Substances for use as a prebiotic or as a medicine

Abstract

The invention relates to the non-therapeutic use of a substance (c) selected from GDL, a gluconate salt, gluconic acid or from a mixture thereof, said substance being intended to be taken orally. The invention also relates to a substance (c) selected from GDL, a gluconate salt, gluconic acid or from a mixture thereof, for use as an oral medicament.

Description

DescriptionTitle: Substances for use as a prebiotic or as a medicineTechnical Field

[0001] The invention pertains to the field of oral prebiotics and medicines that act on gut microbiota.Background Art

[0002] Prebiotics are a group of nutrients that are degraded by gut microbiota. Their relationship with human overall health has been an area of increasing interest in recent years. They can feed the intestinal microbiota, and their degradation products are short-chain fatty acids that are released into blood circulation, consequently, affecting not only the gastrointestinal tracts but also other distant organs. Fructo-oligosaccharides and galactooligosaccharides are the two important groups of prebiotics with beneficial effects on human health. Since low quantities of fructo-oligosaccharides and galacto-oligosaccharides naturally exist in foods, scientists are attempting to produce prebiotics on an industrial scale. Considering the health benefits of prebiotics and their safety, as well as their production and storage advantages compared to probiotics, they seem to be fascinating candidates for promoting human health condition as a replacement or in association with probiotics.Technical Problem

[0003] It was an object of the invention to provide a new prebiotic or medicament acting on gut microbiota.

[0004] It was an object of the invention to provide a medicament for use in preventing or treating diseases or conditions such as inflammation, in particular inflammatory bowel diseases (e.g., Irritable Bowel Syndrome, Crohn disease), cardiovascular diseases, chronic heart failure, kidney failure, depression, anxiety, neurological disorders, cognitive impairment, memory loss, attention disorders, perception disorders, cancer, tumor, muscle dysfunction, and metabolic syndrome.

[0005] It was an object of the invention to provide a prebiotic for a non-therapeutic use of improving or preserving gut health and / or gut barrier function and / or intestinal homeostasis and / or immunity and / or mood, and / or having a neuroprotective effect and / or for modulating gutbrain axis and / or for ameliorating or decreasing the risk of diseases or conditions such as inflammation, in particular inflammatory bowel diseases (e.g., Irritable Bowel Syndrome, Crohn disease), cardiovascular diseases, chronic heart failure, kidney failure, depression, anxiety, neurological disorders, cognitive impairment, memory loss, attention disorders, perception disorders, cancer, tumor, muscle dysfunction, and metabolic syndrome.

[0006] It was an object of the invention to provide a prebiotic for a non-therapeutic use as an anti-aging agent and / or as an antioxidant.Presentation of the Invention

[0007] The inventors successfully developed a composition that has multiple benefits on gut microbiota. The composition includes a substance (a) selected from gluconic acid and / or glucono-delta-lactone (GDL), and a substance (b) selected from indigestible dextrins and / or indigestible maltodextrins. The weight ratio of substance (a) to substance (b) is higher than 10 / 90 and lower than 90 / 10.

[0008] As it is apparent form the Example section herein after, the distinct nature of substance (a) and of substance (b) makes that they are fermented in a different fashion, which lead to distinct set of effects. By combining substance (a) with substance (b) in as specific range of ratios, fermentation pathways are modified, which induce the production of multiple beneficial metabolites at the same time, thus allowing to greatly increase the overall health benefits. Furthermore, the combination according to the disclosure provides both health benefits and good tolerance, regardless of the microbiota variability between individuals.

[0009] As it is apparent from the Example section, such combination exerts strong benefits on fermentation parameters such as SCFAs, barrier integrity, metabolites production with health effects (e.g., butyrate, 3 hydroxy-butyric acid).

[0010] Prebiotic effects highly increased with a weight ratio of substance (a) to substance (b) higher than 25 / 75 and lower than 90 / 10, for example from 50 / 50 to 75 / 25. Where potential toleration of the combination is a key aspect, lowering the substance (a) fraction to a weight ratio lower than 50 / 50 (e.g., of 25 / 75), but still higher than 10 / 90, was found to be also a good option.

[0011] As a result of its remarkable potential effects on health, this combination may thus advantageously be used for improving or preserving gut health and / or gut barrier function and / or intestinal homeostasis and / or immunity and / or mood, and / or to exert a neuroprotective effect and / or for modulating gut-brain axis, and / or for preventing or treating diseases or conditions such as inflammation, in particular inflammatory bowel diseases (e.g., irritable bowel syndrome, Crohn disease), cardiovascular diseases, chronic heart failure, kidney failure, depression, anxiety, neurological disorders, cognitive impairment, memory loss, attention disorders, perception disorders, cancer, tumor, muscle dysfunction, and metabolic syndrome.

[0012] Furthermore, as it is apparent from Example section A-lll, 3, the inventors found that GDL was able to promote the production of 3-HBA. GDL may accordingly be used, alone or in combination with an indigestible (malto)dextrin, for preventing or treating diseases or conditions such as cardiovascular diseases, chronic heart failure, kidney failure, depression, anxiety,neurological disorders, cognitive impairment, memory loss, attention disorders, perception disorders, cancer, muscle dysfunction, inflammation, and metabolic syndrome.Brief Description of the Invention

[0013] The invention first relates to the non-therapeutic use, as a prebiotic, of a combination of a substance (a) selected from glucono-delta-lactone, gluconic acid, or from a mixture thereof, and of a substance (b) selected from an indigestible dextrin, an indigestible maltodextrin, orfrom a mixture thereof; the weight ratio of (a) to (b) being higher than 10 / 90 and lower than 90 / 10.

[0014] Preferably, the weight ratio of (a) to (b) is equal to or higher than 15 / 85, preferably equal to or higher than 20 / 80, preferably equal to or higher than 25 / 75. Preferably, the weight ratio of (a) to (b) is equal to or lower than 85 / 15, preferably equal to or lower than 80 / 20, preferably equal to or lower than 75 / 25.

[0015] Preferably, the non-therapeutic use is for at least one of improving or preserving gut health, improving or preserving gut barrier function, improving or preserving intestinal homeostasis, improving or preserving immunity, improving or preserving mood, modulating gutbrain axis, exerting a neuroprotective effect, ameliorating or decreasing the risk of a disease or a condition selected from at least one of inflammation, in particular inflammatory bowel diseases (e.g., irritable bowel syndrome, Crohn disease), cardiovascular disease, chronic heart failure, kidney failure, depression, anxiety, neurological disorders, cognitive impairment, memory loss, attention disorders, perception disorders, cancer, tumor, muscle dysfunction, and metabolic syndrome.

[0016] The invention also relates to a combination, for use as a medicament, of a substance (a) selected from glucono-delta-lactone, gluconic acid, or from a mixture thereof, and of a substance (b) selected from an indigestible dextrin, an indigestible maltodextrin, or from a mixture thereof; the weight ratio of (a) to (b) being higher than 10 / 90 and lower than 90 / 10.

[0017] Preferably, the weight ratio of (a) to (b) is equal to or higher than 15 / 85, preferably equal to or higher than 20 / 80, preferably equal to or higher than 25 / 75. Preferably, the weight ratio of (a) to (b) is equal to or lower than 85 / 15, preferably equal to or lower than 80 / 20, preferably equal to or lower than 75 / 25.

[0018] Preferably, the combination is for preventing or treating a disease or a condition selected from inflammation, in particular inflammatory bowel diseases (e.g., irritable bowel syndrome, Crohn disease), cardiovascular disease, chronic heart failure, kidney failure, depression, anxiety, neurological disorders, cognitive impairment, memory loss, attention disorders, perception disorders, cancer, tumor, muscle dysfunction, and metabolic syndrome.

[0019] The invention also relates to a substance (c) selected from gluconodeltalactone (GDL), gluconic acid, a gluconate salt, or from a mixture thereof, for use as an oral medicament for the prevention or the treatment of a disease or condition selected from cardiovascular diseases,chronic heart failure, kidney failure, depression, anxiety, neurological disorders, cognitive impairment, memory loss, attention disorders, perception disorders, cancer,

[0020] The invention also relates to the non-therapeutic use of a substance (c) selected from GDL, a gluconate salt, gluconic acid or from a mixture thereof, as an antiaging agent, and / or as an antioxidant, and / or for ameliorating or decreasing the risk of a disease or condition selected from cardiovascular diseases, chronic heart failure, kidney failure, depression, anxiety, neurological disorders, cognitive impairment, memory loss, attention disorders, perception disorders, cancer, muscle dysfunction, inflammation, and metabolic syndrome; said substance being intended to be taken orally.Brief Description of Drawings

[0021] Other features, details and advantages will be shown in the following detailed description and on the figures, on which:Fig. 1

[0022] [Fig. 1] is a scheme of the sampling and analysis performed according to Example section A-ll.Fig. 2

[0023] [Fig. 2] includes graphs showing the impact on pH of a treatment with GDL, NUTRIOSE®, or combinations thereof. The tests were conducted for human adults (n=6) as tested via the SIFR® technology platform, for GDL, NUTRIOSE®, and different combinations thereof, compared to NSC control. Samples were collected after 6h, 24h and 48h of simulated colonic incubations. Statistical differences between treatments and the NSC are indicated with * (0.01 < adjusted p-value < 0.05), ** (0.001 < adjusted p-value < 0.01), *** (adjusted p-value < 0.001).Fig. 3

[0024] [Fig. 3] includes graphs showing the impact on gas production of a treatment with GDL, NUTRIOSE®, or combinations thereof. The were conducted for human adults (n=6) as tested via the SIFR® technology platform, for GDL, NUTRIOSE®, and different combinations thereof compared to NSC control. Samples were collected after 6h, 24h and 48h of simulated colonic incubations. Statistical differences between treatments and the NSC are indicated with * (0.01 < adjusted p-value < 0.05), ** (0.001 < adjusted p-value < 0.01), *** (adjusted p-value < 0.001).Fig. 4

[0025] [Fig. 4] includes graphs showing the impact on total SCFA level of a treatment with GDL, NUTRIOSE®, or combinations thereof. The tests were conducted for human adults (n=6) as tested via the SIFR® technology platform, for GDL, NUTRIOSE®, and different combinationsthereof compared to NSC control. Samples were collected after 6h, 24h and 48h of simulated colonic incubations. Statistical differences between treatments and the NSC are indicated with* (0.01 < adjusted p-value < 0.05), ** (0.001 < adjusted p-value < 0.01), *** (adjusted p-value < 0.001).Fig. 5

[0026] [Fig. 5] includes graphs showing the impact on acetate level of a treatment with GDL, NUTRIOSE®, or combinations thereof. The tests were conducted for human adults (n=6) as tested via the SIFR® technology platform, for GDL, NUTRIOSE®, and different combinations thereof compared to NSC control. Samples were collected after 6h, 24h and 48h of simulated colonic incubations. Statistical differences between treatments and the NSC are indicated with* (0.01 < adjusted p-value < 0.05), ** (0.001 < adjusted p-value < 0.01), *** (adjusted p-value < 0.001).Fig. 6

[0027] [Fig. 6] includes graphs showing the impact on propionate level of a treatment with GDL, NUTRIOSE®, or combinations thereof. The tests were conducted for human adults (n=6) as tested via the SIFR® technology platform, for GDL, NUTRIOSE®, and different combinations thereof compared to NSC control. Samples were collected after 6h, 24h and 48h of simulated colonic incubations. Statistical differences between treatments and the NSC are indicated with* (0.01 < adjusted p-value < 0.05), ** (0.001 < adjusted p-value < 0.01), *** (adjusted p-value < 0.001).Fig. 7

[0028] [Fig. 7] includes graphs showing the impact on butyrate level of a treatment with GDL, NUTRIOSE®, or combinations thereof. The tests were conducted for human adults (n=6) as tested via the SIFR® technology platform, for GDL, NUTRIOSE®, and different combinations thereof compared to NSC control. Samples were collected after 6h, 24h and 48h of simulated colonic incubations. Statistical differences between treatments and the NSC are indicated with* (0.01 < adjusted p-value < 0.05), ** (0.001 < adjusted p-value < 0.01), *** (adjusted p-value < 0.001).Fig. 8

[0029] [Fig. 8] includes graphs showing the impact on valerate level of a treatment with GDL, NUTRIOSE®, or combinations thereof. The tests were conducted for human adults (n=6) as tested via the SIFR® technology platform, for GDL, NUTRIOSE®, and different combinations thereof compared to NSC control. Samples were collected after 6h, 24h and 48h of simulated colonic incubations. Statistical differences between treatments and the NSC are indicated with* (0.01 < adjusted p-value < 0.05), ** (0.001 < adjusted p-value < 0.01), *** (adjusted p-value < 0.001).Fig. 9

[0030] [Fig. 9] includes graphs showing the impact on bCFA level of a treatment with GDL, NUTRIOSE®, or combinations thereof. The tests were conducted for human adults (n=6) as tested via the SIFR® technology platform, for GDL, NUTRIOSE®, and different combinations thereof compared to NSC control. Samples were collected after 6h, 24h and 48h of simulated colonic incubations. Statistical differences between treatments and the NSC are indicated with* (0.01 < adjusted p-value < 0.05), ** (0.001 < adjusted p-value < 0.01), *** (adjusted p-value < 0.001).Fig. 10

[0031] [Fig. 10] includes graphs showing the impact on lactate level of a treatment with GDL, NUTRIOSE®, or combinations thereof. The tests were conducted for human adults (n=6) as tested via the SIFR® technology platform, for GDL, NUTRIOSE®, and different combinations thereof compared to NSC control. Samples were collected after 6h, 24h and 48h of simulated colonic incubations. Statistical differences between treatments and the NSC are indicated with* (0.01 < adjusted p-value < 0.05), ** (0.001 < adjusted p-value < 0.01), *** (adjusted p-value < 0.001).Fig. 11

[0032] [Fig. 11] is a graph showing the impact on bacterial cell density of a treatment with GDL, NUTRIOSE®, or combinations thereof, compared to NSC control, as tested via the SIFR® technology for human adults (n=6). Samples were collected after 48h of incubations. Statistical differences between treatments and the NSC are indicated with * (0.01 < adjusted p-value < 0.05), ** (0.001 < adjusted p-value < 0.01), *** (adjusted p-value < 0.001).Fig. 12

[0033] [Fig. 12] includes graphs showing the impact on observed number of OTUs (A), Chad diversity index (B), reciprocal Simpson diversity index (C) and the Shannon diversity index (D), of a treatment with GDL, NUTRIOSE®, or combinations thereof, compared to NSC control, as tested via the SIFR® technology for human adults (n=6). Samples were collected after 48h of incubations. Statistical differences between treatments and the NSC are indicated with * (0.01 < adjusted p-value < 0.05), ** (0.001 < adjusted p-value < 0.01), *** (adjusted p-value < 0.001).Fig. 13

[0034] [Fig. 13] includes graphs showing the impact on microbial composition at phylum level (based on 16S rRNA gene profiling) as averaged over simulations for human adults (n=6) viathe SIFR® technology, of a treatment with GDL, NUTRIOSE®, or combinations thereof, compared to NSC control. Samples were collected after 48h of incubations and are presented both as proportional (%) (A) and absolute values (cells / mL) (B).Fig. 14

[0035] [Fig. 14] includes graphs showing the impact on the abondance of Actinobacteria (A), Bacteroides (B), Firmicutes (C) and Proteobacteria (D), of a treatment with GDL, NUTRIOSE®, or combinations thereof, compared to NSC control, as tested via the SIFR® technology for human adults (n=6). Samples were collected after 48h of incubation. Statistical differences between treatments and the NSC are indicated with * (0.01 < adjusted p-value < 0.05), ** (0.001 < adjusted p-value < 0.01), *** (adjusted p-value < 0.001).Fig. 15

[0036] [Fig. 15] Figure (15A) shows the impact on bacterial families that were significantly affected by any of the indicated treatments (FDR=0.10), expressed as Iog2(abundance treatment / abundance NSC), averaged over simulations for human adults (n=6) using the SIFR® technology, for GDL, NUTRIOSE®, or combinations thereof. Samples were collected at 48h after initiation of the colonic incubations. Significant differences are indicated by bold and underlining. Figure (15B) shows the regularized Canonical correlation analysis (rCCA) to highlight correlations between fundamental fermentation parameters and significantly affected bacterial families (threshold > 0.4). Specific correlations that are elaborated along the Examples section of the description are highlighted with a rectangle.Fig. 16

[0037] [Fig. 16] includes a figure showing the impact on OTUs that were significantly affected by any of the treatments (FDR=0.10), expressed as Iog2(abundance treatment / abundance NSC), averaged over simulations for human adults (n=6) using the SIFR® technology for GDL, NUTRIOSE®, or combinations thereof. Samples were collected at 48h after initiation of the colonic incubations. Significant differences are indicated by bold and underlining.Fig. 17

[0038] [Fig. 17] includes a figure showing the regularized Canonical correlation analysis (rCCA) to highlight correlations between fundamental fermentation parameters and microbial composition at OTUs level. All OTUs shown in Figures 16; threshold > 0.4). Specific correlations that are elaborated along the Examples section of the description are highlighted with a rectangle.Fig. 18

[0039] [Fig. 18] includes a figure showing the impact on a selection of metabolites (annotated at level 1 , 2a, 2b or 3 and previously linked with the human gut microbiota) as quantified via untargeted LC-MS, tested via the SIFR® technology for human adults (n=6), for a treatment with GDL, NUTRIOSE®, or combinations thereof. Samples were collected at 48h after initiation of the colonic incubations. The reported metabolites were significantly affected by any of the treatment (FDR=0.20). Significant differences are indicated by bold and underlining of the average Iog2 (abundance treatment / abundant blank).Fig. 19

[0040] [Fig. 19] includes graphs showing the impact on gut barrier integrity (TEER) of a treatment with GDL, NUTRIOSE®, or combinations thereof compared to NSC control as measured by the TEER of the Caco-2 epithelial layer after (A) 1 h, (B) 24h or (C) 30h of treatment (=24h in absence of LPS and 6h in the presence of LPS in the basal compartment containing differentiated THP-1 cells), for six human adults using the SIFR® technology platform. Sampled exposed to the cells were collected from the SIFR® model after 48h of simulated colonic incubation. Statistical differences between treatments and the NSC are indicated with * (0.10 < adjusted p-value < 0.20), ** (0.05 < adjusted p-value < 0.10), *** (adjusted p-value < 0.05).Fig. 20

[0041] [Fig. 20] includes graphs showing the correlation analysis between gut barrier integrity (TEER) and (D) acetate, (E) propionate and (F) butyrate levels, after a treatment with GDL, NUTRIOSE®, or combinations thereof / NSC. Gut barrier integrity was measured by the TEER of the Caco-2 epithelial layer after 30h of treatment (= 24h in absence of LPS and 6h in presence of LPS in the basal compartment containing differentiated THP-1 cells).Fig. 21

[0042] [Fig. 21] includes figures showing the regularized Canonical correlation analysis (rCCA) to highlight correlations between gut barrier integrity (TEER) and microbial composition at family (left) and OTUs species (right) levels.Description of EmbodimentsUses of the combination

[0043] The invention first relates the non-therapeutic use, as a prebiotic, of a combination of: a substance (a) selected from glucono-delta-lactone, gluconic acid, or from a mixture thereof, and of a substance (b) selected from an indigestible dextrin, an indigestible maltodextrin, or from amixture thereof; the weight ratio of (a) to (b) being higher than 10 / 90 and lower than 90 / 10.

[0044] This use of the combination as a prebiotic is a non-therapeutic use, preferably intended for healthy subjects (human or animal).

[0045] The invention also relates to a combination, for use as a medicament, of: a substance (a) selected from glucono-delta-lactone, gluconic acid, or from a mixture thereof, and of a substance (b) selected from an indigestible dextrin, an indigestible maltodextrin, or from a mixture thereof; the weight ratio of (a) to (b) being higher than 10 / 90 and lower than 90 / 10.

[0046] The invention also relates to a method for treating a human or an animal, comprising administering an effective amount of a combination of:- a substance (a) selected from glucono-delta-lactone, gluconic acid, or from a mixture thereof, and of- a substance (b) selected from an indigestible dextrin, an indigestible maltodextrin, or from a mixture thereof; the weight ratio of (a) to (b) being higher than 10 / 90 and lower than 90 / 10.

[0047] This use is a therapeutic use. The term “therapeutic” includes both veterinary and human uses for preventing or treating a disease or a condition in a subject in need thereof. Although human uses are preferred.

[0048] The term “combination” means that the substances are used in the same treatment protocol. Substances (a) and (b) may be used in same or separate composition or dosage form. It may be administered or ingested at the same or different time. Preferably, substance (a) and substance (b) are in the same composition or dosage form. Preferably, substance (a) and substance (b) are the sole active ingredients of the composition or dosage form. Preferably, substance (a) and substance (b) are administered or ingested at the same time.

[0049] The term “dosage form” typically refers to a veterinary or pharmaceutical product that is formulated for administration. In the present disclosure, the substances are to be used orally. Therefore, dosage forms comprising substance (a) and / or (b) are oral dosage forms. Such dosage form may be a solid, semi-solid, or liquid form. It is preferably a solid form, for example a powder, a tablet, a hard or soft capsule. In the latter case, substance (a) and / or (b) is typically in the filler of the hard or soft capsule. Such filler may be a solid (e.g., a powder), a semi-solid, or a liquid).

[0050] In an embodiment, especially when the combination is used as a prebiotic (non- therapeutic use), the use if for improving or preserving gut health, and / or gut barrier functionand / or intestinal homeostasis and / or immunity and / or mood. Preferably said uses are in a healthy subject.

[0051] The term “improving or preserving gut health” may include decreasing or preventing abdominal pain, discomfort, diarrhea, constipation, increasing or preserving the quantity of beneficial bacteria in gut microbiota, increasing or preserving gut barrier function, increasing or preserving the production of metabolites that promote the aforementioned effects.

[0052] The term “gut barrier function” refers to the ability of the semipermeable structure of the intestinal wall to allow the uptake of essential nutrients and immune sensing, while being restrictive against pathogenic molecules and bacteria. Both structural and molecular components act together to fulfil this essential function of the gastrointestinal tract. Gut barrier function may for example include intestinal wall permeability.

[0053] The term “gut homeostasis” refers to the complex immune and nonimmune mechanisms that permit the intestinal mucosa to minimize the adverse health effects of commensals even during microenvironmental perturbations. It is maintained by commensal bacteria, gut barrier function and tolerant immune response.

[0054] In an embodiment, especially when it is used as a prebiotic (non-therapeutic use), the use is for modulating gut-brain axis, preferably in a healthy subject.

[0055] The term “gut-brain axis” refers to the bidirectional communication between the gut and brain that contribute to maintain gut homeostasis, and potentially affecting various cognitive functions such as attention, perception, and memory.

[0056] In an embodiment, the non-therapeutic use is for providing neuroprotective effect, preferably in a healthy subject.

[0057] The term “neuroprotection” or “neuroprotective effect” refers to the mechanisms and strategies employed to defend the central nervous system (CNS) against injury due to both acute (e.g. trauma or stroke) and chronic neurodegenerative disorders (e.g. dementia, Parkinson's, Alzheimer's, epilepsy etc.). In the present disclosure, the neuroprotective effect is rather against injuries due to chronic neurodegenerative disorders.

[0058] In an embodiment, the combination according to the disclosure is used non- therapeutically as an antiaging agent and / or as an antioxidant.

[0059] In an embodiment, especially when the combination is used as a medicament (therapeutic use), the use if for preventing or treating a disease or a condition selected from inflammation, in particular inflammatory bowel diseases (e.g., irritable bowel syndrome, Crohn disease), cardiovascular disease, chronic heart failure, kidney failure, depression, anxiety, neurological disorders, cognitive impairment, memory loss, attention disorders, perception disorders, cancer, tumor, muscle dysfunction, and metabolic syndrome.

[0060] In an embodiment, especially when the combination is used as a prebiotic (non- therapeutic use), the use if for ameliorating or decreasing the risk of a disease or a condition selected from inflammation, in particular inflammatory bowel diseases (e.g., irritable bowel syndrome, Crohn disease), cardiovascular disease, chronic heart failure, kidney failure, depression, anxiety, neurological disorders, cognitive impairment, memory loss, attention disorders, perception disorders, cancer, tumor, muscle dysfunction, and metabolic syndrome.

[0061] Preferred examples of cardiovascular diseases include atherosclerosis and nonalcoholic fat liver disease (NAFLD), e.g., nonalcoholic steatohepatitis (NASH). Preferred examples of neurological disorders include Alzheimer disease and Parkinson disease. Preferred examples of cognitive impairments include learning impairments, memory impairments, complex attention, social cognition, language impairments. Preferred examples of muscle dysfunction include sarcopenia and cachexia.

[0062] Typically, the combination is for use in a human or an animal, preferably a mammal, more preferably a human.

[0063] In a preferred embodiment, the combination is for use in a subject or an organism (e.g., a human or an animal) suffering from or being at risk for developing a disease or a condition selected from inflammation, in particular inflammatory bowel diseases (e.g., irritable bowel syndrome, Crohn disease), cardiovascular disease, chronic heart failure, kidney failure, depression, anxiety, neurological disorders, cognitive impairment, attention disorders, perception disorders, memory loss, cancer, tumor, muscle dysfunction, and metabolic syndrome. In a preferred embodiment, especially when use is a non-therapeutic use, the use is for a healthy subject, e.g., a healthy human or animal.

[0064] Substance (a) is selected from GDL and / or gluconic acid. Preferably, substance (a) is GDL. GDL forms gluconic acid in water, for example when it is ingested. Without being bond by any theory, the inventor(s) believe that the acidic nature of gluconic acid is important for the performance of the combination.

[0065] Substance (b) according to the disclosure is selected from an indigestible maltodextrin, an indigestible dextrin, or from a mixture thereof. It is more preferably an indigestible dextrin.

[0066] Preferably, the indigestible maltodextrin according to the disclosure is a branched maltodextrin. Preferably, the indigestible dextrin according to the disclosure is a branched dextrin.

[0067] Preferably, substance (b) is a fiber. The term "fiber" classically refers to saccharides which are not or only partially digested in the intestine by the action of acids or digestive enzymes present in the human upper digestive tract (small intestine and stomach), but which are at least partially fermented by the human intestinal flora. Preferably, the fiber according to the disclosure is a water-soluble fiber. The term “water-soluble” refers to substances which arefrom soluble to very soluble in water at 20°C. This solubility is also sometimes referred to as “cold water-solubility”. This solubility in water is well defined for instance in 11th edition of The International Pharmacopeia (2011), Section “General Notice, Solubility”: substances which are from soluble to very soluble in water at 20°C require 30 mL or less of water to dissolve 1g of said substance.

[0068] Preferably, substance (b) according to the disclosure has from 15 to 35% of 1->6 glucoside linkages, preferably from 20 to 35%, preferably from 25 to 35%. This percentage of 1->6 glucoside linkages may be determined by the person skilled in the art by using the conventional methylation technique described in HAKOMORI, S., 1964, J. Biol. Chem., 55, 205.

[0069] Preferably, substance (b) according to the disclosure has a reducing sugar content, as dry weight, of less than 20%, preferably equal to or lower than 15%, preferably equal or lower than 10%; said percentage being expressed with respect to the total dry weight of substance (b). This reducing sugar content may be determined by the person skilled in the art according to the well-known "Bertrand method" by precipitation with cuprous oxide in reducing media, filtration on a sintered-glass filter, and weighting of the residue.

[0070] Preferably, substance (b) according to the disclosure has a number-average molecular weight (Mn) equal to or lower than 10 000 g / mol, preferably equal to or lower than 8 000 g / mol, preferably equal to or lower than 7 000 g / mol, preferably equal to or lower than 6 000 g / mol, preferably equal to or lower than 5 000 g / mol, preferably equal to or lower than 4 500 g / mol.Preferably, this Mn is equal to or higher than 100 g / mol, preferably equal to or higher than200 g / mol, preferably equal to or higher than 300 g / mol.

[0071] Preferably, substance (b) according to the disclosure has a weight-average molecular weight (Mw) equal to or lower than 20 000 g / mol, preferably equal to or lower than 10 000 g / mol, preferably equal to or lower than 9 000 g / mol, preferably equal to or lower than 8 000 g / mol, preferably equal to or lower than 7 000 g / mol, preferably equal to or lower than 6 000 g / mol. Preferably, this Mw is equal to or higher than 100 g / mol, preferably equal to or higher than 500 g / mol, preferably equal to or higher than 700 g / mol.

[0072] Preferably, substance (b) according to the disclosure has a polymolecularity index (Mw / Mn) lower than 5, preferably lower than 4, preferably lower than 3. It is further in general equal to or higher than 1 , even equal to or higher than 2. It is for example from 1 to 3, or from 1.0 to 3.0.

[0073] The Mn and Mw may be determined by the person skilled in the art by gel-permeation chromatography (GPC) on calibrated chromatographic columns using pullulans standards.

[0074] Preferably, substance (b) according to the disclosure is derived from wheat starch.

[0075] Example of suitable substance (b) according to the disclosure is commercially available. Mentioned can be made for example of NUTRIOSE® FM HF (Roquette Freres).

[0076] Preferably, the weight ratio of substance (a) to substance (b) is equal to or higher than 15 / 85, preferably equal to or higher than 20 / 80, preferably equal to or higher than 25 / 75, preferably higher than 25 / 75, preferably equal to or higher than 30 / 70, preferably equal to or higher than 35 / 65, preferably equal to or higher than 40 / 60, preferably equal to or higher than 45 / 55, preferably equal to or higher than 50 / 50. It is preferably equal to or lower than 85 / 15, preferably equal to or lower than 80 / 20, preferably equal to or lower than 75 / 25.

[0077] In an embodiment, this ratio is:- equal to or higher than 15 / 85, preferably equal to or higher than 20 / 80, preferably equal to or higher than 25 / 75, preferably higher than 25 / 75, preferably equal to or higher than 30 / 70, preferably equal to or higher than 35 / 65, preferably equal to or higher than 40 / 60, preferably equal to or higher than 45 / 55, preferably equal to or lower than 50 / 50, preferably equal to or higher than 55 / 45, preferably equal to or higher than 60 / 40, preferably equal to or higher than 65 / 35, preferably equal to or higher than 70 / 30, and,- equal to or lower than 85 / 15, preferably equal to or lower than 80 / 20.

[0078] In an embodiment, this ratio is:- equal to or higher than 15 / 85, preferably equal to or higher than 20 / 80, preferably equal to or higher than 25 / 75, preferably higher than 25 / 75, preferably equal to or higher than 30 / 70, preferably equal to or higher than 35 / 65, preferably equal to or higher than 40 / 60, preferably equal to or higher than 45 / 55, and,- equal to or lower than 85 / 15, preferably equal to or lower than 80 / 20, preferably equal to or lower than 75 / 25, preferably equal to or lower than 70 / 30, preferably equal to or lower than 65 / 35, preferably equal to or lower than 60 / 40, preferably equal to or lower than 55 / 45.Products comprisinq the combination

[0079] The invention also relates to a product comprising a substance (a) according to the disclosure and a substance (b) according to the disclosure, for the uses as described herein.

[0080] The product may be final products i.e. , a product intended to be orally administered or ingested as is, or a product to be included into a final product, for example after addition of other ingredients or after one or more steps of shaping.

[0081] Preferably, said product is selected from a food product, a nutraceutical product, a prebiotic product, a cosmetic product, or a pharmaceutical product.

[0082] In an embodiment, said product is a composition, preferably selected from a food composition, a nutraceutical composition, a prebiotic composition, a cosmetic composition, or a pharmaceutical composition.

[0083] In an embodiment, said product is a final product, preferably selected from a food, nutraceutical, cosmetic or pharmaceutical product. The latter is also often referred to as “dosage form”. Such dosage from typically includes ingredients other that the substances according to the disclosure. These other ingredients typically are selected depending on the final dosage form desired. They may be selected for example from fillers, binders, (super)disintegrants, flavors, sweeteners, colors.

[0084] In an embodiment, substances (a) and (b) according to the disclosure are the sole active ingredients of the product, in particular of the final product.

[0085] The term "active ingredient" classically refers to any substance of food, nutraceutical, cosmetic or pharmaceutical interest (the latter being classically referred to as “active pharmaceutical ingredient”). A food, nutraceutical or pharmaceutical active ingredient is intended to mean an ingredient that provides health benefit. A cosmetic active ingredient is intended to mean an ingredient that provides cosmetic benefit.

[0086] Preferably, the nature and / or ratio of substances (a) and / or (b) are as described before for the combination according to the disclosure.Effects of the combination or product

[0087] In an embodiment, the combination or product according to the disclosure is for increasing (or increases), the production in the gut of an organism (e.g. a human or an animal) of at least one of: total short-chain fatty acids (Total SCFAs), acetate, propionate, butyrate, lactate.

[0088] In an embodiment, the combination or product according to the disclosure is for decreasing (or decreases) the production in the gut of an organism (e.g. a human or an animal) of branched short-chain fatty acids (bCFA).

[0089] In an embodiment, the combination or product according to the disclosure is for maintaining (or maintains) the production in the gut of an organism (e.g. a human or an animal) of valerate.

[0090] The term “total short-chain fatty acids (Total SCFAs)” refers the sum of unbranched and branched SCFAs (bCFA). The term “unbranched SCFAs” refers to the sum of acetate, propionate, butyrate, valerate and caproate. The term “branched SCFAs (bCFA)” refers to the sum of isobutyrate, isocaproate and isovalerate.

[0091] This ability to increase, decrease or maintain the production of SCFAs and lactate may be determined by the person skilled in the art by incubating a fecal inoculum together with the item to be tested, at 37°C in anaerobic condition during 6h to 48h, and then measuring the molar concentration of SCFAs of interest or lactate (for example after 6h, 24h and 48h of incubation) by gas chromatography (GC) with flame ionization detection, upon diethyl etherextraction. The amounts obtained are compared to those obtained with a control, which is a fecal inoculum that is incubated without the item to be tested. It is possible to proceed for example according to the method given in the Example section herein after.

[0092] In an embodiment, the combination or product according to the disclosure is for increasing (or increases) bacterial cell density in the gut of an organism (e.g. a human or an animal). This ability to increase cell density may be determined by the person skilled in the art by incubating a fecal inoculum together with the item to be tested at 37°C in anaerobic condition for48h, and then measuring cell density using flow cytometry. This cell density is then compared to those obtained with a control, which is a fecal inoculum that is incubated without the item to be tested. It is possible to proceed for example according to the method given in the Example section herein after.

[0093] In an embodiment, the combination or product according to the disclosure is for decreasing (or decreases) bacterial diversity in the gut of an organism (e.g. a human or an animal). This ability to decrease bacterial diversity may be determined by the person skilled in the art by incubating a fecal inoculum together with the item to be tested at 37°C in anaerobic condition for 48h, then by using metagenomic, and then by calculating the reciprocal Simpson diversity index or Shannon diversity index. This bacterial diversity is then compared to those obtained with a control, which is a fecal inoculum that is incubated without the item to be tested. It is possible to proceed for example according to the method given in the Example section herein after.

[0094] In an embodiment, the combination according to the disclosure is for promoting (or promotes) the growth of at least one of the following phyla: Actinobacteriota, Bacteroidota, Firmicutes, Proteobacteria.

[0095] In an embodiment, the combination according to the disclosure is for hindering (or hinders) the growth of at least one of the following phyla: Actinobacteriota, Bacteroidota, Firmicutes, Proteobacteria.

[0096] In an embodiment, the combination according to the disclosure is for promoting (or promotes) the growth of at least one of the following families: Bifidobacteriaceae, Bacteroidaceae, Tannerellaceae, Erysipelotrichaceae, Lachnospiraceae, Ruminococcaceae, Enterobacteriaceae. Preferably, the combination according to the disclosure is for promoting (or promotes) the growth of at least one of Bifidobacteriaceae, Bacteroidaceae, Tannerellaceae, Erysipelotrichaceae, and Lachnospiraceae families.

[0097] In an embodiment, the combination according to the disclosure is for is for hindering (or hinders) the growth of at least one of the following families: Bacteroidaceae, Monoglobaceae, Oscillospiraceae.

[0098] In an embodiment, the combination according to the disclosure is for promoting (or promotes) the growth of at least one of the following species: Bifidobacterium adolescentis, Bacteroides uniformis, Bacteroides xylanisolvens, Parabacteroides distasonis, Parabacteroides merdae, Faecalibacillus intestinalis, Blautia obeum / wexlerae, Fusicatenbacter saccharivorans, Blautia massiliensis, Ruminococcus faecis, Blautia faecis, Oliverpabstia intestinalis, Anaerobutyricum hallii, Clostridium clostridioforme / bolteae, Ruminococcus lactaris, Gallintestinimicrobium propionicum, Roseburia sp., Faecalibacterium prausnitzii, Dialister invisus. Preferably, the combination according to the disclosure is for promoting (or promotes) the growth of at least one of the following species: Bifidobacterium adolescentis, Bacteroides uniformis, Bacteroides xylanisolvens, Parabacteroides distasonis, Parabacteroides merdae, Faecalibacillus intestinalis, Blautia obeum / wexlerae, Fusicatenbacter saccharivorans, Blautia massiliensis, Ruminococcus faecis, Blautia faecis, Anaerobutyricum hallii, Ruminococcus lactaris, Gallintestinimicrobium propionicum, Faecalibacterium prausnitzii.

[0099] In an embodiment, the combination according to the disclosure is for hindering (or hinders) the growth of at least one of the following species: Bacteroides vulgatus / dorei, Bacteroides massiliensis, Bacteroides theta iota micron, Alistipes putredinis, Dorea longicatena, Dorea formicigenerans, Coprococcus comes, butyrate-producing bacterium SS3 / 4, Anaerotignum lactatifermentans, Lachnoclostridium sp., Ruminococcus torques, Lachnospiraceae bacterium Lach-103, Hominilimicola fabiformis, Oscillospiraceae bacterium, Hominicoprocolafusiformis, Ruminococcus bromii. Preferably, the combination according to the disclosure is for hindering (or hinders) the growth of at least one of the following species: Bacteroides massiliensis, Alistipes putredinis, Dorea longicatena, Dorea formicigenerans, Coprococcus comes, Anaerotignum lactatifermentans, Lachnoclostridium sp., Ruminococcus torques, Lachnospiraceae bacterium Lach-103, Hominilimicola fabiformis, Hominicoprocolafusiformis. Preferably, the combination according to the disclosure is for hindering (or hinders) the growth of at least one of the following species: Alistipes putredinis, Dorea longicatena, Dorea formicigenerans, Coprococcus comes, Lachnoclostridium sp., Ruminococcus torques, Lachnospiraceae bacterium Lach-103.

[0100] The term “promoting the growth” means increasing the number and / or the proportion of the considered phylum, family, or species in the gut of an organism (e.g. a human or an animal). It is preferably for increasing the number of the considered phylum, family, or species. The term “hindering the growth” means decreasing the number and / or the proportion of the considered phylum, family, or species in the gut of an organism. It is preferably for decreasing the number of the considered phylum, family, or species.

[0101] This ability to promote or hinder the growth of bacteria phylum, family, or species may be determined by the person skilled first by incubating a fecal inoculum together with the item to be tested at 37°C in anaerobic condition for 48h. Then, upon DNA extraction, librarypreparation and sequencing are performed on a DNA sequencer. The 16S rRNA gene V3-V4 hypervariable regions are amplified using primers 341 F (50 -CCT ACG GGN GGC WGC AG- 30) and 785Rmod (50 -GAC TAC HVG GGT ATC TAA KCC-30). Results are analyzed at different taxonomic levels (phylum, family and OTU level). For taxonomic analysis, the proportional data derived from sequencing (%) are corrected for the total amount of cells present in each sample (detected via flow cytometry). The number and / or proportion of the phylum, family, or species of interest is then compared with a control, which is a fecal inoculum that is incubated without the item to be tested. It is possible to proceed for example according to the method given in the Example section herein after.

[0102] In an embodiment, the combination according to the disclosure is for increasing (or increases) the production in the gut of an organism (e.g. a human or an animal) of at least one of the following metabolites: N-acetyl-alanine, N-acetyl-L-leucine, N-acetyl-tryptophan, 2- aminoadipic acid, 2-hydroxy-4-(methylthio)butanoic acid, 3-(3-hydroxyphenyl)propionic acid, 5- Indolol, acetylcadaverine, hydroxyphenyllactic acid, indole-3-lactic acid, methyl 3-indoleacetate, tyramine, 2-oxobutyric acid, 5-methylcytosine, 2 / 3-hydroxybutiric acid, 3-methylxanthine, trimethylamine N-oxide, pyridoxine, pyridoxamine. Preferably, the combination according to the disclosure is for increasing (or increases) the production of at least one of N-acetyl-alanine, N- acetyl-L-leucine, N-acetyl-tryptophan, 2-hydroxy-4-(methylthio)butanoic acid, 3-(3- hydroxyphenyl)propionic acid, 5-lndolol, acetylcadaverine, hydroxyphenyllactic acid, indole-3- lactic acid, tyramine, 2-oxobutyric acid, 2 / 3-hydroxybutiric acid, pyridoxine, pyridoxamine.

[0103] In an embodiment, the combination according to the disclosure is for decreasing (or decreases) the production in the gut of an organism (e.g. a human or an animal) of at least one of the following metabolites: N-acetyl-glycine, N-acetyl-lysine, 2-hydroxyphenylacetic acid, phenylacetic acid, 4-amino-3-hydroxybenzoic acid, pyridoxine. Preferably, the combination according to the disclosure is for decreasing (or decreases) the production of at least one of 2- hydroxyphenylacetic acid and phenylacetic acid.

[0104] This ability to increase or decrease such metabolites may be determined by the person skilled in the art by incubating a fecal inoculum together with the item to be tested, at 37°C in anaerobic condition for 48h, and then measuring the amount of the metabolite(s) of interest by Liquid Chromatography - Mass Spectrometry (LC-MS). The metabolite amount is then compared to those obtained with a control which is a fecal inoculum that is incubated without the item to be tested. It is possible to proceed for example according to the method given in the Example section herein after.

[0105] In an embodiment, the combination according to the disclosure is for increasing (or increases) gut barrier integrity in an organism (e.g. a human or an animal). This ability to increase gut barrier integrity may be determined by the person skilled in the art by a TEER measurement after exposing Caco-2 and THP-1 cells of a transwell system to samples obtainedfrom a fecal inoculum incubated for48h with the item to be tested, at 37°C in anaerobic condition. The TEER value of the tested item is compared to those obtained with a control, which is sample obtained from a fecal inoculum that is incubated without the item to be tested. It is possible to proceed for example according to the method given in the Example section herein after.

[0106] Preferably, the incubation according to the disclosure is performed by way of a bioreactor-based method that relies on anaerobic bioreactors with supplements that promote bacterial growth, in order to mimic human colonic fermentation, preferably using SIFR® technology.

[0107] Preferably the fecal inoculum used according to the disclosure is a human fecal inoculum.

[0108] It is understood from the Example section herein after that the aforementioned embodiments that recite various effects on fermentation parameters, microbial composition, metabolomic, and gut barrier function can be selected and combined in accordance with the weight ratio (or range of weight ratio) of substance (a) to (b). Accordingly, the instant disclosure is considered to disclose various combination of ratios and effects which can be isolated and extrapolated from the Example section A-lll (in particular from Figures 4-16, 18 and 19). Those effects are those that were identified as significant in the Example section, or those for which a trend was observed.Process for making the Product

[0109] The invention also relates to a process for making a product according to the disclosure, for uses as disclosed herein, comprising blending a substance (a) according to the disclosure with a substance (b) according to the disclosure.Uses of GPL alone, or in combination with a (malto)dextrin

[0110] The invention also relates to a substance (c) selected from glucono-delta-lactone (GDL), a gluconate salt, gluconic acid or from a mixture thereof, for use as an oral medicament for the prevention or treatment of a disease or condition selected from cardiovascular diseases, chronic heart failure, kidney failure, depression, anxiety, neurological disorders, cognitive impairment, memory loss, attention disorders, perception disorders, cancer, muscle dysfunction, inflammation, and metabolic syndrome.

[0111] The invention also relates to a method for preventing or treating a condition or a disease selected from cardiovascular diseases, chronic heart failure, kidney failure, depression, anxiety, neurological disorders, cognitive impairment, memory loss, attention disorders, perception disorders, cancer, muscle dysfunction, inflammation, and metabolic syndrome,comprising administering in an organism in need thereof an effective amount of a substance (c) selected from GDL, a gluconate salt, gluconic acid or from a mixture thereof.

[0112] The term “oral medicament” is intended to mean that the medicament is intended to be taken orally.

[0113] Preferably, substance (c) according to the disclosure is selected from GDL, gluconic acid, or from a mixture thereof. More preferably, said substance (c) comprises (or consists of) GDL.

[0114] Preferably, the medicament is for a human or an animal, preferably a mammal, more preferably a human.

[0115] Preferably, the medicament is for an organism (e.g., a human) suffering from or being at risk for developing a disease or a condition selected from cardiovascular disease, chronic heart failure, kidney failure, depression, anxiety, neurological disorder, cognitive impairment, memory loss, attention disorders, perception disorders, cancer, muscle dysfunction, inflammation, and metabolic syndrome.

[0116] In an embodiment, substance (c) according to the disclosure is the sole active ingredient of the medicament. In another embodiment, it is combined with a substance (b) selected from an indigestible dextrin, an indigestible maltodextrin, or from a mixture thereof. In such a case, such combination is previously as described in the claims and embodiments included in the present specification, except that substance (c) is used instead of substance (a).

[0117] The invention also relates to the non-therapeutic use of a substance (c) selected from GDL, a gluconate salt, gluconic acid or from a mixture thereof, as an antiaging agent, and / or as an antioxidant, and / or for ameliorating or decreasing the risk of a disease or condition selected from cardiovascular diseases, chronic heart failure, kidney failure, depression, anxiety, neurological disorders, cognitive impairment, memory loss, attention disorders, perception disorders, cancer, muscle dysfunction, inflammation, and metabolic syndrome; said substance being intended to be taken orally.

[0118] Preferably, substance (c) according to the disclosure is selected from GDL, gluconic acid, or from a mixture thereof. More preferably, said substance (c) comprises (or consists of) GDL.

[0119] Preferably, the use is for a mammal, more preferably a human.

[0120] In a preferred embodiment, the use is for a human or an animal suffering from or being at risk for developing a disease or a condition selected from cardiovascular disease, chronic heart failure, kidney failure, depression, anxiety, neurological disorder, cognitive impairment, memory loss, attention disorders, perception disorders, cancer, muscle dysfunction,inflammation, and metabolic syndrome. In a preferred embodiment, especially when use is a non-therapeutic use, the use is for a healthy subject, e.g., a healthy human or animal.

[0121] In an embodiment, substance (c) according to the disclosure is the sole active ingredient used. In another embodiment, it is combined with a substance (b) selected from an indigestible dextrin, an indigestible maltodextrin, or from a mixture thereof. In such a case, such combination is previously as described in the claims and embodiments included in the present specification, except that substance (c) is used instead of substance (a).

[0122] The invention also relates to a product comprising substance (c) according to the disclosure, for the uses as described herein.

[0123] Preferably, said product is as described in the claims and embodiment included in the present specification.

[0124] In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for increasing (or increases), the production in the gut of an organism (e.g. a human or an animal) of at least one of: total short-chain fatty acids (Total SCFAs), acetate, propionate, butyrate, lactate.

[0125] In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for decreasing (or decreases) the production in the gut of an organism (e.g. a human or an animal) of branched short-chain fatty acids (bCFA).

[0126] In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for maintaining (or maintains) the production in the gut of an organism (e.g. a human or an animal) of valerate.

[0127] In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for increasing (or increases) bacterial cell density in the gut of an organism (e.g. a human or an animal).

[0128] In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for decreasing (or decreases) bacterial diversity in the gut of an organism (e.g. a human or an animal).

[0129] In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for promoting (or promotes) the growth of at least one of the following phyla: Actinobacteriota, Proteobacteria, preferably Actino ba cteriota.

[0130] In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for hindering (or hinders) the growth of at least one of the following phyla: Bacteroidota, Firmicutes, Proteobacteria. Preferably, substance (c) or the product comprising substance (c) according to the disclosure is for hindering (or hinders) the growth of Bacteroidota phylum.

[0131] In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for promoting (or promotes) the growth of at least one of the following families: Bifidobacteriaceae, Enterobacteriaceae. In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for promoting (or promotes) the growth of Bifidobacteriaceae family.

[0132] In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for is for hindering (or hinders) the growth of at least one of the following families: Bacteroidaceae, Rikenellaceae, Tannerellaceae, Christensenellaceae,Erysipelotrichaceae, Lachnospiraceae, Monoglobaceae, Oscillospiraceae.

[0133] In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for promoting (or promotes) the growth of at least one of the following species: Bifidobacterium adolescentis, Anaerobutyricum hallii, Faecalibacterium prausnitzii. Dialister invisus. Preferably, substance (c) or the product comprising substance (c) according to the disclosure is for promoting (or promotes) the growth of at least one of the following species: Bifidobacterium adolescentis, Anaerobutyricum hallii, Faecalibacterium prausnitzii.

[0134] In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for hindering (or hinders) the growth of at least one of the following species: Bacteroides vulgatus / dorei, Alistipes putredinis, Parabacteroides distasonis, Parabacteroides merdae, Faecalibacillus intestinalis, Fusicatenbacter saccharivorans, Dorea longicatena, Ruminococcus faecis, Lachnoclostridium edouardi, Dorea formicigenerans, Coprococcus comes, butyrate-producing bacterium SS3 / 4, Anaerotignum lactatifermentans, Lachnoclostridium sp., Ruminococcus torques, Gallintestinimicrobium propionicum, Lachnospiraceae bacterium Lach-103, Hominilimicola fabiformis, Oscillospiraceae bacterium, Hominicoprocolafusiformis, Ruminococcus bromii.

[0135] It is reminded that the term “promoting or hindering the growth” means increasing or decreasing the number and / or the proportion of the considered phylum, family, or species in the gut of an organism (e.g. a human or an animal).

[0136] In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for increasing (or increases) the production in the gut of an organism (e.g. a human or an animal) of at least one of the following metabolites: N-acetyl-tryptophan, 2-hydroxy- 4-(methylthio)butanoic acid, 3-(3-hydroxyphenyl)propionic acid, acetylcadaverine, hydroxyphenyllactic acid, indole-3-acetaldehyde, indole-3-lactic acid, methyl 3-indoleacetate, tyramine, 5-methylcytosine, 2 / 3-hydroxybutiric acid, 3-methylxanthine, trimethylamine N-oxide. Preferably, substance (c) or the product comprising substance (c) according to the disclosure is for increasing (or increases) the production of at least one of 3-(3-hydroxyphenyl)propionic acid, hydroxyphenyllactic acid, indole-3-lactic acid, 2 / 3-hydroxybutiric acid. Preferably, substance (c)or the product comprising substance (c) according to the disclosure is for increasing (or increases) the production of 2 / 3-hydroxybutiric acid.

[0137] In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for decreasing (or decreases) the production in the gut of an organism (e.g. a human or an animal) of at least one of the following metabolites: N-acetyl-glycine, N-acetyl- lysine, 2-hydroxyphenylacetic acid, 5-lndolol, phenylacetic acid, pyridoxamine, pyridoxine. Preferably, substance (c) or the product comprising substance (c) according to the disclosure is for decreasing (or decreases) the production of at least one of 2-hydroxyphenylacetic acid and phenylacetic acid.

[0138] In an embodiment, substance (c) or the product comprising substance (c) according to the disclosure is for increasing (or increases) gut barrier integrity.

[0139] The ability of substance (c) or the product comprising substance (c) to exert the above- mentioned effects may be determined by the person skilled in the art by the same methods as described before for the combination of substances (a) and (b).Remarks

[0140] The amounts of ingredients may be expressed in percentages by weight. These weights are amounts of ingredients as such, in their typical powdery or oily form (or liquid form for solvents). All compounds may include small amounts of impurities. Powdery ingredients may also include small amount of water (also referred to as %moisture or as “loss on drying”). With that respect, substance (a) and substance (c) according to the disclosure typically have a %moisture equal to or lower than 0.2%, and substance (b) typically have a %moisture equal to or lower than 6%.

[0141] Also, when referring to amounts or ratio of ingredients in a product such as in the dosage form, it is classically understood that these amounts are those used for making said product. These amounts might differ to a small extent from the amounts actually found in the final product (that may actually not be measurable), for example due to small material losses or due to a small heterogeneity of the blend used for making said product.

[0142] It is reminded that, as used in the specification and in the claims, the term “comprising” may include the embodiments “consisting of’ and “consisting essentially of’. The terms “comprise(s),” “include(s),” “having,” “has,” “can,” “contain(s),” and variants thereof, are intended to be open-ended transitional phrases that require the presence of the named feature(s) / element(s) / step(s) / unit operation(s) and permit the presence of other feature(s) / element(s) / step(s) / unit operation(s). However, such description should be construed as also describing products or methods “consisting of’ and “consisting essentially of’ the enumerated feature(s) / element(s) / step(s) / unit operation(s), which allows the presence of onlythe named feature(s) / element(s) / step(s) / unit operation(s), along with any impurities or moisture that might result therefrom, and excludes other feature(s) / element(s) / step(s) / unit operation(s).

[0143] It is reminded that, as used in the specification and the appended claims, the singular form "a," "an," and "the" comprise plural referents unless the context clearly indicates otherwise and means “at least one” or “one or more”. For example, reference to a component in the singular is intended to comprise a plurality of components. Thus, for example, “an” indigestible dextrin means “one or more” indigestible dextrin.

[0144] It is reminded that, where the specification and / or claims disclose more than one upper and / or lower limits for a value (e.g., the weight ratio of substance (a) to substance (b), such description should be construed as also describing any combination of said upper and lower limits.

[0145] Other characteristics and advantages of the present invention will emerge clearly on reading the examples given hereinafter, which illustrate the invention without however limiting it.ExamplesA- Impact of glucono-delta-lactone (GDL) and indigestible dextrin on human gut microbiota

[0146] The aim of this study was to investigate the impact of glucono-delta-lactone (GDL), an indigestible dextrin (NUTRIOSE® FM HF-dextrin) and different combinations thereof on the human adult gut microbiota. The high throughput of the ex vivo SIFR® technology allowed to include 6 subjects, which covered the broad spectrum of microbial composition that occurs in vivo, in line with the so-called enterotypes, thus ensuring representative findings.A-l Test item

[0147] Product tested were the following:- glucono-delta-lactone, Roquette Freres.- indigestibe dextrin: NUTRIOSE® FM HF-Dextrin, Roquette Freres.

[0148] Test items were tested alone or as blends, as shown in table 1 .

[0149] [Table 11 Test items

[0150] Test items were compared with no substrate control (NSC). All test products were tested at a dose equivalent to 5 g / day.A-ll Protocol

[0151] A kinetic, ex vivo SIFR® study was implemented, simulating the colonic fermentation of test items by the gut microbiota derived from human adults (n = 6). The simulation parameters were as follows:

[0152] Study arms = 8:- No substrate control (NSC) = background medium + microbiota (no product) .- GDL and NUTR tested as such and in 5 different ratios (90G / 10N, 75G / 25N, 50G / 50N, 25G / 75N, 10G / 90N) at a dose equivalent to 5 g / day.- Time points = 4 (Oh (NSC only), 6h, 24h, and 48h) - each time point was performed in an independent reactor.The NSC at 48h was run in technical triplicate to demonstrate the high reproducibility of the SIFR® technology.

[0153] Timeline and analysis (Figure 1):- Fundamental fermentation parameters: pH, gas production, SCFA (acetate, propionate, butyrate and valerate), bCFA (isobutyrate, isovalerate and isocaproate), total SCFA and lactate: Oh (NSC only), 6h, 24h and 48h (all).- Bacterial composition (quantitative 16S rRNA gene profiling): Oh (NSC only) and 48h (all).- Sampling for: Metabolomics (untargeted LC-MS / MS semi-polar analysis): Oh (NSC only) and 48h (all) and Host-microbiome interactions - gut barrier integrity and immune functioning (caco- 2 / THP-1 co-culture model): Oh (NSC only) and 48h (all).A-ll, 1. In derived microbiota

[0154] Based on the data of Lavelle et al. demonstrating minor longitudinal differences along the colon of healthy individuals (Lavelle, A. et al. (2015) Spatial variation of the colonic microbiota in patients with ulcerative colitis and control volunteers. Gut 64, 1553-1561), fecal samples were used as a proxy for the colonic microbiota. The 6 fecal donations scored between 3 and 4 on the Bristol stool score (BSS) scale suggesting they were not subject to long distal transit times (Falony, G. et al. (2016). Population-level analysis of gut microbiome variation. Science 352, 560-564; Muller, et al. (2019). Distal colonic transit is linked to gut microbiota diversity and microbial fermentation in humans with slow colonic transit. Am. J. Physiol. -Gastrointest. Liver Physiol. 318, G361-G369). The donors complied to the following criteria: healthy subjects, age 25-65, no antibiotic / probiotic use in 3 months prior to study, no gastrointestinal complaints nor diagnosed disorders (cancer, ulcers, IBD), non-smoking, alcohol consumption <3 units / day and BMI <30 and >18.5.

[0155] All 6 samples consistently contained taxa belonging to key phyla.

[0156] According to the Microbiota phylogenetic analysis by Quantitative shallow shotgun sequencing (see methods below), the stratification of donors was in line with the stratification of human gut microbiota according to so-called “enterotypes”. Donors 1 , 3 and 6 given the higher levels of Prevotellaceae, a key distinctive feature of the Prevotella enterotype. In contrast, donors 2 and 5 had high levels of Bacteroidaceae, while donor 4 had high levels of Bacteroidaceae, Lachnospiraceae and Ruminococcaceae, so that these three donors classify as Bacteroides-Firmicutes enterotype donors. While the health relevance of gut enterotypes remains to be elucidated, they have been proposed as a useful stratification tool in gut microbiome research. Overall, this stresses that the 6 human adults used during the current study cover the broad range of microbiota composition that occurs in vivo.A-ll, 2. SIFR® Technology

[0157] Individual bioreactors were processed in parallel in a bioreactor management device (Cryptobiotix, Ghent, Belgium). Each bioreactor contained 5 mL of nutritional medium-fecal inoculum blend supplemented with 5 g test compound / L, then sealed individually, before being rendered anaerobic. After preparation, bioreactors were incubated under continuous agitation (140 rpm) at 37°C for48 h (MaxQ 6,000, Thermo Scientific, Thermo Fisher Scientific, Merelbeke, Belgium). Upon gas pressure measurement in the headspace, liquid samples were collected for subsequent analysis.A-ll, 3. Fundamental fermentation parameters

[0158] Unbranched SCFAs (acetate, propionate, butyrate, valerate and caproate), and branched short-chain fatty acids (bCFA; sum of isobutyrate, isocaproate and isovalerate) and lactate were determined via GC with flame ionization detection, upon diethyl ether extraction.

[0159] SCFA were extracted from the samples with diethyl ether, after the addition of 2-methyl hexanoic acid as an internal standard. Extracts were analyzed using a GC with flame ionization detection and a split injector (Trace 1300, Thermo Fisher Scientific, Merelbeke, Belgium), equipped with a capillary fatty acid-free EC-1000 Econo-Cap column (dimensions: 25 mm x 0.53 mm, film thickness 1.2 mM; Alltech, Laarne, Belgium). The injection volume was 1 mL and the temperature profile was set from 110 to 160 °C, with a temperature increase of 6°C min-1. The carrier gas was nitrogen, and the temperature of the injector and detector were 100 and 220°C respectively. The production of unbranched and branched SCFA was calculated by summing the molar concentrations of acetate, propionate, butyrate, valerate and caproate, and summing isobutyrate, isovalerate and isocaproate molar concentrations respectively. The total SCFA production was defined as the sum of unbranched and branched SCFA.

[0160] pH was measured using an electrode (Hannah Instruments Edge HI2002, Temse, Belgium).A-ll, 4. Bacterial composition

[0161] Quantitative data was obtained by correcting abundances [%; shallow shotgun sequencing (3 M reads) with total cell counts for each sample (cells / mL; flow cytometry), resulting in estimated cell counts / mL. Upon DNA extraction, library preparation and sequencing were performed on an Illumina MiSeq platform with v3 chemistry. The 16S rRNA gene V3-V4 hypervariable regions were amplified using primers 341 F (50 -CCT ACG GGN GGC WGC AG- 30) and 785Rmod (50 -GAC TAC HVG GGT ATC TAA KCC-30).

[0162] DNA was extracted via the SPINeasy DNA Kit for Soil (MP Biomedicals, Eschwege, Germany), according to manufacturer’s instructions. Subsequently, DNA libraries were prepared using the Nextera XT DNA Library Preparation Kit (Illumina, San Diego, CA, United States) and IDT Unique Dual Indexes with total DNA input of 1 ng. Genomic DNA was fragmented using a proportional amount of Illumina Nextera XT fragmentation enzyme. Unique dual indexes were added to each sample followed by 12 cycles of PCR to construct libraries. DNA libraries were purified using AMpure magnetic Beads (Beckman Coulter, Brea, CA, United States), eluted in QIAGEN EB buffer, quantified using a Qubit 4 fluorometer and a Qubit dsDNA HS Assay Kit, and sequenced on an Illumina Nextseq 2000 platform 2 x 150 bp. Unassembled sequencing reads were converted to relative abundances (%) using the CosmosID-HUB Microbiome Platform (CosmosID Inc., Germantown, MD, United States). For total cell count analysis, liquid samples were diluted in anaerobic phosphate-buffered saline (PBS), after which cells were stained with SYTO 16 at a final concentration of 1 pM and counted via a BD FACS Verse flow cytometer (BD, Erembodegem, Belgium). Data was analyzed using FlowJo, version 10.8.1.A-ll, 5. Metabolomic

[0163] The Liquid Chromatography - Mass Spectrometry (LC-MS) analysis was carried out using a Thermo Scientific Vanquish LC coupled to Thermo Q Exactive HF MS. An electrosprayionization interface was used as ionization source. Analysis was performed in negative and positive ionization mode.

[0164] LC conditions LC system: ACQUITY UPLC System ; Column: ACQUITY UPLC HSS T3 C18 2.1 x 150 mm, 1.8 pm; Flow rate: 300 pL / min ; Column temp.: 30°C ; Mobile phase A: 10 mM ammonium formate, 0.1% formic acid in water (pH 3.1) ; Mobile phase B: 10 mM ammonium formate, 0.1 % formic acid in methanol ; Gradient: 0.0 min 0% B 2.0 min 0% B 12.0 min 35% B 13.0 min 90% B 14.0 min 90% B 15.0 min 0% B; Injection vol.: 50 pL (partial loop injection mode) ; MS conditions : MS system: Xevo TQ MS Ionization mode: ESI+ Capillary voltage: 3.5 kV Source temp.: 120°C Acquisition mode: MRM MS1 / MS2 mass window: 1 Da (unit mass resolution); Dwell time: 20 ms.

[0165] Peak areas were extracted using Compound Discoverer 3.1 (Thermo Scientific). In addition to the automatic compound extraction by Compound Discoverer 3.1 , a manual extraction of compounds included in an in-house library was performed using Skyline 21.1 (MacCoss Lab Software).

[0166] Identification of compounds were performed at three levels: Level 1 : identification by retention times (compared against in-house authentic standards), accurate mass (with an accepted deviation of 3 ppm), and MS / MS spectra, Level 2a: identification by retention times (compared against in-house authentic standards), accurate mass (with an accepted deviation of 3 ppm). Level 2b: identification by accurate mass (with an accepted deviation of 3 ppm), and MS / MS spectra. Level 3: identification by accurate mass alone (with an accepted deviation of 3 ppm).A-ll, 6. Gut barrier

[0167] This assay involved appropriate differentiation periods for both Caco-2 (epithelial cells) and THP1-cells (immune cells) after which the co-culture experiment was performed during which the in vivo gut wall was recreated by covering the THP-1 cells with the epithelial layer in a transwell system. The actual experiment consisted of (i) a 24h treatment period during which test products were applied on the apical side of the epithelial cells allowing to evaluate the impact on gut barrier integrity.

[0168] Caco-2 cell lines obtained from the ATCC were cultured in MEM media supplemented with 1X NEAA and 1 mM Sodium Pyruvate with 10% FBS. 24-well trans-well inserts were coated with Collagen I Rat Tail Protein and 1 x 105 Caco-2 cells seeded onto the apical chambers. The basal chambers were filled with 200 pL culture media and plates incubated in a 5% CO2 humidified incubator for 14 days. During the differentiation process, media were changed every other day. The TEER was measured to ensure that only transwells with a TEER of more than 300 Q.cm2 were selected for the main experiment.

[0169] THP-1 cells were cultured in RPMI-1640 supplemented with 10% FBS, 1 mM sodium pyruvate and 10mM HEPES at 37°C with 5% CO2. Cultures were initially inoculated at a density of 3 x 105cells / mL and split once density had reached 1 x 106cells / mL. To differentiate THP-1 cells into macrophages, THP-1 cells were centrifuged and resuspended in cell culture medium containing 100 ng / mL PMA. The PMA-treated THP-1 cells were seeded (5 x 105 cells) on transwell-suitable 24-well plates and incubated at 37°C 5% CO2 to induce differentiation. After 48 hours, Caco-2 bearing inserts were moved to the transwell-suitable 24-well plates containing the PMA differentiated THP-1 cells.

[0170] At the start of the main experiment, culture media in the apical chamber were replaced with samples derived from the SIFR® incubations (for 48h), diluted in cell medium. Upon measuring TEER, plates were incubated for24h after which the TEER was again measured and 500 ng / mL of LPS was added to the basal chamber of the transwells containing the THP-1 cells. Upon a 6h LPS challenge, TEER was measured.A-ll, 7. Data analysis

[0171] For the statistical evaluation of the treatment effects on fundamental fermentation parameters, bacterial cell counts, bacterial diversity (4 indices) and bacterial composition (phylum level) across 6 different donors, a repeated measures ANOVA analysis was performed (based on paired t-testing, thus accounting for fact that values are compared between samples of a given donor). The statistical significance of the potential treatment effects was determined via Benjamini-Hochberg post hoc testing (False discovery rate: FDR = 0.05). The latter involves that a correction for multiple comparisons was implemented where p-values were adjusted by multiplying them with the total amount of comparisons divided by the rank of each original p- value (across all p-values). In this specific case, there were 7 comparisons to be considered (control = NSC vs. 7 treatments). In practice, this means that while the largest obtained p-value remained uncorrected (i.e. , multiplied by 1), the lowest p-value was multiplied by 7, thus strongly decreasing the chance of type 1 errors (i.e., false positives). Statistical differences versus the NSC were visualized via * (0.01 < adjusted p-value < 0.05), ** (0.001 < adjusted p-value < 0.01) or *** (adjusted p-value < 0.001).

[0172] For the statistical evaluation of the treatment effects on bacterial composition (family and OTU level), the Benjamini-Hochberg correction was applied within each comparison (control = NSC vs. 7 test products), given the large number of features analyzed.

[0173] As a remark, for statistical analysis of the quantitative 16S rRNA gene profiling, a value below the limit of quantification (LOQ) was equalled to the LOQ. Then, statistics was performed based on log-transformation of the absolute values (to render the data normally distributed). To establish an overall LOQ, first, 1 read was divided by the total amount of reads in each sample, followed by multiplication with the bacterial cell count detected via flow cytometry. This allowedto obtain a LOQ for each sample individually. Then, the highest LOQ was used as overall LOQ of the entire dataset.

[0174] Further, regularized Canonical Correlation Analysis (rCCA) was performed to highlight correlations between metabolites and compositional data (at OTU (Operational Taxonomic Unit) level). Regarding compositional data, log-transformed, absolute phylogenetic data was used as input. rCCA was executed using the mixOmics package with the shrinkage method for estimation of penalisation parameters in R.A-lll Results and discussionA-lll, 1. Fundamental fermentation parameters (Figures 2 to 10)

[0175] At 48h, NUTRIOSE® alone significantly increased acetate, propionate, butyrate (and thus also total SCFA) levels, decreased pH and increased gas production. These changes were highly consistent among the 6 different human donors tested. Particularly the marked increase of acetate / propionate at only very low gas production was remarkable. While significant effects were already observed at 6h, the extent was very small compared to the changes observed at 24 / 48h, stressing the slow initial fermentation of NUTRIOSE®. Further, NUTRIOSE® lowered proteolytic fermentation as followed from the significantly decreased bCFA levels.

[0176] In contrast, GDL alone was rapidly fermented within as short as 6h as followed from the marked pH decrease due to the production of acetate, propionate but especially lactate. The subsequent consumption between 6-24h suggested that lactate was consumed by lactate- consuming, propionate / butyrate-producing gut microbes. At 48h, GDL significantly and markedly increased acetate, propionate, butyrate (and thus also total SCFA) levels, decreased pH and increased gas production. In comparison to NUTRIOSE®, GDL more strongly stimulated butyrate levels and gas production. Further, the effects of GDL were prone to larger interpersonal differences, particularly with respect to propionate and butyrate production.

[0177] With respect to the combinations of NUTRIOSE® and GDL, it followed that higher proportions of NUTRIOSE® or GDL drove the response towards the response of the respective individual ingredient. That is to say that higher amounts of NUTRIOSE® resulted in slower initial fermentation, higher final propionate and lower final levels, while higher GDL levels resulted in higher final butyrate levels.

[0178] While NUTRIOSE®, GDL and combinations thereof all significantly increased the production of health-related acetate, propionate and butyrate (total SCFA), while decreasing bCFA levels, there were marked differences in the kinetics and extent of how NUTRIOSE® and GDL impacted metabolite production. NUTRIOSE® was more gradually fermented and resulted in high final propionate levels at only low gas production. In contrast, GDL was more rapidly fermented within as short as 6h as followed from the marked pH decrease due to the productionof acetate, propionate and especially lactate. At 48h, GDL boosted butyrate (likely via crossfeeding based on acetate / lactate).A-lll, 2. Bacterial compositionA-lll, 2.1. Bacterial cell counts and bacterial diversity (Figures 11 and 12)

[0179] Analysis of the total cell numbers across the different samples demonstrated that there were marked differences in cell numbers between test conditions with all products significantly increasing cell numbers versus the control (NSC) at 48h (Figure 11). Product-specific differences were noted with NUTRIOSE® resulting in much stronger increase of bacterial density compared to GDL.

[0180] Four diversity indices were calculated to obtain optimal insights in bacterial diversity (Figure 12). First, both the observed number of species and the Chaol diversity index were calculated as a measures of species richness (Figure 12A / B). As the Chad diversity index estimates the number of missing species (and thus “counts the uncountable”), this index is likely the most appropriate measure for species richness for the current dataset. Interestingly, while a lower number of observed OTUs (operational taxonomic unit) was noted for all treatments, all treatments maintained a similar species richness compared to the untreated NSC based on the Chad diversity index (except for 50G / 50N).

[0181] Further, two additional indices were based on both species’ richness and evenness (Figure 12C / D; reciprocal Simpson diversity index and Shannon diversity index). These indices are rather based on the dominant community members and have higher values as these dominant members are more evenly distributed. All test products significantly decreased both indices (except reciprocal Simpson diversity index for NUTRIOSE®), suggesting a less evenly distributed community upon treatment. In otherwords, a more limited number of species became dominant, indicating the selective utilization of the different compounds by specific host microorganisms. Such decrease in diversity has been observed upon prebiotic intervention in line with the prebiotic definition that defines prebiotics as substrates that are selectively utilized by specific micro-organisms.A-lll, 2.2. Bacterial composition analysis (Figures 13 to 17)

[0182] The microbial composition results at phylum level, expressed in abundance (%) and in absolute (bacterial cells / mL) calculated from flow cytometry total bacteria counting (Figure 13) demonstrated that the four key phyla detected across the 6 human adults donors were Actinobacteriota, Bacteroidota, Firmicutes and to a lesser extent also Proteobacteria. GDL strongly boosted Actinobacteriota, while NUTRIOSE® strongly stimulated Bacteroidota and Firmicutes. Interestingly, the GDL / NUTRIOSE® ratio resulted in an effect that was more similar to the one of the main ingredients. As a result, a targeted analysis was performed for each of these phyla (Figure 14). This revealed a fundamental difference between GDL and NUTRIOSE®. NUTRIOSE® strongly increased Bacteroidota and Firmicutes in contrast to GDL. As a remark,GDL even significantly decreased Bacteroidota. Adding 10% NUTRIOSE® in the blend already resulted in similar Bacteroidota levels compared to the control (NSC), while this phylum significantly increased from 25% NUTRIOSE® onwards. GDL strongly increased Actinobacteriota in contrast to NUTRIOSE®. Although not significant, GDL tended to increase Proteobacteria which related to an effect observed for a limited number of donors.

[0183] All taxa that were significantly affected (FDR = 0.10) were displayed in a heat map based on the average ratios versus the control (NSC) (Figure 15(A) and Figure 16). Both at family and OTU level, a rCCA was performed to highlight correlations between fundamental fermentation parameters and specific taxa (Figure 15(B) and Figure 17). Owing to the interpersonal differences, correlations could be established between specific metabolites and certain taxa, in line with known metabolic capabilities of these taxa.

[0184] A first key difference between GDL and NUTRIOSE® was that in contrast to NUTRIOSE®, GDL significantly increased the Bifidobacteriaceae family due to the marked stimulation of Bifidobacterium adolescentis (Figures 15(A) and 16). The increase of Bifidobacteriaceae and B. adolescentis was significant from 50% GDL onwards. Both Bifidobacteriaceae and B. adolescentis strongly correlated with butyrate levels (Figures 15(B) and 17), despite the fact that B. adolescentis is unable to produce butyrate but rather produces acetate and lactate as its main end-metabolites.

[0185] Interestingly, two other OTUs correlated with butyrate production and also markedly increased upon GDL treatment:- Faecalibacterium prausnitzii: an acetate-consuming, butyrate-producing species, belonging to the Ruminococcaceae family, increased significantly from 25% GDL onwards (Figure 16).- Anaerobutyricum hallii (formerly known as Eubacterium hallii): an acetate / lactate-consuming, butyrate-producing species, belonging to the Lachnospiraceae family, tended to increase at higher doses of GDL (even if such increases were not significant given the large interpersonal differences) (Figures 16).

[0186] Interestingly, earlier studies have demonstrated cross-feeding between acetate / lactate-producing B. adolescentis and the butyrate-producing F. prausnitzii and A. hallii. Therefore, it is likely that Bifidobacteriaceae thus indirectly stimulated butyrate production upon GDL treatment via such cross-feeding mechanisms.

[0187] In contrast to GDL, NUTRIOSE® significantly increased:-Tannerellaceae due to the marked increases of OTUs related to Parabacteroides distasonis and P. merdae (Figures 15(A), 16). Given the very specific impact, Tannerellaceae and P. distasonis already significantly increased from 10% NUTRIOSE® onwards. P. distasonis levels strongly related with the production of propionate (Figure 17), further suggesting its contribution to the fermentation of NUTRIOSE®.- Bacteroidaceae due to the stimulation of OTUs B. uniformis and B. xylanisolvens (Figures15(A), 16). The increase of Bacteroidaceae was significant from 90% NUTRIOSE® onwards and interestingly, the OTU related to B. uniformis markedly correlated with propionate (Figure 17).- Lachnospiraceae (from 25% NUTRIOSE® onwards) due to the marked increase of a series of OTUs (Figures 15(A), 16). From 25% NUTRIOSE® onwards, OTUs related to following species increased significantly (Figure 16): Fusicatenibacter saccharivorans, Blautia obeum / wexlerae, Blautia faecis, Oliverpabstia intestinalis. The latter remarkably increased up to levels reaching almost 109 cells / mL (data not shown). From 50% NUTRIOSE® onwards, OTUs related to Blautia massiliensis and Galliintestinimicrobium propionicum increased significantly (Figure 16). From 75% NUTRIOSE® onwards, OTUs related to Lachnoclostridium edouardi and Ruminococcus lactaris increased significantly (Figure 16), while from doses of 90% NUTRIOSE® onwards, OTUs related to Ruminococcus faecis, and Roseburia sp. increased (Figure 16).- Erysipelotrichaceae due to the marked increases of an OTU related to Faecalibacillus intestinalis. These increases were significant from 90% NUTRIOSE® onwards (Figures 15(A), 16).

[0188] While NUTRIOSE® stimulated a broad range of species, the key NUTRIOSE®- fermenting species likely involved Parabacteroides species, Bacteroides uniformis, Fusicatenibacter saccharivorans, Blautia species and Oliverpabstia intestinalis. These species generally already strongly increased from 25% NUTRIOSE® onwards.

[0189] In consistency with recent in vivo studies, key contributors to NUTRIOSE® fermentation involved OTUs related to Parabacteroides distasonis, Parabacteroides merdae, Bacteroides uniformis, Blautia species and Fusicatenibacter saccharivorans, while NUTRIOSE® also remarkably increased the recently isolated Oliverpabstia intestinalis. GDL also specifically impacted microbial composition, yet, in a very different way compared to NUTRIOSE®. GDL strongly increased the acetate / lactate-producing Bifidobacterium adolescentis that, likely via cross-feeding mechanisms, boosted the butyrate-producing Faecalibactrium prausnitzii and Anaerobutyricum hallii.

[0190] Altogether, these results show that a combination in a GDL to NUTRIOSE® ratio higher than 10 / 90 and lower than 90 / 10 (in particular from 25 / 75 to 75 / 25), allows to promote the simultaneous growth of beneficial bacteria via the production of metabolites of interest or SCFAs (cf, Example section A-lll, 1 and A-lll, 3). Beneficial bacteria comprise: Bifidobacteriaceae including Bifidobacterium adolescentis which produces lactate and acetate; butyrate-producing bacteria like Faecalibacterium prausnitzii (Ruminococcaceae) and Anaerobutyricum hallii (Lachnospiraceae); propionate- and succinate-producing bacteria such as Parabacteroides distasonis and P. merdae (Tannerellaceae), B. uniformis and B. xylanisolvens (Bacteroidaceae). Some of the promoted bacteria like Fusicatenibacter saccharivorans and Blautia obeum / wexlera (Lachnospiraceae) are able to use complex molecules such as resistant dextrins, and some species from Ruminococcus family are able to degrade complex polysaccharides into a varietyof metabolites having a positive impact on intestinal health. The combination of GDL and NUTRIOSE® also allows to promote the growth of Blautia species, which are described as to be acetate, succinate, lactate, and butyrate producers. Some other promoted species like Faecalibacillus intestinalis have been described as being beneficial in diseases like IBD.

[0191] The simultaneous increase of all those species allows to avoid obtaining an effect that is too specific of some species, and thus decrease the impact of microbiota variability between individuals.

[0192] Furthermore, the combination of GDL and NUTRIOSE® allows to decrease the growth of some species that have been described as being detrimental to health when they overgrow into the colon, such as: Alistipes putredinis and Lachnoclostridum species that are suspected to play a role in colorectal cancer; Dorea longicatena and Dorea formicigenerans that are suspected to play a role in liver cirrhosis in MAFLD, obesity and cardiovascular risks; and Ruminococcus torques which is involved in IBD mechanisms.A-lll, 3. Metabolomics (Figure 18)

[0193] In order to see whether a combination of GDL and NUTRIOSE® could have specific effects on metabolomic, the 25 significantly affected metabolites were presented in a heat map (based on the Iog2 ratios versus the control) (Figure 18).

[0194] First, a series of amino acids was efficiently consumed in presence of NUTRIOSE® and GDL when comparing the 48h samples with those collected at Oh (INO) (data not shown). As a remark, there were minor inter-product differences such as a more pronounced fermentation of isoleucine, leucine and phenylalanine for NUTRIOSE®. Nevertheless, amino acid fermentation was overall highly efficient. An efficient conversion of amino acids is of interest given the potential resulting production of health-related metabolites. In this context, the marked conversion of tryptophan and tyrosine was of special interest and will be elaborated below.

[0195] In view of amino-acid related metabolites, a first finding was that hydroxyphenyllactic acid and indole-3-lactic acid, metabolites from respectively tyrosine and tryptophan, were specifically increased for upon GDL treatment (Figure 18. A Figure showing metabolism of tyrosine, tryptophan and phenylalanine can be found Figure 3 of Dylan Dodd, et al. (2017). A gut bacterial pathway metabolizes aromatic amino acids into nine circulating metabolites. Nature. 551 . 10.1038 / nature24661). Data from literature showed that samples with enhanced indole-3- lactic acid associated with activation of the aryl hydrocarbon receptor (AhR), a receptor important for controlling intestinal homeostasis and immune responses. Further, Hydroxyphenylpropionic acid could be derived from tyrosine catabolism (Figure 3 of Dylan Dodd, et al. (2017)), an amino acid that was indeed efficiently consumed as mentioned above.

[0196] The observation that phenylacetic acid and hydroxyphenylacetic acid were decreased upon treatment compared to the control, further confirmed that GDL steered phenylalanine, tyrosine and tryptophan metabolism from the oxidative to the reductive pathway.

[0197] Another interesting observation for GDL was the increase of hydroxybutyric acid (Figure 18). As described in literature, in mammalian cells, 2 / 3-hydroxybutyric acid (2 / 3-HBA) is formed as a product of fatty acid oxidation and can be used as an energy source in absence of glucose. 2 / 3-HBA is an important signaling molecule that can influence gene expression, lipid metabolism, neuronal function, and the overall metabolic rate. Some of these effects are direct 2 / 3-HBA effects, while others are indirect effects, regulated by the metabolites into which 2 / 3- HBA is converted. 3-Hydroxybutyric acid is able to cross the blood-brain-barrier into the central nervous system. One of the most important regulatory functions of 2 / 3-HBA is the inhibition of the activity of histone deacetylase (HDAC). 2 / 3-HBA may have clinical relevance in the treatment for example of depression, anxiety, and cognitive impairment.

[0198] A class of metabolites that was also strongly affected by particularly NUTRIOSE® were the N-acetylated amino acids (Figure 18). NUTRIOSE® stimulated N-Acetyl-alanine, N-Acetyl- L-leucine and N-acetyl Tryptophan. Stimulation of N-acetylation of amino acids has recently been observed upon treatment with human milk oligosaccharides and was suggested to contribute to early-life immune development and a healthy gut barrier function.

[0199] In terms of vitamins, NUTRIOSE® significantly increased the levels of pyridoxine (= vitamin B6) and particularly pyridoxamine (= vitamin B6) (Figure 18). Vitamin B6 is a water- soluble vitamin that may play a vital role in irritable bowel syndrome (IBS) and participate in inflammatory conditions. The levels of vitamin B6 are linked to a high IBS-symptom score, and the symptoms can be alleviated by increased vitamin B6 intake.

[0200] NUTRIOSE® also exerted remarkable effects on amino-acid derived metabolites such as:- 5-lndolol or 5-hydroxyindole, which is a potent stimulant of intestinal motility via its action on L-type calcium channels. The gut microbiota can metabolize the dietary supplement and antidepressant 5-hydroxytryptophan (5-HTP) to 5-indolol. Interestingly, oral administration of 5- HTP restores gut microbiota dysbiosis in a mouse model of depression.- 2-oxobutyric acid, which is a precursor of propionate in fermentation of methionine and threonine to propionate that was indeed markedly increased upon NUTRIOSE® treatment.- 2-hydroxy-4-(methylthio)butanoic acid, which is a precursor of methionine (it has a hydroxy group instead of amine group) and has been shown to exert beneficial effects on gut barrier.

[0201] Further, several compounds increased upon treatment with both GDL and NUTRIOSE® (Figure 18):- Acetylcadaverine (polyamine): polyamines regulate multiple biological processes, including translation, transcription and cell proliferation and differentiation, and are important intestinalgrowth factors that can contribute to the maintenance of intestinal homoeostasis.- Tyramine is a neurotransmitter that is produced via decarboxylation of tyrosine.

[0202] In conclusion, it was shown that GDL steered fermentation of tyrosine and tryptophan towards the production of aromatic lactic acids such as indole-3-lactic acid, hydroxyphenyllactic acid and its metabolite hydroxyphenylpropionic acid. This may positively impact immune functioning accordingly. GDL also stimulated hydroxybutyric acid, which may positively impact the gut-brain axis. In contrast.

[0203] NUTRIOSE® exerted highly specific effects on:- N-acetylated amino acids, which may positively impact the early-life immune development and gut barrier function;- vitamin B6, which may contribute to protect against inflammatory bowel diseases;- health-related metabolites such as 2-hydroxy-4-(methylthio)butanoic acid which has an impact on barrier integrity, 2-oxobutyric acid and 5-indolol having an impact on gut brain axis.

[0204] This altogether stressed the distinct and various effects of GDL and NUTRIOSE®. Interestingly, specific effects were also identified for the combinations thereof, confirming that combining GDL with NUTRIOSE® in the claimed ratio is an appropriate strategy to obtain improved health benefits. Combinations in weight ratio of GDL to NUTRIOSE® higherthan 10 / 90 and lower than 90 / 10 (in particular of 25 / 75 to 75 / 25) allowed to obtain a gradual increase of hydroxyphenyllactic, indole-3-lactic acid, 2 / 3-HBA, 2-hydroxy-4-(methylthio)butanoic acid, 2- oxobutyric acid, Acetylcadaverine, Tyramine, Triethylamine N-oxide, N-Acetyl-alanine and N- acetyl Tryptophan. Combinations in weight ratio of GDL to NUTRIOSE® higher than 10 / 90 and lower than 75 / 25 (in particular of 25 / 75 to 50 / 50) further allowed to obtain a gradual increase of 5-lndolol, N-Acetyl-L-leucine, pyridoxamine, pyridoxine.A-lll, 4. Gut barrier integrity (Figures 19-21)

[0205] First, the TEER at Oh was on average 739 ± 113 ohm. cm2suggesting reproducible and comparable gut barrier integrity across the different wells used to assess the impact of the various test. A key observation was that upon 1 h and 24h of interaction between the colonic samples (collected at 48h of colonic incubation) and the co-culture of epithelial and immune cells (Caco-2 and THP-1 differentiated macrophages), most test products tended to improve gut barrier integrity as followed from the increased TEER of the epithelial cell layer (Figure 19B). Interestingly, upon stimulation of THP-1 differentiated macrophages with LPS overan additional 6h period (thus total of 30h), it followed that all treatments (except 25G / 75N) significantly increased barrier integrity compared to the control (Figure 19C). Interestingly, it thus seems that the beneficial impact of all treatments on barrier integrity is more profound when cells are under stress (in casu, simulated by an LPS challenge).

[0206] To further scope for correlations between the gut barrier integrity and SCFA production, a correlation analysis was performed between the individual SCFA and TEER. This revealedthat particularly acetate and butyrate correlated with an increase in TEER (Figure 20D / F). While many other metabolites are expected to play a role in the modulation of gut barrier integrity, these data confirm the key role of SCFAs for maintaining a functional gut barrier. Finally, a correlation between bacterial composition (Figure 21) revealed the marked positive correlation between especially Bifidobacterium adolescentis (and to a lesser extent the butyrate-producing Anaerbutyricum hallii) and an improved gut barrier integrity at 30h, suggesting that these species that markedly increased upon GDL or GDL / NUTRIOSE® treatment could be involved in the beneficial impact on barrier integrity.

[0207] Applying samples collected upon 48h of colonic fermentation on human cells (coculture model of epithelial and immune cells) demonstrated that the production of health-related metabolites by health-related species upon treatment with GDL / NUTRIOSE® also translated in a beneficially impacted barrier integrity, especially when the simulated intestinal epithelial layer was under stress ( / .e., challenged with LPS).A-lll, 5. General conclusion

[0208] Altogether, the obtained results suggest distinct effects of GDL and NUTRIOSE® on microbial metabolite production and microbial composition. When combined, they act in synergy, which may result in a series of health benefits for the human host. Prebiotic effects significantly increased with a weight ratio of GDL to NUTRIOSE® from 50 / 50 to 75 / 25. Where potential toleration of the combination is a key aspect, lowering the GDL fraction to GDL to NUTRIOSE® weight ratio lower than 50 / 50 (e.g., of 25 / 75) also appeared to be a good option.

Claims

37Claims

1. Non-therapeutic use of a substance (c) selected from GDL, a gluconate salt, gluconic acid or from a mixture thereof, for ameliorating or decreasing the risk of a disease or condition selected from depression, anxiety, neurological disorders, cognitive impairment, memory loss, attention disorders, and perception disorders; said substance being intended to be taken orally.

2. Non-therapeutic use of a substance (c) selected from GDL, a gluconate salt, gluconic acid or from a mixture thereof, for ameliorating or decreasing the risk of muscle dysfunction; said substance being intended to be taken orally.

3. Non-therapeutic use of a substance (c) selected from GDL, a gluconate salt, gluconic acid or from a mixture thereof, as an antiaging agent, and / or as an antioxidant; said substance being intended to be taken orally.

4. The non-therapeutic use of any of claims 1 to 3, wherein said substance (c) is combined with a substance (b) selected from an indigestible dextrin, an indigestible maltodextrin, or from a mixture thereof.

5. The non-therapeutic use of claim 4, wherein the weight ratio of (c) to (b) is higher than 10 / 90 and lower than 90 / 10.

6. The non-therapeutic use of claim 5, wherein the weight ratio of (c) to (b) is equal to or higher than 20 / 80.

7. The non-therapeutic use of claim 6, wherein the weight ratio of (c) to (b) is equal to or higher than 25 / 75.

8. The non-therapeutic use of any of claims 4 to 7, wherein the weight ratio of (c) to (b) is equal to or lower than 85 / 15.

9. A substance (c) selected from glucono-delta-lactone (GDL), gluconic acid, a gluconate salt, or from a mixture thereof, for use as an oral medicament for the prevention or the treatment of a disease or condition selected from depression, anxiety, neurological disorders, cognitive impairment, memory loss, attention disorders, and perception disorders.

10. A substance (c) selected from glucono-delta-lactone (GDL), gluconic acid, a gluconate salt, or from a mixture thereof, for use as an oral medicament for the prevention or the treatment of muscle dysfunction.

11. The substance (c) of any of claims 9 or 10, wherein said substance (c) is combined with a substance (b) selected from an indigestible dextrin, an indigestible maltodextrin, or from a mixture thereof.

12. The combination of claim 11 , wherein the weight ratio of (c) to (b) is higher than 10 / 90 and lower than 90 / 10.38

13. The combination of claim 12, wherein the weight ratio of (c) to (b) is equal to or higher than 20 / 80.

14. The combination of claim 13, wherein the weight ratio of (c) to (b) is equal to or higher than 25 / 75.

15. The combination of any of claims 11 to 14, wherein the weight ratio of (c) to (b) is equal to or lower than 85 / 15.

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