Composition for treating imbalance of gut microbiota

Abstract

The present invention relates to the use of a tomato extract for modulating the gut microbiota in a subject to confer a health benefit, and to a composition containing a tomato extract formulated for delivery to the large intestine. The tomato extract can be used to improve resistance to infection in the gut, reduce inflammation in the gut, treat or minimize inflammatory bowel disease, or treat or minimize irritable bowel syndrome. The extract can also be used to treat conditions associated with an imbalance of the gut microbiota such as anxiety, depression, type II diabetes, non-alcoholic fatty liver disease, obesity, or cognitive impairment.

Description

【Technical Field】 【0001】 The present invention relates to the use of tomato extracts and compositions containing tomato extracts for delivery to the large intestine for treating conditions associated with an imbalance of the gut microbiota. 【Background Art】 【0002】 New evidence suggests that the gut microbiota plays an important role in maintaining host health and in the development of many diseases or conditions. Targeted modulation of the gut microbiota has been suggested as a preventive and / or novel therapeutic approach for several diseases, including obesity, type 2 diabetes (T2DM), and cardiovascular or intestinal inflammatory diseases (e.g., inflammatory bowel disease or irritable bowel syndrome). Several nutritional therapies, including prebiotics and probiotics, are available today, but their full potential has not been realized, in part due to ongoing debates about the precise definition of prebiotics and probiotics, which have led to misinformation and confusion among consumers, researchers, and industry, particularly in the case of prebiotics. To promote the appropriate use of the term "prebiotics," the International Scientific Association of Probiotics and Prebiotics (ISAPP) published a consensus paper in 2017, which defines prebiotics as "substrates selectively utilized by host microorganisms conferring a health benefit." A few prebiotics that are positively correlated with health benefits have been marketed. These include fructans (fructooligosaccharides (FOS) and inulin) and galactans (galactooligosaccharides or GOS). However, many other compositions are sold as prebiotics and are claimed to affect host microorganisms, but it remains a challenge to clearly demonstrate that these candidates confer health benefits to the host. Accordingly, it is an object of the present invention to provide further compositions having demonstrable prebiotic activity that confer a benefit to health. SUMMARY OF THE INVENTION 【0003】 The inventors have established that water-soluble tomato extract (WSTE) is effective in modulating the gut microbiota in the body of a subject, and thus such extract is useful as a prebiotic for treating conditions caused or affected by the gut microbiota, or an imbalance thereof. According to a first aspect of the present invention, there is provided a water-soluble tomato extract having activity for inhibiting platelet aggregation for use in modulating the gut microbiota in a subject to confer a health benefit. The health benefits conferred by WSTE may directly affect the digestive system. By way of example, WSTE may confer a benefit to the subject in at least one of the following manners: (A) improving digestion; (B) improving resistance to infection in the gut; (C) reducing inflammation in the gut; (D) treating or minimizing inflammatory bowel disease (IBD); or (E) treating or minimizing irritable bowel syndrome (IBS). 【0004】 Alternatively, the health benefits conferred by WSTE may be to address conditions where the condition is caused or exacerbated by compounds released by the gut microbiota into the bloodstream, whereby WSTE modulates the plasma levels of such compounds by modulating the gut microbiota. By way of example, WSTE may also confer a benefit to the subject by: (A) treating or preventing anxiety and / or depression; (B) treating or preventing type II diabetes; (C) treating or preventing non-alcoholic fatty liver disease; (D) treating or preventing obesity; (E) Preventing or reducing inflammaging (or inflamm-aging); (F) Immunodeficiency; and (G) Improving or maintaining cognitive function and brain health. 【0005】 According to a second aspect of the present invention, there is provided a method of modulating the gut microbiota of a subject to provide a health benefit to the subject, the method comprising administering to a subject in need of such treatment a water-soluble tomato extract having activity for inhibiting platelet aggregation. The inventors were surprised to find that WSTE has efficacy for modulating the gut microbiota, and have thus come to recognize that WSTE can be formulated in a novel manner to form a composition beneficial to human health. According to a third aspect of the present invention, there is provided a composition formulated for delivery of a water-soluble tomato extract having activity for inhibiting platelet aggregation to the large intestine of a subject, the composition comprising the water-soluble tomato extract and at least one additive that promotes delivery to the large intestine. According to a fourth aspect of the present invention, there is provided a composition comprising a water-soluble tomato extract having activity for inhibiting platelet aggregation and at least one further prebiotic. 【0006】 According to a fifth aspect of the present invention, there is provided a composition comprising a water-soluble tomato extract having activity for inhibiting platelet aggregation and a probiotic. WSTE can be administered to any mammalian subject and is useful in treating animals for veterinary purposes. However, the subject is preferably a human subject. The above summary and the following detailed description will be better understood when read in conjunction with the accompanying drawings. For the purpose of illustrating the disclosed compositions and methods, exemplary embodiments of the compositions and methods are shown in the drawings; however, the compositions and methods are not limited to the specific embodiments disclosed. BRIEF DESCRIPTION OF THE DRAWINGS 【0007】 【Fig. 1】 It is a flowchart showing the experimental setup of Example 3. 【Fig. 2】 As discussed in Example 3, it is a bar graph of the mean absolute change of (A) plasma and (B) urinary TMAO from baseline to the end of intervention in the Fruitflow (preferred WSTE) group and the placebo group. 【Fig. 3】 As discussed in Example 3, it is a figure representing the principal component analysis (PCoA) based on the (A) Bray Curtis and (B) Jaccard distance matrices at the baseline time point and the end of intervention in the Fruitflow group and the placebo group. The ellipse represents the 80% confidence interval. The lines connect samples from the same participant. The density plot shows the projection of the PCoA points onto the PC1 axis and the PC2 axis. The black circles and white circles represent data from the Fruitflow (WSTE) - treated subjects at the start and end of the trial, respectively. The black triangles and white triangles represent data from the placebo - treated subjects at the start and end of the trial, respectively. 【Fig. 4A】 As discussed in Example 3, it is a Volcano plot display of differentially abundant operational taxonomic units (OUTs) between the baseline time point and the end of intervention within (A) Fruitflow and (B) placebo groups, and between groups at (C) species level and (D) genus level. The X - axis position of each point represents the effect - size difference at the end of intervention. The horizontal line represents the unadjusted p - value cutoff at 0.05. Points exceeding the effect - size cutoff (0.20) and the p - value cutoff have different symbols and / or are annotated in the text as follows: Dots within Box I represent non - significant taxa; dots within Box II in FIGS. 4A, 4B, and 4C represent taxa exceeding the p - value cutoff; dots within Box III in FIG. 4D represent taxa exceeding the effect - size cutoff; triangles represent taxa exceeding both the effect - size and p - value cutoffs. 【Fig. 4B】 The same as above. 【Fig. 4C】 The same as above. 【Fig. 4D】 The same as above. 【Fig. 5A】 As discussed in Example 3, (A) PLS-DA 2D plot of multivariate analysis of plasma samples (triangles) and control samples (circles) collected after Fruitflow intervention (R2 = 0.432, Q2 = -0.766); and (B) A non-targeted metabolomics figure showing the 15 most essential bins that result in discrimination between Fruitflow samples and control samples. 【Fig. 5B】 Ibid. 【Fig. 6】 As discussed in Example 3, a bar graph of the mean absolute change in plasma LPS from baseline to the end of intervention in the Fruitflow group and the placebo group. 【DETAILED DESCRIPTION OF THE INVENTION】 【0008】 The disclosed compositions and methods can be more readily understood by reference to the following detailed description, which is related to the accompanying figures that form a part of this disclosure. It is to be understood that the disclosed compositions and methods are not limited to the specific compositions and methods described and / or shown herein, and that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the claimed compositions and methods. References to particular numerical values, unless otherwise indicated in the context, include at least that particular value. When ranges of values are expressed, other embodiments include from one particular value and / or to the other particular value. Further references to values stated in ranges include any and all values within that range. All ranges are inclusive and combinable. 【0009】 It is to be understood that certain features of the disclosed compositions and methods, which are described herein in the context of separate embodiments, can also be provided in combination in a single embodiment. Conversely, various features of the disclosed compositions and methods, which are described herein in the context of a single embodiment for brevity, can also be provided separately or in any sub-combination. The following abbreviations are used in this specification: Cardiovascular disease (CVD); Active pharmaceutical ingredient (API); Water-soluble tomato extract (WSTE); Trimethylamine-N-oxide (TMAO); Trimethylamine (TMA); Lipopolysaccharide (LPS); Short-chain fatty acid (SCFA); Operational Taxonomic Unit (OTU); Type 2 diabetes (T2DM); Inflammatory bowel disease (IBD); Irritable bowel syndrome (IBS); Non-alcoholic fatty liver disease (NAFLD); and Colony-forming unit (CFU). As used herein, the singular forms "a", "an", and "the" include the plural forms. 【0010】 When values are presented as approximations by use of the antecedent "about", it will be understood that the particular value forms another embodiment. Further, the term "about" when used in connection with a numerical range, cutoff, or specific value is used to indicate that the recited value may vary by up to about 10% from the recited value. Many of the numerical values used herein are experimentally determined and as such the determination can vary between different experiments and often such will be understood by one of ordinary skill in the art. The values used herein should not be regarded as overly limiting due to this inherent variability. Thus, the term "about" is used to encompass variations of ±10% or less, ±5% or less, ±1% or less, ±0.5% or less, or ±0.1% or less from the specifically recited value. As used herein, "treating" and like terms refer to reducing the severity and / or frequency of symptoms, eliminating symptoms and / or the root cause thereof, decreasing the frequency or likelihood of symptoms and / or their root cause, delaying, halting, and / or slowing the progression of a condition, and improving or repairing damage caused directly or indirectly by the condition. 【0011】 As used herein, the phrase "therapeutically effective amount" refers to an amount of a composition, such as a WSTE as described herein, effective to achieve a particular biological or therapeutic result, such as, but not limited to, a biological or therapeutic result disclosed, described, or exemplified herein. The therapeutically effective amount can vary depending on factors such as the disease state, the age, sex, and weight of the individual, and the ability of the composition to elicit the desired response in the subject. Such results include, but are not limited to, reducing the plasma levels of TMAO or LPS. "Pharmaceutically acceptable vehicle" can be any physiological vehicle known to those skilled in the art that is useful in formulating a pharmaceutical composition. WSTE is known to confer several health benefits, including the treatment of cardiovascular conditions (see, e.g., WO2010 / 049707). The mechanism of action of WSTE is thought to be by directly modulating the pathophysiology of the subject being treated, and heretofore, no prebiotic activity has been associated with WSTE. The inventors were triggered by the realization that plant polyphenols have been said to be able to act as prebiotics and the realization that WSTE contains such polyphenols, leading to an investigation of the effect of WSTE on the gut microbiota. 【0012】 An estimated 90–95% of dietary polyphenols are not absorbed in the small intestine and reach the colon where they are understood to undergo extensive biotransformation by the gut microbiota (Clifford (2004) Planta Med 70:1103–11014). Health benefits associated with polyphenol consumption have been hypothesized to depend on microbial utilization and production of metabolites rather than the parent compounds (Duenas et al. (2015) BioMed Res Int 2015:850902). As an example, Chen et al. (2016) (mBio American Society of Microbiology 2016;e02210–e2215) have shown that polyphenols can attenuate trimethylamine-N-oxide (TMAO)-induced atherosclerotic lesions by remodeling the gut microbiota and thus reducing TMAO synthesis. TMAO is derived from trimethylamine (TMA), which is a microbial metabolite mainly produced by various taxa of the gut microbiota from dietary phosphatidylcholine and L-carnitine, which are commonly found in red meat, cheese, and eggs. TMA is absorbed through the intestinal epithelium and further transported to the liver where it is subsequently converted to TMAO. TMAO is known for its pro-inflammatory and pro-atherogenic activities, and higher baseline levels of TMAO have been associated with major adverse cardiovascular events. Accordingly, the inventors have conducted studies as outlined in the Examples and have been surprised to find that WSTE was able to migrate through the stomach and small intestine to the bacterial flora found in the large intestine; was able to modulate TMAO and LPS production mediated by the gut microbiota (discussed below); and also affected the beta diversity of the gut microbiota (discussed below). These surprising findings have led to the recognition that WSTE has prebiotic activity and can be used to confer health benefits in a subject, as outlined in the Summary of the Invention and discussed below. 【0013】 The water-soluble tomato extract used according to the present invention The water-soluble tomato extract (WSTE) used according to the present invention contains substantially thermostable colorless water-soluble compounds having an activity for preventing platelet aggregation and having a molecular weight of less than 1,000 daltons. The extract can be derived from the pulp of peeled tomato fruits and / or the juice around the seeds of tomato fruits. 【0014】 The extract can be derived from the juice of tomato fruits, which is then further processed essentially as discussed herein. The extract can be an active fraction isolable from tomatoes, and preferably it is characterized by the following: (a) It is colorless or light yellow; (b) It contains water-soluble compounds; (c) It consists of components having a molecular weight of less than 1000; and (d) It contains one or more nucleosides having platelet aggregation inhibitory activity. Preferred WSTE contains at least one of the following: (a) Glycosylated phenolic acids or phenolic esters or derivatives thereof; (b) Glycosylated flavonoids; and (c) Nucleosides. The glycosylated phenolic acid or phenolic ester can be glycosylated cinnamic acid or a derivative thereof. Such glycosylated cinnamic acid or a derivative thereof can be selected from the group including caffeoyl-4-O-quinic acid, caffeoyl-4-O-glucoside, coumaroyl-4-O-glycoside (glue / gal), and coumaroyl-4-O-glycoside (disaccharide). 【0015】 The glycosylated phenolic acid or phenolic ester can be selected from the following: glucoside of caffeic acid; hexose / p-dihydrocaffeic acid hexose of p-coumaric acid; glycoside of ferulic acid; and derivatives of p-coumaric acid. The glycosylated flavonoid can be quercetin-3-O-glucoside or rutin. The nucleoside can be selected from the group including AMP, uridine, adenosine, guanosine, or GMP. In a preferred embodiment, the WSTE is any extract having an activity for preventing platelet aggregation, as disclosed in WO99 / 55350 or WO2010 / 049707. Preferred WSTE, including those disclosed in WO99 / 55350 or WO2010 / 049707, is lycopene-free or substantially lycopene-free. By "substantially lycopene-free", the inventors mean that the WSTE has had the lycopene removed that is found in most, if not all, tomato juices. Thus, the inventors mean that in the WSTE there is less than 10% w / w, preferably less than 5% w / w, most preferably less than 1% w / w (e.g., 0.75%, 0.5%, 0.25%, or 0.1%) of the lycopene found in an equivalent mass of tomato fruit or tomato juice. In a preferred embodiment, the WSTE contains no lycopene or only trace amounts of lycopene. 【0016】 WO99 / 55350 and WO2010 / 049707 also disclose preferred methods for producing tomato extracts (WSTE) that can be used according to the present invention. WO2010 / 049707 discloses the most preferred method for producing WSTE that can be used according to the present invention. Such water-soluble extracts have been found to have significant efficacy for preventing or reducing platelet aggregation in response to adenosine diphosphate and collagen in human trials and are on the market as a nutritional supplement having health benefits in the cardiovascular area under the brand name FRUITFLOW® with a nutrition function claim approved by the European Food Safety Authority in Europe. WO2010 / 049707 discloses, in Figures 2 (method for making a liquid / syrup extract from fruit) and 4 (method for treating the extract to make a powder from which sugar has been removed), the most preferred methods for manufacturing WSTE that can be used according to the present invention. These extracts and the methods for manufacturing them are incorporated herein by reference. Figures 2 and 4 of WO2010 / 049707 describe preferred methods for manufacturing two types of WSTE that are commercially available as FRUITFLOW®. Most preferably, the WSTE used according to the present invention is FRUITFLOW®. 【0017】 Optional further active ingredients included in the composition according to the present invention Further prebiotics As described above, ISAPP defines prebiotics as "substrates selectively utilized by host microorganisms that confer health benefits". Various prebiotics have been proposed, and the inventors have now surprisingly found that WSTE has prebiotic properties and beneficial effects as described herein. It will be understood that the health benefits provided by WSTE can be further enhanced by including other prebiotics. According to a fourth aspect of the present invention, the composition can include WSTE combined with further prebiotics. Preferred further prebiotics used according to the fourth aspect of the present invention are resistant to mammalian digestion and are thus, in the subject's large intestine, fiber and starch that can be fermented by the gut microbiota, particularly probiotics. Prebiotic fibers are typically complex polysaccharides, which can include starch resistant to mammalian digestion, as well as the soluble fibers inulin, fructooligosaccharides (FOS), and galactooligosaccharides (GOS). 【0018】 The prebiotics included in the composition according to the fourth aspect of the present invention can be supplied from starch-rich plants that are resistant to mammalian digestion, and examples of such plants include green bananas, plantain powder, oats, white rice, and raw potatoes. The prebiotics included in the composition according to the fourth aspect of the present invention can be supplied from plants rich in inulin (such as chicory roots, Jerusalem artichokes, garlic, onions, leek onions, asparagus, and ripe bananas). The prebiotics included in the composition according to the fourth aspect of the present invention can also be supplied from plants rich in FOS such as garlic, shallots, onions, Jerusalem artichokes, burdock roots, and yacon roots. The prebiotics included in the composition according to the fourth aspect of the present invention can also be supplied from plants rich in GOS such as lentils, chickpeas, green peas, lima beans, and kidney beans. 【0019】 Probiotics In some embodiments, WSTE can be combined with probiotics, and according to the fifth aspect of the present invention, the composition can include WSTE combined with probiotics. The human body acts as a host for a wide variety of microorganisms, including various strains of bacteria, and many of these microorganisms live in the intestine. Most of these are located in the large intestine, and some are considered beneficial to human health. The World Health Organization (WHO) defines probiotics as live microorganisms that provide health benefits when consumed in appropriate amounts. The increased range of health benefits contributes to the consumption of probiotics by healthy humans. These include improved digestion, improved resistance to infection, and better mood and sleep. Furthermore, probiotics can also provide benefits for several clinical conditions including irritable bowel syndrome (IBS); anxiety and depression, obesity, and elevated blood LDL cholesterol. Thus, it will be understood that in some embodiments, the beneficial effects of WSTE can be supplemented by also administering probiotics to the subject. The selection of the bacterial strains used for the probiotic component of the composition according to the present invention depends on the condition to be treated. 【0020】 The inventors have found that the compositions according to the fifth aspect of the present invention yield general health benefits when they contain at least one bacterial strain selected from the following: Ruminococcus albus, Alistipes ihumii, Anaeromassilibacillus senegalensis, Saccharofermentans acetigenes, Oribacterium sinus, Clostridium carnis, Ruthenibacterium lactatiformans, Alistipes obesi, Faecalitalea cylindroides, Clostridium methylpentosum, Ruminococcaceae, Saccharofermentans, Alistipes, Anaeromassilibacillus, and Ruthenibacterium, Oribacterium. 【0021】 The composition according to the fifth aspect of the present invention also results in general health benefits when it contains at least one bacterial strain selected from L. Rhamnosus, B. Longum, B. Lactis, B. Bifidum, L. Acidophilus, L. Plantarum, L. Casei, L. Reuteri, B. Infantis, L. Bulgaris, L. Fermentum, L. Paracasei, S. Boulardii, B. Breve, L. Brevis, L. Gasseri, L. Helveticus, S. Thermophillus, Pediococcus acidilactici, and B. Animalis. In one embodiment, the probiotics are selected such that they provide benefits to intestinal health and, in particular, improve the condition of subjects suffering from digestive disorders such as IBS. 【0022】 When billions of colony forming units (CFU) are referred to herein, the inventors mean the total number of probiotics in the composition according to the fifth aspect of the present invention. This total number may include a single bacterial strain or multiple bacterial strains. It will be understood that a minimum total number of probiotic CFU needs to reach the intestine for the probiotics to be effective. The probiotic industry often recommends that the dosage of probiotics should be CFU per 20 billion. The oral consumption of probiotics requires them to pass through the stomach, which is strongly acidic. The stomach can typically contain 0.1 molar hydrochloric acid and can have a pH of 1 to 2. This gastric acid provides a harsh environment that serves as a chemical barrier and has the function of reducing the risk of gastrointestinal infections. The low pH of gastric acid can inactivate viruses and kill potentially pathogenic yeasts, molds, and bacteria by lysing their cell membranes. However, while these positive effects can increase infection resistance, gastric acid also makes it difficult for orally ingested probiotic bacteria to survive digestion. Once the probiotics reach the small intestine, the near-neutral pH of the intestine (typically in the range of 6.8 to 7.4) is much more favorable for probiotic survival and growth. Thus, it will be understood that the compositions according to the fifth aspect of the invention can be formulated to reduce the destruction of probiotics in the stomach (e.g., by encapsulation with acid-resistant materials). 【0023】 Composition for oral administration The compositions used according to the present invention can contain WSTE (and optionally, prebiotics or probiotics) without any additional components (e.g., used by diluting in a liquid or as a powder of encapsulated WSTE). However, in a preferred embodiment, WSTE is formulated with other active substances as discussed below to improve their commercial properties (e.g., to improve delivery, shelf life, taste, and the like). The compositions of the present invention are preferably formulated as a medicament, nutraceutical, health supplement, or food / drink composition for oral administration. For example, they can be formulated as a gel, solution, suspension, syrup, powder, tablet, capsule, lozenge, snack bar, or drink. Such formulations can be prepared according to methods well known in the art. WSTE can be formulated in a syrup or other solution, for example, as a health drink, for oral administration. One or more additives selected from sugars, vitamins, flavoring agents, coloring agents, preservatives, and thickening agents may be included in such a syrup or solution. Tonicity regulators such as sodium chloride or sugar can be added to provide a solution with a specific osmotic power. One or more pH regulators, such as buffering agents, can be used to adjust the pH to a specific value and preferably maintain it at that value. Examples of buffering agents include sodium citrate / citric acid buffer and phosphate buffer. 【0024】 In another embodiment, the powder form of WSTE is formulated as a tablet for oral consumption. For tablet formation, WSTE can typically be mixed with a diluent, such as sugars (e.g., sucrose and lactose); sugar alcohols such as xylitol, sorbitol, and mannitol; or modified celluloses or cellulose derivatives such as powdered cellulose, crystalline cellulose, or carboxymethyl cellulose. Tablets also typically contain one or more additives selected from granulating agents, binders, lubricants, and disintegrants. Examples of disintegrants include starches and starch derivatives, and other swelling polymers, such as cross-linked carboxymethyl cellulose, cross-linked polyvinylpyrrolidone, and cross-linked polymer disintegrants such as starch glycolic acid. Examples of lubricants include stearates such as magnesium stearate and stearic acid. Examples of binders and granulating agents include polyvinylpyrrolidone. If the diluent is not inherently very sweet, a sweetening agent, such as ammonium glycyrrhizinate, or an artificial sweetener such as aspartame or sodium saccharinate, can be added. 【0025】 In a preferred embodiment, the WSTE is formulated as a powder, granule, gel, or semi-solid for incorporation into capsules. When used in powder form, the WSTE can be formulated with any one or more of the additives defined above for tablets or provided in undiluted form. For provision in gel or semi-solid form, the WSTE can be dissolved or suspended in a liquid carrier such as a viscous liquid or semi-solid medium like polyethylene glycol, or a glycol such as propylene glycol, or glycerol, or an oil selected from vegetable oils and fish oils such as olive oil, sunflower oil, safflower oil, evening primrose oil, soybean oil, cod liver oil, sardine oil, etc. These can then be filled into capsules made from either hard gelatin type or soft gelatin type capsules, or capsules made from hard or soft gelatin equivalents. Soft gelatin or soft gelatin equivalent capsules are preferred for viscous liquid or semi-solid fillings. In one preferred embodiment, the composition according to the invention is provided in powder form, optionally together with preferred solid (e.g., powdered) additives, for incorporation into capsules, e.g., hard gelatin capsules. The composition according to the fifth aspect of the invention containing probiotics is preferably encapsulated such that the probiotic content is protected from gastric acid when passing through the stomach. 【0026】 Preferred compositions are for human consumption and can take the form of gummies, pouch powders, tablets, and other similar items retailed by the food and beverage industry as dietary supplements. According to one embodiment of the invention, the WSTE can be formulated into a powder provided to a subject as a premix for making a beverage (if diluted in water or other similar substances) before consumption or for mixing with food by the subject. Alternatively, the premix can be used by a food or beverage manufacturing company to produce a drink or food (e.g., a snack bar) having health benefits as described herein. The most preferred form is a capsule (either hard or soft) for use as a dietary supplement. By way of example only, the composition can be a size 00 Vegecaps (LGA, La Seyne - sur - Mer, France) capsule containing 150 mg of WSTE and a suitable vehicle / bulking agent for the WSTE. The final mass of each capsule is 600 mg (mass of WSTE + mass of bulking agent (e.g., tapioca starch)). It will be understood that a product for use as a food, beverage, or dietary supplement can be adapted to form a pharmaceutical composition and a nutraceutical composition containing WSTE. 【0027】 Dosage form and administration schedule The amount of WSTE required to be administered to a subject depends on several factors. By way of example, if the subject is human, the age, sex, and weight of the subject. WSTE can be provided in the form of unit dosage forms containing a specified amount of WSTE. Such unit dosage forms can be selected to reach the desired level of biological activity in the large intestine. For the liquid extract produced according to method 1.1 of Example 1 (see below), the recommended daily amount of the fruit extract according to the invention is from 0.5 g to 20 g, more preferably from 2 g to 7 g. The daily amount can be about 3 g. A typical administration schedule for humans can be from about 70 mg to 285 mg, preferably from about 25 mg to 100 mg per kilogram of body weight per day. 【0028】 In a preferred embodiment, a low-sugar type of WSTE is used, and most preferably, a powdered WSTE produced according to Method 1.2 of Example 1 (see below) is used. In this case, the recommended daily dose can be 10 mg to 1,500 mg of WSTE; preferably, the daily dose can be 50 mg to 1,000 mg of WSTE, more preferably about 100 mg to about 500 mg. In one embodiment, a subject can receive about 300 mg of WSTE per day, which can be administered as a 300 mg dosage form or a 2 x 150 mg dosage form. In the most preferred embodiment, the daily dose comprises 2 capsules each containing 150 mg of WSTE. Merely by way of example, a subject will benefit from improved digestion, improved resistance to infection in the gut, or reduced inflammation in the gut if they consume 2 capsules (each containing 150 mg of WSTE) per day. These capsules can be consumed in the morning (e.g., with breakfast). A subject can take the capsules as long as a health benefit is provided from the consumption of WSTE. A subject can take WSTE as a preventative (to prevent the occurrence of a condition, e.g., IBS), or can begin taking WSTE to treat a condition (e.g., IBS). Thus, the composition according to the invention can be taken as part of a morning health supplement routine (e.g., for more than 28 days). 【Examples】 【0029】 (Example 1) Preparation of WSTE WSTE for use in the composition according to the invention was prepared by one of the following protocols: 1.1 The liquid (syrup) extract was prepared according to the protocol of Example 2 and Figure 2 of WO2010 / 049707 and is known as FRUITFLOW® 1. 1.2 The low-sugar content powder extract is essentially prepared according to the protocol of Example 3 and Figure 4 of WO2010 / 049707 and is known as FRUITFLOW® 2. 150 mg of FRUITFLOW® 2 contains up to 9 mg of nucleoside derivatives, up to 10 mg of simple phenolic conjugates (e.g., chlorogenic acid, other caffeic acid / phenolic acid derivatives), and up to 7 mg of flavonoid derivatives, and at least 2.4 mg of the flavonoid derivatives are quercetin derivatives. 【0030】 (Example 2) Composition containing WSTE The WSTE described in 1.2 was mixed with a three-fold amount (by mass) of tapioca starch bulking agent by ordinary means, and the mixture was used to fill size 00 capsules (Vegecaps manufactured by LGA (La Seyne-sur-Mer, France)). Each capsule contained 150 mg of WSTE and 450 mg of tapioca starch bulking agent. (Example 3) The inventors determined to evaluate the effect of the composition of Example 2 on the gut microbiota by conducting a randomized, double-blind, placebo-controlled crossover trial to examine the 4-week supplementation with 2 × 150 mg WSTE capsules on plasma and urinary TMAO and fecal microbiota, in addition to plasma and fecal metabolites (e.g., plasma lipopolysaccharide (LPS), bile acids, short-chain fatty acids (SCFA), and other organic acids), and the effect on gastrointestinal comfort. 【0031】 3.1 Materials and methods 3.1.1 Study population The study population consisted of 40 healthy, overweight and obese adults aged 35 - 65 years (BMI 28 - 35 kg / m 2) consisted of. The main exclusion criteria were as follows: severe acute or chronic diseases; smoking; history of drug and / or alcohol abuse (more than 2 drinks of alcohol / day); pregnancy; use of antibiotics within the past 3 months; major dietary changes in the past 3 months; eating disorders; vegetarian or vegan; enemas; prebiotics, probiotics, or dietary supplements containing fiber within 4 weeks before baseline visit and during the intervention period; long-term drug therapy for active gastrointestinal disorders (except when the product was taken for at least 2 months before screening and the exact dosage was maintained throughout the study), and high habitual intake of tomato or tomato-based products as confirmed by a food frequency questionnaire. Eligible participants were randomly assigned to the intervention sequence on a 1:1 basis, with n = 20 participants allocated to the placebo / WSTE arm (group 1) and n = 20 participants allocated to the WSTE / placebo arm (group 2). Three participants withdrew voluntarily after randomization; the remaining 37 participants completed the study as planned. Before unblinding the data to determine inclusion in the per-protocol population, a third-party committee evaluated all participants individually. Fifteen subjects were excluded from the analysis due to the use of concomitant drug therapy such as antibiotics (n = 9) and less than 80% IP compliance and / or lack of important variables (n = 6). 【0032】 3.1.2 Study Design The trial was designed as a randomized, double - blind, placebo - controlled, crossover trial consisting of five visits: a screening visit (visit 1), the start of intervention period 1 after a 21 - day trial introduction period (visit 2), the end of a 4 - week intervention (visit 3), a 6 - week drug washout, and the start of the second intervention period (visit 4), and the end of a 4 - week intervention (visit 5) (Figure 1). During the screening period, vital signs were recorded and a complete health assessment including medical history and demographic / anthropometric evaluation was performed. In addition, tomato consumption was questioned weekly and fasting venous blood samples were taken for safety profiling. Participants were provided with a stool collection kit, instructions for collecting and storing stool samples, and a Bristol Stool Chart (Lewis & Heaton. (1997) Scand J Gastroenterol Taylor and Francis 32:920 - 924) to be completed at the time of stool collection. At visit 2, participants arrived at the research facility and fasted for at least 10 hours overnight. Participants returned the Bristol Stool Chart, and within 24 hours before the visit, stool samples were collected and stored at - 20 °C in a household freezer. Venous blood samples were taken for safety profiling and analysis of trimethylamine N - oxide (TMAO) and lipopolysaccharide (LPS) in plasma. In addition, urine samples were taken for analysis of urinary TMAO, and participants completed the Gastrointestinal Symptom Rating Scale (GSRS) and the Food Frequency Questionnaire (FFQ) before they were randomized to one of the two intervention groups. Participants were provided with two stool collection kits and Bristol Stool Charts and were instructed to collect stool samples at home after 2 weeks and another sample within 24 hours of their next scheduled visit at week 4. At visit 3, participants arrived at the research facility again, fasted overnight, returned the stool samples and the Bristol Stool Chart. Another blood and urine sample were taken and the Gastrointestinal Symptom Rating Scale was completed. In addition, participants returned any unused study products to evaluate compliance during intervention period 1. Thereafter, participants were sent home during a 6 - week drug washout period before entering intervention period 2, which followed the same experimental setup as visits 2 and 3. 【0033】 3.1.3 Ethical Statement The trial was conducted in accordance with the Helsinki Declaration by Atlantia Food Clinical Trials Ltd. in Cork, Ireland, and approved by the Clinical Research Ethics Committee of Cork Teaching Hospitals. Written informed consent for participation was obtained from all participants. 【0034】 3.1.4 Measurement Blood and urine samples: Blood samples were taken for safety profiling (hematology, chemistry, glucose, high-sensitivity C-reactive protein (hsCRP), and bilirubin), as well as for the analysis of TMAO, lipopolysaccharide (LPS), bile acids (BA), short-chain fatty acids (SCFA), and other organic acids, in addition to non-target metabolomics. Urine samples were taken for the analysis of urinary TMAO. Safety profiling, including hematology, chemistry, and glucose, was performed by standard clinical laboratory methods at Eurofins Biomnis (Sandyford, Dublin, Ireland). TMAO and LPS samples were sent to MS-Omics ApS (Bygstubben 9, 2950 Vedbaek, Denmark) for analysis. Plasma LPS analysis was performed by LC-MS method. 【0035】 For analysis, bilirubin and hsCRP samples were sent to Eurofins Biomnis (Sandyford, Dublin, Ireland). Plasma bile acids were extracted from plasma and quantified using a commercially available bile acid assay (Biocrates Life Sciences AG, Austria). SCFA and other organic acids were first extracted from plasma by protein precipitation. Subsequently, the plasma extract was derivatized and the reaction products were extracted by liquid-liquid extraction using dichloromethane. The resulting extract was finally injected into a UHPLC-MS / MS system for analysis in a mode combining positive and ESI MRM. 【0036】 For non-target metabolomics, plasma samples were prepared using the method described previously (Meilko et al. (2021) Sustain Chem Pharm 22:1000574-) with minor adaptations. Briefly, 100 μl of human plasma was mixed with 100 μl of deuterated phosphate buffer pH = 7.3 prepared by adding PBS tablets (OXOID) to 100 ml of D2O. Maleic acid (0.5 mM) was used as an internal standard, and subsequently, the samples were placed in 3 mm NMR tubes. All spectra were acquired at 298 K on a Bruker Avance III NMR spectrometer operating at a proton Larmor frequency of 600 MHz and equipped with a 5 mm TCI cryoprobe. 1D 1H NMR spectra were acquired using the cpmgpr1d pulse sequence, a waiting time (relaxation delay) of 5 s, and a D20 delay of 0.0003 s; 256 scans were accumulated per spectrum over 36.5 minutes. 【0037】 Fecal samples: Fecal samples were collected into DNA / RNA Shield (trademark) and fecal collection tubes (Zymo Research, Irvine, USA) and shipped on dry ice in an -80°C compatible box to BaseClear BV (Leiden, Netherlands) for microbiome profiling. Briefly, nucleic acids were extracted from fecal samples using the ZymoBIOMICS (trademark) DNA Miniprep (Zymo Research Corp., Irvine, CA, USA) kit as per the manufacturer's instructions. The 16S rRNA gene variable regions V3-V4 were amplified using the synthetic primers 341F (5'-CCTACGGGNGGCWGCAG-3') (SEQ ID NO: 1) and 785R (5'-GACTACHVGGGTATCTAATCC-3' (SEQ ID NO: 2)) and sequenced using the Illumina MiSeq sequencing platform to generate paired-end sequence reads. Subsequently, reads containing PhiX control signals were removed by aligning the sequence reads to the Phix genome (NC_001422.1) using Bowtie2 (2.2.6) (Langmead & Salzberg (2012) Nat Methods Nature Publishing Group 9:357-9) and excluded from further analysis. Additionally, reads containing (partial) adapters were trimmed (up to a minimum read length of 50 bp). A second quality assessment was based on the remaining reads using the FASTQC quality control tool version 0.11.5. The paired-end sequence reads were collapsed into so-called pseudo-reads using sequence dereplication in USEARCH version 9.2 (Edgar (2010) Bioinformatics 26:2460-2461). Classification of these pseudo-reads was against the RDP database for bacterial organisms (Cole et al. (2014) Nucleic Acids Res 42:D633-642) using SNAP version 1.0.23 *It is carried out based on the alignment results in (Zaharia et al. (2022) http: / / arxiv.org / abs / 1111.5572). Alpha diversity indices (observed species, Chao1, Shannon, and Simpson diversity indices) and beta diversity indices (Jaccard and Bray-Curtis) were calculated by running mothur version 1.35.1 (https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC2786419 / ). 【0038】 Stool hardness and gastrointestinal symptoms: Stool hardness was evaluated using the Bristol Stool Chart: Type 1 (hard, lumpy pieces like nuts, separated), Type 2 (sausage-shaped but lumpy), Type 3 (like a sausage but with cracks on its surface), Type 4 (smooth and soft like a sausage or snake), Type 5 (soft, small round pieces with a clear edge), Type 6 (irregular, mushy pieces, diarrhea), Type 7 (watery, no hard pieces, completely liquid) (Lewis & Heaton, supra). Gastrointestinal symptoms were evaluated on a 6-point scale using the GSRS, which uses a 7-point rating scale based on the intensity and frequency of GI symptoms experienced during the previous week. Higher scores indicate more adverse symptoms. 【0039】 3.1.5 Investigational Product 150 mg of the WSTE described in 1.2 was combined with 450 mg of a tapioca starch bulking agent and encapsulated using size 00 Vegecaps (LGA, La Seyne-sur-Mer, France). 600 mg of maltodextrin (encapsulated in the same way) was used as a placebo control (Essential Nutrition Ltd., Brough, UK). Participants were instructed to consume 2 capsules orally with a cup of water at morning breakfast during the 28-day intervention period of each intervention phase. 【0040】 3.1.6 Sample Size Calculation and Statistical Analysis The sample size was determined based on findings from previous studies on nutritional interventions to reduce TMAO in plasma. For 80% power, a 5% significance level, and an expected effect size of 0.5 (mean / SD) for WSTE relative to placebo, it was calculated that 34 participants would be required. To account for potential losses up to follow-up, 40 participants were enrolled. To confirm the randomization method, the effectiveness of the washout was evaluated using a paired-samples t-test comparing within-group changes from baseline in Period 1 (Visit 2) to baseline in Period 2 (Visit 4). There were no statistically significant changes from Period 1 baseline to Period 2 baseline for any primary or secondary objectives, either within the WSTE group or within the placebo group. As a result, the WSTE group is a combination of Group 2 - Period 1 data (Visits 2 - 3) and Group 1 - Period 2 data (Visits 4 - 5), while the placebo group is a combination of Group 1 - Period 1 data (Visits 2 - 3) and Group 2 - Period 2 data (Visits 4 - 5). All data were analyzed using SPSS IBM V26.0 and R software. Unpaired t-tests (or Mann Whitney U tests) were used to compare between-group differences at baseline for each period. For the efficacy analysis, paired t-tests (or Wilcoxon signed-rank tests) were used to determine (1) whether there were statistically significant within-group changes from baseline to the end of the intervention in either the Fruitflow group or the placebo group; or (2) whether there were statistically significant between-group differences (i.e., differences in results between the WSTE group and the placebo group). All data are reported as mean ± SE. All tests were two-sided, and differences were considered statistically significant at p ≤ 0.05. 【0041】 For non-target metabolomics, all spectra were collected and processed using ACD Labs 2012. All spectral regions containing NMR features were grouped by intelligent bucketing (bucket width = 0.02 ppm, bandwidth = 50%). The resulting bins were divided into two groups, namely, subjects who had received WSTE before blood sampling (Group 1, Clinic visits 5; Group 2, Clinic visits 3) and those who had not (Group 1, Clinic visits 2, 3, and 4; Group 2, Clinic visits 2, 4, and 5), and then analyzed using MetaboAnalyst v4.0 (Chong et al. (2018) Nucleic Acids Res 46:W486-494). All data were normalized (quantile normalization) and autoscaled (Pareto scaling) before univariate analysis (t-test) and multivariate analysis (PLS-DA). The Q2 and R2 values for partial least squares discriminant analysis (PLS-DA), as well as the variable importance in projection (VIP) plot, were determined by MetaboAnalyst according to the established workflow (Macias et al. (2019) Metabolites 9:E201). 【0042】 For microbial alpha diversity, the Wilcoxon test with continuity correction was used to determine the statistical significance between and within groups. For beta diversity, Permutational Multivariate Analysis of Variance (PERMANOVA) with 9999 permutations was used to quantify the differences within and between groups (WSTE vs placebo). This test evaluates whether the group or time point significantly affects the overall gut microbiota composition and structure. Intra- and inter-group changes in the relative abundances of microbial taxa (i.e., differences in results between the WSTE group and the placebo group) were analyzed using the Wilcoxon test with continuity correction, followed by multiple testing correction with Benjamini-Hochberg (BH). Additionally, for inter-group changes, to ensure that the data were scale-invariant and compositionally coherent at lower levels, raw abundance counts were normalized using Centroid Log Ratio (CLR). The effect size differences for each taxon were calculated using the Cohen's d function (https: / / easystats.github.io / effectsize / ), an R-based package. 【0043】 3.2 Results 3.2.1 TMAO in Plasma and Urine Fasting plasma and urinary TMAO: In the WSTE group, there were significant intra-group changes in plasma (p = 0.05) and urinary (p = 0.009) TMAO from baseline to the end of the intervention, but not in the placebo group (NS). The inter-group change (i.e., the difference in results between the WSTE group and the placebo group) was significant only for urinary TMAO, but not for plasma TMAO (p = 0.05, Figures 2A and B). 【0044】 3.2.2 Gut Microbiota in Feces Alpha and beta diversity: In both the placebo and WSTE groups, no significant within-group or between-group changes in species diversity and richness were observed using the observed species, Chao1, Shannon, and Simpson diversity indices (data not shown). To gain insights into the temporal dynamics of the microbial community, fecal samples were also subjected to multivariate analysis using the Bray-Curtis and Jaccard distance methods. However, no significant shifts in beta diversity were observed in global or pairwise PERMANOVA analyses within and between groups. In contrast, a substantial shift in Jaccard principal component (PC1) was observed when comparing the end points of the intervention between the WSTE and placebo groups (p = 0.019, Figure 3). 【0045】 Microbial composition: Changes in the relative abundances of species-level OTUs within groups from baseline to the end of the intervention were performed using ALDEx2. In the WSTE group, four species-level OTUs related to Ruminococcus faecis_OTU#1473 (p = 0.0005), Bacteroides uniformis_OTU#406 (p = 0.018), Bacteroides ovatus_OTU#392 (p = 0.025), and Hungatella hathewayi_OTU#1083 (p = 0.036) had lower relative abundances at the end of the intervention compared to baseline (Figure 4A). In contrast, in the placebo group, three species-level OTUs related to Coprococcus catus_OTU#1222 (p = 0.023), Anaeromassilibacillus senegalensis_OTU#1422 (p = 0.028), and Barnesiella intestinihominis_OTU#428 (p = 0.043) were significantly depleted at the end of the intervention compared to baseline (Figure 4B). 【0046】 The inventors calculated the change between groups (i.e., the difference in CLR abundances for various levels of OTUs between the WSTE group and the placebo group) using the Wilcoxon test. They found that the relative abundances of 17 OTUs were significantly different between the groups, and among these 17, 9 OTUs related to Ruminococcus albus_OTU#1468 (p = 0.002), Alistipes ifymi_OTU#528 (p = 0.002), Anaeromassilibacillus senegalensis_OTU#1422 (p = 0.017), Saccharofermentans acetigenes_OTU#1479 (p = 0.017), Olsenella sinensis_OTU#1283 (p = 0.027), Clostridium carnis_OTU#994 (p = 0.035), Lachnobacterium lactatifolmans_OTU#1478 (p = 0.039), Alistipes obesi_OTU#532 (p = 0.046), and Faecalibacterium prausnitzii_OTU#1564 (p = 0.046) were significantly increased in the WSTE group compared to the placebo. Additionally, 4 OTUs related to Parabacteroides goldsteinii_OTU#448 (p = 0.025), Bacteroides acidifaciens_OTU#364 (p = 0.027), Veillonella parvula_OTU#1638 (p = 0.030), and Ruminococcus faecis_OTU#1473 (p = 0.039) had significantly lower abundances in the WSTE group compared to the placebo (Figure 4C). 【0047】 At the genus level, three taxonomic groups closely related to Streptococcus (p = 0.005), Luminococcus (p = 0.021), and Hungateella (p = 0.039) had significantly lower abundances at the end of the intervention compared to the baseline in the WSTE group. In contrast, in the placebo group, Anaeromassilibacillus (p = 0.028), Allistipes (p = 0.038), and Eubacterium (p = 0.038) were significantly depleted at the end of the intervention compared to the baseline (data not shown). Furthermore, comparison of the changes between groups revealed a significant decrease in the relative abundances of Prevotella (p = 0.007) and Veillonella (p = 0.023). At the same time, five taxonomic groups closely related to Saccharofermentans (p = 0.01), Alistipes (p = 0.023), Anaeromassilibacillus (p = 0.033), Lutibacterium (p = 0.039), and Olivibacterium (p = 0.042) were significantly increased in the FRUITFLOW group compared to the changes in the placebo group (Figure 4D). The inventors did not observe any significance in the intra-group or inter-group changes at the phylum level (data not shown). 【0048】 3.2.3 Metabolites in Plasma Plasma non-targeted metabolomics: Principal component analysis (PCA) of NMR data shows a clear distinction between plasma samples collected after the WSTE intervention and control samples (Figure 5A). Using the variable importance in projection (VIP), the top 15 ranking features contributing to this distinction included TMAO, formic acid, valine, glucose, and lactate, with TMAO being the most discriminative metabolite (low in WSTE samples and high in control samples, Figure 5B). Plasma LPS: The plasma LPS concentration was significantly lower at the end of the intervention than at the baseline in the Fruiflow group (p = 0.05), but not in the placebo group. However, there was no significant inter-group change (Figure 6). 【0049】 Some bile acids, including cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), glycocholic acid (GCA), glycochenodeoxycholic acid (GCDCA), glycodeoxycholic acid (GDCA), glycoursodeoxycholic acid (GUDCA), taurochenodeoxycholic acid (TCDCA), taurodeoxycholic acid (TDCA), and ursodeoxycholic acid (UDCA), were detected in plasma in both groups. TCDCA and TDCA increased slightly (P≤0.05) from baseline to the end of the intervention in both groups, WSTE and placebo, while CDCA, GCA, GCDCA, and GDCA increased only in the WSTE group (P≤0.05). However, there were no significant between-group changes (data not shown). SCFAs and other organic acids in plasma: Acetate, lactate, pyruvate, and 3-hydroxybutyrate were detected in plasma. A slight but significant increase within the group from baseline to the end of the intervention was observed for plasma pyruvate in the WSTE group. In contrast, plasma acetate increased slightly in both groups (P≤0.05 each). There were no significant between-group changes (data not shown). 【0050】 3.3.4 Metabolites in Feces Bile acids in feces: Some bile acids, including CA, CDCA, DCA, GCA, GCDCA, GDCA, glycolithocholic acid (GLCA), GUDCA, lithocholic acid (LCA), taurochenodeoxycholic acid (TCDC), and TDCA, were detected in feces in both groups. The inventors observed one significant within-group change; CA in feces increased from baseline to the end of the intervention in the WSTE group but not in the placebo group (P≤0.05). However, there were no significant between-group changes (data not shown). SCFAs and other organic acids in feces: Acetate, propionate, butyrate, valerate, isovalerate, 2-methylbutyrate, isobutyrate, and pyruvate were all detected in feces. However, there was no effect of Fruitflow on SCFAs in feces; only valerate increased from baseline to the end of the intervention in the placebo group, and this difference was also significant between groups (P ≤ 0.05 each, data not shown). 3.3.5 Hardness of feces and gastrointestinal symptoms There were no significant within-group or between-group changes in the hardness of feces or gastrointestinal symptoms. Data not shown. 3.3.6 Adverse events and blood safety profiling During the study period, no serious adverse events (SAEs) or discontinuations due to adverse events (AEs) were observed. All AEs were of mild or moderate intensity, and none were judged by the assigned study clinicians to be related to the investigational product. Standard hematological and biochemical evaluations, including glucose, hsCRP, and bilirubin, showed no significant within-group or between-group effects. Data not shown. 【0051】 3.4 Discussion The inventors found that WSTE significantly decreased fasting plasma (-1.51 μM) and urinary (-19.09 μM) TMAO compared to baseline, and similarly for plasma LPS (-5.33 ng / mL), a marker of intestinal permeability and low-grade inflammation. The effect of WSTE on plasma TMAO was confirmed by non-targeted metabolomics analysis. A clear distinction was observed between plasma samples collected after WSTE intervention and control samples, and TMAO was the top-ranking feature contributing to this distinction. When analyzing the gut microbiota in feces, the inventors found changes in the beta diversity of the microbiota, but not in the alpha diversity, with significant differences in the principal components based on Jaccard distance when comparing WSTE to placebo. Additionally, there were several significant changes in the microbial composition with respect to WSTE, such as a decrease in OTUs related to the genera Bacteroides, Lachnococcus, and Hungateella (known for their involvement in TMA / TMAO metabolism), and an increase in OTUs related to the genus Allistipes and the class Clostridia (the latter being well-known SCFA-producing strains). Finally, the inventors found an increase in the number of bile acids in feces and plasma and pyruvate in plasma, but no change in SCFA in feces. No effect of WSTE on gastrointestinal comfort was observed. 【0052】 3.4.1 Effect of WSTE on reducing TMAO levels TMAO has been established as a non-dependent risk factor for promoting atherosclerosis by stimulating foam cell formation, deregulating enterohepatic cholesterol and bile acid metabolism, and impairing cholesterol reverse transport by macrophages (Wang et al. (2011) Nature 472: p57-63; Koeth et al. (2013) Nat Med 19 p576-585; Canyelles et al. (2018) Int J Mol Sci 19:E3228; and Tang et al. (2013) N Engl J Med 368 p1575-1584). Assuming that the production of TMAO from dietary phosphatidylcholine is dependent on metabolism by the gut microbiota (Wang et al., Koeth et al., and Tang et al., supra), gut microbiota-based therapies have been suggested as a novel strategy for preventing and treating cardiovascular disease (CVD). Oral broad-spectrum antibiotics can suppress TMAO-induced atherosclerosis by inhibiting the gut microbiota (Koeth et al., supra); however, the potential for side effects and resistance limits their usefulness. Thus, the focus is narrowed to natural products that selectively modulate the gut microbiota, inhibit microbial TMA production, and as a result, have the characteristic of reducing the risk of CVD. This profile suggests that WSTE is within the definition of prebiotics and is a selectively utilized substrate for host microorganisms that confer health benefits. 【0053】 Although the inventors do not intend to be bound by any hypothesis, they believe that the prebiotic benefits of WSTE may be due, at least in part, to the polyphenols contained within its extracts. Several polyphenol-rich natural products have recently been shown to reduce plasma TMAO levels in animals and humans. However, there are also conflicting reports that some polyphenol-rich natural products increase plasma TMAO levels. For example, Chen et al. (supra) initially demonstrated in mice that resveratrol attenuates TMAO-induced atherosclerotic lesions by controlling TMAO synthesis through reconfiguration of the gut microbiota. This was confirmed in other studies in experimental rodents and in a pilot study in 20 normal-weight subjects (Annunziata et al. (2019) Digital Publishing Institute 11: 139-139) that showed a significant decrease in serum TMAO (from 1.87 to 0.66 μM) after 4 weeks of supplementation with 300 mg of polyphenol-rich grape pomace. In contrast, a recent study in overweight and obese subjects revealed that consumption of 280 mg / day of raspberries increased plasma TMAO levels (Franck et al. (2022) Nutrients 14:1656). These prior art reports obscure whether those skilled in the art would expect a polyphenol-rich extract such as WSTE to increase or decrease TMAO levels. Franck et al. (supra) suggested that a discriminatory gut microbial signature with a higher relative abundance of Actinobacteriota in participants whose TMAO levels increased after intervention could explain such an effect; however, this required further study. The inventors did not observe an increase in the level of Actinobacteria in this study. Nevertheless, the inventors believe that the data presented herein correlates the decrease in plasma TMAO levels caused by WSTE with a health benefit. 【0054】 The mean decrease in plasma TMAO of 1.51 μM for this study was similar to that observed (mean decrease of 1.2 μM) in the grape extract study by Annunziata and co-workers (above). Furthermore, considering the findings of a recent cross-sectional study including 377 patients with acute ischemic stroke and 50 healthy controls, the observed effect suggests clinical relevance. In this study, plasma TMAO levels were higher in patients with ischemic stroke than in healthy controls (median 5.1 μmol / L vs. 3.0 μmol / L; p < 0.001), and after adjustment for vascular risk factors, for every 1 μmol / L increase in TMAO, the NIH Stroke Scale increased by 1.13 points and infarct volume increased by 1.69 mL. 【0055】 The inventors have noticed that increased plasma TMAO levels are associated with having an adverse effect on the conditions contemplated in the Summary of the Invention section. Based on the data presented herein (illustrating that WSTE decreases plasma TMAO levels), the inventors have come to recognize that WSTE, acting as a prebiotic, can be used to treat, prevent, or alleviate the symptoms of such conditions. By way of example, WSTE is effective in treating inflammaging and age-associated cognitive impairment. Studies in mice and rats suggest that aging-induced intestinal dysbiosis gives rise to higher TMAO, contributing to increased peripheral and central inflammatory tone, resulting in vascular inflammation, oxidative stress, and ultimately age-related endothelial dysfunction and cognitive impairment (Li et al. (2018) Aging Cell 17:e12768). Furthermore, in humans, cerebrospinal fluid TMAO was higher in individuals with mild cognitive impairment and Alzheimer's disease dementia compared to individuals without cognitive impairment (Vogt et al. (2018) Alzheimers Res Ther. 10(1):124). Taken together, this has led the inventors to recognize that if WSTE is hypothesized to decrease plasma TMAO by modulating the gut microbiota, it also represents a novel therapeutic option for treating inflammaging and neurodegenerative diseases. Additionally, WSTE can be used to generally promote brain health in subjects of all ages, particularly the elderly. 【0056】 3.4.2 Effect of WSTE on the gut microbiota The inventors were surprised to find that WSTE also changes the gut microbiota. These changes were not reflected in the microbial alpha diversity index. However, surprising changes were seen in beta diversity, with a significant shift in Jaccard PC1, indicating that WSTE consumption substantially affects the gut microbial composition. Franck et al. (supra), who examined the effect of raspberry consumption on TMAO (contrary to the present invention, they found an overall increase in plasma TMAO), also did not find significant changes in the alpha diversity index; however, they did not report beta diversity. The inventors' observations regarding changes in beta diversity can support the concept that changes in plasma and urinary TMAO are dependent on changes in the gut microbiota induced by WSTE administration. WSTE appears to achieve this by efficiently decreasing the proportion of bacteria that produce TMA and / or increasing the proportion of microorganisms that metabolize TMA. The inventors observed a significant decrease in the relative abundance of Bacteroides uniformis and Bacteroides ovatus within the WSTE group, as well as Bacteroides acidifaciens and the related species Parabacteroides goldsteinii when comparing WSTE to placebo. Bacteroides species have been suggested to reduce TMAO to TMA, and thus the inventors propose that the decrease in the genus Bacteroides by WSTE is one mechanism explaining the observed effect on TMAO. The inventors also observed a decrease in Hungatella hathewayi and Lachnococcus faecis with WSTE. Hungatella hathewayi is known to be a TMA-producing strain, while the genus Lachnococcus has been positively correlated with plasma TMAO in studies in rats. Finally, the inventors found an increase in the genus Alistipes, another member of the phylum Bacteroidetes, with WSTE. 【0057】 3.4.3 Effect of WSTE on bile acids The present inventors have noted that TMAO can promote atherosclerosis through several mechanisms including inhibition of hepatic bile acid synthesis (Wang et al. (2011) Nature 472 p57-63). In a study in mice by Chen et al. (supra), resveratrol attenuated TMAO-induced atherosclerosis by inhibiting microbial TMA production and increasing hepatic bile acid synthesis. This was thought to be because resveratrol increased the relative abundances of the genera Lactobacillus and Bifidobacterium, resulting in an increase in bile salt hydrolase activity, and bile acid deconjugation, as well as an increase in fecal excretion of bile. The resulting decrease in ileal bile acid content and suppression of the intestinal-liver farnesoid X-activated receptor (FXR)-fibroblast growth factor 15 (FGF15) axis increased hepatic bile acid synthesis. The present inventors found that plasma CDCA, GCA, GCDCA, and GDCA were significantly increased in the WSTE group but not in the placebo group. This also suggests an increase in hepatic bile acid synthesis. However, the present inventors did not observe changes in the genera Lactobacillus or Bifidobacterium, nor a decrease in fecal bile acid content. 【0058】 3.4.4 Effect of WSTE on LPS WSTE also reduced plasma LPS, an enteric microbiota-derived factor involved in the onset and progression of chronic inflammation-related diseases such as obesity, T2DM, or non-alcoholic fatty liver disease (NAFLD). Cani et al. ((2012) Gut Microbes 3 p279-288) showed that this effect was directly dependent on the enteric microbiota because a high-fat diet contributed to the disruption of tight junction proteins and antibiotic treatment abrogated diet-induced intestinal permeability. Furthermore, they found that a high-fat diet increased plasma LPS to concentrations sufficient to increase metabolic endotoxemia. Specific modulation of the enteric microbiota composition with prebiotics improved gut barrier integrity, reduced metabolic endotoxemia, and decreased inflammation and glucose intolerance. Interestingly, in patients with T2DM and progressive chronic kidney disease, TMAO showed a positive correlation with zonulin, an intestinal permeability marker, and LPS, as well as inflammatory biomarkers (IL-6, TNFα, and CRP) and an endothelial dysfunction biomarker (ET-1) (Al-Obadie et al. (2017) J Clin Med6:E86), suggesting that upregulation of zonulin results in uncontrolled influx of microbial endotoxins such as TMA and LPS from the gut into the bloodstream, leading to low-grade inflammation and endothelial dysfunction. Therefore, the observed decrease in plasma LPS after WSTE consumption suggests an improvement in gut barrier function, and this data exemplifies the utility of WSTE for treating or preventing chronic inflammation-related diseases such as IBS, IBD, obesity, T2DM, or non-alcoholic fatty liver disease (NAFLD). 【0059】 3.4.5 Summary The inventors found that supplementation with WSTE (a polyphenol-rich aqueous tomato extract containing secondary metabolites) for 4 weeks resulted in the following: (A) reduced fasting plasma and urinary TMAO compared to baseline; (B) reduced plasma LPS, a marker of intestinal permeability and low-grade inflammation; (C) increased plasma levels of bile salts; and (D) These changes were paralleled by several changes in the microbial composition, including an increase in PC1 based on Jaccard distance but no increase in alpha diversity, changes in the fecal microbiota, and a decrease in OTUs closely related to the genera Bacteroides, Lachnococcus, and Hungatella, which are known to be involved in TMA / TMAO metabolism, and an increase in OTUs closely related to the genera Alistipes and Clostridium. 【0060】 These data strongly support the use of WSTE for modulating the gut microbiota to confer health benefits to the host. The cardiovascular health benefits of WSTE are evident from previous publications made by the applicant (see WO99 / 55350, WO2010 / 049707, or WO2018 / 083137). The data presented herein may provide new insights into the mechanisms by which WSTE may contribute to its published effects on cardiovascular health. However, this data also exemplifies that WSTE is unexpectedly useful as a prebiotic. As such, it can be used to promote gut health. From this, the inventors have come to recognize that: (a) WSTE can be advantageously formulated for delivery to the large intestine and may be even more useful when co-administered with other prebiotics or probiotics; and (b) the knowledge that WSTE modulates the gut microbiota, TMAO levels, and LPS levels has revealed new and unexpected uses for WSTE. These include treating chronic inflammation-related diseases, type II diabetes, obesity, and cognitive dysfunction.

Claims

[Claim 1] A composition for use in providing health benefits by regulating the intestinal microbiota in a subject, comprising a water-soluble tomato extract having activity for inhibiting platelet aggregation, wherein the extract is (a) substantially thermally stable; (b) Colorless or pale yellow; (c) Water-soluble; (d) consisting of components having a molecular weight of less than 1000; and (e)(i) Glycosylated phenolic acid or phenol ester or derivative thereof; (ii) Glycosylated flavonoids; and (iii) Nucleoside A compound comprising one or more compounds selected from which platelet aggregation is inhibited, The aforementioned composition. [Claim 2] The composition according to claim 1, wherein the water-soluble tomato extract acts as a prebiotic. [Claim 3] The composition according to claim 1, wherein the water-soluble tomato extract comprises glycosylated phenolic acid or phenol ester or a derivative thereof, glycosylated flavonoid, and nucleoside, respectively. [Claim 4] (a) a water-soluble tomato extract as defined in claim 1; and (b) at least one additive that facilitates the delivery of the water-soluble tomato extract to the large intestine. A composition formulated for delivery to the target colon of a water-soluble tomato extract having activity to inhibit platelet aggregation, comprising: [Claim 5] (a) a water-soluble tomato extract as defined in claim 1; and (b) Prebiotics A composition containing the following: [Claim 6] (a) a water-soluble tomato extract as defined in claim 1; and (b) Probiotics A composition containing the following: [Claim 7] The composition according to any one of claims 1 to 6, in the form of a pharmaceutical, nutritional supplement, health supplement, beverage, food, or food supplement. [Claim 8] The composition according to claim 7, formulated as a gel, powder, food bar, beverage, tablet, or contained within a capsule. [Claim 9] A composition according to any one of claims 1 to 6, for providing a health benefit selected from improving digestion, improving resistance to infection in the intestines, reducing inflammation in the intestines, treating or minimizing inflammatory bowel disease, or treating or minimizing irritable bowel syndrome. [Claim 10] A composition for providing health benefits to subjects having a condition in which the pathological state is caused or aggravated by compounds released into the bloodstream by the intestinal microbiota, wherein the water-soluble tomato extract regulates plasma levels of such compounds by regulating the intestinal microbiota, according to any one of claims 1 to 6. [Claim 11] The composition according to claim 10 for providing health benefits to subjects suffering from anxiety, depression, type II diabetes, non-alcoholic fatty liver disease, obesity, or cognitive impairment. [Claim 12] The composition according to any one of claims 4 to 6 for use as a pharmaceutical.

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