also relates to the use of a composition according to the invention for the manufacture of a medicament or nutritional composition for the prevention or treatment of inflammatory bowel diseases and / or the prevention of colon cancer.

Preferably, the infectious disease is ulcerative colitis or Crohn's disease.

The invention also relates to the use of a composition according to the invention for the manufacture of a medicament or a nutritional composition for stimulating the growth of Bifidobacterium and / or Lactobacillus bacteria and / or inhibiting the growth of at least one organism selected from bacteroids, Clostridium bacteria, enterobacteria, sulfate reducing bacteria.

The invention also relates to the use of a composition according to the invention for the manufacture of a medicament or nutritional composition for the prevention or treatment of infections caused by pathogenic intestinal bacteria.

Compositions containing FOS and GOS will hereinafter be referred to as compositions of the invention.

The composition according to the invention can be used as a drug or clinical product. The composition of the invention in the form of a medicament, e.g. a clinical product, or a nutritional composition can be used to treat or prevent disorders of the gastrointestinal (GI) passage such as e.g. IBD, diarrhea or constipation; chronic bowel disease and / or prolonging periods of remission or relieving symptoms or conditions associated with the disease, such as ulcerative colitis, Crohn's disease, and / or colorectal cancer.

The composition of the invention can also be used in the manufacture of a medicament, e.g. a clinical product or a nutritional composition, to alter the population of gut bacteria towards a healthier composition, e.g. by stimulating the growth and / or metabolism of beneficial intestinal bacteria such as bifidobacteria and / or lactobacilli, and / or inhibiting the growth and / or metabolism of adverse intestinal bacteria such as Bacteroides, coliform bacteria, Clostridium, reducing bacteria sulfur.

The composition of the invention can also be used in the manufacture of a medicament, e.g. a clinical product or a nutritional composition for the prevention or treatment of pathogenic intestinal bacterial infections.

Dietary intake of dietary fiber, particularly fructans and / or non-digestible oligosaccharides, increases the density of lactic acid producing bacteria in the gastrointestinal tract and reduces the number of undesirable bacteria such as Enterobacteriaceae. The latter type includes several pathogens. Thus, dietary intake of dietary fiber such as fructans and / or oligofructose can be used to selectively stimulate the growth of beneficial bacteria in the gastrointestinal tract. In turn, improving the ratio of beneficial to pathogenic bacteria has a beneficial effect on the health of the host.

Soluble fiber as used herein refers to fibers that are capable of being fermented in the large intestine to yield short chain fatty acids (SCFA).

The term oligosaccharides as used herein refers to sugars consisting of at least two to 20 monosaccharide units linked by glycosidic bonds, i.e. having a degree of polymerization (DP) of 2 to 20, most preferably 2 to 15 monosaccharide units, more preferably 2 to 10 monosaccharide units, and more preferably 2 to 7 monosaccharide units or 2 to 6 monosaccharide units.

Fructooligosaccharides (also called oligofructose) (FOS) are non-digestible oligosaccharides that belong to the fructans of the inulin group. In nature, they are found in many plant species, including onions, garlic, shallots, wheat, rye, bananas, asparagus, tomatoes, artichokes, dahlias and chicory root. FOS can be obtained enzymatically, by chemical means or by extraction from natural substances. Short-chain FOS consists of one to three fructose molecules linked to one sucrose molecule: their degree of polymerization (DP) is not higher than 6, and can be synthesized from sucrose by transfructosylating enzymes. Treatment of sucrose with these transfructosylating enzymes produces a FOS mixture containing 2, 3 or 4 fructose units, such as 1-kestose, nystose and fructosylnystose. In vivo human studies have shown that the addition of FOS to the diet increases the amount of fecal bifidobacteria and is a very effective prebiotic.

The term FOS as used herein includes FOS and short chain FOS. According to the invention, the FOS may contain between 2 and 20 saccharide units, more preferably between 2 and 15 saccharide units, even more preferably between 2 and 7 saccharide units and even more preferably between 2 and 6 saccharide units. In one aspect of the invention, the FOS may contain about 95% by weight of heptasaccharide disaccharides, based on the total weight of the FOS.

Fructooligosaccharides are commercially available, e.g. as Actilight 950P®, from Beghin-Meiji (France), which contains about 92 wt.% FOS, or oligofructose (RaftiIose) from Orafti, (Tienen, Belgium), produced in various variants, such as e.g. RAFTILOSE® P95 which comprises 95 wt% FOS composed of chains with a degree of polymerization from 2 to about 7, typically a DP of 3.5 to 4.5, and containing about 5 wt% glucose, fructose and sucrose together.

Galacto-oligosaccharides (GOS) can include di, tri, tetra, penta, and hexasaccharides, mainly consisting of galactose as the carbohydrate component, and are formed by the action of beta-galactosidase on lactose. According to the invention, the GOS may contain between 2 and 15 saccharide units, more preferably between 2 and 10 saccharide units, more preferably between 2 and 7 saccharide units, and more preferably between 2 and 6 saccharide units. In one aspect of the invention, the GOS may contain from about 0 to about 45 wt% disaccharides, more preferably from about 10 to about 40 wt% disaccharides, even more preferably from about 20 to about 35 wt% disaccharides, and even more preferably about 33 wt% disaccharides. relative to the total weight of the GOS. According to the invention, the GOS may contain from about 0 to about 50% by weight of trisaccharides, more preferably from about 10 to about 45% by weight of trisaccharides, even more preferably from about 20 to about 40% by weight of trisaccharides, and even more preferably about 39% by weight of trisaccharides, based on to the total weight of the GOS. According to the invention, the GOS may contain from about 0 to about 50% by weight of tetrasaccharides, more preferably from about 5 to about 45% by weight of tetrasaccharides, even more preferably from about 10 to about 40% by weight of tetrasaccharides, and even more preferably about 18% by weight of tetrasaccharides, based on the total weight of the GOS. According to the invention, the GOS may contain from about 0 to about 30% by weight of pentasaccharides, more preferably from 1 to about 25% by weight of pentasaccharides, even more preferably from 2 to about 10% by weight of pentasaccharides, and even more preferably 7% by weight of pentasaccharides, based on the total weight of the GOS. .

The term GOS as used herein includes GOS as defined above and transgalacto-oligosaccharides, also called tGOS.

GOS are commercially available, e.g. as Elixor® from Borculo Domo Ingredients (The Netherlands), which contains about 58 wt.% Galacto-oligosaccharides, 23 wt.% Lactose, and 19 wt.% Glucose, as Vivinal® GOS 10 (powder) from Borculo Domo Ingredients (Netherlands), which contains about 28 wt.% Galacto-oligosaccharides, 46.5 wt.% Monosaccharides and disaccharides, 17.5 wt.% Protein, 3.1 wt.% Minerals, 1 wt.% Fat and 4 wt.% Moisture, and which is available as a dry spray mixture with the addition of 50% whey protein concentrate (WPC 35), or as Vivinal GOS (syrup) from Borculo Domo Ingredients, which contains 75% by weight dry substance, 41% by weight of which are monosaccharides and disaccharides and 59% by weight are GOS.

The amount of FOS and GOS present in the compositions of the invention can be determined by various factors of importance, including the purpose of administration, the age, sex and weight of the individual patients, the form of the composition of the invention, e.g. a powder or a ready-to-eat composition, e.g. a ready-to-eat composition. drinking or an instant drink, or worsening of symptoms.

The compositions of the invention may contain FOS in an amount of from at least 0.3 g to about 20 g, more preferably from about 0.5 to about 12 g, more preferably from about 0.5 to about 5 g. According to the invention, the amount of GOS in the compositions of the invention is it may be from about 0.3 g to about 20 g, more preferably from about 0.5 to about 12 g, more preferably from about 0.5 to about 5 g.

FOS will be present in suitable amounts in the compositions of the invention, e.g. in solid form, e.g. in powder form, if the amount will be from about 0.1 to about 40% by weight, preferably from about 0.2 to about 30% by weight. , e.g., about 0.5 to about 20% by weight, even more preferably from about 1 to about 15% by weight, and even more preferably from about 2 to about 12% by weight, based on the total weight of the composition.

The amount of GOS in the compositions of the invention, e.g. in solid form, e.g. in powder form, may be from about 0.1 to about 40% by weight, preferably from about 0.2 to about 30% by weight, even more preferably from about 0%. 5 to about 20% by weight, even more preferably from about 1 to about 15% by weight, and even more preferably from about 2 to about 12% by weight, based on the total weight of the composition, e.g. the total weight of the powdered composition.

PL 213 665 B1

According to the invention, the amount of FOS and GOS mixed in the compositions of the invention, e.g. in solid form, e.g. in powder form, may be from about 0.1 to about 60% by weight, preferably from about 0.2 to about 40% by weight, more more preferably from about 0.5 to about 30% by weight, even more preferably from about 1 to about 25% by weight, even more preferably from about 2 to about 20% by weight, and even more preferably from about 2% to about 15% by weight, based on the total weight of the composition, e.g. based on the total weight of the powdered composition.

In another embodiment of the invention, suitable amounts of FOS contained in ready-to-eat compositions of the invention range up to about 20% by weight, or up to about 15% by weight, e.g. from about 0.05 to about 20% by weight, e.g. , 1 to about 10% by weight, e.g., from about 0.2 to about 5% by weight, or from about 0.5 to about 3% by weight, based on the total weight of the ready-to-eat composition.

GOS in ready-to-eat compositions of the invention may be present in amounts up to about 20% by weight, or up to about 15% by weight, e.g. from about 0.05 to about 20% by weight, e.g. from about 0.1 to about 10%. by weight, e.g., from about 0.2 to about 5% by weight, e.g., from about 0.5 to about 3% by weight, based on the total weight of the ready-to-eat composition.

According to the invention, the amount of FOS and GOS mixed in the ready-to-eat compositions of the invention ranges up to about 40% by weight, up to about 30% by weight, e.g. from about 0.05 to about 40% by weight, from about 0.1 to about 30% by weight, from about 0.2 to about 20% by weight, from about 0.5 to about 15% by weight, from about 1 to about 10% by weight, from about 2 to about 5% by weight, e.g. e.g. about 20%, e.g. about 10% by weight, e.g. about 5% by weight, based on the total weight of the composition, e.g., ready-to-eat composition.

The dose, e.g., daily dose, of FOS, e.g. per meal, may be, for example, from about 0.01 g to about 0.5 g / kg body weight, most preferably from about 0.03 g to about 0.3 g / kg body weight. body weight, more preferably from about 0.05 g to about 0.2 g / kg body weight, more preferably from about 0.06 g to about 0.15 g / kg body weight. According to the invention a dose, e.g. a daily dose of GOS, e.g. per meal may, for example, be from about 0.01 g to about 0.5 g / kg body weight, more preferably from about 0.03 g to about 0.3 g / kg body weight, more preferably from about 0.05 g to about 0.2 g / kg body weight, and more preferably from about 0.06 g to about 0.15 g / kg body weight. A suitable dose, e.g., daily dose of a mixture of FOS and GOS, e.g. per meal may, for example, be from about 0.01 g to about 1 g / kg body weight, more preferably from about 0.03 g to about 0.6 g / kg body weight, more preferably from about 0.05 g to about 0, 4 g / kg body weight, and more preferably from about 0.10 g to about 0.3 g / kg body weight.

A suitable dose, e.g., the daily dose of the mixture of FOS and GOS may be, for example, from about 1 to about 20 g, more preferably from about 1 to about 15 g, more preferably from about 2 to about 10 g, e.g. about 15 g, more preferably about 10 g, and more preferably about 5 g.

In one aspect the invention provides a composition according to the invention wherein the FOS / GOS weight ratio is from about 0.01 to about 50, more preferably from about 0.5 to about 20, more preferably from about 0.1 to about 10, more preferably from about 0. , 2 to about 5, more preferably from about 0.3 to about 3, more preferably from about 0.5 to about 2, and more preferably from about 0.6 to about 1.5. The FOS / GOS mass ratio may be around 1.

The relative ratio of the active ingredients of the compositions according to the invention may, of course, vary widely depending on the particular type of composition in question, e.g. whether the composition is in liquid or solid form or whether it is provided in the form of food. All proportions and relative weight ranges indicated herein are therefore to be understood as indicative of preferred or individual instances according to the invention which merely illustrate, but do not limit the invention in its broadest aspect.

According to the invention, the compositions according to the invention may also contain insoluble and / or soluble fiber, such as non-starch polysaccharides, e.g. cellulose, hemicellulose, metabolism-resistant starch (not digested), natural gums, guar gum, hydrolyzed guar gum, e.g. partially hydrolyzed guar gum, available e.g. as Benefibre® (from Novartis Nutrition Corporation), pectin, acacia and mixtures thereof.

The compositions according to the invention may also contain polyunsaturated fatty acids, in particular cis-polyunsaturated fatty acids such as n-3 fatty acids and / or n-6 fatty acids, e.g. alpha-linolenic acid (18: 3), stearidonic acid, eicosapentaenoic acid (EPA) (20: 5), docosapentaenoic acid (22: 5) and docosahexaenoic acid (DHA) (22: 6), linolenic acid (18: 2), gamma-linolenic acid (18: 3), arachidonic acid ( 20: 4), either in free form, or

PL 213 665 B1 in the form of an oil or fat, and mixtures thereof. The use of a combination of eicosapentaenoic acid and docosahexaenoic acid is preferred.

The compositions of the invention may also contain a nitrogen source, e.g. proteins and / or amino acids, and / or a fat source, e.g. lipids, and / or a carbohydrate source. The compositions according to the invention may also contain vitamins and / or minerals.

In another variant of the invention, the compositions according to the invention may also contain other oligosaccharides such as for example lactulose, xylooligosaccharides (XOS), soybean oligosaccharides (SOS), isomaItooligoascharides (IMO), arabinogalactan (ABG), genciooligosaccharides, fructans, partially guhydroligosaccharides (fructans). PHGG), and mixtures thereof, most preferably XOS.

In a further embodiment of the invention, the compositions of the invention consist mainly or exclusively of FOS and GOS as described herein.

In one embodiment, the invention provides a nutritional or pharmaceutical composition comprising an oligosaccharide mixture consisting of FOS and GOS as described above, e.g. a mixture comprising FOS containing 2 to 7 monosaccharide units and GOS containing 2 to 7 monosaccharide units.

Further components of the compositions of the invention may include any biologically active compounds or extracts whose properties are beneficial to health, especially compounds that have a beneficial effect on the gastrointestinal tract, such as glutamine / glutamate or their precursors.

The compositions of the invention may also contain one or more additional substances that inhibit bacterial adhesion to the epithelium of the gastrointestinal tract, including mannans, galacturonic acid oligomers, most preferably of natural origin. The compositions of the invention can be combined with drugs useful in the treatment of ulcerative colitis such as mesalamine, sulfasalazine, 5-ASA agents, corticosteroids such as adrenal cortex hormones, prednisone, hydrocortisone or budesonide; or drugs for pain relieving, antipyretic, infection or IBS treatment, such as a serotonin-4 receptor agonist, e.g. Zelnorm / Zelmac ™. For example, a composition of the invention may be provided in the form of a combined pharmaceutical preparation or kit for administration separately, sequentially or simultaneously with such drugs as described above. These drugs can be conveniently combined with the composition according to the invention in standard dosage forms.

In one embodiment of the invention, the compositions according to the invention do not contain plant species of the Ericaceae family or material derived from wheat leaf.

In another embodiment of the invention, the compositions of the invention are raffinose-free.

In one aspect of the present invention, the compositions of the invention can be readily incorporated into nutritional or pharmaceutical preparations, typically nutritional supplements, dietary supplements, therapeutic or functional foods, and beverages.

In a further aspect of the invention, the compositions according to the invention may be used as a medicament. Thus, the compositions of the invention may be administered in pharmaceutical form or as a dietary supplement, most preferably in combination with at least one pharmaceutically or food acceptable carrier.

The compositions according to the invention in the form of nutritional products, e.g. supplements or pharmaceutical preparations, may consist solely of the compositions according to the invention, or may additionally contain pharmaceutically or food acceptable carriers.

In a further aspect the invention provides a medicament, e.g. a clinical product, nutritional or pharmaceutical preparation, e.g. a food supplement, containing a composition of the invention. The medicament, e.g. the clinical product, nutritional or pharmaceutical composition of the invention may additionally contain pharmaceutically acceptable carriers. Moreover, the invention relates to combined pharmaceutical preparations for simultaneous, separate or sequential use in the treatment or prevention of gastrointestinal diseases, e.g. for controlling and / or restoring the gut microflora, stimulating the growth or multiplication of beneficial gut bacteria, and / or inhibiting the growth or multiplication of unfavorable or pathogenic gut bacteria, for treating or preventing IBD, for prolonging periods of remission, or for alleviating symptoms or conditions associated with the disease , such as ulcerative colitis, Crohn's disease, and / or colorectal cancer, to suppress or prolong periods of remission in IBD patients. especially in patients with ulcerative colitis. In another aspect of the invention, the compositions of the invention, e.g. combined pharmaceutical preparations, can be used in the prevention and treatment of IBS and the symptoms thereof.

PL 213 665 B1

The term preferred gut bacteria as used herein refers to beneficial or potentially beneficial gut bacteria such as lactobacilli and bifidobacteria; the term unfavorable intestinal bacteria refers to unfavorable or pathogenic intestinal bacteria such as strains of the genera Bacteroides or Clostridium, coliform organisms, sulfate reducing bacteria.

According to the invention, the term nutritional compositions refers to nutritional preparations and food products such as nutrients, dietary or nutritional supplements, functional foods, beverages, meal replacement preparations or food additives.

Such nutritional compositions may be nutritionally complete, i.e. may contain vitamins, minerals, trace elements as well as sources of nitrogen, carbohydrate and fatty acids so that they can be used as the sole source of nutrition providing substantially all the necessary vitamins, minerals, carbohydrates. , fatty acids, proteins and the like. Thus, the compositions of the invention may be provided in the form of a nutritionally balanced composition, e.g. a complete dietary preparation or a complete meal, e.g. suitable for oral or tube feeding.

Optionally, the compositions of the invention may be provided as part of a meal, e.g. a food or nutritional supplement, e.g. in the form of a health drink.

It may be desirable to provide the compositions of the invention in the form of a low caloric composition, e.g. a meal replacement formulation. In this case, a nutritional composition is preferred, i.e., a low fat meal replacement formulation, i.e., less than about 10%, or nearly skim, i.e., containing less than about 2.5% fat derived, such as about 2% fat, based on the total caloric content of the composition. Suitably, a single meal of a low caloric nutritional composition, e.g., a meal replacement formulation, will have a caloric value of less than about 1000 calories, and most preferably between about 200 kcal and about 500 kcal. A suitable low calorie nutritional composition may include a non-alcoholic beverage such as juice, milkshake or soy based beverage, or admixed with any type of food, e.g. dairy-based bars, soups, cereals such as cereals, muesli, sweets, drops, biscuits, shortbread cookies, crackers such as rice crackers, and dairy products such as milkshakes, yoghurt drinks, yoghurt, and fruit drinks.

Optionally, the compositions of the invention may be provided as high caloric compositions, e.g., high caloric nutritional supplements or meal replacement preparation, e.g., more than 400 kcal calories, more preferably more than 600 kcal, most preferably more than 800 kcal.

Compositions of the invention, e.g. high calorie compositions, may be protein-rich, i.e. may contain greater than about 1% protein (e.g., 1 g per 100 kcal), more preferably greater than about 2% protein, more preferably greater than about 4% protein. , for example, they may contain from about 1% to about 15% protein, more preferably from about 2% to about 10% protein, more preferably from about 3% to about 5% protein, based on the total caloric content of the composition.

Compositions of the invention, e.g. high calorie compositions, may contain fat, e.g., greater than about 4%, more preferably greater than 5%, more preferably greater than 6%, more preferably greater than 7% and most preferably greater than 8%, of the total caloric content of the composition. .

In one embodiment, the invention provides a composition comprising, e.g., per 100 ml, from about 1 to about 3 g FOS, from about 1 to about 3 g GOS, from about 2 to 20 g protein, from about 1 to about 30 g carbohydrate, from about 0.5 to about 20 g of fat as well as minerals and vitamins, most preferably about 1.25 g FOS, about 1.25 g GOS, about 9 g protein, about 22 g carbohydrate and about 9 g fat as well as minerals and vitamins .

Such a composition can be a complete dietary preparation. Such a composition, e.g. a complete dietary preparation, can be a ready-to-eat composition.

The compositions of the invention may also contain conventional food additives such as emulsifying, stabilizing, sweetening, flavoring, coloring, preserving, chelating, osmotic, buffering or pH adjusting agents, acidulants, thickeners, formers and so on.

Suitable forms of the products of the present invention include ready-to-drink solution compositions, e.g. ready-to-drink compositions, soluble drinks, liquid food products such as soft drinks, juices, sports drinks,

And milkshakes, milkshakes, yoghurt drinks and soups. In a further aspect of the invention, the compositions of the present invention may be produced and sold in the form of a concentrate, powder, or granules, e.g. effervescent granules, which are dissolved in water or other liquid such as milk or fruit juice to make it ready for consumption. a composition, e.g. ready-to-drink compositions or soluble drinks.

In a further aspect the invention relates to the use of the compositions according to the invention as food additives.

The pharmaceutical compositions, e.g. clinical products, and nutritional compositions of the invention, e.g. dietary supplements, may be provided in the form of soft gel capsules, sachets, powders, syrups, liquid suspension, emulsions and solutions in easy-to-dose form. In soft capsules, it is most preferred that the active ingredients are dissolved or suspended in suitable liquids, such as fatty oils, paraffin oil or liquid polyethylene glycols. Optionally, stabilizers can be added.

Oral pharmaceutical or nutritional supplement forms can be obtained by conventional combination methods known in the pharmaceutical art, i.e. by mixing the active ingredients together with pharmaceutically acceptable edible solid or liquid carriers and / or excipients, e.g. fillers such as cellulose, lactose, sucrose, mannitol, sorbitol, and calcium phosphates, and binders such as starch, gelatin, tragacanth, methylcellulose, and / or polyvinylpyrrolidone (PVP). Other possible additives include lubricants and flow aids, e.g. silicic acid, silica, talc, stearic acid, magnesium / calcium stearates, polyethylene glycols (PEG) as solvents, disintegrants e.g. starch, carboxymethyl starch, cross-linked PVP, agar, alginic acid and alginates, coloring agents, flavoring agents, and dissolving agents. Dyes and pigments may be added to the coating layer of the tablets and dragees, for example for identification purposes or to indicate different doses of active ingredient.

The composition of the invention may be in any form suitable for administration to humans, and in particular for administration to any part of the gastrointestinal tract. The present invention encompasses enteral administration of the compositions of the invention, most preferably oral administration, and administration via a tube or catheter.

The amount and dosing regimen of the compositions of the invention to be administered will be assessed in light of a variety of factors that may be relevant, including the purpose of administration, the age, sex and weight of the individual patient, and the severity of the patient's symptoms.

The compositions of the invention may be administered under the supervision of a person skilled in the art or the patient may take them alone.

Compositions of the invention, e.g. pharmaceutical or nutritional compositions, e.g. beverages or foods containing the compositions of the invention can be safely consumed by anyone, and are particularly recommended for mammals, such as humans, e.g. adults suffering from diarrhea and / or constipation, or diseases, conditions and symptoms associated with IBD, in particular ulcerative colitis, Crohn's disease or colorectal cancer, or in IBS and its syndromes. Compositions according to the invention, e.g. dietary supplements are especially recommended for mammals, such as humans, e.g. adults at risk of diarrhea, constipation, or other chronic bowel diseases as described above, e.g. malnourished, hospitalized or long-term patients, and / or to patients with ulcerative colitis during remission. The compositions according to the invention are particularly suitable for the malnourished and / or the elderly.

In one embodiment, the invention relates to a method of treating and / or preventing gastrointestinal disorders such as diseases, conditions and symptoms associated with chronic bowel diseases, e.g. IBD, and in particular ulcerative colitis, Crohn's disease, colorectal cancer, or IBS, and syndromes thereof, as described above, in mammals, including humans, in need of such therapy, the method comprising administering to said mammal an effective amount of a composition of the invention. An effective amount as used herein refers to an amount sufficient to achieve the desired therapeutic effect, such as treating and / or preventing diarrhea, constipation or diseases, conditions and symptoms associated with IBD, particularly ulcerative colitis, Crohn's disease, colorectal cancer, or conditions and symptoms associated with IBS, as described above.

In another embodiment of the invention, the compositions of the invention can be used in the manufacture of a medicament or nutritional formulation for the prevention or treatment of diseases, conditions and symptoms associated with chronic bowel disorders, e.g. IBD, and in particular ulcerative colitis, Crohn's disease, colon cancer, or for preventing or treating IBS or its syndrome10

For the treatment or prevention of gastrointestinal disorders such as diarrhea and constipation, as described above, in mammals, including humans.

In a further embodiment, the invention finds use in a method of maintaining and / or restoring the intestinal flora, e.g. the intestinal microflora, converting the intestinal bacterial population towards a healthier composition, in particular stimulating the growth and / or metabolism of beneficial intestinal bacteria, e.g. bifidobacteria and / or acid bacilli lactic acid (Lactobacillus), and / or inhibiting the growth and / or metabolism of unfavorable or pathogenic intestinal bacteria, e.g. Bacteroides, Clostridium, coliform organisms and / or sulfate reducing bacteria.

In yet another aspect, the invention finds use in a method of preventing or treating infections caused by pathogenic intestinal bacteria.

In a further aspect the invention finds use in a method of preventing and / or treating diseases, conditions and symptoms associated with chronic bowel diseases, e.g. IBD, particularly ulcerative colitis, Crohn's disease, colorectal cancer, or for prolonging the remission period in IBD patients. , particularly in patients with ulcerative colitis.

In a further aspect, the invention finds use in a method of treating or preventing gastrointestinal (GI) disorders, such as diarrhea or constipation.

In addition, the invention finds use in a method of preventing or treating IBS or its syndromes as described above in mammals, including humans.

Depending on the method of administering the compositions of the invention, e.g. as a complete dietary preparation, e.g. a complete meal, or as part of a meal, food additive, beverage, sachets, tablets or capsules, the compositions of the invention may be administered from once a day to six times a day. .

For patients using the compositions according to the invention as a supplement to their normal diet, the daily dose may be in the form of 1 or 2 meals per day. For patients using the compositions of the invention as a whole of their daily food intake, up to six meals per day may be recommended. The compositions according to the invention may be administered independently of the time of day, e.g. together with the main meals.

The compositions according to the invention may be administered under the control of a person skilled in the art or the patient may take them alone

Optimally, the compositions of the invention, e.g. food supplements, are consumed at least weekly or daily, regularly throughout the course of therapy, e.g., until a normal weekly frequency of bowel movements (e.g., 3 to 10 times) and normal stool are observed (e.g. e.g. and / or until the diarrhea or constipation improves or subsides, and / or the patient no longer experiences any discomfort, abdominal pain, gas, gas, and / or blood or excess mucus in the stools has ceased.

In another aspect, the present invention relates to a method of producing a composition of the invention, said method comprising intimately mixing the ingredients of the composition of the invention. Such methods are well known in the art.

The utility of all compositions of the present invention can be observed in standard clinical trials with, for example, the indications described above, for example using dosages of FOS and GOS ranging from about 0.01 to about 1 g / kg body weight / day, most preferably from about 0.01 g / kg body weight / day. 05 to about 0.5 g / kg body weight / day, or using doses of FOS and GOS ranging from about 1 to about 30 g, or up to about 30 g per meal, for a mammal, e.g., an adult, and in standard animal models. The effect of the compositions of the invention on the control and / or restoration of the intestinal flora, e.g., the prevention and treatment of IBD and associated diseases or symptoms, can be controlled by any method known to those skilled in the art, e.g. from the sigmoid / rectal area. The bacterial population of healthy individuals and those with e.g. ulcerative colitis can be compared and differences between colon bacteria populations e.g. by fluorescence in situ hybridization (FISH) / or denaturing gradient gel electrophoresis (DGGE), polymerase chain reaction (PCR) and 16S rDNA sequence analysis. FISH is a culture-independent molecular technique that uses 16S rRNA oligonucleotide probes labeled with fluorescent dyes (see Table 1). The FISH method enables imaging and localization of whole bacterial cells in situ in biological samples. It is obvious that such a method is known to the person skilled in the art.

PL 213 665 B1

Table 1. Oligonucleotide probes used to characterize the intestinal microflora by FISH

<td>Probe</td><td>Sequence</td><td>Destination type</td><td>Temp.</td>

<td>Bac 303</td><td>5'-CCAATGTGGGGGACCTT-3 '</td><td>Bacteroides spp.</td><td>450 ° C</td>

<td>Bif 164</td><td>5'-CATCCGGCATTACCACCC-3 '</td><td>Bifidobacterium spp.</td><td>500 ° C</td>

<td>Erec 482</td><td>5'-CGGUACCUGACUAAGAAGC-3 '</td><td>Group Clostridium coccoidesEubacterium rectale</td><td>500 ° C</td>

<td>Chis 150</td><td>5'-AAAGGAAGAUUAAUACCCGCAUA-3 '</td><td>Clostridium histolyticum group</td><td>500 ° C</td>

<td>Ec1531</td><td>5'-CACCGTAGTGCCTCGTCATCA-3 '</td><td>E.coli</td><td>370 ° C</td>

<td>Lab 158</td><td>5'-GGTATTAGCA (T / C) CTGTTTCCA-3 '</td><td>Lactobacillus / Enterococcus spp.</td><td>450 ° C</td>

<td>Srb 687</td><td>5-TACGGATTTCACTCCT-3 '</td><td>Desulfovibno spp.</td><td>480 ° C</td>

One human clinical trial is conducted as follows: A double-blind randomized study comparing the compositions of the invention, e.g. using doses of FOS ranging from about 0.01 to about 1 g / kg body weight / day, more preferably from about 0.05 to about 0.5 g / kg body weight / day, more preferably from about 0.1 to 0.15 to about 0.2 g / kg body weight / day and doses of GOS ranging from 0.01 to about 1 g / kg body weight / day, more preferably from about 0.05 to about 0.5 g / kg body weight / day , more preferably from about 0.1 or 0.15 to about 0.2 g / kg body weight / day, with a standard food supplement is performed in patients with ulcerative colitis to evaluate the effects on intestinal function, fecal microflora, or intestinal prebiotic fermentation over a 12-month treatment period. 30 patients are examined. The following parameters are assessed weekly: stool frequency, blood in the stool, excess mucus production, stool consistency, gas, and abdominal pain. Fecal microflora is characterized by FISH and compares the number of preferred strains (e.g. Lactobacillus and Bifidobacterium) with the number of strains associated with disease conditions (e.g. Clostridium, Bacteroides and sulfate reducing bacteria). The concentration of short-chain fatty acids (acetic, propionic, butyric, lactic and veraic) in stool samples is also assessed. Testing of total DNA extracts from stools is performed using PCR and DGGE.

The following examples illustrate the invention:

Example 1: In vitro studies of the prebiotic potential of several oligosaccharides and mixtures thereof

The fructooligosaccharides (FOS) used as positive control is Actilight950P®, from Beghin-Meiji Industries, France (containing 92% oligosaccharides).

Galactooligosaccharides (GOS) are Elixor®, FCDF, The Netherlands (containing 58% oligosaccharides, 23% lactose, 19% glucose).

Xylooligosaccharides (XOS) are Xylo-oligo95P®, Suntory Limited, Japan (containing 91% oligosaccharides).

Soy Oligosaccharides (SOS) are Soybean Oligosaccharides Syrup, Soya Oligo Japan Inc., Japan (containing 23% oligosaccharides, 21% sucrose, 31% other saccharides).

Arabinogalactan (ABG) is ClearTracAG-99®, Larex Inc., USA (containing 95% soluble fiber).

Gum Arabic (AG), also known as Senegalese gum, is Fibregum®, Colloides Naturels International, France (containing 85% soluble fiber).

Wheat Germ (WG) is BiogermPB1®, Multiforsa, Switzerland (containing 32% oligosaccharides, 30% protein, 12% fiber, 7% fat).

Isomaltooligosaccharides (IMO) are Isomalto900®, Showa Sangyo Co., Japan (containing 89% oligosaccharides).

1. Way:

The prebiotic potential of FOS, GOS, XOS, SOS, ABG, AG, WG and IMO was assessed using in vitro cultures of stool samples. Similarly, the in vitro prebiotic potential of the mixtures of oligosaccharides FOS + GOS, AG + FOS, FOS + XOS, XOS + GOS and AG + GOS was investigated in equal ratios. FOS was used as a positive control, to which the prebiotic properties of the tested W12 were compared

Of carbohydrates. Cultures of stool samples using test compound (s) as sole carbon source were performed using fecal inoculants from 6 healthy adults who did not suffer from known metabolic diseases and gastrointestinal diseases (e.g. diabetes, ulcerative colitis, Crohn's disease, ulcers). digestive tract, IBS, gastroenteritis, and cancer). Samples were collected on site, stored anaerobically, and used for a maximum of 5 minutes after collection. A 1:10 dilution was prepared in 0.1M anaerobic phosphate buffer (pH 7.4) and the samples were homogenized in a stomacher for 2 minutes.

(a) Culture series

135 ml of sterile chemostatic medium was stored in an anaerobic chamber (continuous O2-free N2 gassing) at 37 ° C in agitated containers. The pH of the culture was maintained at 6.7. Carbon sources were then added, 1% w / v. 15 ml of fresh human faecal suspension (10% w / v) prepared in phosphate buffered anaerobic saline was inoculated into each container. The cultures of the samples were grown for 24 hours.

(b) Counting bacteria

Samples of the batch cultures were taken after 0.5, 10 and 24 hours of culture. Fluorescence in situ hybridization was used to count the bacteria. Total numbers of bacteria, Bacteroides spp., Bifidobacterium spp., Clostridium perfringens / histolyticum and Lactobacillus subgroups were counted using group-specific fluorescently labeled DNA probes complementary with 16S rRNA (see Table 1).

2. Results

The results presented and discussed here represent the combined results of 6 trials for each possible prebiotic.

FOS + GOS were selectively and efficiently fermented by bifidobacteria both after 10 and 24 hours of cultivation. Growth was higher at both time points than when fermented with FOS or GOS separately. The amount of Bacteroides decreased slightly. The combination increased to one of the highest levels of all substrates tested in Lactobacillus during the first 10 hours of fermentation. The high increases did not persist after 24 hours, but their number remained above the baseline level. Although Clostridiums showed a slight increase after 10 hours, their numbers had dropped below baseline levels after 24 hours. After 24 hours, a very slight decrease in the number of Bacteroides was observed. FOS + GOS were quickly consumed by bifidobacteria and lactobacilli, and the increase in their number was much higher than for FOS or GOS separately. This relatively high increase in the number of Lactobacillus after 10 hours of fermentation is very important as prebiotics are usually fermented by bifidobacteria and have little or no effect on Lactobacillus. The combination appears to synergistically increase the number of beneficial bacteria.

3. Discussion

The results clearly showed that the oligosaccharides are fermented by the beneficial bacteria much more selectively, mainly by bifidobacteria and lactobacilli, than the soluble fiber tested in these studies.

The combination of FOS and GOS works synergistically and surprisingly increases the number of bifidobacteria and lactobacilli to a greater extent than when these oligosaccharides are tested separately. This combination does not appear to reduce the number of less beneficial bacteria, but rather slightly increases their number, compared to the two oligosaccharides tested separately. They are particularly good prebiotics due to their selective growth-promoting properties. The studies clearly showed that, of all the ingredients and combinations tested, the combination of FOS and GOS showed the best prebiotic activity over the entire fermentation period.

Example 2: nutritional composition: per 100 g

FOS 1 g

GOS 1 g

Hydrolysed guar gum⁽¹⁾ 0.2 g

Glutamine 1 g

Eicosapentaenoic acid (EPA) 0.43 g

Docosahexaenoic acid (DHA) 0.28 g ⁽¹⁾ Benefiber®, ex Novartis Nutrition Corporation.

Example 3: nutritional composition: per 100 g:

The composition is in the form of a powder, the daily dose (42 g) should be dissolved in about 200 ml of cold water before consumption.

PL 213 665 B1

<td>Vivinal® GOS 10⁽²⁾Actilight 950P FOS⁽³⁾Skimmed milk powder Powdered full-fat milk Coconut flavor MCT oil A chocolate flavoring agent Caramel flavor Sweetener - sodium saccharin, purified vanillin</td><td>42.38 g 12.62 g 21.49 g 9.52 g 6.55 g 1.07 g 0.71 g 0.19 g 0.07 g 0.03 g</td>

⁽²⁾ from Borculo Domo Ingredients (The Netherlands), it contains about 28% by weight of GOS ⁽³⁾ from Beghin-Meiji Industries (France), contains about 92% by weight of FOS.

Example 4: ready-to-drink nutritional supplement, per 100 ml (100 kcal)

<td>_FOS⁽⁴⁾_GOS⁽⁵⁾Protein Carbohydrates Saccharose Lactose Fats Saturated fatty acids. Monounsaturated fatty acids. Polyunsaturated fatty acids Vitamin A Vitamin D Vitamin E Vitamin C Vitamin K. Vitamin B1 Vitamin B2 Vitamin B6 Vitamin B12 Biotin Folic acid Niacin Pantothenic acid Sodium Potassium Calcium Magnesium Phosphorus Chlorine Iron Zinc Copper Molybdenum Manganese Chrome Selenium Fluorine Iodine</td><td>1.25 g 1.25 g 9 g 21.4 g 5 g 0.7 g 8.7 g 0.81 g 4.84 g 2.54 g 80 μg 1 μg 1 mg 6 mg 7 μg 0.14 mg 0.16 mg 0.2 mg 0.14 μg 7.5 μg 20 μg 1.8 mg 0.6 mg 60 mg 160 mg 190 mg 30 mg 90 mg 70 mg 1.5 mg 1.5 mg 200 μg 8 μg 0.3 mg 10 μg 6 μg 150 μg 15 μg</td>

⁽⁴⁾ from Wiwinal GOS z Borculo Domo Ingredients (Netherlands) ⁽⁵⁾ from Raftiloz P95 from Orafti (Belgium).