Combining Microbiome Materials to Reduce Uremic Toxins in Cardiometabolic or Neurodegenerative Conditions

Abstract

The present invention relates to compositions for use in reducing or avoiding the accumulation of uremic toxins in cardiometabolic or neurodegenerative conditions, in particular compositions comprising probiotic bacteria, prebiotics (carbohydrates or fiber), and lipids.

Description

[Technical Field]

[0001]

[0000] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 439,638, filed January 18, 2023, and U.S. Provisional Patent Application No. 63 / 480,729, filed January 20, 2023, the entire disclosures of which, including sequence listings, are incorporated by reference in their entireties. [Background technology]

[0002]

[0001] The present disclosure relates generally to compositions and methods that enable the reduction or avoidance of uremic toxin accumulation, preferably in cardiometabolic or neurodegenerative conditions, more preferably in chronic kidney disease (CKD), comprising a specifically selected prebiotic, a specifically selected probiotic, and a specifically selected lipid, as defined herein.

[0003]

[0002] Uremic toxins can typically be classified into three categories: free water-soluble low molecular weight solutes, protein-bound solutes, and medium molecules, which are usually products of protein / amino acid metabolism by body tissues, particularly the liver. Uremic toxins include, among others, urea, indoxyl sulfate, TMAO, p-cresol sulfate, 3-carboxy-4-methyl-5-propyl-2-furanpropionic acid, p-cresyl glucuronide, and uric acid. The accumulation of such systemic uremic toxins, also known as uremia or the uremic state, is a condition frequently observed in individuals with cardiometabolic and neurological conditions, including, but not limited to, chronic kidney disease (CKD).

[0004]

[0003] For example, chronic kidney disease is a disease characterized by a progressive and gradual decline in kidney function, which can lead to the accumulation of uremic toxins that are normally removed by the kidneys, resulting in uremia. CKD patients are at high risk of developing cardiovascular disease (CVD) due to multiple risk factors specific to CKD. The accumulation of uremic toxins in the circulation and tissues is associated with the progression of CKD and its comorbidities, including CVD (see Yong Jin Lim et al., Toxins 2021, 13, 142).

[0005]

[0004] Elevated levels of uremic toxins are not only manifested in chronic kidney disease (CKD) and CVD, but are also associated with other cardiometabolic and neurological conditions. Furthermore, early indicators of excessive uremic toxin load are often widespread, and at first glance, such cardiometabolic and neurological conditions may not always be clearly identified as the primary cause. In general, accumulation of uremic toxins can lead to the development of syndromes and symptoms, including, but not limited to, fatigue, loss of appetite, and protein energy wasting (PEW), which ultimately affect the quality of life of individuals with high levels of circulating uremic toxins. In this context, several publications have demonstrated that uremic toxins, including indoxyl sulfate, p-cresyl sulfate, TMAO, and urea, act to induce tissue damage and dysfunction in the kidney or other organs, leading to disease progression, organ failure, and death [References: Vanholder R., Schepers E, Pletinck A, Nagler EV, Glorieux G. 2014. J Am Soc Nephrol. 25(9):1897-907. doi:10.1681 / ASN.2013101062; Falconi et al. 2021. Front Physiol. 12:686249].

[0006]

[0005] Therefore, inadequate management of systemic uremic toxins can lead to disease progression, the appearance of associated symptoms and syndromes (e.g., uremic syndrome, anorexia), and a decline in quality of life, and even death of the patient. Therefore, early intervention is very important and essential to effectively treat such diseases and positively influence their outcomes.

[0007] As generally discussed above, two main factors typically contribute to abnormal levels of uremic toxins in the circulation. The first factor is increased accumulation, typically due to poor renal filtration. The second factor is altered production due to several changes in metabolism, including the liver, gut, and microbiome.

[0008]

[0007] There are currently several proposals in the prior art for managing uremic toxins and the causes that lead to abnormal levels of uremic toxins.

[0009]

[0008] Because some uremic toxins can also result from a high protein content in the diet, one currently effective solution for managing uremic toxins is to administer a low-protein diet to patients. However, because patients usually require a balanced diet and good nutrition, a low-protein diet is often not sustainable. For example, protein is needed to manage other complications associated with kidney disease (e.g., PEW). In many cases, a low-protein diet is even harmful, for example, in cases of anorexia and muscular dystrophy, and in elderly people in general. All of these require a minimum or even higher amount of protein to prevent or treat excessive muscle protein breakdown (muscle catabolism).

[0010]

[0009] Another method for addressing uremic toxins is their removal by dialysis, which can be considered the gold standard in the management of kidney disease. However, dialysis requires continuous treatment and is often performed in specialized nursing facilities or hospitals, making it burdensome and expensive. Furthermore, in later stages of disease, such as end-stage renal disease, dialysis is unable to remove uremic toxins. In particular, dialysis is inefficient at removing protein-bound uremic toxins, such as p-cresyl sulfate, indoxyl sulfate, 3-carboxy-4-methyl-5-propyl-2-furanpropionic acid, and p-cresyl glucuronide.

[0011]

[0010] There are also methods that can reduce the amount of gut microbiota-derived metabolites used as substrates for tissue metabolism of uremic toxins, such as (1) AST-120, a charcoal-based solution aimed at removing microbiota-derived precursors at the gut level; (2) the use of probiotics / live therapeutics to correct dysbiosis and degrade uremic toxin precursors; and / or (3) the use of fiber / glycans / oligosaccharides to correct dysbiosis.

[0012]

[0011] However, such solutions have the drawback that they only partially address the problem of uremic toxin production and do not eliminate the primary cause, which may still result in or not prevent the accumulation of uremic toxins in the body and organs. There have also been reports that the accumulation of uremic toxins in the systemic circulation may be at least partly independent of the production of uremic toxin precursors by the intestinal microflora.

[0013]

[0012] In the prior art, drugs such as SGLT2 inhibitors have also been developed that are intended to help improve glomerular hemodynamic function. Such SGLT2 inhibitors are thought to improve other local and systemic mechanisms involved in the pathogenesis of CKD. However, these drugs have not been sufficiently shown to improve the levels of uremic toxins in the circulation and to provide a reliable basis for the treatment of conditions caused by such uremic toxins, for example, in individuals with cardiometabolic and neurological conditions.

[0014]

[0013] Therefore, there is a need in the art to provide alternative, preferably improved, therapeutic concepts to more effectively address such problems and to avoid and / or treat the accumulation of uremic toxins in patients, preferably individuals with cardiometabolic and neurological conditions, particularly chronic kidney disease.

[0015] The invention described herein addresses such needs and provides compositions, uses and corresponding methods of treatment to prevent and / or treat the accumulation of uremic toxins in such patients. The invention also provides a multi-part kit for such purposes. Summary of the Invention

[0016]

[0015] The above-mentioned problems are solved by the subject matter of the independent claims and are described in more detail in the following specification, its embodiments and aspects, and the dependent claims. The embodiments and aspects disclosed in this specification can be combined with each other as needed and unless expressly stated otherwise.

[0017]

[0016] The present invention is particularly directed to novel compositions that target multiple direct and indirect mechanisms that contribute to avoiding the production of uremic toxins and prevent their accumulation or at least reduce their levels.

[0018]

[0017] A first embodiment is a composition suitable for use in reducing and / or avoiding the accumulation of uremic toxins, preferably in cardiometabolic or neurodegenerative conditions. The composition comprises specifically selected probiotic bacteria, specifically selected prebiotics, and specifically selected lipids. More precisely, the composition comprises: (a) The following: probiotic bacteria lacking genes that produce at least one of urea, uric acid, p-cresol, p-cresyl sulfate, indoxyl sulfate, indole, indoleacetic acid, indoxyl, trimethylamine, TMAO, dimethylglycine, betaine, and / or glutarate; and / or probiotic bacteria deficient in at least one bacterial enzyme selected from urease, carnitine monooxygenase and / or reductase, tryptophanase, and / or hydroxyphenylacetate; and / or probiotic bacteria expressing at least one bacterial enzyme selected from α-galactosidase, β-galactosidase, glucan 1,4-β-glucosidase, cellulase, β-fructofuranosidase and / or licheninase; and (b) a prebiotic selected from carbohydrates, wherein the carbohydrates can be hydrolyzed by at least one bacterial enzyme selected from α-galactosidase, β-galactosidase, glucan 1,4-β-glucosidase, cellulase, β-fructofuranosidase, and / or licheninase; and (c) lipids selected from triglycerides, short-chain fatty acids, and / or medium-chain fatty acids, wherein the triglycerides are butyrates, short-chain fatty acids, medium-chain fatty acids, long-chain fatty acids, or any of these; lipids, including mixtures; and

[0019]

[0018] According to a first aspect, the composition defined herein comprises probiotic bacteria. According to a first criterion, such probiotic bacteria may be selected from probiotic bacteria lacking genes for producing at least one of urea, uric acid, p-cresol, p-cresyl sulfate, indoxyl sulfate, indole, indoleacetic acid, indoxyl, trimethylamine, TMAO, dimethylglycine, betaine, and / or glutarate. According to a second criterion, such probiotic bacteria may also be selected from probiotic bacteria lacking at least one bacterial enzyme selected from urease, carnitine monooxygenase and reductase, tryptophanase, and / or hydroxyphenylacetate. According to the third criterion, such probiotic bacteria may be selected from probiotic bacteria that express at least one bacterial enzyme selected from α-galactosidase, β-galactosidase, glucan 1,4-β-glucosidase, cellulase, β-fructofuranosidase and / or licheninase. Any of these three criteria may be applied alone or in combination with the other two criteria. Preferably, all three criteria are met.

[0020] Preferably, the probiotic bacteria meeting all three criteria are selected from the following species or subspecies: Bifidobacterium animalis subspecies lactis, Bifidobacterium longum subspecies infantis, Bifidobacterium longum subspecies longum, Enterococcus faecium, Lactobacillus johnsonii, Lactococcus lactis, Lacticaseibacillus paracasei (formerly Lactobacillus paracasei), paracasei), Limosilactobacillus reuteri (formerly classified as Lactobacillus reuteri), Lacticaseibacillus rhamnosus (formerly classified as Lactobacillus rhamnosus), Staphylococcus carnosus and / or Streptococcus thermophiles, preferably Lactobacillus johnsonii. johnsonii), or a probiotic bacterium having a genome with at least 95% average nucleotide identity (ANI) to the genome sequence of any one of the heretofore defined probiotic bacteria, preferably 96% ANI, more preferably at least 97% ANI, even more preferably at least 98% ANI, and most preferably at least 99% ANI, or even 99.5% ANI, or 99.9% ANI.

[0021] Even more preferably, such probiotic bacteria, preferably meeting all three criteria, are the following strains: a. Bifidobacterium animalis subsp. lactis NCC 2818, which was deposited at the CNCM [Institut Pasteur National Collection of Microorganisms (Paris, France)] on June 7, 2005, and assigned the accession number CNCM I-3446); b. Bifidobacterium longum subsp. infantis NCC 341 (ATCC 15697(T)), available at https: / / www.atcc.org / products / 15697); c. Bifidobacterium longum subsp. longum NCC 2705 (CNCM I-2618) (NCBI refseq; GCA_000007525.1) [NCC 2075 was deposited at the CNCM on January 29, 2001, and assigned accession number CNCM I-2618]; d. Enterococcus faecium NCC 2768 (NCIMB 10415 - available from Cerbios-Pharma SA Barbengo Switzerland (cerbios.swiss / e-faecium-sf68-a-model-for-efficacy-safety-for-pharmaceutical-probiotics / ); e. Lactobacillus johnsonii NCC 533 [originally known as La 1, deposited at the CNCM on June 30, 1992, and assigned accession number CNCM I-1225 (see also NCBI refseq; GCA_000008065.1, and GenBank AE017198.1 (included as SEQ ID NO: 1)]; f. Lactococcus lactis NCC 2287 (CNCM I-4154) [NCC 2287 was deposited with the CNCM on April 24, 2009, and assigned the accession number CNCM I-4154]; g. lacticasei Bacillus paracasei NCC 2461 (CNCM I-2116) [NCC 2461 was deposited with the CNCM on January 12, 1999 and assigned accession number CNCM I-2116]; h. lacticasei Bacillus rhamnosus NCC 4007 (CGMCC 1.3724) [NCC 4007 was deposited in October 2004 at the CGMCC [China General Microbiological Culture Collection Center (CGMCC)] Institute of Microbiology, Chinese Academy of Sciences, PO Box 2714, Beijing 100080, China] under the identification number CGMCC 1.3724; i. Staphylococcus carnosus NCC 1052 (CNCM I-5400) [NCC 1052 was deposited with the CNCM on February 1, 2019, and assigned the accession number CNCM I-5400]; j. Staphylococcus carnosus NCC 971 (CNCM I-5398) [NCC 971 was deposited with the CNCM on February 1, 2019, and assigned accession number CNCM I-5398]; and / or k. Streptococcus thermophilus NCC 2496 (CNCM I-3915) [NCC 2496 was deposited with the CNCM on February 5, 2008, and assigned the accession number CNCM I-3915]; or probiotic bacteria having a genome with at least 95% average nucleotide identity (ANI) to the respective genome sequences set forth in any one of a. to k. defined above, preferably at least 96% ANI, more preferably at least 97% ANI, even more preferably at least 98% ANI, and most preferably at least 99% ANI, or even 99.5% ANI or 99.9% ANI. For identification of CNCM, please refer to the Collection Nationale de Cultures de Microorganismes, Institut Pasteur, 22 rue du docteur Roux, 75724 Paris, France. For identification of CGMCC, please refer to the China General Microbiological Culture Collection Center (CGMCC), Institute of Microbiology, Chinese Academy of Sciences, PO Box 2714, Beijing 100080, China.

[0022] Even more preferably, the probiotic bacteria of the compositions defined herein are selected from Lactobacillus johnsonii NCC 533 (CNCM I-1225) or probiotics having a genome with an ANI of at least 95%, preferably at least 96%, more preferably at least 97%, even more preferably at least 98%, and most preferably at least 99%, or even 99.5%, or 99.9%, relative to the genome sequence of Lactobacillus johnsonii NCC 533 (CNCM I-1225). The NCBI reference sequence for Lactobacillus johnsonii (NCC 533 (CNCM I-1225)) is GCA_000008065.1 (SEQ ID NO: 1).

[0023] According to a second aspect, the composition defined herein also comprises a prebiotic. Such prebiotic is selected from carbohydrates that can be hydrolyzed by at least one bacterial enzyme selected from α-galactosidase, β-galactosidase, glucan 1,4-β-glucosidase, cellulase, β-fructofuranosidase and / or licheninase. Such carbohydrates are preferably prebiotics selected from α-galactooligosaccharides, raffinose, β-galactooligosaccharides, and cellooligosaccharides, or combinations thereof. Even more preferably, such prebiotics are selected from pea galactooligosaccharides (α-GOS), soy galactooligosaccharides (α-GOS), β-galactooligosaccharides (β-GOS), bovine milk oligosaccharides (BMOS) (β-GOS), Vivinal GOS (β-GOS), cellobiose, cellotriose, cellotetraose, human milk oligosaccharides (HMO), soluble hydrolyzed wheat, soluble hydrolyzed oats, and / or β-glucan hydrolysates containing cellobiose or cellotriose or cellotetraose, or combinations thereof. Preferred HMOs include 2'-fucosyllactose (2'FL), 3-fucosyllactose (3'FL), lacto-N-neotetraose (LNnT), lacto-N-tetraose (LNT), 3'-sialyllactose (3SL), and / or 6'-sialyllactose (6SL), or a combination thereof. Particularly preferred HMOs include LNT, 2'FL, and / or LNnT, or a combination thereof.

[0024] According to a third aspect, the composition defined herein also comprises a lipid (c). Such lipid (c) is selected from triglycerides, short-chain fatty acids and / or medium-chain fatty acids, wherein the triglycerides comprise short-chain fatty acids, medium-chain fatty acids, long-chain fatty acids, or mixtures thereof. Preferred triglycerides include butyrate-containing triglycerides, butyrate and caprylate-containing triglycerides, and butyrate and oleate-containing triglycerides. More preferably, such lipid (c) is selected from a. triglycerides (TG) composed of a mixture of butyrate and medium-chain fatty acids (MCFA); b. triglycerides (TG) composed of a mixture of short-chain and medium-chain fatty acids (SMCFA); c. triglycerides (TG) composed of a mixture of butyrate and long-chain fatty acids (e.g., a mixture of butyrate and oleate); d. triglycerides composed of medium-chain fatty acids; e. triglycerides (TG) composed of short-chain fatty acids; f. short-chain fatty acids that can be metabolized into ketone bodies; and / or g. short-chain and / or medium-chain fatty acids that can be metabolized into ketone bodies, preferably C4 and / or C8 fatty acids (e.g., butyric acid (C4:0) and / or caprylic acid (C8:0)). Any combination is possible, e.g., a., b., c., d., a. / b., a. / c., a. / d., a. / e., a. / f., a. / g., b. / c., b. / d., b. / e., b. / f., c. / d., c. / e., c. / f., a. / b. / c., a. / d. / c.; a. / e. / c., a. / f. / c., b. / c. / d., a. / b. / c. / d., etc. In one embodiment, the composition defined herein comprises a triglyceride composed of butyrate and caprylate. In one embodiment, the composition defined herein comprises a triglyceride composed of butyrate and oleate.

[0025] According to a further aspect, the probiotics are preferably present in the composition as defined herein in an effective amount, preferably 10 per day 3 cfu~10 12 cfu, typically 10 per daily dose 4 cfu~10 11cfu, preferably 10 per daily dose 5 cfu~10 10 cfu or 10 per daily dose 5 cfu~10 9 cfu, also preferably 10 per daily dose 6 cfu~10 9 cfu, 10 per daily dose 6 cfu~10 8 cfu or 10 per daily dose 8 cfu~10 10 cfu, more preferably about 10 per daily dose 7 cfu~10 9 The preferred daily dose is about 10 total cfu per daily dose. 8 cfu, 10 per daily dose 7 ~10 9 cfu, or 10 per daily dose 8 ~10 9 cfu.

[0026]

[0025] Furthermore, according to another aspect, the prebiotics are preferably contained in the composition defined herein in an amount of 0.1 g to 30 g per daily dose, preferably in an amount of 2 g to 15 g per daily dose.

[0027]

[0026] According to a further aspect, the lipid is preferably contained in the composition defined herein in an amount of 0.1 to 30 g per daily dose, preferably in an amount of 2 g to 15 g per daily dose.

[0028] According to a preferred embodiment, the composition defined herein may comprise a probiotic, a prebiotic and a lipid as defined above, (a) The probiotic bacterium is selected from the probiotic bacterium defined above in a. to k., or a probiotic bacterium having a genome with an ANI of at least 95%, preferably at least 96%, more preferably at least 97%, even more preferably at least 98%, and most preferably at least 99%, or even 99.5%, or 99.9%, relative to any one of the genome sequences of the probiotic bacterium defined above in a. to k., or a combination thereof, and more preferably is selected from Lactobacillus johnsonii NCC 533 (CNCM I-1225) or Lactobacillus johnsonii NCC 533 (CNCM I-1225). The probiotics are selected from at least one species of probiotic having a genome having at least 95% ANI relative to the genome sequence (SEQ ID NO: 1) of I-1225, preferably at least 96% ANI, more preferably at least 97% ANI, even more preferably at least 98% ANI, and most preferably at least 99% ANI, or even 99.5% ANI, or 99.9% ANI. (b) the prebiotic is selected from α-galactooligosaccharides, raffinose, β-galactooligosaccharides, and cellooligosaccharides, or a combination thereof, preferably selected from pea galactooligosaccharides (pea GOS, α-GOS), soybean galactooligosaccharides (soybean GOS, α-GOS), β-galactooligosaccharides (β-GOS), milk oligosaccharides (e.g., Vivinal GOS, β-GOS), cellobiose, cellotriose, cellotetraose, soluble hydrolyzed wheat, soluble hydrolyzed oats, human milk oligosaccharides (HMO), β-glucan hydrolysates containing cellobiose or cellotriose or cellotetraose, or a combination thereof; (c) The lipid is selected from a triglyceride, a short chain fatty acid, and / or a medium chain fatty acid, wherein the triglyceride comprises a short chain fatty acid, a medium chain fatty acid, or a mixture thereof, and further, in one embodiment, the triglyceride comprises butyrate and / or caprylate. Preferably, the lipid is a. Triglycerides (TG) composed of a mixture of butyrate and medium-chain fatty acids (MCFA); b. Triglycerides (TG) composed of a mixture of short- and medium-chain fatty acids (SMCFA); c. Triglycerides (TG) composed of a mixture of butyrate and long-chain fatty acids (e.g., a mixture of butyrate and oleate); d. Triglycerides composed of medium-chain fatty acids; e. Triglycerides (TG) composed of short-chain fatty acids; f. short-chain fatty acids that can be metabolized into ketone bodies; and g. Short- and / or medium-chain fatty acids that can be metabolized into ketone bodies, preferably C4 and / or C8 fatty acids (e.g., butyric acid (C4:0) and / or caprylic acid (C8:0)); h. Triglycerides (TG) composed of butyrate and caprylate;

[0029]

[0028] According to another aspect, the composition defined herein is selected from a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a small-volume liquid supplement, a meal replacement beverage, and combinations thereof.

[0030]

[0029] A second embodiment is the use of a composition as defined herein for reducing or avoiding the accumulation of uremic toxins, preferably in the treatment of a cardiometabolic or neurodegenerative condition. The composition is preferably used in accordance with the preceding for the reduction of uremic toxins in cardiometabolic or neurodegenerative conditions and associated comorbidities, for slowing the progression of such cardiometabolic or neurodegenerative conditions and comorbidities, and / or for managing the symptoms and syndromes associated with the toxic effects of uremic solutes in such cardiometabolic or neurodegenerative conditions and associated comorbidities.

[0031]

[0030] The cardiometabolic or neurodegenerative conditions and associated comorbidities that can be treated by using the composition are particularly relevant in the following cases: Treatment or prevention of kidney disease, including chronic and acute kidney disease; dialysis and pre-dialysis kidney disease; rare kidney diseases, including genetically and metabolically induced kidney diseases; Treatment or prevention of PEW syndrome, uremic syndrome including bone loss, severe anorexia, fatigue, or inflammation; Delayed complications of progressive kidney disease, including anemia, hyperphosphatemia, secondary hyperparathyroidism, bone disease, and neurological conditions; Delayed renal comorbidities, including cardiovascular disease Preventing or managing the risk of malnutrition; Slowing the progression of cardiometabolic diseases; Prevention or management of the risk of cardiovascular disease and comorbidities, such as diabetes; and / or Prevention or management of the risk of neurodegenerative and neurological conditions.

[0032]

[0031] A third embodiment is a method for treating a cardiometabolic or neurodegenerative condition as defined above, preferably comprising, as a first step, (a) preparing and providing a composition as discussed above comprising a specifically selected probiotic, a specifically selected prebiotic, and a specifically selected lipid as defined above, and (b) administering such composition to a patient in need thereof, typically suffering from increased uremic toxins in the context of a cardiometabolic or neurodegenerative condition as defined herein.

[0033]

[0032] A fourth embodiment is a (multi-part) kit suitable for use in reducing and / or avoiding the accumulation of uremic toxins in a cardiometabolic or neurodegenerative condition as defined herein, comprising a composition as described herein, for example comprising a specifically selected probiotic, a specifically selected prebiotic and a specifically selected lipid as defined above; a kit for use in separate containers, for example as two or more liquid solutions or dry powders, for forming one or more of the compositions disclosed herein and / or for use in one or more of the methods disclosed herein. [Brief explanation of the drawings]

[0034] [Figure 1] The biochemical pathways for the formation of enterobacterial uremic toxins and some of their precursors are illustrated. In this example, the production of urea (NH), p-cresyl sulfate (PCS), indoxyl sulfate (IS), trimethylamine-N-oxide (TMAO), and other catabolic enzymes are shown, along with the corresponding enzyme codes (ECs) of the enzymes. Uremic toxin precursors, such as p-cresol, indole, and TMA, are by-products of microbial amino acid metabolism, such as tryptophan, tyrosine, carnitine, and choline. These metabolic by-products then undergo further chemical modification in the liver before being converted to uremic toxins, which then reach the circulatory system and affect target organs. [Figure 2-1] Figure 2A shows a schematic diagram of the production and retention of uremic toxins in the body, illustrating how microbiome dysbiosis can contribute to exacerbation of symptoms, comorbidities, and disease progression due to accumulation of uremic toxins in the systemic circulation. [Figure 2-2] Figure 2B shows a schematic diagram of the production and storage of uremic toxins in the body. Uremic toxins adversely affect multiple organs, leading to clinical outcomes and symptoms. Figure 2B is adapted from Rosner et al., Clin J Am Soc Nephrol. 2021. [Figure 3]An exemplary selection of probiotic strains for the compositions and therapeutic purposes disclosed herein is shown. As can be seen, the probiotic strains were preselected in an in silico screening process to lack different genes and key bacterial enzymes involved in uremic toxin metabolism. Lactobacillus johnsonii NCC 533 lacks the majority of relevant enzymes to avoid the production or accumulation of uremic toxins, proving to provide an optimal basis for the proposed treatment. White cells: enzyme absent. Gray cells: enzyme presence unknown. Black cells: enzyme present. Tra: tyramine; Trp: tryptophan; Tyr: tyrosine; indole-3-pyruvate: IPA; IAM: indole-3-acetamide; IAN: indole-3-acetonitrile; 4-HPPA: 4-hydroxyphenylpyruvic acid. [Figure 4] The growth profile of Lactobacillus johnsonii NCC 533 during 48 hours of incubation with different carbohydrate sources (α-galactooligosaccharide, galactomannan, and fructan, which require the probiotic-encoded enzymes α-galactosidase and β-fructofuranosidase, respectively) is shown. As can be seen, the addition of pea GOS and soybean GOS was most effective, while the addition of PHGG, fenugreek, ScFOS, and inulin, while suitable, resulted in less growth of Lactobacillus johnsonii NCC 533 (from top to bottom: soybean GOS, pea GOS, inulin, sFOS, PHGG, and fenugreek). The growth test using Lactobacillus johnsonii NCC 533 as the strain is provided for illustrative purposes only and can be applied to other selected probiotics. Data were normalized to the negative control. [Figure 5]The growth profile of Lactobacillus johnsonii NCC 533 during 48 hours of incubation with different carbohydrate sources (β-galactooligosaccharides, which require β-galactosidase as the probiotic-encoded enzyme) is shown. As can be seen, the addition of BMOs and Vivinal® GOS results in particularly efficient growth of Lactobacillus johnsonii NCC 533 (from top to bottom: BMO and Vivinal® GOS). The growth test using Lactobacillus johnsonii NCC 533 as the strain is provided purely as an example and can also be applied to selected other probiotics. Data were normalized to the negative control. [Figure 6] The growth profile of Lactobacillus johnsonii NCC 533 during 48 hours of incubation with different carbohydrate sources (cellooligosaccharides, requiring the probiotic-encoded enzymes glucan 1,4-β-glucosidase, cellulase, and licheninase). As can be seen, the addition of cellobiose and cellotriose resulted in particularly efficient growth of Lactobacillus johnsonii NCC 533, followed by cellotetraose, soluble hydrolyzed wheat, and soluble hydrolyzed oats (from top to bottom: cellobiose, cellotriose, cellotetraose, soluble hydrolyzed wheat, and soluble hydrolyzed oats). The growth study using Lactobacillus johnsonii NCC 533 as a strain is provided merely as an example and can be applied to other selected probiotics. [Figure 7]This paper outlines the in vitro / ex vivo experimental setup to evaluate differences in microbiome profiles between healthy and chronic kidney disease (CKD) donors using Prodigest's short-term single-stage colonic simulation technology. Figure 7A shows a schematic diagram of the conditions and donor group samples. For each donor group, two conditions were run in parallel: (1) a standard condition consisting of the experimental timeline and sample analysis of basal nutrient medium samples containing minimal amino acids, mimicking the amino acid content of a low-protein diet; and (2) an AA-spiked condition consisting of a nutrient medium containing a mixture of L-tryptophan, L-tyrosine, L-carnitine, choline, and L-phenylalanine. The AA mixture was selected to contain substrates that are converted by gut bacteria into uremic toxins and their precursors. The AA concentration mimics the daily amino acid intake required by an adult. A total of nine healthy donors and eight CKD donors were used in this example. Figure 7B shows the timeline for the experiment and sample analysis. On day 0, the fecal microbiome was inoculated into the ProDigest short-term, single-stage colonic system. From day 1 to day 2, the system was fed basal nutrient medium with or without the AA mixture. The standard group received basal nutrient medium, and the AA-spiked group received basal nutrient medium plus an amino acid mixture. Samples were taken at different time points to measure overall fermentation activity, microbial community activity, and microbiome composition.The readouts were: (1) Overall fermentation activity: acid / base consumption; (2) Microbial community activity: lactate; Short-chain fatty acids (SCFA): butyrate, propionate, acetate; Markers of proteolytic activity: ammonium and branched SCFA (isobutyric acid, isovaleric acid, and isocaproic acid); Urea toxins and precursors: p-cresol, p-cresyl sulfate, indole, indole-3-triacetic acid, betaine, trimethylamine, trimethylamine-N-oxide, indoxyl, indoxyl sulfate, semialdehyde, glutaric acid, uric acid, urea; and (3) Microbial community composition: quantitative deep shotgun sequencing. [Figure 8] An example of the difference in microbiome metabolic capacity between the microbiomes of chronic kidney disease (CKD) patients and healthy donors is shown. After 48 hours of incubation in the Prodigest short-term, one-step colon simulation system, the CKD microbiome exhibited higher production of uremic toxin precursors, such as p-cresol, than the healthy microbiome, especially in the presence of excess amino acid substrates (Figure 8(A)), suggesting dysregulation of amino acid metabolism in the CKD microbiome. Furthermore, the CKD microbiome exhibited increased production of branched-chain fatty acids (BCFAs) compared with the healthy microbiome (Figure 8(B)), suggesting greater proteolytic activity by the CKD microbiome. As expected, byproducts of protein metabolism, such as BCFAs, were not affected by excess amino acid substrates. Data represent mean ± SEM. *p<0.05 by Fisher's least significant difference LSD method (α=5%). [Figure 9]This figure shows an outline of the in vitro / ex vivo experimental setup using a modified Prodigest SHIME® system to evaluate the impact of novel nutritional / synbiotic materials on the CKD microbiome and fecal microbiota of chronic kidney disease (CKD) patients. Specifically, a sample schematic of the modified SHIME® system, consisting of an upper GIT vessel functioning as the stomach and small intestine and a colon vessel simulating the transverse colon, is shown in Figure 9(A). Three parallel conditions, consisting of two treatment groups and one control group, were performed for each donor. A total of eight CKD donors were used in this example. The experimental timeline and sample analysis of the sample are shown in Figure 9(B). On day 0, fecal microbiomes from CKD donors were inoculated into the ProDigest SHIME system. From days 1 through 10, the system was fed a basal nutrient medium (containing minimal amino acids to mimic the amino acid content of a low-protein diet) with or without a nutrient / synbiotic blend intervention. In this example, two nutrient / synbiotic blend combinations (P1 and P2) were tested. The P1 intervention consisted of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, and short-medium chain triglycerides containing butyrate and caprylate. The P2 intervention consisted of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, pea GOS, and short-medium chain triglycerides containing butyrate and caprylate. During the final two days (days 8 to 10), the system was further supplemented with an additional amino acid (AA) mixture containing L-tryptophan, L-tyrosine, L-carnitine, choline, and L-phenylalanine. The AA mixture was selected to contain substrates that can be converted by gut bacteria into uremic toxins and their precursors. Furthermore, the AA concentration was formulated to mimic the daily amino acid intake required by an adult. Samples were collected at different time points to measure overall fermentation activity, microbial community activity, and microbiome composition.The readouts were: (1) Overall fermentation activity: acid / base consumption; (2) Microbial community activity: lactate; Short-chain fatty acids (SCFA): butyrate, propionate, acetate; Markers of proteolytic activity: ammonium and branched SCFA (isobutyric acid, isovaleric acid, and isocaproic acid); Urea toxins and precursors: p-cresol, p-cresyl sulfate, indole, indole-3-triacetic acid, betaine, trimethylamine, trimethylamine-N-oxide, indoxyl, indoxyl sulfate, semialdehyde, glutaric acid, uric acid, urea; (3) Microbial community composition: quantitative shotgun deep sequencing. [Figure 10]This figure shows an example of the effect of a novel nutritional or synbiotic blend on in vitro / ex vivo microbiota-derived uremic toxin precursor production by the fecal microbiota of chronic kidney disease (CKD) patients. As shown, the interventions designated P1 and P2 significantly reduced the increased production of clinically relevant uremic toxin precursors by the CKD microbiome compared to untreated controls. Indole (A), p-cresol (B), and trimethylamine (C) are by-products of amino acid metabolism by the gut microbiota. Therefore, production will be higher under conditions where additional amino acid substrates are available. Notably, the intervention was effective in conditions with normal and minimal amino acid levels, thus suggesting the potential for broader benefits for CKD patients with different dietary requirements, e.g., low- to high-protein diets. Overall, this data highlights the benefit of the present invention in correcting amino acid metabolic abnormalities in the microbiome of CKD patients, resulting in less uremic toxin accumulation and associated clinical consequences. The P1 intervention consisted of treating the CKD microbiome with a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, and short-medium chain triglycerides containing butyrate and caprylate. The P2 intervention consisted of treating the CKD microbiome with a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, pea galactooligosaccharides, and short-medium chain triglycerides containing butyrate and caprylate. The control represents the CKD microbiome left untreated. Data represent the mean ± SEM. *p<0.05 by Student's t-test. [Figure 11]This figure shows an example of the effect of a novel nutritional or synbiotic blend on in vitro / ex vivo microbiota-derived uremic toxin production by the fecal microbiota of patients with chronic kidney disease (CKD). As shown, the interventions designated P1 and P2 significantly reduced the increase in urea production by the CKD microbiome compared to untreated controls. Urea is a by-product of protein metabolism. Therefore, there would be little effect on urea levels after the addition of additional amino acids. Overall, this data highlights the benefit of the present invention in helping to ameliorate the accumulation of uremic toxins and associated clinical outcomes, particularly in CKD patients. The P1 intervention consisted of treating the CKD microbiome with a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, and short-medium chain triglycerides containing butyrate and caprylate. The P2 intervention consisted of treating the CKD microbiome with a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, pea galactooligosaccharides, and short-medium chain triglycerides containing butyrate and caprylate. The control represents the CKD microbiome left untreated. Data represent the mean ± SEM. *p<0.05 by Student's t-test. [Figure 12]This figure shows the effect of a novel nutritional or synbiotic blend on in vitro / ex vivo protein metabolism abnormalities by the CKD microbiome in chronic kidney disease (CKD) patients. As shown, CKD microbiomes treated with P1 and P2 interventions exhibited significantly lower levels of branched-chain fatty acids (A) and ammonium (B), suggesting amelioration of excessive proteolytic activity that contributes to higher uremic toxin accumulation. Proteolytic activity by the gut microbiome is not significantly affected by the addition of additional amino acids. Overall, this data highlights the benefit of the present invention in correcting dysbiosis in the CKD microbiome, which may therefore help ameliorate uremic toxin accumulation. The P1 intervention consisted of treating the CKD microbiome with a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, and short-medium chain triglycerides containing butyrate and caprylate. The P2 intervention consisted of treating the CKD microbiome with a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, pea galactooligosaccharides, and short-medium chain triglycerides containing butyrate and caprylate. The control represents the CKD microbiome left untreated. Data represent the mean ± SEM. *p<0.05 by Student's t-test. [Figure 13]This study demonstrates the effects of novel nutritional or synbiotic blends on glycolytic activity and the production of beneficial microbiota-derived metabolites, particularly short-chain fatty acids (SCFAs), by the fecal microbiota of patients with chronic kidney disease (CKD) in vitro and ex vivo. The gut microbiota can influence host metabolic health through microbial metabolites. The balance between the production of microbial metabolites by glycolytic fermentation and that by proteolytic fermentation may be an important determinant of metabolic health. Indeed, dysbiosis in patients with chronic kidney disease (CKD) is characterized by a higher ratio of proteolytic to glycolytic activity dominated by specific bacterial groups. Among the most studied glycolytic microbial metabolites are the short-chain fatty acids (SCFAs) acetate (A), propionate (B), and butyrate (C). SCFAs have been shown to have multiple benefits for the host, including, but not limited to, improved intestinal epithelial barrier function and inflammation. Increased intestinal permeability has been suggested to contribute to the increased availability of gut-derived uremic toxins and precursors in the systemic circulation. Henceforth further reported that increased production of metabolites may help improve the integrity of the intestinal lining and indirectly prevent the accumulation of excess uremic toxins in the systemic circulation, particularly for CKD patients. As shown in the graph, the P1 and P2 interventions demonstrated significantly higher SCFA levels compared to the untreated control. This suggests that the present invention has the potential to correct dysbiosis in the CKD microbiome and provide promising metabolic health benefits. The P1 intervention consisted of treating the CKD microbiome with a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, and short- and medium-chain triglycerides containing butyrate and caprylate. The P2 intervention consisted of treating the CKD microbiome with a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, pea galactooligosaccharides, and short- and medium-chain triglycerides containing butyrate and caprylate. Control represents the CKD microbiome left untreated. Data represent mean ± SEM. *p<0.05 by Student's t-test. [Figure 14]This paper outlines the setup of an animal study to evaluate the effects of novel nutritional or synbiotic materials on uremic toxin production and chronic kidney disease (CKD) progression. The animal model used was a 5 / 6 nephrectomy model. This model is the gold standard in CKD research, including renal pharmacological studies, and is one of the most frequently used rodent models. Kidneys were removed in a two-step surgical procedure, resulting in reduced renal function (reminiscent of human CKD stage 3b or higher). Additional animals underwent sham surgery and served as non-CKD controls. From weeks 3 to 10, CKD animals were fed a diet with or without a nutritional or synbiotic blend intervention. In this example, two nutritional or synbiotic blend combinations (P1 and P2) were tested. The P1 intervention consisted of Lactobacillus johnsonii NCC533 and a material blend containing cellobiose and short-medium chain triglycerides containing butyrate and caprylate. The P2 intervention consisted of a material blend containing Lactobacillus johnsonii NCC533, cellobiose, pea GOS, and short-medium chain triglycerides containing butyrate and caprylate. Metabolic parameters were assessed beginning at week 8, including a glucose tolerance test. At week 9, mice were placed in metabolic cages and urine collected for uremic toxin and renal parameter analysis. At week 10, mice were sacrificed and multiple tissues were harvested for subsequent analysis. Tissues collected included plasma and serum, urine, liver, heart, eWAT, scWAT, gastrocnemius, soleus, kidney, all intestinal segments, and cecum. Physiological, behavioral, and food and water intake monitoring was performed throughout the study.Endpoint Analyses Blood and urine concentrations of uremic toxins (e.g., urea, indoxyl sulfate, p-cresyl sulfate, TMAO, uric acid, CMPF, IAA, PCG) Assessment of renal function - blood and urine renal markers (e.g., proteinuria, creatinine, albumin, cystatin C), and renal histopathology analysis Metabolic inflammatory markers (blood and tissue cytokines IL6, TNFa, IL1b); blood CRP; fecal calprotectin and albumin) Intestinal function (epithelial function and permeability markers in blood, e.g., citrulline; intestinal histological analysis; lipid / steatorrhea levels) Improvement in CKD comorbidities (glucose regulation assessed during oral glucose tolerance test, fasting glucose, insulin) Improvement in other metabolic parameters (plasma triglycerides, AST, ALT, cholesterol, LDL or HDL) Improvement in CKD comorbidities (fat, liver, heart, intestine and muscle will be sampled for protein or RNA analysis) Cecal microbiome analysis - composition and function (metagenomics) Untargeted metabolomic analysis of feces, urine and serum. [Figure 15-1]This study demonstrates the effect of a novel nutritional or synbiotic blend on clinically relevant plasma uremic toxin levels in an animal model of chronic kidney disease (CKD). As shown, animals undergoing nephrectomy (CKD group, mimicking animals with CKD and reminiscent of human CKD stage 3b or higher) exhibited higher uremic toxin production compared with non-CKD control animals (sham-operated group, reminiscent of healthy individuals without CKD). Group P1 represents CKD animals treated with an intervention consisting of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, and short- and medium-chain triglycerides containing butyrate and caprylate. Group P2 represents CKD animals treated with an intervention consisting of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, pea GOS, and short- and medium-chain triglycerides containing butyrate and caprylate. CKD animals treated with the P1 and P2 interventions for 7 weeks had significantly lower plasma levels of uremic toxins, such as p-cresyl sulfate (PCS) (A), indoxyl sulfate (IS) (B), p-cresyl glucuronide (PCG) (C), indoleacetic acid (IAA) (D), 3-carboxy-4-methyl-5-propyl-2-furanpropionate (CMPF) (E), and uric acid (F), compared with untreated CKD animals. The data clearly demonstrated the benefit of the interventions to ameliorate the accumulation of uremic toxins in the systemic circulation, especially in individuals with impaired renal function. Each data point represents one animal. The line represents the mean. Multiple comparisons were performed using ANOVA followed by Fisher's LSD without correction. *p<0.05, **p<0.01, ***p<0.001. [Figure 15-2]This study demonstrates the effect of a novel nutritional or synbiotic blend on clinically relevant plasma uremic toxin levels in an animal model of chronic kidney disease (CKD). As shown, animals undergoing nephrectomy (CKD group, mimicking animals with CKD and reminiscent of human CKD stage 3b or higher) exhibited higher uremic toxin production compared with non-CKD control animals (sham-operated group, reminiscent of healthy individuals without CKD). Group P1 represents CKD animals treated with an intervention consisting of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, and short- and medium-chain triglycerides containing butyrate and caprylate. Group P2 represents CKD animals treated with an intervention consisting of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, pea GOS, and short- and medium-chain triglycerides containing butyrate and caprylate. CKD animals treated with the P1 and P2 interventions for 7 weeks had significantly lower plasma levels of uremic toxins, such as p-cresyl sulfate (PCS) (A), indoxyl sulfate (IS) (B), p-cresyl glucuronide (PCG) (C), indoleacetic acid (IAA) (D), 3-carboxy-4-methyl-5-propyl-2-furanpropionate (CMPF) (E), and uric acid (F), compared with untreated CKD animals. The data clearly demonstrated the benefit of the interventions to ameliorate the accumulation of uremic toxins in the systemic circulation, especially in individuals with impaired renal function. Each data point represents one animal. The line represents the mean. Multiple comparisons were performed using ANOVA followed by Fisher's LSD without correction. *p<0.05, **p<0.01, ***p<0.001. [Figure 16]This study demonstrates the effects of a novel nutritional or synbiotic blend on renal parameters / markers of renal function in an animal model of chronic kidney disease (CKD). The sham-operated group represents non-CKD animals reminiscent of healthy individuals without CKD. The CKD group represents animals with CKD reminiscent of human CKD stage 3b or higher. Group P1 represents CKD animals treated with an intervention consisting of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, and short-medium chain triglycerides containing butyrate and caprylate. Group P2 represents CKD animals treated with an intervention consisting of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, pea GOS, and short-medium chain triglycerides containing butyrate and caprylate. As shown, animals that underwent nephrectomy (CKD group) showed impaired renal function, as evidenced by increased proteinuria (A), as evidenced by a higher protein-to-creatinine ratio in urine, and increased concentrations of urea in plasma (B), compared with non-CKD control animals (sham-operated group, reminiscent of healthy individuals without CKD disease). CKD animals treated with P1 and P2 interventions for 7 weeks showed significantly better proteinuria and plasma urea levels compared with untreated CKD animals. The data demonstrate the benefits of the present invention in stabilizing renal function and may therefore be useful in preventing the vicious cycle of uremic toxin accumulation and slowing the progression of kidney damage and associated clinical outcomes. For proteinuria, each data point represents two animals, whereas for urea levels, each data point represents one animal. Lines represent means. Multiple comparisons were performed using ANOVA followed by Fisher's LSD without correction. *p<0.05, **p<0.01, ***p<0.001. [Figure 17-1]This study demonstrates the effects of a novel nutritional or synbiotic blend on kidney tissue structure in an animal model of chronic kidney disease (CKD). The sham-operated group represents non-CKD animals resembling healthy individuals without CKD. The CKD group represents animals with CKD resembling human CKD stage 3b or higher. Group P1 represents CKD animals treated with an intervention consisting of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, and short-medium chain triglycerides containing butyrate and caprylate. Group P2 represents CKD animals treated with an intervention consisting of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, pea GOS, and short-medium chain triglycerides containing butyrate and caprylate. As shown, the remaining kidneys of animals that underwent nephrectomy (CKD group) exhibited microscopic damage characterized by increased fibrosis (A) and impaired glomerular size (B) and volume (C) compared with non-CKD control animals (sham-operated group, reminiscent of healthy individuals without CKD disease). CKD animals treated with P1 and P2 interventions for 7 weeks exhibited fibrosis and decreased glomerular size and volume. The data demonstrate the benefit of the present invention in reducing kidney microscopic damage and may therefore be useful in preventing the vicious cycle of uremic toxin accumulation and helping to slow the progression of kidney damage and associated clinical outcomes. Each data point represents one animal. Multiple comparisons were performed using ANOVA followed by Fisher's LSD without correction. *p<0.05, **p<0.01, ***p<0.001. HES: Hematoxylin and eosin staining. [Figure 17-2]This study demonstrates the effects of a novel nutritional or synbiotic blend on kidney tissue structure in an animal model of chronic kidney disease (CKD). The sham-operated group represents non-CKD animals resembling healthy individuals without CKD. The CKD group represents animals with CKD resembling human CKD stage 3b or higher. Group P1 represents CKD animals treated with an intervention consisting of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, and short-medium chain triglycerides containing butyrate and caprylate. Group P2 represents CKD animals treated with an intervention consisting of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, pea GOS, and short-medium chain triglycerides containing butyrate and caprylate. As shown, the remaining kidneys of animals that underwent nephrectomy (CKD group) exhibited microscopic damage characterized by increased fibrosis (A) and impaired glomerular size (B) and volume (C) compared with non-CKD control animals (sham-operated group, reminiscent of healthy individuals without CKD disease). CKD animals treated with P1 and P2 interventions for 7 weeks exhibited fibrosis and decreased glomerular size and volume. The data demonstrate the benefit of the present invention in reducing kidney microscopic damage and may therefore be useful in preventing the vicious cycle of uremic toxin accumulation and helping to slow the progression of kidney damage and associated clinical outcomes. Each data point represents one animal. Multiple comparisons were performed using ANOVA followed by Fisher's LSD without correction. *p<0.05, **p<0.01, ***p<0.001. HES: Hematoxylin and eosin staining. [Figure 18]The effects of novel nutritional or synbiotic blends on (A) body weight change, (B) food intake, and energy reserve wasting (ERW) / PEW in an animal model of chronic kidney disease (CKD). The adverse consequences of uremic toxin accumulation in kidney disease are anorexia and loss of energy reserves, including muscle and fat reserves. As shown in the figure, after surgery, animals undergoing nephrectomy (CKD group, representing animals with CKD and reminiscent of human CKD stage 3b or higher) showed slower weight gain compared to the non-CKD control group (sham-operated group, reminiscent of healthy individuals without CKD disease). Furthermore, CKD animals showed significantly lower food intake compared to the sham-operated group, starting from week 2 and continuing through week 10. At the end of the study, (C) CKD animals also showed a significant decrease in epididymal white adipose tissue (eWAT), suggesting a decrease in energy reserves reminiscent of that seen in human CKD. CKD animals treated with the P1 and P2 interventions showed significantly improved weight change and normalized food intake to the same levels as non-CKD sham-operated animals. The P1 group consisted of CKD animals treated with an intervention consisting of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, and short-medium chain triglycerides containing butyrate and caprylate. The P2 group consisted of CKD animals treated with an intervention consisting of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, pea GOS, and short-medium chain triglycerides containing butyrate and caprylate. Furthermore, treated animals, particularly those treated with P1, showed better fat reserve capacity compared to untreated CKD animals, as seen by improvements in epididymal white adipose tissue (eWAT). The data demonstrated promising benefits of the intervention for alleviating uremia-related symptoms, including anorexia, which leads to weight loss and PEW. For body weight graphs, the line represents the mean across all animals in each group, with error bars showing SEM. For food intake, each data point represents one animal. Multiple comparisons were performed using ANOVA followed by Fisher's LSD without correction. *p<0.05, **p<0.01, ***p<0.001. [Figure 19] This study demonstrates the effect of a novel nutritional or synbiotic blend on intestinal barrier dysfunction in an animal model of chronic kidney disease (CKD). As shown, nephrectomized animals (CKD group, representing animals with CKD, reminiscent of human CKD stage 3b or higher) exhibited impaired intestinal barrier function, as evidenced by lower protein expression of occludin in the tight junctions in the ileum compared with non-CKD control animals (sham-operated group, reminiscent of healthy individuals without CKD). Tight junctions are specialized connections between two adjacent cell membranes and, in the case of the intestinal lining, are important structures that prevent excessive amounts of intestinal-derived molecules, such as uremic toxin precursors, from entering the systemic circulation. CKD animals treated with P1 and P2 interventions for 7 weeks showed significantly higher protein expression of occludin compared with untreated CKD animals. Group P1 represents CKD animals treated with an intervention consisting of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, and short-medium chain triglycerides containing butyrate and caprylate. Group P2 represents CKD animals treated with an intervention consisting of a blend of ingredients containing Lactobacillus johnsonii NCC533, cellobiose, pea GOS, and short-medium chain triglycerides containing butyrate and caprylate. This data demonstrates the benefit of the present invention in improving intestinal dysfunction, particularly in those suffering from renal dysfunction. Each data point represents one animal. The line represents the mean. Multiple comparisons were performed using ANOVA followed by Fisher's LSD without correction. *p<0.05, **p<0.01, ***p<0.001. [Figure 20-1]1 shows the effect of novel nutritional or synbiotic blends on (A) urinary albumin, (B) urinary creatine, (C) urinary protein (albumin) to creatine ratio, and (D) LOG urinary protein (albumin) to creatine ratio at week 7 in a rat animal model of CKD. From left to right: sham-operated, 5 / 6 vehicle, 5 / 6 Nx P1-rat intervention (P1-rat intervention consisted of a material blend containing 10g Lactobacillus johnsonii NCC533, 1% cellobiose, and 1% short-medium chain triglycerides containing butyrate and caprylate), 5 / 6 Nx P3-intervention (intervention consisted of a material blend containing 10g Lactobacillus johnsonii NCC533, 0.3% cellobiose, and 0.3% short-medium chain triglycerides containing butyrate and caprylate), and 5 / 6 Nx lisinopril 20 mg / kg. (A) Values ​​are expressed as mean + SEM for n = 12–18. Dunnett's test, one-factor linear model. **: P < 0.01 compared to 5 / 6 Nx vehicle. (B) Values ​​are expressed as mean + SEM for n = 12–18 mice. Dunnett's test, one-factor linear model. **: P<0.01, ***: P<0.001 compared to 5 / 6 Nx vehicle. Sham-operated mice had reduced urinary albumin levels at week 7 when compared to 5 / 6 Nx vehicle. Sham-operated mice had reduced urinary albumin levels at week 7 when compared to 5 / 6 Nx vehicle. (C) Values ​​are expressed as mean + SEM for n = 12–18 mice. Dunnett's test, one-factor linear model. *: P<0.05, **: P<0.01 compared to 5 / 6 Nx vehicle. (D) Values ​​are expressed as mean + SEM for n = 12–18 mice. Dunnett's test, one-factor linear model. *: P<0.05, ***: P<0.001 compared to 5 / 6 Nx vehicle. Sham and 5 / 6 Nx P1 had reduced urinary ACR when compared to 5 / 6 Nx vehicle at week 7. Sham and 5 / 6 Nx lisinopril 20 mg / kg had reduced LOG urinary ACR when compared to 5 / 6 Nx vehicle at week 7. [Figure 20-2]1 shows the effect of novel nutritional or synbiotic blends on (A) urinary albumin, (B) urinary creatine, (C) urinary protein (albumin) to creatine ratio, and (D) LOG urinary protein (albumin) to creatine ratio at week 7 in a rat animal model of CKD. From left to right: sham-operated, 5 / 6 vehicle, 5 / 6 Nx P1-rat intervention (P1-rat intervention consisted of a material blend containing 10g Lactobacillus johnsonii NCC533, 1% cellobiose, and 1% short-medium chain triglycerides containing butyrate and caprylate), 5 / 6 Nx P3-intervention (intervention consisted of a material blend containing 10g Lactobacillus johnsonii NCC533, 0.3% cellobiose, and 0.3% short-medium chain triglycerides containing butyrate and caprylate), and 5 / 6 Nx lisinopril 20 mg / kg. (A) Values ​​are expressed as mean + SEM for n = 12–18. Dunnett's test, one-factor linear model. **: P < 0.01 compared to 5 / 6 Nx vehicle. (B) Values ​​are expressed as mean + SEM for n = 12–18 mice. Dunnett's test, one-factor linear model. **: P<0.01, ***: P<0.001 compared to 5 / 6 Nx vehicle. Sham-operated mice had reduced urinary albumin levels at week 7 when compared to 5 / 6 Nx vehicle. Sham-operated mice had reduced urinary albumin levels at week 7 when compared to 5 / 6 Nx vehicle. (C) Values ​​are expressed as mean + SEM for n = 12–18 mice. Dunnett's test, one-factor linear model. *: P<0.05, **: P<0.01 compared to 5 / 6 Nx vehicle. (D) Values ​​are expressed as mean + SEM for n = 12–18 mice. Dunnett's test, one-factor linear model. *: P<0.05, ***: P<0.001 compared to 5 / 6 Nx vehicle. Sham and 5 / 6 Nx P1 had reduced urinary ACR when compared to 5 / 6 Nx vehicle at week 7. Sham and 5 / 6 Nx lisinopril 20 mg / kg had reduced LOG urinary ACR when compared to 5 / 6 Nx vehicle at week 7. [Figure 21](From left to right) Sham-operated group, 5 / 6 vehicle groups, 5 / 6 Nx P3 intervention group (intervention consisted of a material blend containing 10g Lactobacillus johnsonii NCC533, 0.3% cellobiose, and 0.3% short-medium chain triglycerides containing butyrate and caprylate), 5 / 6 Nx P1 rat intervention group (shown as "5 / 6 Nx P1" in Figures 21(A)-(C)) (P1 rat intervention consisted of a material blend containing 10g Lactobacillus johnsonii NCC533, 1% cellobiose, and 1% short-medium chain triglycerides containing butyrate and caprylate), and 5 / 6 of the 5 / 6 Nx lisinopril 20mg / kg group. Plasma levels of (A) indoxyl sulfate (IS), (B) p-cresyl sulfate (PCS), and (C) p-cresol glucuronide (PCG) in Nx rats are shown. 5 / 6 Nx P1 rat intervention reduced both IS and pCS plasma levels (approximately 36% for IS and 77% for pCS compared to the 5 / 6 Nx vehicle group). DETAILED DESCRIPTION OF THE INVENTION

[0035] definition Some definitions provided below are applicable to the present specification and the invention described. However, definitions may be located in the "Background," "Detailed Description of the Invention" sections, the "Examples" section below, and the "Definitions" section below.

[0036] All percentages stated herein are by weight of the total composition unless otherwise specified.

[0037]

[0035] As used herein, "about," "approximately," and "substantially" are understood to refer to a number within a range of numerical values, for example, within -10% to +10% of a referenced numerical value, preferably within -5% to +5% of a referenced numerical value, more preferably within -1% to +1% of a referenced numerical value, and most preferably within -0.1% to +0.1% of a referenced numerical value.

[0038]

[0036] All numerical ranges herein should be understood to include every integer, whole, or fractional number within that range. Furthermore, these numerical ranges should be construed to support claims directed to any number or subset of numbers within that range. For example, a disclosure of 1 to 10 should be construed to correspond to ranges of 1 to 8, 3 to 7, 1 to 9, 3.6 to 4.6, 3.5 to 9.9, etc.

[0039] As used in this disclosure and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context dictates otherwise. Thus, for example, reference to "a component" or "the component" includes two or more components.

[0040] The terms "comprise," "comprises," and "comprising" should be construed as inclusive rather than exclusive. Similarly, the terms "include," "including," and "or" should all be construed as inclusive unless such interpretation is clearly prevented by the context. However, the compositions disclosed herein may not include elements not specifically disclosed herein. Accordingly, disclosure of an embodiment using the term "comprising" includes disclosure of embodiments "consisting essentially of" and "consisting of" the specified component. A composition "consisting essentially of" means that the referenced component comprises at least 50% by weight, preferably at least 75% by weight, more preferably at least 85% by weight, and most preferably at least 95% by weight of the referenced component.

[0041] The term "and / or" used in the context of "X and / or Y" should be interpreted as "X" or "Y" or "X and Y". Similarly, "at least one of X or Y" should be interpreted as "X" or "Y" or "both X and Y".

[0042] As used herein, the terms "example" and "such as," particularly when followed by a list of terms, are merely exemplary and illustrative and should not be considered exclusive or inclusive. As used herein, a condition "associated with" or "linked with" another condition means that the conditions occur simultaneously, preferably that the conditions are caused by the same underlying symptom, and most preferably that one of the specified conditions is caused by the other specified condition.

[0043] The terms "food," "food product," and "food composition" refer to a product or composition intended for consumption by an individual, such as a human, and providing at least one nutrient to such an individual. Food products typically contain at least one of protein, lipid, and carbohydrate, and optionally one or more vitamins and minerals. The compositions of the present disclosure, including the many embodiments described herein, may comprise, consist of, or essentially consist of the elements disclosed herein as well as any additional or optional ingredients, components, or elements described or not described herein that are useful in a diet.

[0044] "Prevention" includes reducing the risk and / or severity of a condition or disease. The terms "treatment," "treating," and "alleviating" include both preventative or prophylactic treatment (treatment that prevents and / or delays the onset of the targeted pathological condition or disorder) and curative, therapeutic, or disease-modifying treatment, including therapeutic measures to cure, delay, reduce symptoms, and / or halt the progression of a diagnosed pathological condition or disorder; including treatment of patients at risk of or suspected of having a disease, as well as patients diagnosed with a disease or condition. The terms do not necessarily imply that a subject is treated until cured. The terms "treatment" and "treating" also refer to maintaining and / or promoting the health of individuals who are not afflicted with a disease but who may be susceptible to an ill-health condition. The terms "treatment / therapy," "treat / treating," and "alleviating" are also intended to include synergistic or otherwise potentiating effects of one or more primary preventative or therapeutic measures. The terms "treatment / therapy," "treat / treating," and "alleviating" are further intended to include dietary management of a disease or condition, or dietary management for the prophylaxis or prevention of a disease or condition. Treatment / therapy may be patient-related or physician-related.

[0045] As used herein, the term "unit dosage form" refers to a physically discrete unit suitable as a dosage unit for human and animal subjects, each unit containing a predetermined amount of a composition disclosed herein sufficient to produce a desired effect, together with a pharmaceutically acceptable diluent, carrier, or vehicle. The specifications for the unit dosage form depend on the particular compound used, the effect to be achieved, and the pharmacodynamics associated with each compound in the host.

[0046] A "subject" or "individual" is a mammal, preferably a human. The term "elderly" in the human context means an age of at least 60 years, preferably greater than 63 years, more preferably greater than 65 years, and most preferably greater than 70 years. The term "older adult" in the human context means a postnatal age of 45 years or older, preferably greater than 50 years, and more preferably greater than 55 years, and includes elderly people.

[0047]

[0045] As used herein, an "effective amount" is an amount that prevents a deficiency, treats a disease or medical condition in an individual, or more generally, alleviates symptoms, manages the progression of a disease, or provides a nutritional, physiological, or medical benefit to an individual. The relative terms "improved," "increased," "enhanced," and the like refer to the effect of the compositions disclosed herein, i.e., compositions comprising at least one probiotic bacterium, at least one prebiotic, and at least one lipid, all as defined herein. As used herein, "promoting" refers to enhancing or inducing compared to levels prior to administration of the compositions disclosed herein.

[0048]

[0046] The term "probiotic bacteria" refers to bacteria that are viable (live bacteria) and that, upon ingestion, generally provide a health benefit by improving or restoring the intestinal flora. The "probiotic bacteria" are preferably present in the composition in an effective amount. The manufacturing procedure for probiotic bacteria is typically standardized and involves fermenting the bacteria in a growth medium containing a carbohydrate source, such as a sugar, for example glucose, fructose, sucrose, lactose, or dextrose. After fermentation, the probiotic bacteria are usually cryoprotected, frozen, or lyophilized, and packaged into the final product used in the composition. The "probiotic bacteria" used according to the present invention are specifically selected taking into account the requirement for reducing uremic toxins, preferably, as already indicated above, making it possible to avoid or reduce the accumulation of such uremic toxins in the circulation, tissues, and organs.

[0049]

[0047] The term "prebiotic" should be understood to include indigestible fiber compounds that typically pass undigested through the upper gastrointestinal tract and stimulate the growth or activity of beneficial bacteria in the colon by acting as a substrate for such bacteria. The prebiotics of the present invention are specifically selected to enable stimulation of the growth or activity of the "probiotic bacteria" used in accordance with the present invention. The "prebiotics" are preferably present in the composition in an effective amount. Procedures for producing probiotic bacteria are typically standardized and well known to those skilled in the art.

[0050]

[0048] The term "average nucleotide identity (ANI)" is a measure of genomic similarity at the nucleotide level between the coding regions of two genomes. ANI can be easily determined by those skilled in the art using general knowledge and available tools, which are well described in the literature. For example, ANI can be evaluated as described in: Yoon SH, Ha SM, Lim J, Kwon S, Chun JA. Large-scale evaluation of algorithms to calculate average nucleotide identity. Antonie Van Leeuwenhoek. 2017 Oct;110(10):1281-1286.

[0051]

[0049] As used herein, the term "cardiometabolic condition" refers to any condition that includes a spectrum of conditions that are associated with or share risk factors, such as overweight and obesity, dyslipidemia, and hypertension.

[0052] As used herein, the term "neurodegenerative condition" refers to any condition involving the progressive loss of functional neurons in the central nervous system. In one embodiment, the neurodegenerative disease is associated with age-related cell death. Non-limiting examples of such neurodegenerative conditions include cardiometabolic or neurodegenerative conditions, particularly those related to the treatment or prevention of kidney disease, preferably chronic and acute; dialysis and pre-dialysis kidney disease; rare kidney diseases, genetically and metabolically induced kidney diseases; treatment or prevention of uremic syndromes, including PEW, bone loss, severe anorexia, fatigue, or inflammation; delaying progressive kidney disease complications, including anemia, hyperphosphatemia, secondary hyperparathyroidism, bone disease, and neurological conditions; delaying kidney disease comorbidities, including cardiovascular disease; risk prevention or management of malnutrition; delaying the progression of cardiometabolic disease; risk prevention or management of cardiovascular disease and comorbidities, such as diabetes; and / or risk prevention or management of neurodegenerative and neurological conditions.

[0053]

[0051] Neurodegenerative diseases include Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (also known as ALS and Lou Gehrig's disease), AIDS dementia complex, adrenoleukodystrophy, Alexander disease, Alpers disease, ataxia-telangiectasia, Batten disease, bovine spongiform encephalopathy (BSE), Canavan disease, corticobasal degeneration, Creutzfeldt-Jakob disease, dementia with Lewy bodies, and fatal familial insomnia. may further include neuroleptic malignant syndrome, frontotemporal lobar degeneration, Kennedy disease, Krabbe disease, Lyme disease, Machado-Joseph disease, multiple sclerosis, multiple system atrophy, neuroacanthocytosis, Niemann-Pick disease, Pick's disease, primary lateral sclerosis, progressive supranuclear palsy, Refsum's disease, Sandhoff's disease, diffuse myelinating sclerosis, spinocerebellar ataxia, subacute combined spinal degeneration, tabes dorsalis, Tay-Sachs disease, toxic encephalopathy, transmissible spongiform encephalopathy, and wobbly hedgehog syndrome.

[0054]

[0052] Sarcopenia is defined as the age-related decline in muscle mass and muscle function (including muscle strength and walking speed). As used herein, "frailty" is defined as a clinically identifiable state of increased vulnerability resulting from age-related declines in capacity and function across multiple body systems, resulting in an impaired ability to cope with routine or acute stressors. In the absence of established quantitative criteria, Fried et al. operationally define frailty as meeting three of the following five phenotypic criteria indicative of impaired energetics: (1) muscle weakness (grip strength in the bottom 20% of the population at baseline, adjusted for sex and body mass index), (2) poor endurance and energy (self-reported fatigue related to VO2 max), (3) bradykinesia (baseline in the bottom 20% of the population based on 15-foot walking time, adjusted for sex and height), (4) low physical activity (weighted score of kilocalories expended per week at baseline, lowest physical activity quintile identified for each sex; e.g., <383 kcal / week for men, <270 kcal / week for women), and / or unintentional weight loss (10 pounds in the past year). Fried LP, Tangen CM, Walston J et al., "Frailty in older adults: evidence for a phenotype." J. Gerontol. A. Biol. Sci. Med. Sci. 56(3):M146-M156 (2001). The presence of one or two of these criteria identifies the pre-frailty stage as being at high risk for progression to frailty.

[0055] "Cachexia" is a severely debilitating condition characterized by significant weight loss, anorexia, asthenia, and anemia. Cachexia is a common feature of many diseases, including cancer, sepsis, chronic heart failure, rheumatoid arthritis, and acquired immune deficiency syndrome (AIDS).

[0056] "Overweight" is defined in humans as a body mass index (BMI) of 25-30 kg / m 2"Obesity" is defined in humans as a BMI of at least 30 kg / m 2 , e.g., 30 to 39.9 kg / m 2 "Weight loss" is defined as a loss of total body weight. Weight loss can refer to a loss of total body weight to improve one or more of, for example, health, fitness, or appearance.

[0057] "Diabetes" includes both types I and II of the disease. Non-limiting examples of risk factors for diabetes include a waistline greater than 40 inches for men or 35 inches for women, blood pressure greater than 130 / 85 mmHg, triglycerides greater than 150 mg / dL, fasting blood glucose greater than 100 mg / dL, or high density lipoprotein less than 40 mg / dL for men or less than 50 mg / dL for women.

[0058]

[0056] As used herein, the term "metabolic syndrome" refers to a combination of medical abnormalities that, when occurring together, increase the risk of developing cardiovascular disease and diabetes. Metabolic syndrome affects one in five people in the United States, and prevalence increases with age. Some studies estimate the prevalence in the United States to be 25% of the population. According to the International Diabetes Foundation's consensus worldwide definition (2006), metabolic syndrome is central obesity in combination with any two of the following: Elevated triglycerides: greater than 150 mg / dL (1.7 mmol / L) or treatment directed at this lipid abnormality; Lowering HDL cholesterol: less than 40 mg / dL (1.03 mmol / L) in men and less than 50 mg / dL (1.29 mmol / L) in women, or treatment directed at this lipid abnormality; Elevated blood pressure: systolic BP > 130 or diastolic BP > 85 mmHg, or treatment of previously diagnosed hypertension; and Elevated fasting plasma glucose (FPG) >100 mg / dL (5.6 mmol / L) or previously diagnosed type 2 diabetes.

[0059]

[0057] Embodiment The present disclosure provides, according to a first embodiment, a composition suitable for use in reducing and / or avoiding the accumulation of uremic toxins, preferably in a cardiometabolic or neurodegenerative condition, the composition comprising a specifically selected prebiotic, a specifically selected probiotic, and a specifically selected lipid, as defined herein.

[0060] The compositions provide specific health benefits to treated patients. Such patients are typically those suffering from a cardiometabolic or neurodegenerative condition and are typically experiencing, or at least at risk of experiencing, an increase and accumulation of uremic toxins. The health benefits that may be advantageously provided to such patients by administering the compositions are preferably a delay in the progression of the disease and comorbidities, and also the possibility of managing the symptoms and syndromes associated with the toxic effects of such uremic solutes, both of which typically result from a reduction in the amount of uremic toxins in such patients that would otherwise accumulate in the patient.

[0061] Uremic toxins within the context of the present invention are typically selected from, but not limited to, urea, trimethylamine (TMA), triethylamine oxide (TMAO), indoxyl and indoxyl sulfate, p-cresol and p-cresol sulfate, p-cresyl glucorinate, uric acid, 3-carboxy-4-methyl-5-propyl-2-furanpropionate (CMPF), and the like, and all metabolic compounds or their precursors (see, e.g., FIG. 1). Such uremic toxins primarily originate from providing patients with excess amino acids, e.g., tryptophan or tyrosine, increased amounts of L-carnitine or choline, and increased amounts of urea. Such systemic uremic toxin accumulation is a condition frequently observed in individuals with cardiometabolic and neurological conditions, including, but not limited to, chronic kidney disease (CKD), as well as the diseases and conditions described herein. Such systemic uremic toxin accumulation also imposes a significant burden on the treatment of such diseases, especially when a high-protein diet is required. This is because, for example, in muscle wasting conditions such as cachexia or sarcopenia and in various neurological conditions, adding more protein is rather harmful due to the addition of more protein, especially the addition of more tryptophan and tyrosine.

[0062] In a first embodiment, the composition represents a novel nutritional and / or synbiotic blend that makes it possible to target multiple direct and indirect mechanisms that may contribute to a beneficial reduction in the development and / or accumulation of such uremic toxins, in particular a specifically selected probiotic as defined herein (e.g., L. johnsonii NCC 533); specifically selected prebiotics, as defined herein, preferably fiber and oligosaccharides, such as cellobiose and pea GOS, as defined herein; Compositions are provided that make this possible by applying compositions that preferably represent a specific blend composed of specifically selected lipids as defined herein, preferably triglycerides composed of short and medium chain fatty acids as defined herein.

[0063]

[0061] The compositions, for example in the form of nutritional and / or synbiotic blends, actively target and improve the microbiome (function), gut function, liver metabolism, provide a mechanism to stabilize kidney function, and help reduce the systemic generation and amount of uremic toxins.

[0064]

[0062] Preferably, the microbiome (function) can be improved as follows: The microbiome normally contributes to the production of certain metabolites that form precursors of uremic toxins. When such metabolites are available in body tissues, particularly the liver and circulatory system, they can be converted into uremic toxins. Importantly, patients with kidney disease have been shown to have dysbiosis, characterized by impaired microbiome function that converts certain amino acids from dietary proteins into uremic toxin precursor metabolites. Thus, administration of the composition improves dysbiosis, particularly the excessive availability of uremic toxin precursors in the circulatory system, by improving microbiota function. In other words, the imbalance in the microbiome is improved, thereby enabling the metabolic processing of such uremic toxin precursor metabolites to also be improved.

[0065]

[0063] The composition further advantageously makes it possible to enhance or improve intestinal function. In this context, it is important to recognize that the microbiota of both healthy and diseased individuals continuously and normally produces uremic toxin precursors without any obvious disease outcome. However, patients with cardiometabolic diseases, particularly kidney disease, may have impaired intestinal function, including reduced intestinal motility, impaired intestinal epithelial barrier function, and increased inflammation, which may contribute to dysregulation of the accumulation of uremic toxin precursors in the intestine and increased permeability of uremic toxin precursors across the intestinal barrier. The composition thereby improves the availability of uremic toxin precursors in the systemic circulation by improving intestinal motility and epithelial barrier function, thereby significantly contributing to avoiding the accumulation of such uremic toxins in tissues and organs, particularly the kidneys.

[0066] The composition also provides significant benefits to liver metabolism. The liver is a key tissue involved in the production of uremic toxins. With regard to production, liver enzymes use substrates derived from endogenous sources or from other tissues, diet, and the intestinal microflora to produce such uremic toxins. In this particular context, the composition makes it possible to improve the metabolism leading to uremic toxin conversion, particularly when substrate availability is in excess, by specific preselection of the administered probiotics, prebiotics, and lipids. As a result, the composition improves hepatic metabolism, thereby improving uremic toxin accumulation in the circulation and effectively avoiding the undesirable accumulation of these uremic toxins in the circulation and liver.

[0067] The composition also provides significant benefits for stabilizing renal function. Under normal, healthy conditions, the kidneys filter uremic toxins, thereby reducing or preventing their accumulation in the blood circulation. However, in situations where proper renal filtration is compromised, uremic toxins can accumulate, potentially initiating or contributing to a vicious cycle of disease exacerbation and progression, including further induction of dysbiosis or liver or intestinal dysfunction. The composition can stabilize renal function and prevent further decline. Thus, the composition directly ameliorates the accumulation of uremic toxins and indirectly ameliorates the production of uremic toxins.

[0068]

[0066] Thus, the present invention is based on a novel combination of specifically selected probiotics, specifically selected prebiotics, and specifically selected lipids, which in combination, in a synbiotic manner, make it possible to improve the levels and avoid the accumulation of uremic toxins (e.g., indoxyl sulfate, p-cresyl sulfate, PCG, CMPF, uric acid) in a variety of cardiometabolic (e.g., kidney disease) and neurodegenerative conditions.

[0069] In the context of the present invention, such avoidance of the accumulation of uremic toxins particularly relates to the compounds urea, uric acid, p-cresol, p-cresyl sulfate, indoxyl sulfate, indole, indoleacetic acid, indoxyl, trimethylamine, TMAO, dimethylglycine, betaine, PCG, CMPF, glutarate and / or other protein-bound uremic toxins, preferably indoxyl sulfate, p-cresyl sulfate, MCPF, PCG, urea and / or uric acid. The measurement of such compounds in tissues, organs and body fluids is well known to those skilled in the art.

[0070]

[0068] In this context, the inventors have surprisingly found that a composition comprising all of the ingredients required herein, i.e., a specifically selected prebiotic, a specifically selected prebiotic, and a specifically selected lipid, not only makes it possible to address individual aspects, e.g., dysbiosis or improving microbiome function, but also represents a holistic approach to alleviating multiple targets known to contribute to both the production and accumulation of uremic toxins, in particular to improve the microbiome (function), gut function, liver metabolism, and preferably provides a mechanism that helps to stabilize kidney function and reduce the systemic appearance and amount of uremic toxins in cardiometabolic or neurodegenerative conditions as defined herein.

[0071] The composition comprises a probiotic as already defined above. Probiotics, as defined herein, are considered to be live bacteria that contribute beneficially to intestinal and microbiome function. The probiotic bacteria of the composition have a significant impact on this treatment and are selected according to the following three criteria, which may be applied separately or in combination, preferably in combination:

[0072]

[0070] According to a first criterion, such probiotic bacteria are selected from at least one species of probiotic bacteria that are deficient in at least one bacterial enzyme that produces urea, uric acid, p-cresol, p-cresyl sulfate, indoxyl sulfate, indole, indoleacetic acid, indoxyl, trimethylamine, TMAO, dimethylglycine, betaine and / or glutarate.

[0073]

[0071] The deficiency of at least one bacterial enzyme is typically achieved by partial or complete deletion or at least inactivation of the corresponding gene coding sequence of such enzyme in the genome of the probiotic bacterium, resulting in the lack of expression of such enzyme in vivo. In cases where multiple copies of the bacterial enzyme are encoded by the genomic sequence, only some or all, preferably all, of the coding sequences for the bacterial enzyme are deleted or at least inactive.

[0074]

[0072] By avoiding the expression of enzymes required for the production of such uremic toxins, it is possible to reduce the amount of uremic toxins in the body, tissues, and organs, which would otherwise become excessively proliferated and overloaded, potentially leading to irreversible accumulation of such toxins. The selected probiotic bacteria are bacteria generally considered to be beneficial in the gut health system, but the inventors surprisingly recognized that uremic toxins can be effectively reduced or at least limited by synbiotic selection of specific types of probiotics, prebiotics, and lipids. The probiotic bacteria used in the composition are specifically selected for certain properties, such as probiotic bacteria lacking certain bacterial enzymes that may contribute to the excessive production of uremic toxins (first and second alternatives below), and / or probiotic bacteria that contribute to improved probiotic bacterial growth by promoting the expression of certain enzymes beneficial for processing certain prebiotics (fiber and carbohydrates).

[0075] According to the second criterion, and preferably in addition to the first criterion, the probiotic bacterium is selected from at least one probiotic bacterium lacking at least one bacterial enzyme selected from urease, carnitine monooxygenase & reductase, tryptophanase, hydroxyphenylacetate. Such enzymes may preferably be selected from one of the following classes of enzymes: EC1.14.13.239、EC:1.14.12.17、EC:1.14.13.25、EC:1.14.99.-、EC:1.3.3.4、EC:1.3.8.2、EC:1.3.98.1、EC:1.4.1.13、EC:1.4.1.14、EC:1.4.3.2、EC:2.1.1.12、EC:2.3.1.54、EC:2.6.1.-、EC:2.6.1.1、EC:2.6.1.2、EC:2.6.1.2、EC:2.6.1.4、EC:2.6.1.44、EC:2.6.1.5、EC:2.6.1.57、EC:2.6.1.58、EC:2.6.1.78、EC:2.6.1.79、EC:2.6.1.9、EC:3.2.1.172、EC:3.5.1.111、EC:3.5.1.3、EC:3.5.1.4、EC:3.5.5.1、EC:3.6.1.3、EC:4.1.1.105、EC:4.1.1.16、EC:4.1.1.25、EC:4.1.1.25、EC:4.1.1.28、EC:4.1.99.1、EC:4.3.99.4、EC:4.4.1.8、EC:5.4.99.9、EC:6.3.5.-、EC:6.3.5.1、EC:6.3.5.7、EC:1.13.11.63、EC:1.13.12.3、EC:1.14.13.148、EC:1.14.13.239、EC:1.18.1.2、EC:1.3.3.4、EC:1.4.1.20、EC:1.4.1.4、EC:1.4.3.2、EC:1.5.1.34、EC:1.8.1.19、EC:1.97.1、EC:2.6.1.-、EC:2.6.1.1、EC:2.6.1.39、EC:2.6.1.57、EC:2.6.1.78、EC:2.6.1.79、EC:2.6.1.9、EC:3.5.1.3、EC:3.5.2.12、EC:4.1.1.15、EC:4.1.1.83、EC:4.4.1.8

[0076] As already outlined for the first criterion, the deficiency of at least one such bacterial enzyme is typically achieved by partial or complete deletion or at least inactivation of the corresponding gene coding sequence of such enzyme in the genome of the probiotic bacterium, resulting in the lack of in vivo expression of such enzyme. In cases where multiple copies of the bacterial enzyme are encoded by the genomic sequence, only some or all, preferably all, of the coding sequences for the bacterial enzyme are deleted or at least inactive.

[0077]

[0075] Furthermore, the deficiency of at least one of the bacterial enzymes urease, carnitine monooxygenase and reductase, tryptophanase, and hydroxyphenylacetate effectively contributes to avoiding further accumulation of uremic toxins that would otherwise be produced in excess and further accumulate beyond tolerable levels in the patient's body, tissues, and organs.

[0078]

[0076] Alternatively, or in addition to either of the above two criteria, preferably further, such probiotic bacteria are preferably selected according to the third criterion from at least one probiotic bacterium having at least one of the following bacterial carbohydrate enzymes: α-galactosidase, β-galactosidase, glucan 1,4-β-glucosidase, cellulase, β-fructofuranosidase and / or licheninase. Such enzymes may be selected from one of the following enzyme classes: EC3.2.1.22, EC3.2.1.23, EC3.2.1.74, EC3.2.1.4, EC3.2.1.26, EC3.2.1.58, EC3.2.1.73, EC2.4 .1.9, EC3.2.1.84, EC3.2.1.33, EC3.2.1.70, EC3.2.1.135, EC3.2.1.3, EC3.2.1.20, EC3.2.1.10

[0079]

[0077] The expression of at least one of these bacterial carbohydrate-degrading enzymes via specifically selected probiotics specifically contributes to the processing of the prebiotics administered herein, providing synbiotic support (in terms of the selection of probiotics and prebiotics) for the growth of the administered probiotics, improvement of the microbiome and gut function, and stabilization of liver metabolism and kidney function, thereby helping to reduce and / or avoid the accumulation of uremic toxins.

[0080]

[0078] The probiotic bacteria of the composition meet at least one, preferably at least two, more preferably at least three, or all of the above criteria for the probiotics of the present invention.

[0081] As already outlined, probiotic bacteria according to any of these criteria, preferably meeting all three criteria, are preferably selected from the following species: Bifidobacterium animalis subsp. lactis, Bifidobacterium longum subsp. infantis, Bifidobacterium longum subsp. longum, Enterococcus faecium, Lactobacillus johnsonii, Lactococcus lactis, Lacticaseibacillus paracasei (formerly classified as Lactobacillus paracasei), Lacticaseibacillus reuteri (formerly classified as Lactobacillus reuteri), Lacticaseibacillus rhamnosus ( The probiotic bacteria may be selected from Lactobacillus rhamnosus (formerly classified as Lactobacillus rhamnosus), Staphylococcus carnosus and / or Streptococcus thermophilus, preferably Lactobacillus johnsonii, or a probiotic bacterium having a genome which has at least 95% average nucleotide identity (ANI) to the genome sequence of any one of the heretofore defined probiotic bacteria, preferably at least 96% ANI, more preferably at least 97% ANI, even more preferably at least 98% ANI, and most preferably at least 99% ANI, or even 99.5% ANI, or 99.9% ANI.

[0082] Even more preferably, such probiotic bacteria, preferably meeting all three criteria, may be selected from any of the following strains: a. Bifidobacterium animalis subsp. lactis NCC 2818, deposited at the CNCM on June 7, 2005, and assigned accession number CNCM I-3446; b. Bifidobacterium longum subsp. infantis NCC 341 ATCC 15697(T) - available at https: / / www.atcc.org / products / 15697); c. Bifidobacterium longum subsp. longum NCC 2705 CNCM I-2618? (NCBI refseq; GCA_000007525.1) [NCC 2075 was deposited at the CNCM on January 29, 2001, and assigned accession number CNCM I-2618]; d. Enterococcus faecium NCC 2768 (NCIMB 10415 available from Cerbios-Pharma SA Barbengo Switzerland (cerbios.swiss / e-faecium-sf68-a-model-for-efficacy-safety-for-pharmaceutical-probiotics / ); e. Lactobacillus johnsonii NCC 533 [originally known as La 1, deposited at the CNCM on June 30, 1992, and assigned accession number CNCM I-1225 (see also NCBI refseq; GCA_000008065.1, and GenBank AE017198.1 (included as SEQ ID NO: 1)]; f. Lactococcus lactis NCC 2287 (CNCM I-4154) [NCC 2287 was deposited with the CNCM on April 24, 2009, and assigned the accession number CNCM I-4154]; g. lacticasei Bacillus paracasei NCC 2461 (CNCM I-2116) [NCC 2461 was deposited with the CNCM on January 12, 1999 and assigned accession number CNCM I-2116]; h. lactisasei Bacillus rhamnosus NCC 4007 (CGMCC 1.3724) [NCC 4007 was deposited in October 2004 at the CGMCC [China General Microbiological Culture Collection Center (CGMCC) Institute of Microbiology, Chinese Academy of Sciences, PO Box 2714, Beijing 100080, China] under the identification number CGMCC 1.3724]; i. Staphylococcus carnosus NCC 1052 (CNCM I-5400) [NCC 1052 was deposited with the CNCM on February 1, 2019, and assigned the accession number CNCM I-5400]; j. Staphylococcus carnosus NCC 971 (CNCM I-5398) [NCC 971 was deposited with the CNCM on February 1, 2019, and assigned accession number CNCM I-5398]; and / or k. Streptococcus thermophilus NCC 2496 (CNCM I-3915) [NCC 2496 was deposited with the CNCM on February 5, 2008, and assigned the accession number CNCM I-3915]; or a. to k., a probiotic bacterium having a genome having at least 95% average nucleotide identity (ANI) to each of the genome sequences set forth in any one of a. to k., preferably at least 96% ANI, more preferably at least 97% ANI, even more preferably at least 98% ANI, and most preferably at least 99% ANI, or even 99.5% ANI or 99.9% ANI, as defined above.

[0083]

[0081] Most preferably, the probiotic bacteria of the compositions defined herein are selected from Lactobacillus johnsonii NCC 533 (CNCM I-1225) or probiotics having a genome with an ANI of at least 95%, preferably at least 96%, more preferably at least 97%, even more preferably at least 98%, and most preferably at least 99%, or even 99.5% or 99.9%, relative to the genome sequence of Lactobacillus johnsonii NCC 533 (CNCM I-1225). The NCBI reference sequence for Lactobacillus johnsonii NCC 533 (CNCM I-1225) is GCA_000008065.1 (SEQ ID NO: 1).

[0084] Typically, an "effective amount" of "probiotic bacteria" as defined herein is typically 10 per daily dose. 3 cfu~10 12 cfu, typically 10 per daily dose 4 cfu~10 11 cfu, preferably 10 per daily dose 5 cfu~10 10 cfu, or 10 per daily dose 5 cfu~10 9 cfu, also preferably 10 per daily dose 6 cfu~10 9 cfu, 10 per daily dose 6 cfu~10 8 cfu, or 10 per daily dose 8 cfu~10 10 cfu per daily dose, more preferably about 10 7 cfu~10 9 A preferred daily dose may contain a total of about 10 cfu of such probiotic bacteria per daily dose. 8 cfu, 10 per daily dose 7 ~10 9 cfu, or 10 per daily dose 8 ~10 9cfu. Thus, the daily dose can be achieved by a single daily administration of the composition or by multiple administrations of the composition, for example, 2, 3, 4, or 5 administrations, preferably 1 to 5 or less, more preferably 1 to 4 or less, and even more preferably 1 to 3 or less. When the daily dose is achieved by a single daily administration or 1 to 5 administrations of the composition, the amount per composition is recalculated based on the required daily dose. In such multiple daily administrations, it is preferred that any administration or each composition contains the same amount of ingredients, thus probiotics, prebiotics, and lipids, and preferably the same volume.

[0085] According to a second aspect, the composition defined herein also comprises a prebiotic as already generally defined above, said prebiotic being selected from carbohydrates, preferably fibres and oligosaccharides, the carbohydrates being hydrolysable by at least one bacterial enzyme selected from α-galactosidase, β-galactosidase, glucan 1,4-β-glucosidase, cellulase, β-fructofuranosidase and / or licheninase, preferably Selected from EC3.2.1.22, EC3.2.1.23, EC3.2.1.74, EC3.2.1.4, EC3.2.1.26, EC3.2.1.58, EC3.2.1.73, EC2.4.1.9, EC3.2.1.84, EC3.2.1.33, EC3.2.1.70, EC3.2.1.135, EC3.2.1.3, EC3.2.1.20, EC3.2.1.10.

[0086]

[0084] Similarly, such prebiotics are selected from the family of α-galactooligosaccharides / raffinose (e.g., pea GOS, soybean GOS), β-galactooligosaccharides (e.g., β-GOS, Vivinal GOS, cow's milk oligosaccharides (BMOS)), human milk oligosaccharides (HMO), cellooligosaccharides (COS) (e.g., cellobiose, cellotriose, cellotetraose, soluble hydrolyzed wheat, soluble hydrolyzed oats, and / or β-glucan hydrolysates containing cellobiose or cellotriose or cellotetraose), or combinations thereof.

[0087]

[0085] The specific selection of prebiotics in the composition acts synbiotically in conjunction with the specific selection of probiotic bacteria of the composition defined above. Such prebiotics are preferably and advantageously specific carbohydrates, preferably fiber and oligosaccharides, that support the growth of the specifically selected probiotic bacteria. More specifically, the specific carbohydrates act as substrates for the specifically selected probiotic bacteria and, depending on the specific selection of probiotic bacteria, can be hydrolyzed by at least one bacterial enzyme selected from α-galactosidase, β-galactosidase, glucan 1,4-β-glucosidase, cellulase, β-fructofuranosidase, and / or licheninase, which are produced by the probiotic bacteria of the composition defined above.

[0088]

[0086] According to a further aspect, the prebiotic is preferably contained in an effective amount in the composition as defined herein. More preferably, such effective amount of prebiotic preferably contained in the composition as defined herein is an amount of 0.1 g to 30 g per daily dose, preferably an amount of 2 g to 15 g per daily dose.

[0089]

[0087] According to a third aspect, the composition comprises, in addition to the specifically selected probiotic and the specifically selected prebiotic as described above, also a specifically selected lipid.

[0090]

[0088] Such lipids include at least one lipid selected from triglycerides, short-chain fatty acids, and / or medium-chain fatty acids, where the triglycerides include short-chain fatty acids, medium-chain fatty acids, long-chain fatty acids, or mixtures thereof. Preferred triglycerides include triglycerides containing butyrate and / or caprylate, triglycerides consisting of butyrate and caprylate, and triglycerides consisting of butyrate and oleate. In one embodiment, the triglycerides are composed of butyrate and caprylate. The inventors have surprisingly found that these specifically selected lipids advantageously support the improvement of the microbiome, intestinal function, liver metabolism, and stabilization of kidney function in a symbiotic manner, thereby significantly contributing to the avoidance of both the production and accumulation of uremic toxins.

[0091]

[0089] In this context, "short-chain triglycerides" (SCTs) are preferably understood as triglycerides having two or three fatty acids each having 1 to 5 carbon atoms, preferably 2 to 5 carbon atoms. Such fatty acids having 1 to 5 carbon atoms, preferably 2 to 5 carbon atoms, are generally understood herein as short-chain fatty acids (SCFAs). Particularly preferred short-chain fatty acids are C4 fatty acids, most preferably butyric acid (C4:0). Short-chain triglycerides containing butyric acid are preferably SCTs containing at least one, two, or even three butyric acids.

[0092] Similarly, in this context, "medium-chain triglycerides" (MCTs) are understood as triglycerides having preferably two or three fatty acids each having 6 to 12 carbon atoms. Such fatty acids having 6 to 12 carbon atoms are generally understood as medium-chain fatty acids (MCFAs). Preferred short-chain fatty acids are C8 fatty acids, for example, caprylic acid (C8:0). A medium-chain triglyceride containing caprylic acid may be an MCT containing at least one, at least two, or even three caprylic acids.

[0093] In the context of triglycerides containing a mixture of butyrate and long-chain fatty acids, such triglycerides are understood to be triglycerides having preferably one or two butyrate moieties and one or two long-chain fatty acids, where long-chain fatty acids are generally understood to be fatty acids having 13 to 21 carbon atoms. Exemplary triglycerides composed of butyrate and containing long chain fatty acids include 1,3-dibutyryl-2-linoleoylglycerol, 1,3-dibutyryl-2-stearoylglycerol, 1-butyryl-2-oleoyl-3-palmitoylglycerol, 1-palmitoyl-2-oleoyl-3-butyrylglycerol, 1-butyryl-2-oleoyl-3-linoleoylglycerol, 1-linoleoyl-2-oleoyl-3-butyrylglycerol, and 1-oleoyl-2-butyryl-3-linoleoylglycerol. , 1-linoleoyl-2-butyryl-3-oleoylglycerol, 1-butyryl-2-linoleoyl-3-oleoylglycerol, 1-oleoyl-2-linoleoyl-3-butyrylglycerol, 1-butyryl-2-stearoyl-3-oleoylglycerol, 1-oleoyl-2-stearoyl-3-butyrylglycerol, 1-butyryl-2-oleoyl-3-stearoylglycerol, and / or 1-stearoyl-2-oleoyl-3-butyrylglycerol, and mixtures of two or more thereof. In the context of triglycerides comprising a mixture of butyrate and long chain fatty acids, a preferred long chain fatty acid is oleic acid (C18:1).

[0094]

[0092] More preferably, the lipid in the context of the composition is selected from a. triglycerides (TG) composed of a mixture of butyrate and medium-chain fatty acids (MCFA); b. triglycerides (TG) composed of a mixture of short-chain fatty acids and medium-chain fatty acids (SMCFA); c. triglycerides (TG) composed of a mixture of butyrate and long-chain fatty acids (e.g., a mixture of butyrate and oleate); d. triglycerides (TG) composed of medium-chain fatty acids; e. triglycerides (TG) composed of short-chain fatty acids; f. short-chain fatty acids that can be metabolized into ketone bodies; and / or g. medium-chain fatty acids that can be metabolized into ketone bodies, preferably C4 and / or C8 medium-chain fatty acids (e.g., butyric acid (C4:0) and / or caprylic acid (C8:0)). Exemplary triglycerides in the context of the composition are triglycerides comprising butyrate and / or caprylate, triglycerides consisting of butyrate and caprylate, and triglycerides consisting of butyrate and oleate. In one aspect, the triglyceride is composed of butyrate and caprylate.

[0095] Particularly preferred as lipids in the context of the composition are triglycerides containing butyrate, short chain fatty acids, medium chain fatty acids, or mixtures thereof. Triglycerides containing at least one short chain fatty acid C1 to C5, preferably C2 to C5; preferably triglycerides containing at least one butyric acid (C4:0), or Triglycerides containing at least one medium chain fatty acid C6 to C12, for example caprylic acid (C8:0), preferably at least one caprylic acid (C8:0), or It may also be a triglyceride containing at least one short-chain fatty acid C1 to C5, preferably C2 to C5, such as butyric acid (C4:0), and at least one medium-chain fatty acid C6 to C12, such as caprylic acid (C8:0).

[0096] In an even more preferred embodiment, the lipid in the context of the composition can be selected from triglycerides containing either butyric acid (C4:0) (BBB, tributyrin) or caprylic acid (C8:0) (CCC, tricaprylin), preferably a mixture of triglycerides containing either butyric acid (C4:0) or caprylic acid (C8:0), or a triglyceride containing both butyric acid (C4:0) and caprylic acid (C8:0) in the same triglyceride. The latter triglycerides can be prepared by interesterification of a mixture of triglycerides containing either butyric acid (C4:0) or caprylic acid (C8:0). BBB, tributyrin, is commercially available (e.g., from Sigma-Aldrich). CCC, tricaprylin, is commercially available, for example, as Neobee 895 (e.g., from Stepan Specialty).

[0097]

[0095] Methods for preparing triglycerides composed of a mixture of butyrate and long-chain fatty acids are known in the art, as described, for example, in International Publication No. 2019228851, which is incorporated by reference in its entirety. Exemplary triglycerides composed of butyrate and containing long-chain fatty acids include 1,3-dibutyryl-2-linoleoylglycerol, 1,3-dibutyryl-2-stearoylglycerol, 1-butyryl-2-oleoyl-3-palmitoylglycerol, 1-palmitoyl-2-oleoyl-3-butyrylglycerol, 1-butyryl-2-oleoyl-3-linoleoylglycerol, 1-linoleoyl-2-oleoyl-3-butyrylglycerol, and 1-oleoyl-2-butyryl-3-linoleoylglycerol.

[0013] Examples of triglycerides include 1-linoleoyl-2-butyryl-3-oleoylglycerol, 1-butyryl-2-linoleoyl-3-butyrylglycerol, 1-oleoyl-2-linoleoyl-3-butyrylglycerol, 1-butyryl-2-stearoyl-3-oleoylglycerol, 1-oleoyl-2-stearoyl-3-butyrylglycerol, 1-butyryl-2-oleoyl-3-stearoylglycerol, and / or 1-stearoyl-2-oleoyl-3-butyrylglycerol, and mixtures of two or more thereof. In the context of triglycerides comprising a mixture of butyrate and long-chain fatty acids, a preferred long-chain fatty acid is oleic acid (C18:1). A preferred triglyceride composed of a mixture of short-chain and long-chain fatty acids is a triglyceride composed of a mixture of butyrate and oleate.

[0098]

[0096] In the context of the composition, preferred are triglycerides comprising fatty acids that can be metabolized into ketone bodies, preferably C4 and / or C8 short-chain and medium-chain fatty acids, more preferably triglycerides comprising both butyric acid (C4:0) and caprylic acid (C8:0) in the same triglyceride; or triglycerides comprising either butyric acid (C4:0) or caprylic acid (C8:0) in the same triglyceride; or mixtures of triglycerides, one triglyceride comprising butyric acid (C4:0) and one triglyceride comprising caprylic acid (C8:0).

[0099] Alternatively, but less preferably, the lipids may not be triglycerides and may be selected from short-chain fatty acids (SCFAs) and medium-chain fatty acids (MCFAs). Thus, such lipids may be selected from at least one short-chain fatty acid C1-C5, preferably C2-C5, more preferably butyric acid (C4:0), or from at least one medium-chain fatty acid C6-C12, such as caprylic acid (C8:0), or from a mixture of at least one short-chain fatty acid C1-C5, preferably C2-C5, such as butyric acid (C4:0), with at least one medium-chain fatty acid C6-C12, such as caprylic acid (C8:0).

[0100]

[0098] According to a further aspect, lipids are preferably contained in the compositions defined herein in an amount of from 0.1 g to 30 g per daily dose, preferably in an amount of from 2 g to 15 g per daily dose.

[0101] According to a preferred embodiment, the composition defined herein comprises (a) probiotic bacteria, preferably probiotic bacteria lacking genes that produce at least one of urea, uric acid, p-cresol, p-cresyl sulfate, indoxyl sulfate, indole, indoleacetic acid, indoxyl, trimethylamine, TMAO, dimethylglycine, betaine, and / or glutarate; probiotic bacteria deficient in at least one bacterial enzyme selected from urease, carnitine monooxygenase & reductase, tryptophanase, and / or hydroxyphenylacetate; and / or a bacterium expressing at least one bacterial enzyme selected from α-galactosidase, β-galactosidase, glucan 1,4-β-glucosidase, cellulase, β-fructofuranosidase, and / or licheninase; and a probiotic bacterium selected from the group consisting of: (b) a prebiotic selected from carbohydrates, wherein the carbohydrates are selected from α-galactosidase, β-galactosidase, glucan 1,4-β-glucosidase, cellulase β-fructofuranosidase and / or licheninase, preferably EC 3.2.1.22, EC 3.2.1.23, EC 3.2.1.74, EC 3.2.1.4, EC 3.2.1.26, EC 3.2.1.58, EC 3.2.1.73, EC 2 EC 3.4.1.9, EC 3.2.1.84, EC 3.2.1.33, EC 3.2.1.70, EC 3.2.1.135, EC 3.2.1.3, EC 3.2.1.20, EC 3.2.1.10, or preferably the carbohydrates are selected from α-galactooligosaccharides / raffinose (e.g., pea GOS, soybean GOS), β-galactooligosaccharides (e.g., b-GOS, Vivinal GOS), a prebiotic selected from the family of GOS, bovine milk oligosaccharides (BMOS)), human milk oligosaccharides (HMO) (e.g., 2'-fucosyllactose (2'FL), 3-fucosyllactose (3'FL), lacto-N-neotetraose (LNnT), lacto-N-tetraose (LNT), 3'-sialyllactose (3SL), and / or 6'-sialyllactose (6SL)), cellooligosaccharides (COS) (e.g., cellobiose, cellotriose, cellotetraose, soluble hydrolyzed wheat, soluble hydrolyzed oats, and / or β-glucan hydrolysates containing cellobiose, cellotriose, or cellotetraose), or combinations thereof; (c) a lipid selected from a triglyceride, a short chain fatty acid, and / or a medium chain fatty acid, wherein the triglyceride comprises a short chain fatty acid, a medium chain fatty acid, or a mixture thereof, and further, in one embodiment, the triglyceride comprises butyrate and / or caprylate.

[0102] According to a more preferred embodiment, the composition defined herein may comprise probiotic bacteria, prebiotics and lipids as defined above, (a) The probiotic bacteria are the probiotic bacteria described in a. to k. as defined below: a. Bifidobacterium animalis subsp. lactis NCC 2818 (CNCM I-3446); b. Bifidobacterium longum subsp. infantis NCC 341 (ATCC 15697(T)); c. Bifidobacterium longum subsp. longum NCC 2705 (CNCM I-2618) (NCBI refseq; GCA_000007525.1); d. Enterococcus faecium NCC 2768 (NCIMB 10415); e. Lactobacillus johnsonii NCC 533 (CNCM I-1225) (NCBI refseq; GCA_000008065.1); f. Lactococcus lactis NCC 2287 (CNCM I-4154); g. Lacticasei Bacillus paracasei NCC 2461 (CNCM I-2116); h. lacticasei Bacillus rhamnosus NCC 4007 (CGMCC 1.3724); i. Staphylococcus carnosus NCC 1052 (CNCM I-5400); j. Staphylococcus carnosus NCC 971 (CNCM I-5398); and / or k. Streptococcus thermophilus NCC 2496 (CNCM I-3915); or a probiotic bacterium having a genome with an ANI of at least 95% relative to any one of the genome sequences of the probiotic bacteria described in a. to k. above, preferably at least 96% ANI, more preferably at least 97% ANI, even more preferably at least 98% ANI, most preferably at least 99% ANI, or even 99.5% ANI, or 99.9% ANI; or a combination of one or more of the above probiotic bacteria; (b) the prebiotic is selected from the family of α-galactooligosaccharides / raffinose (e.g., pea GOS, soybean GOS), β-galactooligosaccharides (e.g., β-GOS, Vivinal GOS, milk oligosaccharides), cellooligosaccharides (COS) (e.g., cellobiose, cellotriose, cellotetraose, soluble hydrolyzed wheat, soluble hydrolyzed oats, and / or β-glucan hydrolysates containing cellobiose, cellotriose, or cellotetraose), human milk oligosaccharides (HMO) (e.g., 2'-fucosyllactose (2'FL), 3-fucosyllactose (3'FL), lacto-N-neotetraose (LNnT), lacto-N-tetraose (LNT), 3'-sialyllactose (3SL), and / or 6'-sialyllactose (6SL)), or a combination thereof; (c) the lipid is selected from a triglyceride, a short chain fatty acid, and / or a medium chain fatty acid, wherein the triglyceride comprises a short chain fatty acid, a medium chain fatty acid, or a mixture thereof, and further, in one embodiment, the triglyceride comprises butyrate and / or caprylate. Preferably, the lipid is a. Triglycerides (TG) composed of a mixture of butyrate and medium-chain fatty acids (MCFA); b. Triglycerides (TG) composed of a mixture of short- and medium-chain fatty acids (SMCFA); c. Triglycerides (TG) composed of a mixture of butyrate and long-chain fatty acids (e.g., triglycerides composed of a mixture of butyrate and oleate); d. Triglycerides composed of medium-chain fatty acids; e. Triglycerides (TG) composed of short-chain fatty acids; f. short-chain fatty acids that can be metabolized into ketone bodies; and / or g. Selected from short- and medium-chain fatty acids that can be metabolized to ketone bodies, preferably C4 and / or C8 short- and medium-chain fatty acids (e.g., butyric acid (C4:0) and / or caprylic acid (C8:0)).

[0103] According to an even more preferred embodiment, the composition defined herein may comprise a probiotic, a prebiotic and a lipid as defined above, (a) The probiotic bacterium is selected from Lactobacillus johnsonii NCC 533 (CNCM I-1225) or a probiotic having a genome with at least 95% ANI relative to the genome sequence of Lactobacillus johnsonii NCC 533 (CNCM I-1225) (SEQ ID NO: 1), preferably at least 96% ANI, more preferably at least 97% ANI, even more preferably at least 98% ANI, and most preferably at least 99% ANI, or even 99.5% ANI, or 99.9% ANI. (b) the prebiotic is selected from the family of α-galactooligosaccharides / raffinose (e.g., pea GOS, soybean GOS), β-galactooligosaccharides (e.g., β-GOS, Vivinal GOS, milk oligosaccharides), cellooligosaccharides (COS) (e.g., cellobiose, cellotriose, cellotetraose, soluble hydrolyzed wheat, soluble hydrolyzed oats, and / or β-glucan hydrolysates containing cellobiose or cellotriose or cellotetraose), or combinations thereof; (c) the lipid is selected from triglycerides, short chain fatty acids and / or medium chain fatty acids, wherein the triglycerides comprise short chain fatty acids, medium chain fatty acids, or mixtures thereof, and further, in one embodiment, the triglycerides comprise butyrate and / or caprylate. More preferably, the lipid is a. Triglycerides (TG) composed of a mixture of butyrate and medium-chain fatty acids (MCFA); b. Triglycerides (TG) composed of a mixture of short- and medium-chain fatty acids (SMCFA); c. Triglycerides (TG) composed of a mixture of butyrate and long-chain fatty acids (e.g., triglycerides composed of a mixture of butyrate and oleate); d. Triglycerides composed of medium-chain fatty acids; e. Triglycerides (TG) composed of short-chain fatty acids; f. short-chain fatty acids that can be metabolized into ketone bodies; and g. Selected from short and medium chain fatty acids that can be metabolized into ketone bodies, preferably C4 and / or C8 short and medium chain fatty acids.

[0104] According to a further preferred embodiment, the composition defined herein may comprise a probiotic, a prebiotic and a lipid as defined above, (a) The probiotic bacterium is selected from Lactobacillus johnsonii NCC 533 (CNCM I-1225) or a probiotic having a genome with at least 95% ANI relative to the genome sequence of Lactobacillus johnsonii NCC 533 (CNCM I-1225) (SEQ ID NO: 1), preferably at least 96% ANI, more preferably at least 97% ANI, even more preferably at least 98% ANI, and most preferably at least 99% ANI, or even 99.5% ANI, or 99.9% ANI. (b) The prebiotic is selected from pea GOS, soy GOS, β-GOS, Vivinal GOS, cow's milk oligosaccharides, human milk oligosaccharides, cellobiose, cellotriose, cellotetraose, soluble hydrolyzed wheat, soluble hydrolyzed oats, and / or β-glucan hydrolysates containing cellobiose or cellotriose or cellotetraose, or combinations thereof. (c) Lipids are a. Triglycerides (TG) composed of a mixture of butyrate and medium-chain fatty acids (MCFA); b. Triglycerides (TG) composed of a mixture of short- and medium-chain fatty acids (SMCFA); c. Triglycerides (TG) composed of a mixture of butyrate and long-chain fatty acids (e.g., triglycerides composed of a mixture of butyrate and oleate); d. Triglycerides composed of medium-chain fatty acids; e. Triglycerides (TG) composed of short-chain fatty acids; f. short-chain fatty acids that can be metabolized into ketone bodies; and g. Selected from short- and medium-chain fatty acids that can be metabolized to ketone bodies, preferably C4 and / or C8 short- and medium-chain fatty acids (e.g., butyric acid (C4:0) and / or caprylic acid (C8:0)).

[0105]

[0103] According to another aspect, the composition as defined herein is selected from a food product or nutritional composition, a dietary supplement, a Food for Special Medical Purposes (FSMP), a nutritional supplement, a dairy-based beverage, a small volume liquid supplement, a functional food product, a functional beverage product, a meal replacement beverage, and combinations thereof. Further, without being limited thereto, the composition may be in an administrable form, preferably selected from a pharmaceutical formulation.

[0106]

[0104] In this context, it is particularly preferred that the composition may be solid or liquid and may be in the form of a powder, tablet, capsule or may be in the form of an oil formulation, emulsion, oil-in-water emulsion (o / w emulsion) or water-in-oil emulsion (w / o emulsion).

[0107] Each compound of the composition may be administered simultaneously with the other compounds (e.g., as a single unit) or separated by time intervals (e.g., in separate units). Preferably, the compounds are provided in single unit form.

[0108]

[0106] The present invention provides a nutrition-based solution, particularly for the management of uremic toxins associated with disease states and their associated complications. Typical applications of such compositions may be in the form of medical foods or FSMP (foods for special medical purposes) supplements, as ingredients in medical nutritional products, as an adjunct / combination with standard treatments, as an adjunct / combination with kidney disease medications such as SGLT inhibitors, as an adjunct / combination with nutritional materials targeting kidney mitochondrial dysfunction, as an adjunct / combination with protein meals, as dietary supplements, as dairy-based beverages, as small-volume liquid supplements, as meal replacement beverages, and combinations thereof.

[0109] According to a second embodiment, the present invention is directed to the use of a composition as defined herein for reducing or avoiding the accumulation of uremic toxins, preferably in the treatment of a cardiometabolic or neurodegenerative condition, or for preventing the accumulation of uremic toxins, preferably in the treatment of a cardiometabolic or neurodegenerative condition. Thus, the composition is preferably used for the reduction of uremic toxins in cardiometabolic or neurodegenerative conditions and associated comorbidities, for slowing the progression of such cardiometabolic or neurodegenerative conditions and associated comorbidities, and / or for managing symptoms and syndromes associated with the toxic effects of uremic solutes in such cardiometabolic or neurodegenerative conditions and associated comorbidities, and / or for preventing the accumulation of uremic toxins in the treatment of a cardiometabolic or neurodegenerative condition and associated comorbidities.

[0110]

[0108] Cardiometabolic or neurodegenerative conditions and associated comorbidities that can be treated by using the composition preferably include the following: Treatment or prevention of kidney disease, including chronic and acute kidney disease; dialysis and pre-dialysis kidney disease; rare kidney diseases, including genetically and metabolically induced kidney diseases; Treatment or prevention of PEW syndrome, uremic syndrome including bone loss, severe anorexia, fatigue, or inflammation; Delayed complications of progressive kidney disease, including anemia, hyperphosphatemia, secondary hyperparathyroidism, bone disease, and neurological conditions; Delayed renal comorbidities, including cardiovascular disease Preventing or managing the risk of malnutrition; Slowing the progression of cardiometabolic diseases; Prevention or management of the risk of cardiovascular disease and comorbidities, such as diabetes; and / or Prevention or management of the risk of neurodegenerative and neurological conditions.

[0111]

[0109] Generally, the subject of any such treatment or the patient in need of such treatment is a mammal, preferably a human or a pet, such as a companion animal, suffering from any of the above-mentioned cardiometabolic or neurodegenerative conditions and associated co-morbidities. Preferably, the subject for any such treatment or the patient in need of such treatment may be a child, a toddler or infant, an elderly person, a companion animal, but may also be a companion pet, such as a cat or a dog, and the subject is preferably at risk of developing or already has such a disease.

[0112]

[0110] According to a third embodiment, there is provided a method for treating a cardiometabolic or neurodegenerative condition as defined above, preferably comprising as a first step (a) preparing and providing a composition as discussed above, comprising a specifically selected probiotic, a specifically selected prebiotic, and a specifically selected lipid as defined above, and (b) administering such composition to a patient in need thereof, typically suffering from increased uremic toxins, typically in the context of a cardiometabolic or neurodegenerative condition as defined herein.

[0113] According to a fourth embodiment, there is provided a (multi-part) kit suitable for use in reducing and / or avoiding the accumulation of uremic toxins in a cardiometabolic or neurodegenerative condition as defined herein, comprising a composition as described herein, comprising specifically selected probiotics, bacteria, specifically selected prebiotics, and specifically selected lipids as defined above, for use in separate containers, for example as two or more liquid solutions or dry powders, to form one or more of the compositions disclosed herein and / or for use in one or more of the methods disclosed herein. The kit may also include instructions for use.

[0114] It should be understood that the various aspects and embodiments of the detailed description as disclosed herein are illustrative of specific ways to make and use the invention, and do not limit the scope of the invention when considered in conjunction with the claims and detailed description. It should also be understood that features from aspects and embodiments of the invention may be combined with additional features from the same or different aspects and embodiments of the invention.

[0115] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of chemistry, biochemistry, molecular biology, microbiology, and immunology which are within the capabilities of those skilled in the art and are explained in the literature. For example, Sambrook, J., Fritsch, EFand Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press; Ausubel, FMet al. (1995 and periodic supplements) Current Protocols in Molecular Biology, Ch.9,13 and 16, John Wiley & Sons; Roe, B., Crabtree, J. and Kahn, A. (1996) DNA Isolation and Sequencing: Essential Techniques, John Wiley & Sons; Polak, JMand McGee, J.O'D. (1990) In Situ Hybridization: Principles and Practice, Oxford University Press; Gait, MJ (1984) Oligonucleotide Synthesis: A Practical Approach, IRL Press and Lilley, DMand. Dahlberg, J.E. (1992) Methods in Enzymology: DNA Structures. See Part A: Synthesis and Physical Analysis of DNA, Academic Press. Each of these general texts is incorporated herein by reference.

[0116]

[0114] Preferred features and embodiments of the present invention will now be described by way of non-limiting examples.

[0117]

[0115] The present invention includes the following embodiments. Embodiment 1 1. A composition for use in reducing or avoiding the accumulation of uremic toxins in a cardiometabolic or neurodegenerative condition, comprising: (a) the following probiotic bacteria: probiotic bacteria lacking genes that produce at least one of urea, uric acid, p-cresol, p-cresyl sulfate, indoxyl sulfate, indole, indoleacetic acid, indoxyl, trimethylamine, TMAO, dimethylglycine, PCG, CMPF, betaine, and / or glutarate; probiotic bacteria deficient in at least one bacterial enzyme selected from urease, carnitine monooxygenase & reductase, tryptophanase, and / or hydroxyphenylacetate; and / or probiotic bacteria selected from probiotic bacteria expressing at least one bacterial enzyme selected from α-galactosidase, β-galactosidase, glucan 1,4-β-glucosidase, cellulase, β-fructofuranosidase and / or licheninase; (b) a prebiotic selected from a carbohydrate or fiber, wherein the carbohydrate or fiber can be hydrolyzed by at least one bacterial enzyme selected from α-galactosidase, β-galactosidase, glucan 1,4-β-glucosidase, cellulase, β-fructofuranosidase, and / or licheninase; (c) a lipid selected from triglycerides, short chain fatty acids and / or medium chain fatty acids, wherein the triglycerides comprise short chain fatty acids, medium chain fatty acids, long chain fatty acids, or mixtures thereof, and further wherein in one aspect the triglycerides comprise butyrate and / or caprylate, and in one aspect the triglycerides consist of butyrate and caprylate or butyrate and oleate, and further preferably the triglycerides consist of butyrate and caprylate. Embodiment 2 The probiotic bacteria (a) are selected from at least one of the following a. to k.: a. Bifidobacterium animalis subsp. lactis NCC 2818 (CNCM I-3446); b. Bifidobacterium longum subsp. infantis NCC 341 (ATCC 15697(T)); c. Bifidobacterium longum subsp. longum NCC 2705 (CNCM I-2618) (NCBI refseq; GCA_000007525.1); d. Enterococcus faecium NCC 2768 (NCIMB 10415); e. Lactobacillus johnsonii NCC 533 (CNCM I-1225) (NCBI refseq; GCA_000008065.1); f. Lactococcus lactis NCC 2287 (CNCM I-4154); g. Lacticasei Bacillus paracasei NCC 2461 (CNCM I-2116); h. lacticasei Bacillus rhamnosus NCC 4007 (CGMCC 1.3724); i. Staphylococcus carnosus NCC 1052 (CNCM I-5400); j. Staphylococcus carnosus NCC 971 (CNCM I-5398); and / or k. Streptococcus thermophilus NCC 2496 (CNCM I-3915); Or the composition for use according to embodiment 1, wherein the composition is selected from probiotics having a genome with at least 95% average nucleotide identity (ANI) to any one of the genome sequences of the probiotic bacteria according to any one of a. to k. defined above, preferably at least 96% ANI, more preferably at least 97% ANI, even more preferably at least 98% ANI, and most preferably at least 99% ANI, or even 99.5% ANI, or 99.9% ANI. Embodiment 3 3. The composition for use according to embodiment 2, wherein the composition comprises two or more species of probiotic bacteria as defined in a. to k. Embodiment 4 A composition for use according to embodiment 2, wherein the composition comprises two or more species of probiotic bacteria having genomes that have an ANI of at least 99% relative to the genome sequences of the probiotic bacteria according to a. to k. Embodiment 5 3. The composition for use according to embodiment 2, wherein the probiotic bacterium (a) is Lactobacillus johnsonii NCC 533 (CNCM I-1225) (NCBI refseq; GCA_000008065.1) or a probiotic bacterium having a genome with at least 95% ANI, preferably at least 96% ANI, more preferably at least 97% ANI, even more preferably at least 98% ANI, and most preferably at least 99% ANI, or even 99.5% ANI, or 99.9% ANI relative to the genome sequence of Lactobacillus johnsonii NCC 533 (CNCM I-1225) (NCBI refseq; GCA_000008065.1, SEQ ID NO: 1). Embodiment 6 6. The composition for use according to any one of embodiments 1 to 5, wherein the prebiotic (b) is selected from α-galactooligosaccharides (α-GOS) / raffinose, β-galactooligosaccharides (β-GOS), and cellooligosaccharides (COS), human milk oligosaccharides (HMO), or combinations thereof. Embodiment 7 7. The composition for use according to any one of embodiments 1 to 6, wherein the prebiotic (b) is selected from soybean GOS (α-GOS), pea GOS (α-GOS), bovine milk oligosaccharides (BMOS) (β-GOS), Vivinal GOS (β-GOS), human milk oligosaccharides (HMO) (2'-fucosyllactose (2'FL), 3-fucosyllactose (3'FL), lacto-N-neotetraose (LNnT), lacto-N-tetraose (LNT), 3'-sialyllactose (3SL), and / or 6'-sialyllactose (6SL)), cellobiose, cellotriose, cellotetraose, and soluble hydrolyzed wheat, soluble hydrolyzed oats, and / or β-glucan hydrolysates containing cellobiose, cellotriose, or cellotetraose, or combinations thereof. Embodiment 8 The lipid (c) a. Triglycerides (TG) composed of a mixture of butyrate (C4:0) and medium-chain fatty acids (MCFA); b. Triglycerides (TG) composed of a mixture of short- and medium-chain fatty acids (SMCFA), preferably C4 and / or C8 fatty acids; c. Triglycerides (TG) composed of a mixture of butyrate and long-chain fatty acids (e.g., triglycerides composed of a mixture of butyrate and oleate); d. Triglycerides composed of medium-chain fatty acids; e. Triglycerides (TG) composed of short-chain fatty acids; f. short-chain fatty acids that can be metabolized into ketone bodies; and g. A composition for use according to any one of embodiments 1 to 7, wherein the composition is selected from short- and medium-chain fatty acids that can be metabolized into ketone bodies, preferably C4 and / or C8 fatty acids (e.g., butyric acid (C4:0) and / or caprylic acid (C8:0)). Embodiment 9 a. the probiotic bacterium is selected from the probiotic bacterium as defined above in a. to k., or a probiotic bacterium having a genome with an ANI of at least 90%, preferably at least 95%, relative to any of the genomes of the probiotic bacteria as defined above in a. to k., or a combination thereof, preferably Lactobacillus johnsonii NCC 533 CNCM I-1225, or at least one probiotic having a genome with an ANI of at least 90%, preferably at least 95%, relative to the genome sequence of Lactobacillus johnsonii NCC 533 (CNCM I-1225); b. the prebiotic is selected from α-galactooligosaccharides / raffinose, β-galactooligosaccharides, or cellooligosaccharides, or a combination thereof, preferably soybean GOS (α-GOS), pea GOS (α-GOS), milk oligosaccharides (BMOS) (β-GOS), Vivinal GOS (β-GOS), cellobiose, cellotriose, cellotetraose, soluble hydrolyzed wheat, soluble hydrolyzed oats, and / or β-glucan hydrolysates containing cellobiose or cellotriose or cellotetraose, or a combination thereof; c. the lipid is selected from triglycerides, short chain fatty acids and / or medium chain fatty acids, wherein the triglycerides comprise short chain fatty acids, medium chain fatty acids, or mixtures thereof, and further, in one embodiment, the triglycerides comprise butyrate and / or caprylate, preferably a. Triglycerides (TG) composed of a mixture of butyrate and medium-chain fatty acids (MCFA); b. Triglycerides (TG) composed of a mixture of short- and medium-chain fatty acids (SMCFA), preferably C4 and / or C8 short- and medium-chain fatty acids; c. Triglycerides (TG) composed of a mixture of butyrate and long-chain fatty acids (e.g., triglycerides composed of a mixture of butyrate and oleate); d. Triglycerides composed of medium-chain fatty acids; e. Triglycerides (TG) composed of short-chain fatty acids; f. short-chain fatty acids that can be metabolized into ketone bodies; and g. The composition for use according to any one of embodiments 1 to 8, wherein the fatty acids are selected from short and medium chain fatty acids that can be metabolized into ketone bodies, preferably C4 and / or C8 short and medium chain fatty acids (e.g., butyric acid (C4:0) and / or caprylic acid (C8:0)). Embodiment 10 10. The composition for use according to claim 9, wherein the probiotic bacterium (a) is selected from Lactobacillus johnsonii NCC 533 (CNCM I-1225) (NCBI refseq; GCA_000008065.1) or a probiotic having a genome with an ANI of at least 90%, preferably at least 95%, to the genome sequence of Lactobacillus johnsonii NCC 533 (CNCM I-1225) (NCBI refseq; GCA_000008065.1). Embodiment 11 9. The composition for use according to embodiment 7 or embodiment 8, wherein the prebiotic (b) is selected from soybean GOS (α-GOS), pea GOS (α-GOS), milk oligosaccharides (BMOS) (β-GOS), Vivinal GOS (β-GOS), cellobiose, cellotriose, cellotetraose, soluble hydrolyzed wheat, soluble hydrolyzed oats, and / or β-glucan hydrolysates containing cellobiose or cellotriose or cellotetraose, or combinations thereof. Embodiment 12 The lipid (c) a. Triglycerides (TG) composed of a mixture of butyrate and medium-chain fatty acids (MCFA); b. Triglycerides (TG) composed of a mixture of short- and medium-chain fatty acids (SMCFA); c. Triglycerides (TG) composed of a mixture of butyrate and long-chain fatty acids (e.g., triglycerides composed of a mixture of butyrate and oleate); d. Triglycerides composed of medium-chain fatty acids; e. Triglycerides (TG) composed of short-chain fatty acids; f. short-chain fatty acids that can be metabolized into ketone bodies; and g. The composition for use according to any one of embodiments 9 to 11, wherein the medium chain fatty acids are selected from medium chain fatty acids that can be metabolized into ketone bodies, preferably C4 and / or C8 medium chain fatty acids (e.g., butyric acid (C4:0) and / or caprylic acid (C8:0)). Embodiment 13 The probiotic bacteria is 10 per daily dose 3 cfu~10 12 cfu, typically 10 per daily dose 4 cfu~10 11 cfu, preferably 10 per daily dose 5 cfu~10 10 cfu, or 10 per daily dose 5 cfu~10 9 cfu, also preferably 10 per daily dose 6 cfu~10 9 cfu, 10 per daily dose 6 cfu~10 8 cfu, or 10 per daily dose 8 cfu~10 10 cfu per daily dose, more preferably about 10 7 cfu~10 9 13. The composition for use according to any one of embodiments 1 to 12, wherein the composition contains an amount of 0.5% or more of the serovar ... Embodiment 14 14. The composition for use according to any one of embodiments 1 to 13, wherein the prebiotic is contained in an amount of 0.1 g to 30 g per daily dose, preferably in an amount of 2 g to 15 g per daily dose. Embodiment 15 15. The composition for use according to any one of embodiments 1 to 14, wherein the lipid is contained in an amount of 0.1 g to 30 g per daily dose, preferably in an amount of 2 g to 15 g per daily dose. Embodiment 16 16. The composition for use according to any one of embodiments 1 to 15, wherein the lipid is a triglyceride consisting of butyrate and caprylate, or a triglyceride consisting of butyrate and oleate. Embodiment 17 16. The composition for use according to any one of embodiments 1 to 15, wherein the lipid is a triglyceride consisting of butyrate and caprylate. Embodiment 18 18. The composition for use according to any one of embodiments 1 to 17, in the form of a food product or nutritional composition, a dietary supplement, a Food for Special Medical Purposes (FSMP), a nutritional supplement, a dairy-based beverage, a small volume liquid supplement, a functional food product, a functional beverage product, a meal replacement beverage, and combinations thereof. Embodiment 19 19. The composition for use according to any one of embodiments 1 to 18, wherein the composition is provided in the form of a powder, a tablet, a capsule, or may be in the form of an oil formulation, an emulsion, an oil-in-water emulsion (o / w emulsion), or a water-in-oil emulsion (w / o emulsion). Embodiment 20 20. The composition for use according to any one of embodiments 1 to 19, wherein the reduction of uremic toxins in said cardiometabolic or neurodegenerative conditions is for slowing the progression of such diseases and comorbidities and / or for managing symptoms and syndromes associated with the toxic effects of uremic solutes in such diseases and comorbidities. Embodiment 21 The reduction of uremic toxins in the cardiometabolic or neurodegenerative condition comprises: Treatment or prevention of kidney disease, including chronic and acute kidney disease; dialysis and pre-dialysis kidney disease; and rare kidney disease caused by genetic and metabolic factors; Treatment or prevention of uremic syndrome, including PEW, bone loss, severe anorexia, fatigue, or inflammation; Delayed complications of progressive kidney disease, including anemia, hyperphosphatemia, secondary hyperparathyroidism, bone disease, and neurological conditions; delayed renal comorbidities, including cardiovascular disease; Preventing or managing the risk of malnutrition; Slowing the progression of cardiometabolic diseases; Prevention or management of the risk of cardiovascular disease and comorbidities (diabetes); and / or 20. The composition for use according to any one of embodiments 1 to 19, including preventing or managing the risk of neurodegenerative and neurological conditions. Embodiment 22 18. A kit of parts suitable for use in reducing or avoiding the accumulation of uremic toxins in a cardiometabolic or neurodegenerative condition, comprising the composition of any one of embodiments 1 to 17, and comprising probiotics, prebiotics and lipids in two or more separate containers, and optionally further comprising instructions for use. Embodiment 23 18. A method for treating a cardiometabolic or neurodegenerative condition as defined above, comprising as a first step: (a) preparing and providing a composition according to any one of embodiments 1 to 17, comprising said probiotic, said prebiotic and said lipid; and (b) administering such composition to a patient in need thereof, who is suffering from an increase in uremic toxins in the context of said cardiometabolic or neurodegenerative condition. Embodiment 24 A method of treating a cardiometabolic or neurodegenerative condition as defined in embodiment 21, comprising administering to a patient a composition according to any one of embodiments 1 to 17. Embodiment 25 Use of a composition as defined in any one of embodiments 1 to 17 in the manufacture of a medicament for the treatment of a cardiometabolic or neurodegenerative condition as defined in embodiment 21. Embodiment 26 The composition contains Lactobacillus johnsonii NCC533 in 10 9 1% cellobiose; and 1% short-medium chain triglycerides containing butyrate and caprylate. Embodiment 27 The composition contains Lactobacillus johnsonii NCC533 in 10 9 1% cellobiose, 1% pea GOS, and 1% short-medium chain triglycerides containing butyrate and caprylate. Embodiment 28 The composition for use according to any one of embodiments 1 to 19, 26 and 27, the kit of parts according to embodiments 20, 26 and 27, the method according to any one of embodiments 21, 22, 26 and 27, or the use according to any one of embodiments 23, 26 and 27, wherein the composition further comprises one or more HMOs, preferably one or more of 2'-fucosyllactose (2'FL), 3-fucosyllactose (3'FL), lacto-N-neotetraose (LNnT), lacto-N-tetraose (LNT), 3'-sialyllactose (3SL) or 6'-sialyllactose (6SL). Embodiment 29 The composition for use according to any one of embodiments 1 to 19, 26 and 27, the kit of parts according to embodiments 20, 26 and 27, the method according to any one of embodiments 21, 22, 26 and 27, or the use according to any one of embodiments 23, 26 and 27, wherein the composition further comprises one or more of 2'-fucosyllactose (2'FL), 3-fucosyllactose (3'FL), lacto-N-neotetraose (LNnT), or lacto-N-tetraose (LNT). Embodiment 30 a. Bifidobacterium animalis subsp. lactis NCC 2818 (CNCM I-3446); b. Bifidobacterium longum subsp. infantis NCC 341 (ATCC 15697(T)); c. A composition, kit of parts, method, or use for use according to embodiment 28 or embodiment 29, further comprising one or more of Bifidobacterium longum subsp. longum NCC 2705 (CNCM I-2618) (NCBI refseq; GCA_000007525.1). Embodiment 31 The composition for use according to any one of embodiments 1 to 15 and 18 to 21, the kit of parts according to embodiments 20, 26 and 27, the method according to any one of embodiments 21, 22, 26 and 27, or the use according to any one of embodiments 23, 26 and 27, wherein the composition comprises triglycerides composed of butyrate and comprising long chain fatty acids. Embodiment 32 The triglycerides composed of butyrate and containing long chain fatty acids include 1,3-dibutyryl-2-linoleoylglycerol, 1,3-dibutyryl-2-stearoylglycerol, 1-butyryl-2-oleoyl-3-palmitoylglycerol, 1-palmitoyl-2-oleoyl-3-butyrylglycerol, 1-butyryl-2-oleoyl-3-linoleoylglycerol, 1-linoleoyl-2-oleoyl-3-butyrylglycerol, 1-oleoyl-2-butyryl-3-linoleoylglycerol, 1-linoleoyl-2-butyryl-3-oleoylglycerol, 1-butyryl 32. The composition, kit of parts, method, or use for use according to embodiment 31, wherein the triglyceride is one or more of a triglyceride comprising butyrate and a long chain fatty acid, including 2-linoleoyl-3-oleoylglycerol, 1-oleoyl-2-linoleoyl-3-butyrylglycerol, 1-butyryl-2-stearoyl-3-oleoylglycerol, 1-oleoyl-2-stearoyl-3-butyrylglycerol, 1-butyryl-2-oleoyl-3-stearoylglycerol, and / or 1-stearoyl-2-oleoyl-3-butyrylglycerol, and mixtures of two or more thereof. Embodiment 33 32. The composition, kit, method, or use for use according to embodiment 31, wherein the triglyceride composed of butyrate and comprising a long chain fatty acid is a mixture of butyrate and a long chain fatty acid, which is oleic acid (C18:1). Embodiment 34 The composition, kit of parts, method or use for use according to any one of embodiments 31 to 33, wherein the composition further comprises one or more HMOs, preferably one or more of 2'-fucosyllactose (2'FL), 3-fucosyllactose (3'FL), lacto-N-neotetraose (LNnT), lacto-N-tetraose (LNT), 3'-sialyllactose (3 SL) or 6'-sialyllactose (6 SL). Embodiment 35 The composition, kit of parts, method, or use for use according to any one of embodiments 31 to 33, wherein the composition further comprises one or more of 2'-fucosyllactose (2'FL), 3-fucosyllactose (3'FL), lacto-N-neotetraose (LNnT), or lacto-N-tetraose (LNT). [Example]

[0118] Method 1: Material Selection In silico selection of probiotic strains with reduced capacity to produce clinically relevant uremic toxins Using Nestlé's in-house bacterial genomics platform WallGene (containing all genomes of the Nestlé Culture Collection), complemented by the in-house BlastP (Basic Local Alignment Search Tool for protein sequences, (Altschul et al., 1990)), we selected probiotic strains from NCC with a reduced ability to contribute to uremic toxin production.

[0119] Specifically, a Blast database containing all predicted proteins of probiotic strains was created. Bacterial and host metabolic pathways involved in the metabolism of uremic toxins were collected from public databases such as the Kyoto Encyclopedia of Genes and Genomes (KEGG pathway) and literature. The amino acid sequences of key enzymes involved in the metabolism of uremic toxins were further extracted from public databases such as KEGG, UniProt, and SwissProt according to EC number and compiled into FASTA files (lists of reference protein sequences). These FASTA files containing the protein sequences of key enzymes were used as reference sequences to perform BLASTp against the protein profiles of probiotic strains available from NCC. When the similarity to the reference proteins indicated by pBLAST was low (e.g., identity <30%), InterProScan (EMBL-EBI) was further used to predict the presence of functional domains (Quevillon et al., 2005).

[0120] In silico selection of carbohydrate substrates capable of supporting the growth of selected probiotic strains with low uremic toxin-producing potential Carbohydrate-associated enzymes (CAZy) of candidate probiotics (Lombard et al., 2014) were annotated in silico using dbCAN (Yin et al., 2012). Based on the CAZy annotation of the probiotic, various carbohydrates exhibiting appropriate biochemical structures were selected using BRENDA - The Comprehensive Enzyme Information System. The list of substrates for each CAZy was then matched with carbohydrate materials composed of polysaccharide and / or oligosaccharide structures corresponding to the preferred substrates.

[0121] In vitro growth studies to validate probiotic and carbohydrate candidate combinations in silico Candidate probiotics are obtained from the Nestlé Culture Collection (NCC) or publicly available deposited strain and culture collections (ATCC, NCIMB, NCBI). Carbohydrate materials are provided by commercial suppliers or by Nestlé or its subsidiaries. A complete list of exemplary carbohydrate sources can be found in Table 1. Table 1 lists representative genes and carbohydrate-activating enzymes (CAZymes) of Lactobacillus johnsonii NCC 533, the Enzyme Code (EC) numbers of the functional enzymes, their functions, relevant related carbohydrate families, recommended fiber materials in terms of preclinical testing of the present invention, and the availability of such compounds. As can be seen, carbohydrates were previously selected in silico based on their biochemical structural characteristics that can be fermented, metabolized, utilized, broken down, degraded, or transformed by the carbohydrate-activating enzymes (CAZymes) present in the genome of Lactobacillus johnsonii NCC 533.

[0122] [Table 1]

[0123] In vitro growth tests of probiotic and carbohydrate candidates are performed using BioLector™ (m2p-labs, Baesweiler, Germany). Each run is tested in 48-well flower plates (m2p-labs, Baesweiler, Germany) with a volume of 1 mL per well. The strains are cultured in sugar-free MRS medium (MRSapi), to which various carbohydrates are added (final concentration 1%). The plates are incubated anaerobically under a CO2 gas phase with shaking at 300 rpm for 48 hours. All cultures are inoculated with 2% of a fresh overnight culture. During the incubation period, the biomass of each well is measured by optical density (OD) at a gain of 30, and pH changes are recorded. After the test, all data are compiled in an Excel spreadsheet and provided for further analysis.

[0124] Preparation of material combinations for preclinical and clinical trials The probiotic and lipid materials are supplied by Nestlé or its subsidiaries (e.g., Sofinol SA, Konolfingen Nestlé Factory). The carbohydrate materials are supplied by various food suppliers, e.g., Olygose (France), AIDP (USA), and Savanna Ingredients GmbH (Germany). The microbiome material combination is produced through the following multi-step procedure:

[0125] Step 1: Encapsulation of fatty oil material: BiPro is hydrated in water with stirring (Ystram) and fatty oil is slowly added (Polytron). The resulting mixture is heated to 82°C for 10 minutes with stirring to fix the encapsulation.

[0126] Step 2: Spray drying of lipid-prebiotic material: The prebiotic is suspended in water and mixed with the lipid suspension. The mixture is then homogenized (Ystram), pasteurized at 72°C for 2 minutes, and then spray dried at 140°C. The final product is a white powder.

[0127] Step 3: Addition of probiotics: The probiotics are added to the spray dried prebiotic-lipid powder mixture. A Turbula is then used to ensure that the probiotics are evenly dispersed in the powder.

[0128] Method 2: In vitro / ex vivo investigation of the relevance of microbiome materials in ameliorating uremic toxin production Preservation of fecal samples Feces were collected from eight CKD donors and nine healthy adult donors. Fecal suspensions were prepared and mixed with ProDigest's optimized cryoprotectant, a modified version of the cryoprotectant developed by Hoefman et al. (2013). The resulting suspensions were aliquoted, flash-frozen, and stored at -80°C (cryostock). Immediately prior to the experiment, aliquots were thawed and immediately added to the reactor.

[0129] Short-term colonic experiments to assess differences in metabolic profiles between the microbiomes of CKD and healthy individuals Short-term colonic experiments were performed in a single reactor to investigate the overall fermentation activity, including glycolytic and proteolytic activity, and changes in microbial metabolite production by fecal microbiomes from healthy and CKD individuals. The short-term colonic experiments represent a simplified simulation of a continuous simulator of the human microbial ecosystem (SHIME®, ProDigest).

[0130] At the beginning of the experiment, an amino acid mixture composed of L-tryptophan, L-tyrosine, L-carnitine, choline, and L-phenylalanine was added to the reactor along with a sugar-depleted nutrient background medium (containing colonic basal nutrients including peptone, yeast extract, mucin, and L-cysteine). A 10% (v / v) cryostock containing 7.5% fecal inoculum from each donor studied (serving as a microbial source) was then added, bringing the total volume in the reactor to 70 mL. For each donor, a reference condition (or negative control) containing only nutrient medium (without spiked AA) was included.

[0131] The reactor was incubated for 48 hours at 37°C under an anaerobic atmosphere with continuous gentle shaking (90 rpm). The incubation was carried out in a completely separate reactor, which not only ensured robust microbial fermentation but also had a sufficiently high volume to allow for the collection of multiple samples over time (0, 24, and 48 hours).

[0132] Samples from multiple time points were evaluated for changes in pH, gases, short chain fatty acids, ammonium, lactate, uremic toxins and precursors.

[0133] Tailored SHIME configuration to assess the impact of microbiome materials on the microbiome derived from CKD patients To optimally address the intervention's ability to correct microbiome dysbiosis in CKD patients, the Prodigest SHIME® formulation was tailored by combining the upper gastrointestinal tract with a single colonic region. This colonic region simulated the conditions of the transverse colon (TC), with a pH of 6.2-6.6 and a residence time of 32 hours. This approach allowed for the maintenance of a diverse microbial community within the system, enabling both carbohydrate- and proteolytic fermentation processes. During this particular example study, the effects of two test products on the microbiome composition and metabolic activity of eight CKD donors were evaluated compared with a negative control for each donor tested. This resulted in 24 test conditions.

[0134] The SHIME® experiment for this study consisted of two phases: (1) Inoculation Period: On day 1, colon reactors were inoculated with appropriate fecal samples (frozen and preserved as part of Phase 1 of this study) and allowed to grow and colonize the reactor. After this overnight incubation, colon reactors were fed with a basal nutrient matrix for an additional 2 days to maximize the maintenance of the diversity of the gut microbiota originally present in the fecal inoculum. This process allowed the microbial community to differentiate within each reactor in response to local environmental conditions while retaining its CKD characteristics.

[0135] (2) Control / Treatment Period: During this 11-day period, the SHIME reactors were operated under nominal conditions and fed SHIME nutrient medium three times daily. On the first day of this period (=day 0), all groups were operated under the nominal conditions, and samples collected on this first day served as baseline parameters. Starting on the second day of this period (=day 1), the test product was added to the nutritional supply of each donor's treatment group, while the nominal conditions were maintained in each donor's control group. On the eighth day of this period, an amino acid mixture was also added to all SHIME groups. During this period, samples were taken from the colon reactors to investigate the specific effects of the test product on the composition and activity of the resident microbial community, compared with the negative control.

[0136] During each period, the model was fed with SHIME nutrient medium three times a day. The feeding schedule is shown in the table below (Table 2).

[0137] [Table 2]

[0138] Samples were collected at multiple time points (days 0, 1, 2, 4, 7, 8, and 10) and assessed for changes in pH, gases, short-chain fatty acids, ammonium, lactate, uremic toxins, and precursors.

[0139] Analysis of microbial community composition and activity The large volume of the colonic region allows for sufficient fluid volume to be collected daily without disturbing the microbial community or adversely affecting the remainder of the experiment. Several microbial parameters are monitored throughout the short-term colonic and SHIME experiments.

[0140] (a) Overall fermentation activity: pH: The degree of acidification during the experiment is an indicator of the intensity of bacterial metabolism and is used as a parameter to control the process. The pH of the incubation provides a rough indication of the fermentation rate of the test product.

[0141] Gas production: The closed incubation allowed for manometric measurement of gas accumulation in the headspace. Gas production is an indicator of microbial activity and, therefore, the rate of fermentation. H2 and CO2 are the first gases produced. These gases can then be used as substrates for CH4 production, reducing the gas volume. H2 can also be used to reduce sulfates generated by proteolytic fermentation to H2S. As a result, N2, O2, CO2, H2, and CH4 constitute 99% of the volume of intestinal gas. The remaining 1% is composed of NH3, H2S, volatile amino acids, and short-chain fatty acids. 3 Each measurement was performed in a single repetition.

[0142] Acid / base consumption: The production of microbial metabolites in the colon reactor changes the pH. Without continuous pH control (by adding acid or base), the pH will exceed the set range. Acid / base consumption is continuously monitored throughout the experiment.

[0143] (b) Microbial community activity: Short-chain fatty acids (SCFAs): Patterns of SCFA production can be assessed by microbial carbohydrate metabolism (acetate, propionate, and butyrate) or protein metabolism (branched CFAs) and compared with typical fermentation patterns of normal GI microbiota. Quantitative analysis of SCFAs is performed by capillary gas chromatography coupled with a flame ionization detector (FID). SCFA separation is performed by liquid-liquid extraction (De Boever et al. 2000).

[0144] Lactate: Both lactate-producing and lactate-consuming bacteria exist in the human intestine. Lactate is produced by lactic acid bacteria and also acts as an antibacterial agent by lowering the pH of the environment. Lactate can also be rapidly converted by other microorganisms to propionate and butyrate. Lactate concentration was measured using the Enzytec™ kit (R-Biopharm).

[0145] Ammonium: Ammonium is a product of protein degradation. Proteolytic fermentation leads to the production of potentially toxic or carcinogenic compounds, such as p-cresol and p-phenol. The ammonium concentration in the samples was determined colorimetrically using the indophenol blue spectrophotometric (IPB) method.

[0146] Branched-chain SCFAs (BCFAs; isobutyric acid, isovaleric acid, and isocaproic acid) are markers of proteolytic fermentation. Quantitative analysis of BCFAs is performed by capillary gas chromatography coupled with a flame ionization detector (FID). Separation of BCFAs is achieved by liquid-liquid extraction (De Boever et al. 2000).

[0147] Uremic toxins: Concentration levels of uremic toxins (e.g., p-cresol, p-cresyl sulfate, indole, indole-3-acetic acid, betaine, trimethylamine, trimethylamine-N-oxide, indoxyl, indoxyl sulfate, semialdehyde glutaric acid, uric acid, and urea) were measured using ultra-performance liquid chromatography coupled with high-resolution mass spectrometry and fluorescence detection.

[0148] Untargeted metabolomics: Prodigest's Metakey platform was used to measure a panel of microbial metabolites, including but not limited to bile acids and amino acids

[0149] Microbial community composition: Population composition was determined using quantitative shotgun deep sequencing.

[0150] Statistical explanation Statistical comparisons between conditions and treatment groups within each donor were performed using two-tailed paired t-tests. Differences were considered statistically significant if p-values ​​were less than 0.05.

[0151] Method 3: Animal testing Animal models Mouse model Animal experiments were performed at the CarMeN Laboratory (Direction Départementale des Services Veterinaires du Rhone). All experiments were conducted in accordance with the guidelines for the care and use of laboratory animals established by the French Ministry of Agriculture and the Council of the European Union. C57BL / 6J mice were purchased from Janvier SA (Le Genest-Saint-Isle, France) and housed in an air-conditioned room at 21°C ± 0.5°C and 60%-70% humidity, with a 12-h light / dark cycle (lights on 07:00-19:00) and free access to food and water. Moderate CKD was induced by a 5 / 6 nephrectomy in a two-step surgical procedure. Additional animals underwent sham surgery and served as controls. See Figure 14 for an overview of the in vivo animal experiments.

[0152] Animal feed and treatment From week 0 to week 3, all animals were fed a standard rodent diet (SAFE A04 standard diet for rodents). Subsequently, animals were fed either a standard diet or a customized standard diet containing 1% (w / w) of the nutritional synbiotic blends P1 and P2 of the present invention for a total of 7 weeks. The P1 intervention consisted of an ingredient blend containing Lactobacillus johnsonii NCC533, cellobiose, and short-medium chain triglycerides containing butyrate and caprylate. The P2 intervention consisted of an ingredient blend containing Lactobacillus johnsonii NCC533, cellobiose, pea GOS, and short-medium chain triglycerides containing butyrate and caprylate. Custom diets containing the nutritional or synbiotic blends were made by dry-mixing powdered SAFE A04 with the blends. Successful incorporation was verified, including measuring probiotic viability. After validation, the custom feed was pelleted according to the standard SAFE procedure. During the custom feed pelleting process, probiotic viability decreased by 3 logs. Therefore, to ensure sufficient viable probiotics were administered to the animals, additional probiotics were administered to the treated animals by dissolving powdered probiotic bacterial stock in sterile distilled water to a final concentration of approximately 1.8 ± 8 CFU / mL. Probiotic viability in the sterile drinking water was confirmed by plating. The probiotic solution was replaced every two days.

[0153] Intraperitoneal glucose tolerance test (IPGTT) After an overnight fast, an intraperitoneal glucose tolerance test (glucose, 1 g / kg body weight) was performed. Blood glucose levels were measured from a drop of blood taken from the tail end using an automated blood glucose meter (Accu-Check Performa, Roche, Meylan, France).

[0154] Urinary parameters Urine was collected in metabolic cages for 24 hours to assess urine volume and markers of renal function, such as protein, creatinine, and albumin levels, as well as for metabolomic analysis. Markers of renal function were measured using commercially available assays. Urinary metabolomics was measured using reverse-phase HPLC coupled to a fluorescence detector or using Prodigest's MetaKey® platform.

[0155] Biochemical and metabolomic measurements Uremic toxins were quantified in plasma and urine using reverse-phase HPLC coupled to a fluorescence detector or by using Prodigest's MetaKey® technology. Plasma or serum concentrations of creatinine, cholesterol, triglycerides, free fatty acids, adiponectin, insulin, cystatin C, and urea were measured using commercially available assays. Serum metabolomics was performed using Prodigest's MetaKey® platform.

[0156] Kidney Histology In both sham-operated and nephrectomized mice, the remaining kidney was removed at the time of sacrifice. The kidney was harvested and the capsule was removed. Renal histological lesions were analyzed after hematoxylin and eosin (HES) and Sirius red staining. Briefly, kidneys were fixed in 4% formalin for 24 hours and then routinely processed and embedded in paraffin. Histological slides were independently examined on a blinded basis for the levels of interstitial inflammation, interstitial fibrosis, and glomerular sclerosis.

[0157] Ileum Histology Ileum samples were collected at the end of the experiment. The ileum was fixed in 4% formalin for 24 hours and then routinely processed and embedded in paraffin. Sections were then immunofluorescently stained with a rabbit antibody against occludin, followed by a goat anti-rabbit secondary antibody. Images were acquired using a confocal microscope, and the relative fluorescence intensity of occludin was quantified using Image J software.

[0158] statistical analysis For each experiment, multiple mice were analyzed as biological replicates. Dot plots on a linear scale represent arithmetic means. Bar graphs are presented as mean ± standard error of the mean (SEM). Statistical analysis was performed using GraphPad Prism version 9. For comparisons between two groups, significance was assessed using a two-tailed Student's t-test or the nonparametric Mann-Whitney test. For comparisons between three or more groups, one-way analysis (ANOVA) followed by Fisher's LSD test without correction was performed. Differences were recorded as significant at p ≤ 0.05.

[0159] Rat model Rat model experiments were performed at Grubra, a fully AAALAC-accredited facility. All animal experiments were conducted in accordance with Grubra's bioethical guidelines, which fully comply with internationally recognized guidelines for the care and use of laboratory animals. All experiments were approved by the Danish Council for Animal Experimentation. A 5 / 6 nephrectomy (Nx) was performed in a two-step surgical procedure under isoflurane anesthesia in Wistar RjHan (WI) rats (Janvier, France).

[0160] Animal feed and treatment Animals were treated as follows: - On week 4, a two-step nephrectomy (Nx) procedure was initiated. - From day 2, animals were fed a standard rodent diet (SAFE A04 standard diet for rodents). On day 1, animals were fed either a standard rodent diet or a diet customized according to the animal group. The customized diet contained cellobiose and short-medium chain triglycerides containing butyrate and caprylate, and was supplemented with Lactobacillus johnsonii NCC533 (10 8 CFU) was administered orally by gavage. This regimen was administered once daily for 8 weeks.

[0161] Example 1 - Identification of optimal synbiotics to ameliorate uremic toxin accumulation In silico screening and identification of candidate probiotics based on the absence of uremic toxin-related enzymes encoded in the genome of bacterial strains.

[0162] The European Uremic Toxin Work Group lists 90 compounds that are considered to be uremic toxins (Yavuz et al., 2005). In this example, gut-derived and plasma-bound uremic toxins were examined, including urea, trimethylamine N-oxide (TMAO), indole-3-acetic acid (IAA), indoxyl, and p-cresol. The bacterial metabolic pathways that produce these uremic toxins were compiled from public databases such as the Kyoto Encyclopedia of Genes and Genomes (KEGG pathway) and literature.

[0163] The different uremic toxin biosynthetic pathways were summarized, and each enzyme potentially catalyzing the different steps was labeled with its EC number (Enzyme Code Number) (Figure 1). The conversion of urea to ammonia (NH3) and carnitine or choline to TMAO requires only one or two steps. In contrast, the conversion of tryptophan to IAA and indoxyl, and tyrosine to p-cresol require several steps. The first and last reactions of each pathway were analyzed in detail to assess the ability of each bacterium to synthesize the selected uremic toxins. In this example, the enzymes involved in these pathways are referred to as key enzymes in the following text.

[0164] The suitable probiotic strains analyzed in this example were: Bifidobacterium animalis subsp. lactis NCC 2818 (CNCM I-3446); Bifidobacterium longum subsp. infantis NCC 341 (ATCC 15697(T)); Bifidobacterium longum subsp. longum NCC 2705 (CNCM I-2618) (NCBI refseq;GCA_000007525.1); Enterococcus faecium NCC 2768 (NCIMB 10415); Lactobacillus johnsonii NCC 533 (CNCM I-1225) (NCBI refseq; GCA_000008065.1); Lactococcus lactis NCC 2287 (CNCM I-4154; Lacticasei Bacillus paracasei NCC 2461 (CNCM I-2116); lacticasei Bacillus rhamnosus NCC 4007 (CGMCC 1.3724); Staphylococcus carnosus NCC 1052 (CNCM I-5400); Staphylococcus carnosus NCC 971 (CNCM I-5398); Streptococcus thermophilus NCC 2496 (CNCM I-3915);

[0165] The presence of key enzymes catalyzing the synthesis of uremic toxins in the selected strains is shown in Figure 3. When creating the protein sequence reference file, we considered both sequences from SwissProt and proteins from bacteria that are as phylogenetically related as possible to the probiotic strains described above. As shown in Figure 3, L. johnsonii NCC 533 is an exception, as its genome does not encode any of the key enzymes and is therefore predicted to be the least likely to produce any of the uremic toxins of concern. This strain was also selected for in silico prebiotic carbohydrate selection and in vitro growth testing, described below.

[0166] In silico screening and identification of prebiotic carbohydrates based on the presence of CAZy encoded in the genome of candidate probiotics. Candidate prebiotic carbohydrates were selected in silico based on their structure and potential for degradation, metabolism, fermentation, or decomposition by candidate probiotics. As an example, we analyzed candidate prebiotic carbohydrates (NCBI refseq; GCA_000008065.1) that can be degraded by the probiotic Lactobacillus johnsonii, NCC 533 (CNCM I-1225), as shown below. Because the enzymatic activities are very similar, this knowledge can be transferred to additional suitable probiotic strains, such as those analyzed above.

[0167] The genome sequence of the probiotic L. johnsonii was examined to identify all carbohydrate-associated enzymes (CAZy) contained in the genome sequence. Table 1 summarizes the CAZy-related coding regions, CAZy family annotation, EC number of the encoded enzyme, function, identified fiber material, and carbohydrate substrate availability.

[0168] Lactobacillus johnsonii has been shown to possess all the carbohydrate-associated enzymes (CAZY) required to be able to degrade the following fibers: α-galactooligosaccharides (raffinose family), galactomannans, β-galactooligosaccharides, cellooligosaccharides, cellooligosaccharides, α-galactooligosaccharides (raffinose family), and scFOS / inulin.

[0169] In vitro growth studies of candidate probiotics and carbohydrates. In this example, growth studies were performed and evaluated using the probiotic strain L. johnsonii to confirm the in silico selections and define the optimal synbiotic to help ameliorate toxin accumulation in uremia.

[0170] Several carbohydrates were tested. Fiber polymers and oligomers containing galactose and glucose as the primary components were selected. For example, both α-linked and β-linked galactooligosaccharides (GOS) and mannans containing galactose as side chains were tested. Furthermore, as glucose polymers with β-linked structures, cellooligosaccharides, purified β-glucans from barley with three molecular weights (high, medium, and low), and the soluble fraction of enzymatically hydrolyzed wheat fiber were tested. To better understand which portions could be metabolized by the probiotic strains, the constituent monomers (glucose, fructose, and galactose) of each fiber material were also analyzed in the experiment.

[0171] As shown in Figure 4, L. johnsonii NCC 533 was also able to grow on carbohydrates belonging to the α-galactooligosaccharide, galactomannan, or fructan families. In particular, it was able to grow on commercially available fiber AlphaGOS® derived from pea or soybean. A decrease in pH was observed with all of these carbohydrates, indicating that L. johnsonii NCC 533 grew well on these substrates. Similarly, L. johnsonii NCC 533 grew equally well in the presence of the commercially available fiber Vivinal® GOS and BMOs (Figure 5). On the other hand, inulin, ScFOS, and PHGG provided minimal support for the growth of L. johnsonii NCC 533 (Figure 4).

[0172] Furthermore, L. johnsonii NCC 533 was able to grow efficiently on different carbohydrates belonging to the cellooligosaccharide family (disaccharides to tetrasaccharides), especially cellobiose and cellotriose, as shown in Figure 6. It was also able to grow effectively on cellotetraose and soluble hydrolyzed wheat and oat containing cellobiose and cellotriose, although to a lesser extent or at later time points.

[0173] result A total of 14 probiotic strains, including those listed above and in Figure 3, were investigated in this example. L. johnsonii NCC 533 was shown to lack any of the key enzymes required for the production of uremic toxins such as ammonia, TMAO, IAA, indoxyl, and p-cresol. Furthermore, L. johnsonii NCC 533 encodes a large repertoire of enzymes for metabolizing different carbohydrates. Based on the CAZy profile, 15 fibers were proposed for growth studies. L. johnsonii NCC 533 was shown to grow on several commercially available fibers, such as AlphaGOS®, Vivinal® GOS, and BMOS, derived from soybeans or peas. Furthermore, the results confirmed that L. johnsonii NCC 533 actually metabolized all monomers (e.g., glucose, fructose, and galactose) that form these fibers. Therefore, it could be concluded that this strain can completely hydrolyze these fibers and use the constituent monomers as substrates for growth. β-Galactooligosaccharides (e.g., Vivinal® GOS) are widely used in infant nutrition today, and they have consistently been shown to have significant bifidobacterial growth effects. This type of fiber is one of the few materials recognized as a prebiotic by the International Scientific Association of Prebiotics and Probiotics (ISAPP). α-Galactooligosaccharides, on the other hand, are a new material. α-Galactooligosaccharides are a by-product of the plant protein industry and are naturally found in legume seeds. Although there are fewer studies on α-galactooligosaccharides, they have also consistently been shown to have a positive impact on the abundance of sachorrylutic and short-chain fatty acid-producing bacteria, such as bifidobacteria and lactobacilli.

[0174] L. johnsonii NCC 533 was also able to grow on cellobiose and related oligosaccharides (cellotriose and cellotetraose). While cellobiose is commercially available in feed grade today, this is not the case for cellooligosaccharides. Cellooligosaccharides are dimeric and trimeric components of cellulosic fibers and / or mixed-linkage β-glucans in oats or barley. Therefore, cellooligosaccharide-rich materials can be produced by applying optimized enzymatic hydrolysis to cellulosic fibers / grains or mixed-linkage β-glucans. When combined with other Lactobacillus strains, cellobiose and / or cellooligosaccharides in the form of β-glucan hydrolysates have been shown to have the potential to exert synbiotic effects, particularly in increasing intestinal glycolytic metabolism.

[0175] Other fibers tested (e.g., inulin, ScFOS, PHGG, and high / medium / low molecular weight β-glucans) minimally supported the growth of L. johnsonii NCC 533, indicating that this strain is likely unable to completely hydrolyze these polymers into their constituent monomers. However, based on the CAZy annotation of L. johnsonii NCC 533, it is possible that it may be able to partially hydrolyze these polymers into smaller fragments or oligomers. If such strains are indeed able to partially hydrolyze these substrates, this suggests that cross-feeding with other inhabiting microorganisms within the same ecosystem may occur in a complex environment such as the gut.

[0176] Overall, these results suggest a beneficial role for the co-administration of L. johnsonii NCC 533 with carbohydrates belonging to several families, including α-galactooligosaccharides, e.g., pea GOS, and cellooligosaccharides, e.g., cellobiose.

[0177] Example 2 - In vitro (ProDigest) studies research design Several publications have documented the dysbiotic profile of the microbiota in CKD patients. Specifically, CKD patients are characterized by altered microbial composition and function compared to healthy individuals. Therefore, in this example, we tested the ability of the present invention to correct microbiome dysbiosis in CKD patients.

[0178] To conduct the study, fecal microbiota were collected from healthy individuals (controls) and patients with chronic kidney disease (CKD). Table 3 provides a summary of the characteristics of eight CKD patients and nine healthy adult donors who provided fecal microbiomes for in vitro / ex vivo experiments. The patients were participating in a clinical trial registered under ClinicalTrials.gov Identifier NCT04768309. Table 3 shows the characteristics of these donors. These fecal microbiota were then used in experimental configurations using Prodigest's in vitro / ex vivo technology.

[0179] [Table 3]

[0180] In this example, two studies were performed sequentially.

[0181] First, we used Prodigest's short-term, single-stage colon simulation technology to assess differences in fecal microbiome profiles between healthy adult donors and adult donors with chronic kidney disease (CKD) (see Figure 7 and its accompanying text for further experimental details).

[0182] Second, a modified Prodigest SHIME® technology was used to evaluate the impact of novel nutritional or synbiotic materials on fecal microbiota from patients with chronic kidney disease (CKD) (see Figure 9 and its description for further experimental details).

[0183] In the second study, eight CKD donors were used and two material blends (groups P1 and P2, see below; the compositions of P1 and P2 are shown below) and a control were tested. Thus, for each CKD microbiota, three conditions were implemented:

[0184] Control (CTRL) group: SHIME units were supplied with a basal nutrient medium that was used to establish baseline parameters for each donor for comparison with the treatment groups.

[0185] Group P1: The SHIME unit was fed with a basal nutrient medium in one cycle (administered to the colon at 1 h) once daily. During the other two cycles, the SHIME unit was fed with a carbohydrate-depleted nutrient medium supplemented with a nutrient or synbiotic material composed of cellobiose, butyric acid, and caprylic acid, and the probiotic C. johnsonii NCC 533.

[0186] Group P2: The SHIME unit was fed with a basal nutrient medium in one cycle per day. During the other two cycles, the SHIME unit was fed with a carbohydrate-depleted nutrient medium supplemented with cellobiose, pea GOS, butyric and caprylic acids, and the probiotic L. johnsonii NCC 533.

[0187] Additionally, an amino acid mixture was added to reach a colonic concentration of 2.5 g / L to provide a substrate for testing whether the present invention can ameliorate the overproduction of uremic toxins and precursors. The amino acid mixture consisted of 23% L-phenylalanine, 20% L-tryptophan, 23% L-tyrosine, 10% L-carnitine, and 23% choline. The amino acid mixture was added in one feeding cycle per day on days 8 (administered to the colon at 1 o'clock), 9 (administered to the colon at 1 o'clock), and 10 (administered to the colon at 1 o'clock) of the experiment. This addition was performed in both the control and treatment groups of each donor.

[0188] result Fecal microbiota from CKD patients with abnormalities in amino acid and protein metabolism To verify whether the in vitro / ex vivo model is a reliable model for testing the efficacy of the present invention in modulating the CKD microbiome, fecal microbiota from CKD patients and healthy donors were inoculated into the Prodigest system.

[0189] As seen in Figure 8, the CKD microbiota showed higher production of uremic toxin precursors, such as p-cresol, especially in the presence of excess amino acid substrates. With regard to proteolytic activity, the CKD microbiota also showed higher concentrations of proteolytic markers, such as branched-chain fatty acids.

[0190] Nutritional or synbiotic blends reduced the production of uremic toxin precursors by microbiota from CKD patient donors The ability of the present invention to ameliorate the overproduction of uremic toxins and their precursors was evaluated in the presence of low amino acid (AA: days 0-7) and additional AA substrates (days 8-10). In this example, three uremic toxin precursors were shown, including indole, p-cresol, and trimethylamine (TMA) (see Figure 10). It is hypothesized that the addition of AA will induce a further increase in the production of uremic toxins and their precursors by the fecal microbiota.

[0191] Indole can be metabolized by the gut microbiota using tryptophan as a precursor molecule. This converts tryptophan first to tryptamine and indolepyruvate, which is then converted to indole. Indole is known to have profound effects on gut microbial composition, microbial metabolism, the host immune system, host-microbiome interactions, and host immune system-gut microbiota interactions. Once produced by gut bacteria, indole is absorbed into the portal circulation and enters the liver. Hydroxylation of indole in the liver produces 3-hydroxyindole (indoxyl), most of which is then sulfonated to indoxyl sulfate. Indoxyl sulfate is considered a uremic toxin and is most frequently implicated in the progression of kidney disease and cardiovascular complications. Indoxyl sulfate is also thought to be involved in adverse effects on bone and the central nervous system. On the day before administration of the AA formulation (day 7), significantly lower indole concentrations were detected in both P1 and P2 compared to the untreated control group (Figure 10). A strong increase in indole concentrations was observed in all donors following administration of the AA formulation. This observation is consistent with the fact that tryptophan was provided as the indole precursor molecule.

[0192] The phenolic compound p-cresol is a microbial metabolite derived from tyrosine, producing the intermediates 4-hydroxyphenylpropionic acid and 4-hydroxyphenylacetic acid. In the liver, p-cresol is sulfated and converted to the toxic metabolite p-cresyl sulfate. Mechanistic studies have shown that this particular metabolite is associated with oxidative stress, endothelial dysfunction, proximal tubule injury, and insulin resistance. In this context, a positive relationship between p-cresyl sulfate levels and the progression of all-cause mortality, cardiovascular disease, and CKD has been demonstrated. As seen in Figure 10, before the AA spike (days 0–7), p-cresol showed a defined reduction in P1 and P2 treatments compared to the untreated control. As expected, administration of the AA formulation resulted in an increase in p-cresol concentration levels compared to the preceding period in all donors in the blank control group. Interestingly, both P1 and P2 were able to ameliorate p-cresol overproduction compared to the blank control.

[0193] Trimethylamine (TMA) is a metabolite produced by the gut microbiota in the intestinal lumen, with various dietary quaternary amines serving as precursor molecules. These precursor molecules primarily include choline and carnitine, but also betaine, γ-butyrobetaine, and other choline-containing compounds. In vivo, the generated TMA is rapidly absorbed into the portal circulation by passive diffusion and subsequently oxidized to trimethylamine-N-oxide by the action of flavin-containing monooxygenases in the liver. Trimethylamine-N-oxide (TMAO) is involved in oxidative stress, inflammation, myocardial fibrosis, endothelial injury, and platelet inactivation. Thus, TMAO has been implicated in several chronic non-communicable diseases, including CKD. As shown in Figure 10, an increase in TMA concentration was observed in the blank control group from days 0 to 7. P1 and P2 showed lower concentration levels compared to the blank control group, with the greatest change observed at P22. Administration of the AA formulation resulted in a clear increase in trimethylamine concentrations across all donors and experimental conditions. This increase is associated with increased availability of the substrate for generating TMA. A significant effect of P2 was calculated compared to the blank control when statistical analysis was performed across all donors at day 10.

[0194] Overall, this data highlights the effectiveness of the present invention in correcting amino acid metabolic abnormalities in the microbiota of CKD patients. In particular, the present invention is effective in conditions with low and normal levels of amino acids, suggesting that the benefits of the present invention can be used by different patients with different dietary restrictions, such as low-protein diets.

[0195] Nutritional or synbiotic blends reduced urea production Urea is a metabolic product produced via the urea cycle, a metabolic pathway that removes excess endogenous and exogenous nitrogen from the body by detoxifying ammonia into urea. Typical nitrogen sources in this context include amino acids such as ornithine, arginine, aspartate, and glutamate. The urea cycle primarily operates in the liver, but the intestinal microbiota also operates the mitochondrial urea cycle. High urea concentrations are typically observed in CKD patients, and such urea concentrations are known to significantly alter the intestinal microbial community, reducing bacterial strains that produce anti-inflammatory and energy molecules and increasing bacterial strains that can metabolize urea but also produce uremic toxins, including indoxyl sulfate and p-cresol sulfate. Furthermore, high urea concentrations can increase intestinal permeability and create a toxic environment that induces the colonization of bacteria that express urease and uricase, reducing urea to ammonia. Ammonia increases intestinal pH, promoting the proliferation of pathogenic bacteria. Because amino acid preparations contain various nitrogen sources, spiking these preparations may induce an increase in the level of urea concentration during colonic incubation. However, it should be noted that the amino acids included in such preparations have not been reported to be specifically involved in the urea cycle. In this regard, the overall change in microbial composition due to amino acid spiking, especially the change in the bacterial strains that metabolize urea, may determine the final outcome of the observed urea concentration level.

[0196] As shown in Figure 11, there was a tendency for the concentration profile to increase during the period from day 0 to day 7 and day 10. A lowering effect on the level of urea concentration was observed for both P1 and P2, with P1 having the strongest effect. As expected, administration of the AA formulation generally had only a limited effect on the level of urea concentration.

[0197] This data highlights the effectiveness of the present invention in reducing the production of uremic toxins by the gut microbiota. Furthermore, the present invention may provide relief from intestinal changes and symptoms, including intestinal permeability and inflammation, associated with excessive urea production.

[0198] Nutritional or synbiotic blends increased or decreased markers of proteolytic fermentation Both ammonium and branched SCFAs (the sum of isobutyrate, isovalerate, and isocaproate) are produced by proteolysis and reflect the proteolytic activity of the gut microbiota. The latter has been associated with direct and indirect adverse health effects (e.g., colon carcinogenesis), so reducing ammonium / branched SCFA production may be beneficial.

[0199] As shown in Figure 12, branched SCFA levels increased in all groups within each donor throughout the experiment. Addition of both material blends (P1 and P2, see above) systematically reduced branched SCFA production in all donors compared to the control (see also above). This reduction was also observed in the average effects across the eight donors at days 7 and 10, where branched SCFA production was significantly reduced in the treatment groups compared to the blank control, with the strongest effect observed for P2.

[0200] A similar effect was observed for ammonium levels (Figure 12), with both P1 and P2 strongly reducing ammonium production throughout the study. This reduction was also observed on days 7 and 10, with both treatments significantly lowering ammonium levels compared to the blank control when averaged across the eight selected donors. The strongest effect was observed after repeated administration of P2, which produced an even stronger significant reduction compared to P1 on days 7 and 10.

[0201] This data demonstrates the efficacy of the present invention in correcting dysbiosis, particularly protein metabolic abnormalities, in the CKD microbiota.

[0202] Nutritional or synbiotic blends increased the production of short-chain fatty acids (SCFAs) The production of SCFAs results from carbohydrate metabolism in the colon and is associated with various health benefits. The most abundant SCFAs are acetate, propionate, and butyrate. SCFAs are well known to play an important role in intestinal health. Acetate can be used as an energy source for the host and as a potential substrate for lipid synthesis in the body. Furthermore, acetate is an important by-product in the synthesis of butyrate, which can exert antibacterial effects against pathogens. However, the health-promoting effects are primarily due to propionate and butyrate. Propionate and butyrate serve as the main energy source for the intestinal epithelium and have shown protective effects against inflammation and colon cancer. The former is also known to be transported to the liver, where it has a plasma cholesterol-lowering effect and a positive impact on glycemic control. Therefore, in summary, the beneficial effects of the investigated substrates on SCFA production include increased production of acetate, propionate, and / or butyrate. For optimal interpretation, SCFA levels from both test conditions are presented for each of the different SCFAs.

[0203] Acetate is one of the important metabolic products in the human gut and is therefore produced by a wide range of gut microorganisms, including Bacteroidetes (phylum Bacteroidetes) and Bifidobacteria. As seen in Figure 13, administration of both ingredient blends (P1 and P2) increased overall acetate production compared to the blank control, averaged across all donors, compared to the control. P2 exerted the strongest effect. P1, P2, and the control were as defined above.

[0204] Propionate can be produced by a wide range of gut microorganisms, with the most abundant propionate-producing bacteria being Bacteroides species (phylum Bacteroidetes), Veillonella (phylum Firmicutes), and Akkermansia muciniphila (phylum Verrucomicrobia). The effect of treatment on propionate production was subsequently shown to be both donor- and product-dependent. Increased levels of propionate were observed following treatment with P1 and P2, but the levels did not reach significance due to some variability in response among the eight donors.

[0205] Butyrate is produced by members of the Clostridium cluster IV and XIVa (phylum Firmicutes). In a process called cross-feeding, these microorganisms convert acetate and / or lactate (along with other substrates) into health-associated butyrate. Both P1 and P2 were observed to have a strong stimulatory effect on butyrate production in certain donors. When averaged across eight selected donors, only treatment with P2 significantly enhanced butyrate production toward the end of the control / treatment period (day 7) before AA spiking, although a similar trend was observed with P1. Finally, after AA spiking during the final three days of the experiment, P1 also significantly enhanced butyrate levels compared to the blank control when averaged across different donors, reaching similar levels compared to P2 treatment.

[0206] Overall, the above examples support the effectiveness of the present invention in improving the glycolytic activity of the CKD microbiota and enhancing the production of beneficial metabolites, such as SCFAs.

[0207] Example 3 - In vivo animal studies: research design To test the efficacy of the present invention in ameliorating uremic toxin accumulation and associated symptoms and clinical outcomes, a 5 / 6 nephrectomy animal model was used. Kidneys were removed during a two-step surgical procedure, resulting in reduced renal function (reminiscent of human CKD stage 3b or higher). Additional animals were sham-operated and served as non-CKD animal controls (see Figure 14 and accompanying text for an overview of the experimental design and analysis).

[0208] From week 3 to week 10, CKD animals were fed diets with or without nutritional or synbiotic blend interventions. In this example, two nutritional / synbiotic blend combinations (P1 and P2) were tested. The P1 intervention consisted of 10 doses of Lactobacillus johnsonii NCC533. 8 (This was corrected from the concentrations reported in U.S. Provisional Patent Applications Nos. 63 / 439,638 and 63 / 480,729, which reported inaccurate concentrations.) 1% cellobiose and 1% short-medium chain triglycerides containing butyrate and caprylate. P2 intervention consisted of a blend of ingredients containing Lactobacillus johnsonii NCC533 for 10 min. 8 (This was corrected from the concentrations reported in U.S. Provisional Patent Applications Nos. 63 / 439,638 and 63 / 480,729, which reported inaccurate concentrations), and a blend of ingredients containing 1% cellobiose, 1% pea GOS, and 1% short-medium chain triglycerides containing butyrate and caprylate.

[0209] At week 8, a glucose tolerance test was performed. At week 9, mice were placed in metabolic cages and urine was collected for analysis of uremic toxins and renal parameters. At week 10, mice were sacrificed and multiple tissues were collected for subsequent analysis.

[0210] result Treatment of uremic mice with nutritional or synbiotic blends improved plasma concentrations of clinically relevant uremic toxins To demonstrate the importance of the nutritional or synbiotic blend of the present invention in ameliorating uremic toxins, a 5 / 6 nephrectomy or remnant kidney model was used. This model is one of the most commonly used animal models of progressive renal failure due to a reduction in nephron number and mimics the condition seen in CKD patients. As seen in Figure 15, animals that underwent nephrectomy (CKD group) showed significantly higher plasma levels of clinically relevant uremic toxins, such as p-cresyl sulfate (PCS), indoxyl sulfate (IS), p-cresyl glucuronide (PCG), indoleacetic acid (IAA), 3-carboxy-4-methyl-5-propyl-2-furanpropionate (CMPF), and uric acid, compared with animals with intact renal function (sham-operated group). This demonstrates that CKD animals are in a uremic state and therefore a good model for investigating the effects of uremic treatment.

[0211] As also seen in Figure 15, elevated plasma uremic toxins were significantly reduced in CKD animals treated with the nutritional or synbiotic blend (groups P1 and P2, see above) compared to CKD animals, thus highly supporting the effectiveness of the blend for its intended use.

[0212] Nutritional or synbiotic blends stabilize renal function in nephrectomized animals The inventors further investigated the benefits of the nutritional or synbiotic blend on kidney function and structure. Proteinuria and blood urea levels were measured to assess the progression of functional damage in the remaining kidney. Furthermore, the progression of microscopic damage was also assessed by histology.

[0213] Proteinuria is a typical symptom of conditions that affect the kidneys. Excess protein in the urine means that the kidney's filtering mechanism (glomeruli) is not functioning properly and is leaking excess protein into the urine. Proteinuria is measured by calculating the ratio of protein to creatinine in the urine.

[0214] Blood urea nitrogen level in blood is an indicator of kidney function. Urea is produced as a by-product in the liver when proteins are metabolized. Healthy kidneys filter urea out of the body through urine. High urea levels generally indicate acute or chronic kidney disease or kidney failure. Urea is also considered one of the clinically relevant uremic toxins.

[0215] As can be seen in Figure 16, nephrectomized animals (CKD group) showed significantly higher proteinuria compared to non-CKD animals (sham-operated), confirming the progression of kidney damage. Interestingly, CKD animals treated with the nutritional or synbiotic blend (groups P1 and P2, see above) showed significant improvements in proteinuria and blood urea levels compared to non-treated CKD animals.

[0216] CKD animals showed progressive damage to kidney histology compared to sham-operated animals, as further shown in Figure 17. Animals treated with the nutritional or synbiotic blend (Groups P1 and P2, see above) showed significantly better histology, as indicated by less fibrosis and improved glomerular size and volume, compared to untreated CKD animals.

[0217] Overall, the data demonstrates promising benefits of the blend not only in reducing levels of uremic toxins in the blood, but also in helping to stabilize kidney function, as indicated by a reduced appearance of markers of progression of kidney damage.

[0218] Nutritional or synbiotic blend reduces appetite loss, weight loss, and body fat mass associated with uremic toxins and chronic kidney disease Uremic toxicity adversely affects multiple organ systems and metabolic pathways, resulting in organ damage and the development of symptoms, including neurological conditions and PEW. Thus, we have tested the ability of nutritional or synbiotic blends to ameliorate some of the deleterious effects of uremic toxin accumulation, including food intake and depletion of energy reserves such as muscle and fat.

[0219] As shown in Figure 18, CKD animals treated with the P1 and P2 interventions (see above) showed significantly improved body weight change to the same level as non-CKD sham-operated animals and normalized food intake. Energy intake was not significantly different between the two groups. Furthermore, treated animals, particularly those treated with P1, showed a reduction in the loss of fat mass observed in CKD compared to untreated CKD animals, as indicated by better fat reserve capacity, as seen by improvements in epididymal white adipose tissue (eWAT).

[0220] Overall, the data demonstrated promising benefits of the intervention to reduce uremia-related symptoms, including anorexia, weight loss, and PEW.

[0221] The nutritional or synbiotic blend improved intestinal barrier dysfunction. One of the detrimental effects of dysbiosis and the accumulation of uremic toxins in the intestine is to induce changes in intestinal barrier function. Alterations in intestinal permeability may also trigger a vicious cycle of accumulation of uremic toxins in the systemic circulation.

[0222] Therefore, we further examined the potential of the novel material blend to ameliorate some of the reported intestinal barrier dysfunction in CKD. As seen in Figure 19, CKD animals showed impaired protein expression of occludin, a key tight junction, compared to non-CKD animals. Intact tight junctions are important for preventing excessive intestinal contents and molecules from entering the systemic circulation.

[0223] Thus, this data demonstrated promising benefits of interventions to not only prevent the accumulation of uremic toxins but also to alleviate some of the intestinal barrier dysfunction associated with this disease.

[0224] Example 4 - In vivo animal testing: research design To further confirm the renal protective effect demonstrated by the mouse model, 5 / 6 nephrectomized rats were treated with a material blend containing Lactobacillus johnsonii NCC533, cellobiose, and short-medium chain triglycerides containing butyrate and caprylate.

[0225] Nine-week-old Wistar RjHan:WI rats (n=84) underwent two-thirds right nephrectomy at week -4 and total left nephrectomy at week -2. An additional 12 rats underwent sham surgery. Rats were randomized into the following treatment groups based on plasma urea, plasma creatinine, and body weight measured 1 week before the start of the study: (1) Sham surgery (n=12); (2) 5 / 6 Nx vehicle (n = 18); (3) 5 / 6 Nx P1-rat diet (P1-rat intervention was a diet containing a blend of ingredients containing 1% cellobiose and 1% short-medium chain triglycerides containing butyrate and caprylate, administered in the diet for 10 days) 8 (n = 18); (4) 5 / 6 Nx P3 diet (P3 intervention is a dietary blend containing 0.3% cellobiose and 0.3% short-medium chain triglycerides containing butyrate and caprylate, administered in the diet for 10 days) 8 (n = 18); (5) 5 / 6Nx lisinopril 20mg / kg.

[0226] Lisinopril is an angiotensin-converting enzyme inhibitor (ACEi), the standard of care for renal protection.

[0227] The diets were administered for 8 weeks. Drinking water was measured on days 9, 10, 11, 25, 26, 27, 39, 40, and 41. Urine was collected for albumin and creatinine analysis at week 7. Plasma was collected at termination for urea, creatinine, indoxyl sulfate, and p-cresyl sulfate analysis.

[0228] result Lactobacillus johnsonii NCC533 10 8 A synbiotic composition containing 1% cellobiose and 1% short-medium chain triglycerides containing butyrate and caprylate was able to reduce plasma uremic toxin concentrations, particularly indoxyl sulfate and p-cresyl sulfate (see Figure 21).

[0229] As can be seen in Figure 20(C), nephrectomized animals (Group 5 / 6Nx Vehicle) showed higher proteinuria compared to non-CKD animals (Sham-operated), confirming the progression of kidney damage. 5 / 6Nx animals treated with nutritional or synbiotic blends, especially the P1 blend, showed significant improvement in proteinuria levels compared to non-treated CKD animals.

[0230] Example 5 - Clinical Trial Overview (Summary) Below, we disclose an overview of the clinical trials that form the basis for human clinical trials.

[0231] [Table 4] JPEG2026503447000005.jpg98147 JPEG2026503447000006.jpg182149 JPEG2026503447000007.jpg104147

[0232]

[0117] All publications mentioned in the above specification are incorporated herein by reference. Various modifications and variations of the compositions, uses, and methods disclosed herein will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been disclosed in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the disclosed modes for carrying out the invention which are obvious to those skilled in the art are intended to be within the scope of the following claims.

Claims

1. 1. A composition for use in reducing or avoiding the accumulation of uremic toxins in a cardiometabolic or neurodegenerative condition, comprising: (a) the following probiotic bacteria: a probiotic bacterium lacking a gene that produces at least one of urea, uric acid, p-cresol, p-cresyl sulfate, indoxyl sulfate, indole, indoleacetic acid, indoxyl, trimethylamine, TMAO, dimethylglycine, betaine, and / or glutarate; probiotic bacteria deficient in at least one bacterial enzyme selected from urease, carnitine monooxygenase & reductase, tryptophanase, and / or hydroxyphenylacetate; and / or probiotic bacteria expressing at least one bacterial enzyme selected from α-galactosidase, β-galactosidase, glucan 1,4-β-glucosidase, cellulase, β-fructofuranosidase and / or licheninase; (b) a prebiotic selected from a carbohydrate or fiber, wherein the carbohydrate or fiber can be hydrolyzed by at least one bacterial enzyme selected from α-galactosidase, β-galactosidase, glucan 1,4-β-glucosidase, cellulase, β-fructofuranosidase, and / or licheninase; (c) a lipid selected from a triglyceride, a short chain fatty acid, and / or a medium chain fatty acid, wherein the triglyceride comprises a short chain fatty acid, a medium chain fatty acid, or a mixture thereof.

2. The probiotic bacteria (a) are selected from the following a to k: Bifidobacterium animalis subspecies lactis NCC 2818 (CNCM I-3446); b. Bifidobacterium longum subspecies infantis NCC 341 (ATCC 15697(T)); c. Bifidobacterium longum subspecies longum NCC 2705 (CNCM I-2618) (NCBI refseq; GCA_000007525.1); d. Enterococcus faecium NCC 2768 (NCIMB 10415); e. Lactobacillus johnsonii NCC 533 (CNCM I-1225) (NCBI refseq; GCA_000008065.1); f. Lactococcus lactis NCC 2287 (CNCM I-4154); g. Lacticaseibacillus paracasei NCC 2461 (CNCM I-2116); h. Lacticaseibacillus rhamnosus NCC 4007 (CGMCC 1.3724); i. Staphylococcus carnosus NCC 1052 (CNCM I-5400); j. Staphylococcus carnosus NCC 971 (CNCM I-5398); and / or k. Streptococcus thermophilus NCC 2496 (CNCM I-3915); or a probiotic having a genome having at least 95% average nucleotide identity (ANI), preferably at least 96% ANI, more preferably at least 97% ANI, even more preferably at least 98% ANI, and most preferably at least 99% ANI, or even 99.5% ANI, or 99.9% ANI, to any one of the genome sequences of the probiotic bacteria according to any one of a. to k. defined above.

3. 3. The composition for use according to claim 2, wherein the probiotic bacterium (a) is Lactobacillus johnsonii NCC 533 (CNCM I-1225) (NCBI refseq; GCA_000008065.1) or a probiotic bacterium having a genome with an ANI of at least 95%, preferably at least 96%, more preferably at least 97%, even more preferably at least 98%, and most preferably at least 99%, or even 99.5%, or even 99.9%, relative to the genome sequence of Lactobacillus johnsonii NCC 533 (CNCM I-1225) (NCBI refseq; GCA_000008065.1, SEQ ID NO: 1).

4. 4. The composition for use according to any one of claims 1 to 3, wherein the prebiotic (b) is selected from α-galactooligosaccharides (α-GOS) / raffinose, β-galactooligosaccharides (β-GOS), and cellooligosaccharides (COS), or a combination thereof.

5. 5. The composition for use according to any one of claims 1 to 4, wherein the prebiotic (b) is selected from soybean GOS (α-GOS), pea GOS (α-GOS), bovine milk oligosaccharides (BMOS) (β-GOS), vivinal GOS (β-GOS), cellobiose, cellotriose, cellotetraose, and soluble hydrolyzed wheat, human milk oligosaccharides (HMO), soluble hydrolyzed oats and / or β-glucan hydrolysates containing cellobiose or cellotriose or cellotetraose, or combinations thereof.

6. The lipid (c) a. Triglycerides (TG) composed of a mixture of butyrate (C4:0) and medium chain fatty acids (MCFA); b. Triglycerides (TG) composed of a mixture of short- and medium-chain fatty acids (SMCFA), preferably C4 and / or C8 fatty acids; c. Triglycerides (TG) composed of a mixture of butyrate and long chain fatty acids (e.g., triglycerides composed of a mixture of butyrate and oleate); d. Triglycerides composed of medium-chain fatty acids; e. Triglycerides (TG) composed of short-chain fatty acids; f. short-chain fatty acids that can be metabolized into ketone bodies, and g. The composition for use according to any one of claims 1 to 5, wherein the fatty acids are selected from short and medium chain fatty acids, preferably C4 and / or C8 fatty acids, that can be metabolized into ketone bodies.

7. a) the probiotic bacterium is selected from the probiotic bacterium described in a. to k. above, or a probiotic having a genome with an ANI of at least 90%, preferably at least 95%, relative to any of the genome sequences of the probiotic bacterium described in a. to k. above, or a combination thereof, preferably Lactobacillus johnsonii NCC 533 (CNCM I-1225) or at least one probiotic having a genome with an ANI of at least 90%, preferably at least 95%, relative to the sequence of Lactobacillus johnsonii NCC 533 (CNCM I-1225); b) the prebiotic is selected from α-galactooligosaccharides / raffinose, β-galactooligosaccharides, or cellooligosaccharides, or a combination thereof, preferably selected from soybean GOS (α-GOS), pea GOS (α-GOS), milk oligosaccharides (BMOS) (β-GOS), Vivinal GOS (β-GOS), cellobiose, cellotriose, cellotetraose, soluble hydrolyzed wheat, soluble hydrolyzed oats, and / or β-glucan hydrolysates containing cellobiose, cellotriose, or cellotetraose, or a combination thereof; c. the lipids are selected from triglycerides, short chain fatty acids and / or medium chain fatty acids, the triglycerides comprising butyrates, short chain fatty acids, medium chain fatty acids, or mixtures thereof, preferably a. Triglycerides (TG) composed of a mixture of butyrate and medium chain fatty acids (MCFA); b. Triglycerides (TG) composed of a mixture of short and medium chain fatty acids (SMCFA), preferably C4 and / or C8 short and medium chain fatty acids; c. Triglycerides (TG) composed of a mixture of butyrate and long chain fatty acids (e.g., triglycerides composed of a mixture of butyrate and oleate); d. Triglycerides composed of medium-chain fatty acids; e. Triglycerides (TG) composed of short-chain fatty acids; f. short-chain fatty acids that can be metabolized into ketone bodies, and g. The composition for use according to any one of claims 1 to 6, wherein the fatty acids are selected from short and medium chain fatty acids that can be metabolized into ketone bodies, preferably C4 and / or C8 short and medium chain fatty acids.

8. 8. The composition for use according to claim 7, wherein the probiotic bacterium (a) is selected from Lactobacillus johnsonii NCC 533 (CNCM I-1225) (NCBI refseq; GCA_000008065.1) or a probiotic having a genome with an ANI of at least 90%, preferably at least 95%, to the sequence of Lactobacillus johnsonii NCC 533 (CNCM I-1225) (NCBI refseq; GCA_000008065.1).

9. 9. The composition for use according to claim 7 or 8, wherein the prebiotic (b) is selected from soybean GOS (α-GOS), pea GOS (α-GOS), bovine milk oligosaccharides (BMOS) (β-GOS), vivinal GOS (β-GOS), cellobiose, cellotriose, cellotetraose, soluble hydrolyzed wheat, soluble hydrolyzed oats, and / or β-glucan hydrolysates containing cellobiose or cellotriose or cellotetraose, or combinations thereof.

10. The lipid (c) a. Triglycerides (TG) composed of a mixture of butyrate and medium chain fatty acids (MCFA); b. Triglycerides (TGs) composed of a mixture of short- and medium-chain fatty acids (SMCFA); c. Triglycerides (TG) composed of a mixture of butyrate and long chain fatty acids (preferably triglycerides composed of a mixture of butyrate and oleate); d. Triglycerides composed of medium-chain fatty acids; e. Triglycerides (TG) composed of short-chain fatty acids; f. short-chain fatty acids that can be metabolized into ketone bodies, and g. The composition for use according to any one of claims 7 to 9, wherein the medium chain fatty acids are selected from C4 and / or C8 medium chain fatty acids that can be metabolized into ketone bodies.

11. The probiotic bacteria is 10 per daily dose ３ cfu ~ 10 １２ Amount of cfu, typically 10 per daily dose ４ cfu ~ 10 １１ cfu, preferably 10 per daily dose ５ cfu ~ 10 １０ cfu, or 10 per daily dose ５ cfu ~ 10 ９ cfu, also preferably 10 per daily dose ６ cfu ~ 10 ９ cfu, 10 per daily dose ６ cfu ~ 10 ８ cfu, or 10 per daily dose ８ cfu ~ 10 １０ cfu, more preferably about 10 per daily dose ７ cfu ~ 10 ９ The composition for use according to any one of claims 1 to 10, wherein the composition contains in an amount of cfu.

12. 12. The composition for use according to any one of claims 1 to 11, wherein the prebiotic is contained in an amount of 0.1 g to 30 g per daily dose, preferably in an amount of 2 g to 15 g per daily dose.

13. The composition for use according to any one of claims 1 to 12, wherein the lipid is contained in an amount of 0.1 g to 30 g per daily dose, preferably in an amount of 2 g to 15 g per daily dose.

14. A composition for use according to any one of claims 1 to 13, wherein the lipid is a triglyceride composed of butyrate and caprylate or a triglyceride composed of butyrate and oleate.

15. A composition for use according to any one of claims 1 to 13, wherein the lipid is a triglyceride composed of butyrate and caprylate.

16. 16. The composition for use according to any one of claims 1 to 15, in the form of a food product or nutritional composition, a dietary supplement, a food for special medical purposes (FSMP), a nutritional supplement, a dairy-based beverage, a small volume liquid supplement, a functional food product, a functional beverage product, a meal replacement beverage, and combinations thereof.

17. 17. The composition for use according to any one of claims 1 to 16, wherein the composition is provided in the form of a powder, a tablet, a capsule or may be in the form of an oil formulation, an emulsion, an oil-in-water emulsion (o / w emulsion) or a water-in-oil emulsion (w / o emulsion).

18. 18. The composition for use according to any one of claims 1 to 17, wherein the reduction of uremic toxins in said cardiometabolic or neurodegenerative conditions is for slowing the progression of such diseases and co-morbidities and / or for managing the symptoms and syndromes associated with the toxic effects of uremic solutes of such diseases and co-morbidities.

19. The reduction of uremic toxins in the cardiometabolic or neurodegenerative condition comprises: Treatment or prevention of chronic and acute kidney disease; including dialysis and pre-dialysis kidney disease, rare genetically and metabolically induced kidney disease; Treatment or prevention of uremic syndrome, including PEW, bone loss, severe anorexia, fatigue, or inflammation; Delayed complications of progressive kidney disease, including anemia, hyperphosphatemia, secondary hyperparathyroidism, bone disease, and neurological conditions delayed renal comorbidities, including cardiovascular disease; Preventing or managing the risk of malnutrition; Slowing the progression of cardiometabolic diseases, Prevention or management of risk of cardiovascular disease and comorbidities (diabetes), and / or 19. A composition for use according to any one of claims 1 to 18, including the prevention or management of the risk of neurodegenerative and neurological conditions.

20. 18. A multi-part kit suitable for use in reducing or avoiding the accumulation of uremic toxins in a cardiometabolic or neurodegenerative condition comprising the composition of any one of claims 1 to 17, comprising the probiotic, the prebiotic and the lipid in two or more separate containers, and optionally further comprising instructions for use.

21. 18. A method for treating a cardiometabolic or neurodegenerative condition as defined above, comprising as a first step: (a) preparing and providing a composition according to any one of claims 1 to 17, comprising said probiotics, said prebiotics and said lipids; and (b) administering such composition to a patient in need thereof, who is suffering from an increase in uremic toxins in the context of a cardiometabolic or neurodegenerative condition.

22. A method for treating a cardiometabolic or neurodegenerative condition as defined in claim 18, comprising administering to a patient a composition according to any one of claims 1 to 17.

23. Use of a composition according to any one of claims 1 to 17 in the manufacture of a medicament for the treatment of a cardiometabolic or neurodegenerative condition as defined in claim 18.

24. 24. The composition for use according to any one of claims 1 to 13 and 16 to 19, the kit of parts according to claim 20, the method of treatment according to claim 21 or 22, or the use according to claim 23, wherein the lipid is a triglyceride comprising butyrate and / or caprylate.

25. 20. A composition for use according to any one of claims 1 to 13 and 16 to 19, a kit of parts according to claim 20 or claim 24, a method of treatment according to any one of claims 21, 22 and 24 or a use according to claim 23 or claim 24, wherein the lipid is a triglyceride composed of butyrate and caprylate.

26. 24. The composition for use according to any one of claims 1 to 14 and 16 to 19, the kit of parts according to claim 20, the method of treatment according to claim 21 or claim 22, or the use according to claim 23, wherein the lipid is a triglyceride comprising butyrate and oleate.

27. 24. The composition for use according to any one of claims 1 to 14 and 16 to 19, the kit of parts according to claim 20, the method according to claim 21 or 22, or the use according to claim 23, wherein the composition comprises Lactobacillus johnsonii NCC533, cellobiose, and a short-long chain triglyceride containing butyrate and oleate.

28. 24. The composition for use according to any one of claims 1 to 14 and 16 to 19, the kit of parts according to claim 20, the method according to claim 21 or 22, or the use according to claim 23, wherein the composition comprises Lactobacillus johnsonii NCC533, cellobiose, pea GOS, and short-long chain triglycerides containing butyrate and oleate.

29. 24. The composition for use according to any one of claims 1 to 13 and 16 to 19, the kit of parts according to claim 20, the method according to claim 21 or 22, or the use according to claim 23, wherein the composition comprises Lactobacillus johnsonii NCC533, cellobiose, and short-medium chain triglycerides containing butyrate and caprylate.

30. 24. The composition for use according to any one of claims 1 to 13 and 16 to 19, the kit of parts according to claim 20, the method according to claim 21 or 22, or the use according to claim 23, wherein the composition comprises Lactobacillus johnsonii NCC533, cellobiose, pea GOS, and short-medium chain triglycerides containing butyrate and caprylate.

31. The composition contains Lactobacillus johnsonii NCC533 in an amount of 10 ９ 24. The composition for use according to any one of claims 1 to 13 and 16 to 19, the kit of parts according to claim 20, the method according to claim 21 or 22, or the use according to claim 23, comprising: 1% cellobiose; and 1% short-medium chain triglycerides containing butyrate and caprylate.

32. The composition contains Lactobacillus johnsonii NCC533 in an amount of 10 ９ 1% of cellobiose, 1% of pea GOS, and 1% of short-medium chain triglycerides containing butyrate and caprylate, a composition for use according to any one of claims 1 to 13 and 16 to 19, a kit of parts according to claim 20, a method according to claim 21 or 22, or a use according to claim 23.

33. The lipid is a triglyceride composed of butyrate and containing a long-chain fatty acid, and one or more of the triglycerides containing butyrate and a long-chain fatty acid are selected from the group consisting of 1,3-dibutyryl-2-linoleoylglycerol, 1,3-dibutyryl-2-stearoylglycerol, 1-butyryl-2-oleoyl-3-palmitoylglycerol, 1-palmitoyl-2-oleoyl-3-butyrylglycerol, 1-butyryl-2-oleoyl-3-linoleoylglycerol, 1-linoleoyl-2-oleoyl-3-butyrylglycerol, 1-oleoyl-2-butyryl-3-linoleoylglycerol, and 1-linoleoyl-2-butyryl-3-oleoylglycerol.

24. The composition for use according to any one of claims 1 to 14 and 16 to 19, the kit of parts according to claim 20, the method of treatment according to claim 21 or 22, or the use according to claim 23, comprising 1-butyryl-2-linoleoyl-3-oleoylglycerol, 1-oleoyl-2-linoleoyl-3-butyrylglycerol, 1-butyryl-2-stearoyl-3-oleoylglycerol, 1-oleoyl-2-stearoyl-3-butyrylglycerol, 1-butyryl-2-oleoyl-3-stearoylglycerol, and / or 1-stearoyl-2-oleoyl-3-butyrylglycerol, and mixtures of two or more thereof.

34. 34. A composition for use, kit of parts, method or use according to any one of claims 1 to 33, wherein the composition further comprises one or more HMOs, preferably one or more of 2'-fucosyllactose (2'FL), 3-fucosyllactose (3'FL), lacto-N-neotetraose (LNnT), lacto-N-tetraose (LNT), 3'-sialyllactose (3SL) or 6'-sialyllactose (6SL).

35. 34. A composition, kit of parts, method or use for use according to any one of claims 1 to 33, wherein the composition further comprises one or more of 2'-fucosyllactose (2'FL), 3-fucosyllactose (3'FL), lacto-N-neotetraose (LNnT), or lacto-N-tetraose (LNT).

36. Bifidobacterium animalis subsp. lactis NCC 2818 (CNCM I-3446); b. Bifidobacterium longum subsp. infantis NCC 341 (ATCC 15697(T)); c. The composition for use, kit of parts, method, or use according to any one of claims 1 to 35, further comprising one or more of Bifidobacterium longum subsp. longum NCC 2705 (CNCM I-2618) (NCBI refseq; GCA_000007525.1).

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