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WO2025229000A1 - Composition nutritionnelle et utilisations - Google Patents

Composition nutritionnelle et utilisations

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Publication number
WO2025229000A1
WO2025229000A1 PCT/EP2025/061734 EP2025061734W WO2025229000A1 WO 2025229000 A1 WO2025229000 A1 WO 2025229000A1 EP 2025061734 W EP2025061734 W EP 2025061734W WO 2025229000 A1 WO2025229000 A1 WO 2025229000A1
Authority
WO
WIPO (PCT)
Prior art keywords
nutritional composition
pectin
composition according
pectin polysaccharides
subject
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/EP2025/061734
Other languages
English (en)
Inventor
Sebastian TIMS
Lotte Hendrika Johanna DOPHEIDE
Noortje IJSSENNAGGER
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nutricia NV
Original Assignee
Nutricia NV
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nutricia NV filed Critical Nutricia NV
Publication of WO2025229000A1 publication Critical patent/WO2025229000A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
    • A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/40—Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula

Definitions

  • the invention relates to a nutritional composition comprising non-digestible oligosaccharides, pectin polysaccharides and 2’-fucosyllactose.
  • the invention further relates to the use of the nutritional composition for improving gut health.
  • Prebiotic fibres can be utilized to shape the human gut microbiota.
  • modulatory and reparatory activity of prebiotics which is highly dependent on their inherent structural features.
  • polymeric pectin with a complex structure has been described as a promoter of proliferation of their utilizing microbes (Cantu-Jungles & Hamakera, New View on Dietary Fiber Selection for Predictable Shifts in Gut Microbiota. mBio. 2020;11 (1):e02179-19.) whereas some, but not all, oligomeric pectin has been reported to stimulate gut bifidogenic effects.
  • WO2013/032674 describes the use of human milk oligosaccharides for preventing injury and/or improving the healing of the gastrointestinal tract.
  • W02013/016111 describes the use of a nutritional composition for preventing injury and/or improving the healing of the gastrointestinal tract via the enhancement of the expression of various mucin- associated proteins.
  • US 2023/0020348 describes a method for enhancing healing of the gastrointestinal tract of an individual by administration of a composition comprising galacto-oligosaccharides which may further comprise a human milk oligosaccharide.
  • the overall health of an individual is linked to an optimal gut microbiota state.
  • Individuals undergoing various therapies or having various diseases and/or conditions are generally more susceptible to an imbalance microbiota than healthy individuals.
  • the optimization of fibre within our diet represents an important health strategy to improve metabolic and overall health. Therefore, the search for nutritional compositions capable of optimizing gut health is important for the development of microbiome-based personalized nutrition. There remains a need in the art for nutritional compositions that specifically promote gut health.
  • SCFA short chain fatty-acids
  • Increased colonic propionate levels are known to lead to a decrease in appetite due to the regulation of the hormones PYY and GLP-1 (Chambers et al. “Effects of targeted delivery of propionate to the human colon on appetite regulation, body weight maintenance and adiposity in overweight adults” Gut., 2015, 11 ; 1744-1754 and Morrison et al. “Formation of short chain fatty acids by the gut microbiota and their impact on human metabolism” Gut Microbes, 7, 189- 200).
  • a decrease in PYY and GLP-1 levels is desired in subjects with obesity, however, is an undesired side effect in subjects with disease-related malnutrition which causes them to lose (additional) weight. Decreasing the appetite of a patient which needs to eat to strengthen and/or recover from an illness is not desired.
  • the present invention overcomes the above problems and relates to a nutritional composition
  • a nutritional composition comprising non-digestible galactooligosaccharides and non-digestible fructooligosaccharides, pectin polysaccharides and human milk oligosaccharide 2’-fucosyllactose, wherein the pectin polysaccharides have a degree of polymerisation of at least 260.
  • the inventors have surprisingly found that the combination of non-digestible oligosaccharides being galactooligosaccharides (GOS) and fructooligosaccharides (FOS) and pectin polysaccharides and 2’-fucosyllactose (2’-FL) has an improved effect on gut health.
  • the inventors have also surprisingly found that the combination of galactooligosaccharides and non-digestible fructooligosaccharides, pectin polysaccharides and human milk oligosaccharide 2’- fucosyllactose synergistically improves the gut barrier function by means of improved integrity of the cellular barriers by measuring the Transepithelial/transendothelial electrical resistance (TEER) in vitro in Caco-2 cells. More specifically, improving gut barrier integrity and function is supportive of promoting gut health and in particular supports promoting food absorption and uptake of nutrients from the Gl tract in subjects in need thereof.
  • TEER Transepithelial/transendothelial electrical resistance
  • the invention relates to the use of the nutritional composition
  • the nutritional composition comprising non- digestible galactooligosaccharides and non-digestible fructooligosaccharides, pectin polysaccharides and human milk oligosaccharide 2’-fucosyllactose wherein the pectin polysaccharides have a degree of polymerisation of at least 260 for use in improving gut health in a subject, in particular, for use in one or more of:
  • a further aspect of the invention is the non-therapeutic use of the nutritional composition according to the invention for supporting one or more of gut health, increased appetite, healthy physical growth in a subject, wherein said use comprises administering the nutritional composition according to the invention to said subject.
  • the inventors surprisingly found that the combination of non-digestible galactooligosaccharides (GOS) and non-digestible fructooligosaccharides (FOS), pectin polysaccharides and 2’-fucosyllactose (2’-FL) decreased propionate levels compared to the combination of GOS/FOS alone. This decrease in propionate levels is beneficial because this inherently means a decrease in the amount of PYY and/or GLP-1 which is produced and thereby decreasing appetite.
  • GOS non-digestible galactooligosaccharides
  • FOS non-digestible fructooligosaccharides
  • 2’-FL 2’-fucosyllactose
  • composition according to the invention further led to improved gut barrier function as shown by the TEER data in Figure 2 and 3. It is known that improved gut barrier function leads to improved food absorption by maintaining the integrity of the gut barrier and reducing gut permeability thereby facilitating the efficient uptake of nutrients from digested food.
  • Figure 1 Propionic acid finding after 72 hours of anaerobic fermentation with pectin polysaccharides, 2’-FL, GOS/FOS 9:1 and the combination of GOS/FOS/pectin polysaccharides/2’-FL (9:1 :2:1).
  • the dotted line represents the expected theoretical value for the combination.
  • Figure 2 TEER values after 22h treatment of the Caco-2 cells for the 24h fermented samples of GOS/FOS/2’-FL (9:1 :1) and GOS/FOS/LvP/2’-FL (9:1 :2:1). Values are normalized for the blanc control. * P ⁇ 0.05 with a T-test.
  • Figure 3 TEER values after 23h treatment of the Caco-2 cells for the 48h fermented samples of LV pectin, GOS/FOS/2’-FL (9:1 :1) and GOS/FOS/LvP/2’-FL (9:1 :2:1). Values are normalized for the blanc control. * P ⁇ 0.05 by a one-way ANOVA with Tukey’s multiple comparisons test.
  • a nutritional composition comprising non-digestible galactooligosaccharides (GOS) and non- digestible fructooligosaccharides (FOS), pectin polysaccharides and human milk oligosaccharide 2’- fucosyllactose (2’-FL) wherein the pectin polysaccharides have a degree of polymerisation of at least 2.
  • the nutritional composition according to embodiment 1 wherein the pectin polysaccharides have a degree of polymerisation of less than 6000 and/or a molecular weight of less than 150 kDa, preferably has a molecular weight of 50 - 150 kDa.
  • pectin polysaccharides comprise a substantially unbranched backbone of galacturonic acid, preferably the pectin polysaccharides comprise, by weight of the polysaccharide, less than 20% of branched galacturonic acid residues, more preferably less than 15% of branched galacturonic acid residues.
  • the pectin polysaccharides comprise at least 65% molar weight of 1 ,4-linked a-D-galacturonic acid residues, preferably 70% molar weight of 1 ,4-linked a-D-galacturonic acid residues, most preferably at least 80% molar weight of 1 ,4-linked a-D-galacturonic acid residues.
  • a non-therapeutic method for supporting one or more of gut health, increased appetite, healthy physical growth in a subject comprising administering a nutritional composition according to any one of embodiments 1 - 10 to said subject, wherein the subject is a healthy adult.
  • the present invention relates to a nutritional composition
  • a nutritional composition comprising non-digestible galactooligosaccharides (GOS) and non-digestible fructooligosaccharides (FOS), pectin polysaccharides and 2’-fucosyllactose (2’-FL), wherein the pectin polysaccharides have a degree of polymerisation of at least 260.
  • the present invention relates to a combination of non-digestible galactooligosaccharides and non-digestible fructooligosaccharides, pectin polysaccharides and human milk oligosaccharide 2’-fucosyllactose wherein the pectin polysaccharides have a degree of polymerisation of at least 260.
  • the invention further relates to the use of the nutritional composition of the invention comprising non-digestible galactooligosaccharides (GOS) and non-digestible fructooligosaccharides (FOS), pectin polysaccharides and 2’-fucosyllactose (2’-FL), wherein the pectin polysaccharides have a degree of polymerisation of at least 260 is for the preparation of a medical nutritional product, preferably an oral nutritional supplement for adults.
  • the use of the nutritional composition of the invention relates to improving gut health in a subject, in particular the use relates to one or more of: - stimulating intestinal barrier functions, stimulating integrity of the gut barrier;
  • the invention pertains to a composition for use in improving gut health in a subject, in particular for use in one or more of:
  • composition comprises non-digestible galactooligosaccharides (GOS) and non-digestible fructooligosaccharides(FOS), pectin polysaccharides and 2’-fucosyllactose (2’-FL), wherein the pectin polysaccharides have a degree of polymerisation of at least 260.
  • the second aspect of the invention can be defined as the use of the nutritional composition for the manufacture of a composition for improving gut health in a subject, in particular the use relates to one or more of:
  • composition comprises non-digestible galactooligosaccharides (GOS) and non-digestible fructooligosaccharides(FOS), pectin polysaccharides and 2’-fucosyllactose (2’-FL), wherein the pectin polysaccharides have a degree of polymerisation of at least 260.
  • the invention may be defined as a method of improving gut health in a subject, in particular the method relates to one or more of:
  • the method comprises administration of the nutritional composition to said subject and wherein the composition comprises non-digestible galactooligosaccharides and non- digestible fructooligosaccharides, pectin polysaccharides and 2’-FL, wherein the pectin polysaccharides have a degree of polymerisation of at least 260.
  • the present invention also encompasses the non-therapeutic method for supporting one or more of gut health, increased appetite, healthy physical growth in a subject, said method comprising administering a nutritional composition according to the invention to said subject, wherein the subject is a healthy adult and wherein the nutritional composition comprises non-digestible galactooligosaccharides and non-digestible fructooligosaccharides, pectin polysaccharides and 2’-FL, wherein the pectin polysaccharides have a degree of polymerisation of at least 260.
  • the concentration of pectin polysaccharides and their monosaccharide composition can be determined by analytical techniques known to the skilled person. After acid hydrolysis, the monosaccharide composition can suitably be determined by High Performance Anion Exchange Chromatography combined with Pulse Amperometric Detection (HPAEC-PAD).
  • HPAEC-PAD High Performance Anion Exchange Chromatography combined with Pulse Amperometric Detection
  • the molecular size distribution can be determined by High Performance Size-Exclusion Chromatography using refractive index (Rl) detection (concentration), light scattering detection (molecular mass detection), UV detection (indicative for presence of proteins) and differential pressure detection (intrinsic viscosity detection).
  • Rl refractive index
  • UV detection indicative for presence of proteins
  • differential pressure detection intrinsic viscosity detection
  • Nutritional composition means a substance or formulation that satisfies at least a portion of a subject's nutrient requirements.
  • the terms “nutritional(s)”, “nutritional formula(s)”, “enteral nutritional(s)”, and “nutritional supplement(s)” are used as non-limiting examples of nutritional composition ⁇ ) throughout the present disclosure.
  • “nutritional composition (s)” may refer to liquids, powders, gels, pastes, solids, concentrates, suspensions, or ready-to-use forms of enteral formulas, oral formulas, formulas for infants, formulas for pediatric subjects, formulas for children, growing-up milks and/or formulas for adults.
  • degree of hydrolysis refers to the extent to which peptide bonds are broken by a hydrolysis method.
  • Degree of polymerization refers to the number of monomeric saccharide units in a (poly- or oligo-)saccharide that are linked by glycosidic bonds.
  • partially hydrolyzed means having a degree of hydrolysis which is greater than 0 percent but less than about 50 percent.
  • fiber or ‘dietary fiber’ means the edible part of plants or analogous carbohydrates that are resistant to digestion and absorption in the human small intestine with complete, partial or no fermentation in the large intestine.
  • SCFA short chain fatty acid(s) that are produced in the intestinal tract as an end-product of bacterial fermentation and refers to aliphatic carboxylic acids with a hydrocarbon chain with a hydrocarbon chain with less than six carbon atoms.
  • LvP stands for low viscosity pectin and refers to a pectin that is partially hydrolyzed.
  • the terms “nutritional composition” and “composition” are used interchangeably.
  • the nutritional composition of the invention comprises non-digestible galactooligosaccharides and non-digestible fructooligosaccharides, pectin polysaccharides and 2’-fucosyllactose (2’-FL).
  • the invention relates to a combination of non-digestible galactooligosaccharides (GOS) and non-digestible fructooligosaccharides (FOS), pectin polysaccharides and 2’-fucosyllactose (2’-FL).
  • the invention also relates to a nutritional composition according to the invention and macro ingredients selected from digestible carbohydrates, fats, proteins.
  • the nutritional composition comprises a weight ratio of the sum of non-digestible oligosaccharides to pectin polysaccharides of 2 : 1 to 30 : 1 , more preferably 3 : 1 to 20 : 1 , even more preferably 4 : 1 to 10 : 1 .
  • the non-digestible oligosaccharides are galactooligosaccharides and fructooligosaccharides in a weight ratio of 2:1 to 15:1 , more preferably 9:1 .
  • the weight ratio of galactooligosaccharides to fructooligosaccharides to pectin polysaccharides is 5 - 15 : 0.5 - 5 : 0.5 - 10, more preferably 5 - 13 : 0.7 - 2 : 0.75 - 4, even more preferably 7 - 10 : 0.8 - 1 .5 : 1 - 3, most preferably 8.5 - 9.5 : 0.8 - 1 .2 : 1 .5 - 2.2.
  • the weight ratio of galactooligosaccharides to fructooligosaccharides to pectin polysaccharides preferably is 5 - 15 : 0.5 - 5 : 0.25 - 8, more preferably 5 - 13 : 0.7 - 2 : 0.5 - 4, most preferably 8.5 - 9.5 : 0.8 - 1 .2 : 0.75 - 2.
  • the nutritional composition comprises galactooligosaccharides, fructooligosaccharides, pectin polysaccharides and 2’-FL.
  • the weight ratio of galactooligosaccharides to fructooligosaccharides to pectin polysaccharides to 2’-FL is 6 - 12 : 0.1 - 3 : 0.5 - 3: 0.1 - 3, more preferably 8 - 10 : 0.5 - 2 : 0.75 - 2.5 : 0.5 - 2, most preferably 8.5 - 9.5 : 0.8 - 1.2 : 1 - 2.2 : 0.8 - 1.2.
  • the weight ratio of galactooligosaccharides to fructooligosaccharides to pectin polysaccharides to 2’FL preferably is 6 - 12 : 0.1 - 3 : 0.25 - 3: 0.1 - 3, more preferably 8 - 10 : 0.5 - 2 : 0.5 - 2.5 : 0.5 - 2, most preferably 8.5 - 9.5 : 0.8 - 1 .2 : 0.75 - 2 : 0.8 - 1.2.
  • the nutritional composition according to the invention may be used as a pharmaceutical product or preferably a nutritional product.
  • the present nutritional composition is a solid (typically a powder or tablet, preferably a powder) which is reconstitutable with a liquid, preferably with water, to obtain a liquid composition, more preferably, the nutritional composition is a liquid composition.
  • the composition is administered orally.
  • the dry weight of the nutritional composition is based on the total amount of protein, carbohydrates and fat present in the composition.
  • the composition comprises 40 - 55 wt.% dry weight based on total weight of the composition.
  • the dry weight of the nutritional composition is in the range of 35 - 55 wt.% based on total weight of the composition.
  • the nutritional composition according to the invention may be used as a pharmaceutical product comprising one or more pharmaceutically acceptable carrier materials.
  • Any conventional carrier material can be utilized.
  • the carrier material can be organic or inorganic inert carrier material suitable for oral administration. Suitable carriers include water, gelatine, gum Arabic, lactose, starch, magnesium stearate, talc, vegetable oils, and the like. Additionally, additives such as thickeners, flavouring agents, preservatives, stabilizers, emulsifying agents, buffers and the like may be added in accordance with accepted practices of pharmaceutical compounding. While the individual active ingredients are suitably administered in a single composition, they may also be administered in individual dosage units.
  • the nutritional composition according to the invention may be used as a nutritional product, for example as a nutritional supplement, e.g. as an additive to a normal diet, as a fortifier, to add to a normal diet, or as a complete nutrition.
  • the nutritional product preferably comprises at least one component, preferably all components, selected from the group of fats, proteins, and carbohydrates. It is understood that a nutritional product differs from a pharmaceutical product by the presence of nutrients which provide nutrition to the subject to whom the composition is administered, in particular, the presence of protein, fat, and digestible carbohydrates. It may further contain ingredients such as minerals, vitamins, organic acids, and flavouring agents.
  • the term "nutraceutical product” is often used in literature, it denotes a nutritional product with a pharmaceutical component or pharmaceutical purpose.
  • the nutritional composition according to the invention may also be used in a nutraceutical product.
  • the nutritional composition is preferably in compliance with food for special medical services, such as according to Food for Special Medical Purposes (FSMP) directive 1999/21 /EC of 25 March 1999 and as defined in Article 2.2(g) in Commission Regulation (EU) 609/2013.
  • FSMP Food for Special Medical Purposes
  • Administration of the nutritional composition occurs preferably at least one time per day, more preferably 1 - 3 times per day, although alternative dosage regimes can be determined from these numbers.
  • the nutritional composition is used as sole source of nutrition, preferably the nutritional composition is administered at least 4 times per day, more preferably 5 times per day to a subject in need thereof.
  • NDO Non-digestible oligosaccharides
  • non-digestible oligosaccharides refers to oligosaccharides which are not digested in the intestine by the action of acids or digestive enzymes present in the human upper digestive tract, e.g. small intestine and stomach, but which are preferably fermented by the human intestinal microbiota.
  • acids or digestive enzymes present in the human upper digestive tract, e.g. small intestine and stomach, but which are preferably fermented by the human intestinal microbiota.
  • glucose, galactose, sucrose, lactose, maltose and maltodextrins are considered digestible.
  • the nutritional composition comprises at least 300 mg of non-digestible oligosaccharides comprising galactooligosaccharides (GOS) and fructooligosaccharides (FOS) per 100 kcal based on total energy content of the composition.
  • the nutritional composition may comprise for instance 350 - 2000 mg per 100 kcal, of non-digestible oligosaccharides, preferably 400 - 1600 mg per 100 kcal of non-digestible oligosaccharides comprising GOS and FOS, based on total energy content of the composition.
  • the non-digestible oligosaccharide is selected from the group consisting of GOS and FOS, and combinations thereof.
  • the composition comprises non-digestible oligosaccharides with a degree of polymerization (DP) of 2 to 250, more preferably 3 to 60.
  • DP degree of polymerization
  • Suitable galactooligosaccharides include, but are not limited to, beta-galactooligosaccharides and/or alpha galactooligosaccharides.
  • the galactooligosaccharides preferably are beta-galacto- oligosaccharides.
  • the present nutritional composition comprises betagalacto-oligosaccharides ([galactose]n-glucose; wherein n is an integer ranging from 2 to 60, i.e. 2, 3, 4, 5, 6, ...., 59 ,60; preferably n is selected from 2, 3, 4, 5, 6, 7, 8, 9, and 10, wherein the galactose units are in majority linked together via a beta linkage.
  • Beta-galactooligosaccharides are also referred to as trans-galacto-oligosaccharides (TOS).
  • the galactooligosaccharide has beta-(1 ,4), beta-(1 ,3) and/or beta-(1 ,6) glycosidic bonds and a terminal glucose.
  • galacto-oligosaccharides comprise at least 80% beta-1 ,4 and beta-1 ,6 linkages based on total linkages, more preferably at least 90%.
  • the galacto-oligosaccharides comprise at least 50% beta-1 ,3 linkages based on total linkages, more preferably at least 60% based on total linkages.
  • the composition comprises non-digestible galacto- oligosaccharide comprising beta-(1 ,4) linkages, in particular beta-(1 ,4) linkages between the galactose units, and having a degree of polymerization of at least 4, preferably with a degree of polymerization from 2 to 10, more preferably of 3 to 7.
  • GOS VivinalOGOS from FrieslandCampina Domo (Amersfoort, The Netherlands) or Bimuno® GOS (Clasado Biosciences, Berkshire, United Kingdom).
  • the composition according to the present invention comprises at least 200 mg GOS per 100 kcal of the composition, more preferably from 300 - 1800 mg, most preferably from 400 - 1400 mg GOS per 100 kcal based on total energy content of the composition.
  • the nutritional composition comprises at least 1 .2 wt.% GOS based on dry weight of the composition, more preferably 1.5 - 5 wt.%, most preferably 1 .8 - 3.5 wt% GOS.
  • the composition according to the invention comprises 600 - 4000 mg GOS per 100 ml, more preferably 800 - 3500 mg, most preferably 1000 - 3200 mg GOS per 100 ml.
  • the composition according to the invention comprises fructo-oligosaccharides (FOS).
  • Fructooligosaccharides are a non-digestible oligosaccharide (NDO) comprising a chain of beta-linked fructose units with a degree of polymerization (DP) or average DP of 2 to 250, more preferably 2 to 100, even more preferably 10 to 60.
  • NDO non-digestible oligosaccharide
  • DP degree of polymerization
  • Fructo-oligosaccharide includes inulin, levan and/or a mixed type of polyfructan.
  • An especially preferred fructo-oligosaccharide is inulin.
  • Fructooligosaccharide suitable for use in the compositions is also commercially available, e.g., Raftiline®HP (Orafti, Beneo GmbH, Mannheim Germany).
  • the fructo-oligosaccharide has an average DP above 20.
  • the composition according to the present invention comprises at least 20 mg FOS per 100 kcal of the composition, more preferably from 30 - 250 mg, most preferably from 40 - 200 mg FOS per 100 kcal based on total energy content of the composition.
  • the composition comprises at least 0.1 wt.% FOS based on dry weight of the composition, more preferably 0.1 - 1.0 wt.%, most preferably 0.15 - 0.5 wt.% FOS.
  • the composition according to the invention comprises 60 - 450 mg FOS per 100 ml, more preferably 80 - 400 mg, most preferably 100 - 350 mg FOS per 100 ml of the composition.
  • the composition according to the invention comprises 800 - 4000 mg non-digestible oligosaccharides comprising galactooligosaccharides (GOS) and fructooligosaccharides (FOS) per 100 ml of the composition, more preferably 1000 - 3800 mg, most preferably 1100 - 3500 mg non-digestible oligosaccharides comprising galactooligosaccharides (GOS) and fructooligosaccharides (FOS) per 100 ml of the composition.
  • GOS galactooligosaccharides
  • FOS fructooligosaccharides
  • the combination of GOS and FOS is present in a weight ratio of from 1 :99 to 99:1 , more preferably from 1 :19 to 19:1 , more preferably from 1 :1 to 19:1 , more preferably from 2:1 to 15:1 , more preferably from 5:1 to 12:1 , even more preferably from 8:1 to 10:1 , even more preferably in a ratio of about 9:1 .
  • Pectins also known as pectin polysaccharides, are rich in galacturonic acid. Several distinct polysaccharides have been identified and characterised within the pectin group. Homogalacturonans are linear chains of a-(1-4)-linked D-galacturonic acid. Pectin fibers are obtainable from fruits and vegetables and are generally rich in galacturonic acid groups. Pectins are used in food as stabilizer or for the gelling properties of pectin.
  • pectins are classified as high- vs. low-methoxy pectins (short HM-pectins vs. LM-pectins), with more or less than 50 percent of all the galacturonic acids esterified respectively.
  • the ratio of esterified to non-esterified galacturonic acid determines the behaviour of pectin in food applications.
  • pectin is not used as stabilizer or thickener.
  • pectin polysaccharides can have a non-agglomerated or agglomerated format.
  • agglomeration refers to a minimal coating around the granules which upon reconstitution is not present anymore.
  • the difference is achieved by the presence of an agglomeration step after processing to make the pectin more dispersible.
  • pectin powder is rewetted with a media, allowing the fine particles to stick together to create a larger particle.
  • the particle’s surface increases its mixability, allowing it to dissolve faster. This can be an advantage when cold dissolution is aimed for.
  • dissolution is done at an elevated temperature, such as e.g. at about 70-80 °C, nonagglomerated pectin commonly also dissolves easily.
  • the agglomeration is not believed to have an impact on the physiological function of the pectin.
  • Pectins can be hydrolyzed to decrease the molecular weight.
  • partially hydrolyzed pectins are preferred to prevent undesired increases in viscosity.
  • they can be sometimes referred to as low viscosity pectins or LvP.
  • the pectin according to the present invention is a partially hydrolysed pectin, also called a low viscosity pectin, that will have a limited effect on viscosity but still is a good source of fermentable fiber.
  • Preferred pectin polysaccharides according to the invention are derived from fruits, preferably derived from citrus, apple, grapes, berries, more preferably derived from apple.
  • the pectin is mildly hydrolysed to prevent increasing the viscosity of the end product.
  • the low viscosity pectin has a degree of polymerisation of less than 6000 or a molecular weight of less than 150 kDa.
  • the pectin polysaccharides according to the invention have a preferred molecular weight of 60-1000 kDa, more preferably 70-500 kDa, more preferably 75 - 250 kDa, more preferably less than 150 kDa, even more preferably 50 - 150 kDa, most preferably 80 - 120 kDa.
  • the pectin polysaccharide of the invention has a degree of polymerisation (DP) of less than 6000, and/or a molecular weight of less than 150 kDa.
  • the DP is preferably at least 260, more preferably at least 300, even more preferably at least 450.
  • the pectin polysaccharides have a DP of less than 6000, more preferably less than 4000, more preferably less than 2000, even more preferably less than 1000.
  • pectins examples include, but are not limited to, Herbapekt SF 50-LV from Herbafood Ingredients GmbH, Germany, and Pectin GENU® from CP Kelco, Europe .
  • the pectin polysaccharides optionally can refer to optionally branched polysaccharides comprising a backbone that substantially consists of galacturonic acid residues.
  • backbone chain and “backbone” are synonyms.
  • the pectin polysaccharides according to the invention comprise at least 65% weight of 1 ,4-linked a-D-galacturonic acid residues, preferably 70% weight of 1 ,4-linked a-D- galacturonic acid residues, most preferably at least 80% weight of 1 ,4-linked a-D-galacturonic acid or at least 90% molar weight of 1 ,4-linked a-D-galacturonic acid residues.
  • the pectin polysaccharides preferably comprises at least 50% by weight of galacturonic acid residues, more preferably at least 55% by weight, even more preferably 60% by weight, most preferably at least 65% by weight of galacturonic acid residues.
  • the pectin polysaccharides comprise a substantially unbranched backbone of galacturonic acid residues.
  • the pectin polysaccharides preferably comprise less than 20% by weight of the polysaccharide of branched galacturonic acid residues, more preferably less than 15% by weight of the polysaccharide of branched galacturonic acid residues.
  • the 1 ,4-linked a-D-galacturonic acid residues as present in the pectin polysaccharide which would comprise further branched chains or side chains.
  • the pectin polysaccharides of the invention comprise less than 30% by weight of the polysaccharide of rhamnose residues, preferably less than 20%, more preferably less than 10%, even more preferably less than 5%, most preferably less than 4% by weight of the polysaccharide of rhamnose residues.
  • the rhamnose residues are in the backbone of the pectin polysaccharides.
  • the low abundance of rhamnose residues in the polysaccharides of the invention may also be expressed by the ratio between galacturonic acid residues and rhamnose residues.
  • the pectin polysaccharides comprise galacturonic acid residues and rhamnose residues in a molar ratio of more than 10:1 , preferably more than 20:1 .
  • the carboxyl groups of the galacturonic acid residues within the backbone of the pectin polysaccharides of the invention are typically esterified.
  • the pectin polysaccharides according to the invention have a high degree of esterification.
  • the esterification of the pectin polysaccharides is more than 50%, preferably more than 60% even more preferably more than 65%, most preferably more than 70%.
  • the esterification of the pectin polysaccharides is preferably 50 - 80%, more preferably 50 - 75%, even more preferably 60 - 72%.
  • Esterified galacturonic acid may occur in the form of the methyl ester or acetyl ester.
  • the pectin polysaccharides according to the invention have a high degree of methylation (DM).
  • the DM of the pectin polysaccharides in the present composition is of at least 45%, more preferably of at least 50%, even more preferably at least 55 or at least 58%.
  • the degree of acetylation (DA) of the pectin polysaccharides in the present composition is typically very low. Accordingly, the preferred DA is less than 18%, more preferably less than 15%, even more preferably less than 10%, most preferably less than 5%, most preferably less than 3%.
  • Pectin polysaccharides are formed by domains.
  • domain refers to the backbone with any sidechains that may attached thereto.
  • Backbone refers to the sequence of glycosidically linked monosaccharides within the backbone of a polysaccharide, excluding any sidechains that are attached thereto.
  • the pectin polysaccharides of the invention are rich in a domain called homogalacturonan (HG).
  • HG domains do not contain any sidechains.
  • the pectin polysaccharides of the present invention may contain small amounts of one or more of the following domains:
  • the backbone of the pectin polysaccharides of the invention is majorly comprised of homogalacturonan (HG) domains, preferably more than 35%, more preferably more than 40% by weight of the polysaccharide is HG domain.
  • HG domains are substantially free of rhamnose residues in the polysaccharide backbone and are substantially unbranched.
  • the sum of XG, AG, RG-I and RG-II domain represent no more than 45 wt.% on dry basis of the polysaccharide, preferably no more than 30 wt%, more preferably no more than 20 wt.% on dry basis of the polysaccharide.
  • the composition according to the invention preferably comprises at least 30 mg pectin polysaccharides per 100 kcal, more preferably 50 - 350 mg, even more preferably 60 - 300 mg pectin polysaccharides per 100 kcal of the composition.
  • the composition comprises at least 0.1 wt.% pectin polysaccharides based on dry weight of the composition, more preferably 0.1 - 1 wt.%, most preferably 0.2 - 0.8 wt% pectin polysaccharides.
  • the composition according to the invention comprises at least 100 mg pectin polysaccharides per 100 ml of the composition, even more preferably 200 - 800 mg, most preferably 250 - 700 mg based on 100 ml of the composition.
  • the composition comprises at least 0.05 wt.% pectin polysaccharides based on dry weight of the composition, more preferably 0.05 - 1 wt.%, most preferably 0.1 - 0.6 wt% pectin polysaccharides.
  • the composition according to the invention comprises at least 50 mg pectin polysaccharides per 100 ml of the composition, even more preferably 75 - 600 mg, most preferably 100 - 450 mg based on 100 ml of the composition.
  • the present nutritional composition may comprise fucosyllactose.
  • Fucosyllactose FL
  • FL is a non- digestible oligosaccharide present in human milk. It is not present in bovine milk. It consists of three monose units, fucose, galactose and glucose linked together.
  • Galactose linked to glucose via a beta 1 ,4 linkage is called lactose.
  • a fucose unit linked to a galactose unit of lactose via an alpha 1 ,2 linkage is 2’-fucosyllactose (2’-FL).
  • the nutritional compositions according to the invention comprise 2’-FL.
  • DP degree of polymerization
  • fucosylated oligosaccharides may be produced by Chemical synthesis from lactose and free fucose. Fucosylated oligosaccharides are also available for example from Kyowa, Hakko, Kogyo of Japan, Friesland Campina, The Netherlands, Glycom DSM, Denmark and Chr. Hansen, Denmark. 2’-FL, preferably a-L-Fuc-(1 a2)-p-D-Gal(1 a4)-D- Glc is commercially available from for instance Sigma-Aldrich. Alternatively, they can be isolated from human milk, for example as described in Andersson & Donald, 1981 , J. Chromatogr.
  • the nutritional composition according to the invention comprises 2’-FL in a concentration of at least 20 mg/100kcal, preferably of 30 mg/100 kcal - 250 mg/100 kcal, more preferably of 40mg/100 kcal - 180 mg/100 kcal.
  • the composition comprises at least 0.1 wt.% 2’-FL based on dry weight of the composition, more preferably 0.1 - 1 .0 wt.%, most preferably 0.15 - 0.5 wt% 2’-FL.
  • the composition according to the invention comprises at least 70 mg 2’-FL per 100 ml of the composition, more preferably 80 - 400 mg, most preferably 100 - 350 mg per 100 ml of the composition.
  • 2’-FL may be present in an amount of at least 0.5 mg/ml of the composition, more preferably in an amount from 0.75 - 4 mg/ml. In a particular embodiment, 2’-FL is present in an amount from 1 - 3.5 mg per ml of the composition based on total weight.
  • the nutritional composition according to the invention may in a preferred aspect be used as a nutritional supplement, i.e., as an additive to a diet.
  • the supplement preferably for enteral application, may be a solid or liquid galenical formulation.
  • solid galenical formulations are tablets, capsules (e.g. hard or soft shell gelatine capsules), pills, sachets, powders, granules and the like which contain the active ingredient together with conventional galenical carriers.
  • Any conventional carrier material can be utilized.
  • the carrier material can be organic or inorganic inert carrier material suitable for oral administration. Suitable carriers include water, gelatine, gum arabic, lactose, starch, magnesium stearate, talc, vegetable oils, and the like.
  • the nutritional composition a ready-to-use liquid composition.
  • the nutritional composition of the invention is a nutritional supplement.
  • the nutritional supplement comprises non-digestible galactooligosaccharides and non-digestible fructooligosaccharides, pectin polysaccharides and 2’-FL.
  • the nutritional supplement also comprises macro ingredients selected from digestible carbohydrates, fats, proteins, and, optionally, vitamins and minerals.
  • the nutritional supplement also comprises micronutrients selected from vitamins, trace elements and/or minerals.
  • the nutritional composition according to the invention comprising non-digestible GOS and non- digestible FOS, pectin polysaccharides and 2’-FL wherein the pectin polysaccharides have a degree of polymerisation of at least 260, is for use in improving gut health in a subject, preferably for use in one or more of:
  • the second aspect of the invention can be defined as the use of the nutritional composition comprising non-digestible GOS and non-digestible FOS, pectin polysaccharides and 2’-FL wherein the pectin polysaccharides have a degree of polymerisation of at least 260, for the manufacture of a composition for improving gut health in a subject, preferably for one or more of:
  • the invention may be defined as a method of improving gut health in a subject, preferably for one or more of:
  • the method comprising administration of the nutritional composition to said subject, the composition comprising GOS, FOS, pectin polysaccharides and 2’-FL and wherein the pectin polysaccharides have a degree of polymerisation of at least 260.
  • the nutritional compositions of the present invention are suitable for achieving beneficial effects in a subject, preferably in a human subject, wherein the human subject is selected from a toddler, a child and/or an adult.
  • the human subject is preferably older than 1 year, more preferably older than 2 years, even more preferably older than 18 years. In a further preferred embodiment, the human subject is an adult of 50 years or older.
  • the composition of the invention is for treating a subject suffering from or at risk of developing one or more selected from undesired rapid weight loss, muscle wasting, sarcopenia, cachexia, frailty, compromised microbiota or is suffering from malnutrition, preferably disease-related malnutrition, or is a subject in need of recovery after illness.
  • the subject has injury to the gastrointestinal tract or is a subject who is more susceptible to or at risk of injury to the gastrointestinal tract by having undergone surgery, by having chronic stress, by having one or more diseases or disorders which may include enteric infection, inflammatory bowel disease, colitis and bowel obstruction and/or intestinal mucosa injury.
  • the subject is suffering from cancer-related malnutrition.
  • the invention concerns a method, in particular a non-therapeutic method for supporting one or more of gut health, increased appetite, healthy physical growth in a subject, said method comprising administering to said subject a nutritional composition comprising GOS, FOS, pectin polysaccharides and 2’-FL, wherein the pectin polysaccharides have a degree of polymerisation of at least 260, and wherein said subject is preferably a healthy adult.
  • the subject is an athlete, a bodybuilder, an underweight individual, is in need of rejuvenation, is recovering from weight loss or has an increased physically active lifestyle compared to an average individual.
  • a subject with an increased physically active lifestyle is a person in need of more than 2000 or 2500 kcal per day being a female or male, respectively.
  • the nutritional composition comprises per 100 kcal of energy content of the composition:
  • pectin polysaccharides • at least 30 mg pectin polysaccharides, more preferably 50 - 350 mg, most preferably 60 - 300 mg pectin polysaccharides;
  • the nutritional composition comprises, when in liquid form, per 100 ml:
  • pectin polysaccharides • at least 100 mg pectin polysaccharides, more preferably 200 - 800 mg, most preferably 250 - 700 mg pectin polysaccharides;
  • the nutritional composition comprises per 100 g dry weight:
  • pectin polysaccharides • 50 - 1200 mg pectin polysaccharides, preferably 50 - 1000 mg, most preferably 100 - 800 mg pectin polysaccharides;
  • a composition according to the invention was prepared by dry blending commercially available GOS, FOS, pectin polysaccharides and 2’-FL in a weight ratio of 9:1 :2:1 .
  • the monosaccharide composition of pectin polysaccharides was analysed by methanolysis and is provided in Table 1 .
  • Fresh faecal samples were collected from 5 healthy Caucasian adults, 2 male and 3 female, aged 23- 48 years. The volunteers were without gastrointestinal problems and did not use antibiotics for the last 4 months. Faecal samples were pooled, homogenized, divided in smaller aliquots, and mixed with glycerol (10%) in an anaerobic cabinet. Subsequent aliquot storage was at -80°C.
  • the negative control was a blanc experiment performed without any substrate at pH 6.3. Faecal samples fed with glucose were performed as positive control. In the fermentation run a negative control was included. Fibres and glucose were added at a concentration of 200 mg dietary fibre (DP>3) per 6 ml of faeces suspension.
  • the faecal pool was defrosted in a water bath for 20 minutes at 37°C.
  • Fermentation medium (Me Bain and MacFarlane) contains buffered peptone water 3,0 g/l, Yeast Extract 2,5 g/l, Tryptone 3,0 g/l, L-Cysteine-HCI 0,4 g/l, Bile salts 0,05 g/l, K2HPO4.3H2O 2,6 g/l, NaHCO 3 0,2 g/l, NaCI 4,5 g/l, MgSC JFW 0,5 g/l, CaCh. 2H2O 0,3 g/l, FeSC JFhO 0,005 g/l.
  • Dialysis medium contains K2HPO4.3H2O 2,6 g/l, NaHCCh 0,2 g/l, NaCI 4,5 g/l, MgSC>4.7H2O 0,5 g/l, CaCl2.2H2O 0,3 g/l, FeSC>4.7H2O 0,005 g/l. pH was adjusted to 6.3 ⁇ 0.1 with K2HPO4 or NaHCCh. Medium was not sterilized because of forming of sediment. The medium was put in the anaerobic cabinet at least 16 hours before use.
  • the pH was measured by immersing a 423 pH-electrode (Mettler Toledo, Columbus, OH, USA), connected to a Handy-lab pH meter (Schott Gias, Mainz, Germany), directly in a sample.
  • the SCFA propionic acid value was quantitatively determined at 3 different time points using a Shimadzu- GC2025 gas chromatograph with a flame ionization detector. Hydrogen was used as mobile phase.
  • the levels of SCFA were determined using 2-ethylbutyric acid as an internal standard. 2-Ethylbutyric acid is used as internal standard because it is chemically synthesized and is not present in the original faecal samples. From the peak area a calibration curve was constructed and the concentration in the samples was calculated.
  • TEER epithelial barrier function trans epithelial resistance
  • Fecal samples were fermented with a composition comprising GOS/FOS/2’-FL (ratio 9:1 :1), GOS/FOS/LvP/2’-FL (ratio 9:1 :2:1) or LvP.
  • Samples were collected after 24 hours (condition A) and 48 hours (condition B) of fermentation.
  • 50 pl of fecal slurry was diluted 1/20 with Caco- 2 medium (DMEM) without FCS and filtered (0.2um) and subsequently was added to the apical upper left channel A1 , and 50 pl was added to apical channel A3.
  • DMEM Caco- 2 medium
  • dialysates (1/20 diluted in medium and filtered (0.2 urn) were added, 50 pl in C1 and 50 pl in C3.
  • the epithelial tubules in the Caco-2 plates were seeded in the upper channel of the plate (day 0) as described in Nicolas et al. (2021).
  • day 4 the plates were incubated horizontally and static for 1 h at 37 °C, whereafter the transport medium was removed and fresh Caco-2 medium (DMEM with FCS) was added according to standard protocols provided by Mimetas and as described in Nicolas et al. (2021).
  • the plates were placed on a rocker in the incubator for 24 h at 37 °C. On day 5 the medium was removed and the samples/dialysates were added to the plate.
  • the effect on TEER values after 22 hours of Caco-2 treatment with samples from condition A are shown in Figure 2.
  • the effect on TEER values after 23 hours of Caco-2 treatment with samples from condition B are shown in Figure 3.
  • the TEER data was normalized according to the blanc control (DMEM without FCS).
  • dialysates derived from the fecal samples treated with GOS/FOS/LvP/2’-FL resulted in a significant increase in TEER compared to dialysates derived from the fecal samples treated with the composition comprising GOS/FOS/2’-FL ( Figure 2).
  • Figure 3 shows that the combination of all four ingredients resulted in an unexpectedly increased TEER compared to the expected effects of LvP alone + combination of GOS/FOS/2’-FL.

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Abstract

L'invention concerne une composition nutritionnelle comprenant des oligosaccharides non digestibles, des polysaccharides de pectine et du 2'-fucosyllactose. L'invention concerne en outre l'utilisation de la composition nutritionnelle pour améliorer la santé intestinale.
PCT/EP2025/061734 2024-04-30 2025-04-29 Composition nutritionnelle et utilisations Pending WO2025229000A1 (fr)

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