EA043686B1 - PHARMACEUTICAL COMPOSITION FOR REDUCING THE LEVEL OF PROTEIN-BOUND UREMIC TOXINS - Google Patents

Description

Field to which the invention relates

The present invention relates to compositions/formulations for reducing the level of protein-bound uremic toxins in chronic kidney disease (CKD). More specifically, the present invention relates to a pharmaceutical composition/formulation comprising a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof for reducing the level of protein-bound uremic toxins. The present application also presents various compositions/formulations and a method for their preparation.

Prerequisites for creating the invention

In the last two decades, there has been renewed interest in uremic syndrome. Uremic syndrome can be thought of as the inappropriate removal and subsequent accumulation of organic products that are normally metabolized or excreted by the kidneys in patients with chronic kidney disease (CKD).

Uremic syndrome is the result of a progressive decline in kidney function, which leads to the accumulation of organic waste products. These waste products are commonly referred to as uremic toxins or uremic retained solutes. Typically, protein-bound uremic toxins are produced by the metabolism of amino acids in the intestines.

In CKD, the influx of urea and other retained toxins causes changes in the gut microbiome. The number of beneficial bacteria that produce short-chain fatty acids, essential nutrients for the colonic epithelium, decreases, while the number of bacteria that produce uremic toxins such as indoxyl sulfate (IS), para-cresyl sulfate (PCS) and trimethylamine K-oxide (TMAO) increases ).

Cardiovascular disease (CVD) is highly prevalent in patients with CKD. Deaths from cardiovascular disease account for approximately half of all deaths among patients with CKD. Protein-bound uremic toxins, associated with the progression of CKD, are independent cardiovascular risk factors in both non-dialysis and dialysis patients.

There are currently five different gut-derived uremic toxins that are associated with cardiovascular disease and mortality in CKD, as well as other target organ toxicity: indoxyl sulfate (IS), indole-3-acetic acid (IAA), p-cresyl sulfate ( PCS), trimethylamine K-oxide (TMAO) and phenylacetylglutamine (PAG).

PCS and IS are major protein-bound uremic toxins that have been reported to not only reduce endothelial proliferation but also inhibit endothelial repair mechanisms. In addition, increasing evidence suggests that they are a valuable predictor of cardiovascular events, infections, and all-cause mortality in hemodialysis patients. However, there is also a significant association of serum PCS with CVD in patients with CKD. PCS and IS appear to be a new and important surrogate in patients with CKD.

Existing treatment and disadvantages

Efforts have been made to understand the mechanisms responsible for such uremic toxicity and to develop therapeutic interventions that can reduce the side effects of uremic toxins. An alternative means of reducing their plasma levels is to reduce their production.

Dialysis processes were developed to separate elements in solution by diffusion through a semipermeable membrane (solute diffusion) down a concentration gradient. Hemodialysis allows CKD patients with loss of kidney function to survive for a longer period, and the advent of dialysis therapy has been good news for many patients. Hemodialysis uses the patient's blood to remove waste, toxins, and excess water from the patient's body. However, unless a kidney transplant is performed, dialysis therapy, which entails chronic complications such as itching and anemia, must be continued throughout life and causes great mental and physical stress in patients. It is often reported that the accumulation of uremic substances in the body leads to the development of complications of dialysis, and therefore the problem is how to significantly and quickly reduce harmful substances that cannot be removed at all or sufficiently by dialysis from the body. Another disadvantage of hemodialysis is the need to use an anticoagulant during treatment, which can inevitably increase the risk of internal bleeding. More importantly, in CKD, even with hemodialysis, protein-bound uremic toxins cannot be excreted by the kidneys and accumulate in the plasma. Moreover, dialysis is very expensive, inconvenient, time-consuming, and can sometimes cause one or more side effects. After a successful kidney transplant, the patient can live a more normal life with fewer long-term costs. However, there are also high costs associated with the transplant surgery, recovery period, and ongoing need for anti-rejection medications. In addition, there is often a shortage of suitable donors.

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Among other possible attempts, nutritional control or dietary/food intake control has been recognized as one of the significant management options for CKD treatment. In addition, a low protein diet is often considered as a possible dietary approach to reduce serum concentrations of uremic toxins. However, some studies suggest that the risk of malnutrition on a low-protein diet is even higher in CKD populations.

In addition to the above, attempts have also been made to remove uremic toxins in CKD patients by adsorbing such toxic substances into the gastrointestinal tract using activated charcoal. However, it also adsorbs enzymes, vitamins, minerals, etc. In addition, activated charcoal treatment involves a large number of pills and is associated with constipation and gastrointestinal distress.

In view of the above disadvantages and side effects associated with traditional treatments, it is desirable to develop a formulation that not only effectively reduces protein-bound uremic toxins but also does not have any side effects in patients with CKD. Moreover, it is also desirable to have a composition/formulation that is cost effective and suitable for patients of all age groups.

One possible way to suppress the production of such solutes (uremic toxins) is to increase dietary fiber intake. The term fiber includes a variety of carbohydrates and related substances that are resistant to digestion in the small intestine. Colon microbes can break down fiber into short-chain fatty acids, which provide energy to the host and promote microbial growth. Increased delivery of fiber to the colon may cause microbes to use amino acids for growth rather than convert them into uremic solutes. In addition, fibers can alter the microbial population in the colon in a way that reduces the production of unwanted solutes. Fiber may also reduce the transit time through the colon available for microbial metabolism.

Dietary fibers such as inulin may play a special role in kidney disease because they have the potential to reduce colon microbial production of protein-derived uremic solutes without protein limitation. It has been suggested that fiber may influence the production of nitrogenous solutes by providing energy to colon microbes. Inulin is also a type of oligosaccharide called fructan. Fructans are a chain of fructose (sugar) molecules linked together. Inulin is fermented by bacteria that normalize the functioning of the colon and is considered a prebiotic. Inulin and oligofructose have lower caloric content than regular carbohydrates because they are not digested by human intestinal enzymes. Thus, inulin and oligofructose pass through the mouth, stomach and small intestine without being metabolized. Some studies show that almost all inulin or oligofructose ingested ends up in the colon, where it is completely fermented by colonic microflora, leading to increased utilization of the amino acids tyrosine and tryptophan, which leads to decreased production of p-cresol and indoles.

Betaines are fully N-methylated amino acids. Betaines are natural products that play an important role in the metabolism of plants and animals. Betaine, also known as glycine betaine or trimethylglycine, is a glycine derivative in which three methyl groups are bonded to the nitrogen atom of the glycine molecule. Betaine has a bipolar structure and contains several reactive methyl groups and can thus donate them in enzyme-catalyzed reactions. Betaine is involved in the metabolism of homocysteine. High concentrations of homocysteine in the blood are associated with an increased risk of cardiovascular disease. Plasma total homocysteine (tHcy) increases as kidney function declines, and more than 80% of people with end-stage renal disease have hyperhomocysteinemia. Betaine is used, among other things, as a feed additive and crop improver under stress conditions, and in the cosmetic, pharmaceutical and food industries.

Related Prior Art Applications

CN 1504229 discloses an immune detoxifying agent in the form of a powder for the treatment of uremia, where the powder includes astragalus root, aralia sinensis root, white atractylis rhizome, red sparrow root, corinth pink and rhubarb.

WO 2005056040 relates to targeting uremic toxins in vivo in kidney disease or dysfunction using enzymes targeting uremic toxins. The composition disclosed herein may include one, two or more enzymes that act on uremic toxins, such as urease, uricase or creatininase.

EP 2754446 discloses a probiotic composition for reducing levels of uremic toxins. The probiotic composition disclosed herein includes: at least one of Lactobacillus plantarum BCRC 12251, Lactobacillus paracasei BCRC 12188, Streptococcus thermophilus BCRC 13869 and Enterococcus faecalis. The composition can be used to remove uremic toxins, such as protein-bound uremic toxins, from the blood.

CN 104740611 discloses a traditional Chinese medicine composition for the treatment of uremia,

- 2 043686 including lyophilized powder of the polypeptide hormone of the brain, milkweed grass, psoralea fruits and wormwood paniculata. This composition is non-toxic, has no side effects and is effective in improving uremia.

US20160051600 discloses the use of a gel-like mixture of probiotics and prebiotics with a synergistic symbiotic effect for the treatment of chronic kidney disease, to reduce the concentration of uremic toxins, improve the renal function of a patient with increased levels of urea, creatinine, uric acid, para-cresols or indoles in the blood. This reference document discloses a gel-like mixture of probiotics and prebiotics containing water, cane sugar, liquid glucose, protein element, xanthan gum, plant-derived prebiotic fiber, vitamins, citrus seed extract, citric acid, malic acid, bifidobacteria, lactobacilli, color, which is a mixture of fine powder pigment with water, and flavor.

It is clear from the above description and the prior art that although there are options for reducing levels of uremic toxins, including protein-bound uremic toxins in CKD, many such treatment options are associated with unpleasant or harmful side effects. Therefore, there is still a need for compositions/formulations that reduce the levels of protein-bound uremic toxins in patients with CKD without any side effects in humans or animals, and also have good tolerability at an effective dose and a good safety profile.

The essence of the invention

In the present invention, it has been found that reduction in the level of protein bound uremic toxins in CKD patients (human or animal) without any side effects, accompanied by good tolerability at an effective dose and a good safety profile, is achieved by administering a stable composition/formulation comprising synergistic combination of inulin with betaine.

Accordingly, the present application provides compositions/formulations for reducing the level of protein-bound uremic toxins.

In a preferred aspect, the present application provides pharmaceutical compositions/formulations for reducing the level of protein-bound uremic toxins in chronic kidney disease (CKD).

The combination of inulin and betaine of the present invention is capable of providing a safe pharmaceutical composition/formulation with enhanced and/or synergistic effect compared to the individual use of inulin or betaine or pharmaceutically acceptable salts thereof for reducing the level of protein-bound uremic toxins in CKD.

The present invention provides pharmaceutical compositions/formulations comprising a synergistic combination of inulin or pharmaceutically acceptable salts thereof and betaine or pharmaceutically acceptable salts thereof.

One aspect of the present invention is to provide a composition/formulation for reducing the level of protein-bound uremic toxins in CKD, wherein the composition/formulation includes a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof.

Another aspect of the present invention is to provide a composition/formulation comprising a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof to reduce the level of protein-bound uremic toxins, such as p-cresyl sulfate (PCS) and indoxyl sulfate (IS), in CKD.

Another aspect of the present invention is to provide a pharmaceutical composition/formulation comprising a combination of inulin and betaine or pharmaceutically acceptable salts thereof in an optimized and/or intelligently selected synergistic ratio.

It is also an aspect of the present invention to provide a suitable dosage form comprising a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof for reducing the level of protein-bound uremic toxins in CKD.

Another aspect of the present invention is to provide a suitable dosage regimen of a composition/formulation comprising a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof to reduce the level of protein-bound uremic toxins in CKD.

In a preferred aspect, the present invention provides pharmaceutical compositions/formulations comprising a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof, together with pharmaceutically acceptable excipients.

In one aspect, the composition/formulation of the present invention further includes resistant starch, omega-3 fatty acid, short chain fatty acid(s) (SCFA), or a combination thereof.

In another aspect, the present invention provides a composition comprising inulin or pharmaceutically acceptable salts thereof, betaine or pharmaceutically acceptable salts thereof, and additional active ingredients selected from resistant starch, omega-3 fatty acid, short chain fatty acid(s) (SCFA). ) or combinations thereof.

In a preferred aspect, the present invention provides pharmaceutical compositions/formulations comprising a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof with additional active ingredients selected from resistant starch, omega-3 fatty acid, short chain fatty acid(s) (SCFA), together with pharmaceutically acceptable excipients.

In yet another preferred aspect, the present invention provides any suitable dosage form for the composition/formulation of the present invention. Preferably, the composition/formulation of the present invention is formulated in an oral dosage form.

Brief description of the figures

Fig. 1 - effect of the tested composition/formulation on the level of indoxyl sulfate (IS) compared to disease control in 5/6 nephrectomy-induced chronic kidney disease in rats;

fig. 2 - effect of the tested composition/formulation on the level of p-cresyl sulfate (PCS) compared to disease control in 5/6 nephrectomy-induced chronic kidney disease in rats.

Detailed Description of the Invention

Although the invention is susceptible to various modifications and alternative forms, specific embodiments are described in detail in the Examples section below. However, it should be understood that they are not intended to limit the invention to the specific forms disclosed, but rather, the invention is intended to cover all modifications, equivalents and alternatives falling within the scope of the invention.

In addition, it should be understood that all terminology used in this application is intended to describe specific embodiments only and is not intended to limit the scope of the invention in any way.

Unless otherwise indicated, all technical and scientific expressions used in this application have the same meaning as commonly understood by one skilled in the art to which the embodiments of the invention relate.

In describing embodiments of the present invention and in the claims, the following terminology will be used in accordance with the definitions set forth below, which are known in the art.

Singular forms include the plural unless the context clearly dictates otherwise.

Unless otherwise indicated, all percentages and quantities stated in this section and elsewhere in the description should be understood to refer to percentages by weight. The amounts given are based on the active mass of the substance.

The term composition includes pharmaceutical compositions, nutraceutical compositions, dietary supplement compositions, drug compositions, dietary supplement compositions, medical specialty foods, and any other suitable compositions.

The term formulation includes pharmaceutical formulations, nutraceutical formulations, dietary supplement formulations, drug formulations, dietary supplement formulations, medical specialty foods, and any other suitable formulations. The terms composition and composition are used interchangeably unless the context requires otherwise.

The term pharmaceutically acceptable salts as used herein means those salts that are medically suitable for use in contact with tissues of humans and animals without undue toxicity, irritation, allergic reaction, or the like. and correspond to a reasonable benefit/risk ratio. In particular, the term pharmaceutically acceptable salts means relatively non-toxic, inorganic and organic acid addition salts of compounds, as well as solvates, co-crystals, polymorphs, derivatives and the like. salts

The present invention is directed to a pharmaceutical composition/formulation comprising a synergistic combination of inulin or its pharmaceutically acceptable salts and betaine or its pharmaceutically acceptable salts for reducing the level of protein-bound uremic toxins such as p-cresyl sulfate (PCS) and indoxyl sulfate (IS), while chronic kidney disease (CKD). In this regard, the inventors conducted extensive research along with preclinical studies and found that the combination of inulin and betaine or pharmaceutically acceptable salts thereof provides a synergistic effect to reduce the level of protein-bound uremic toxins such as PCS and IS in CKD.

Some prior art studies show an effect of inulin to reduce plasma concentrations of IS and PCS in CKD. However, the role of betaine in reducing the levels of protein-bound uremic toxins such as IS and PCS in CKD is not yet known. Moreover, the effects of the composition/formulation of the present invention, comprising a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof, have not been reported to dramatically/significantly improve the reduction of protein-bound uremic toxins.

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The synergistic combination of the present invention is capable of providing a safe pharmaceutical composition/formulation comprising inulin with betaine or their pharmaceutically acceptable salts with enhanced and/or synergistic effects in reducing the level of protein-bound uremic toxins in CKD compared to inulin or betaine or their pharmaceutically acceptable salts in separately.

Within the scope of the present invention, it has been discovered that the synergistic combination/formulation of inulin with betaine or pharmaceutically acceptable salts thereof has unexpected and particularly beneficial effects. This makes them particularly suitable for reducing levels of protein-bound uremic toxins in CKD.

Inulin can be taken in any suitable form. More specifically, inulin can be selected from its pharmaceutically acceptable salts, esters, polymorphs or derivatives. In particular, the present invention uses inulin propionate ester (IPE), oligofructose-enriched inulin, inulin sulfate or inulin.

In one embodiment, the pharmaceutical composition/formulation of the present invention includes inulin or pharmaceutically acceptable salts thereof, wherein the amount of inulin or pharmaceutically acceptable salts thereof in the composition/formulation of the present invention is at least 90% by weight, based on the weight of the composition. In another embodiment, the amount of inulin or pharmaceutically acceptable salts thereof in the pharmaceutical composition/formulation of the present invention is less than or equal to 99% by weight, based on the weight of the composition. In yet another embodiment, the amount of inulin or pharmaceutically acceptable salts thereof that can be used in the pharmaceutical composition/formulation of the present invention is from 90 to 99% by weight, based on the weight of the composition/formulation. In another embodiment, the amount of inulin or pharmaceutically acceptable salts thereof is from 90 to 95% by weight, based on the weight of the composition. In yet another embodiment, the amount of inulin or pharmaceutically acceptable salts thereof is from 95 to 99% by weight, based on the weight of the composition.

In another embodiment, the pharmaceutical composition/formulation of the present invention includes inulin or pharmaceutically acceptable salts thereof, wherein the amount of inulin or pharmaceutically acceptable salts thereof in the composition/formulation of the present invention is at least 1 g per dosage unit. In another embodiment, the composition/formulation of the present invention includes an amount of inulin or pharmaceutically acceptable salts thereof of less than or equal to 20 g per dosage unit. In yet another embodiment, the amount of inulin or pharmaceutically acceptable salts thereof that can be used in the pharmaceutical composition/formulation of the present invention is from 1 to 20 g per dosage unit. In a preferred embodiment, the amount of inulin or pharmaceutically acceptable salts thereof is from 5 to 20 g per dosage unit. In a preferred embodiment, the intake of inulin or pharmaceutically acceptable salts thereof is from about 1 to 20 g per day.

The composition of the present invention also includes betaine in synergistic combination with inulin, wherein betaine can participate in the metabolism of homocysteine. Betaine can be taken in any suitable form. In particular, betaine may be selected from its pharmaceutically acceptable salts, esters, polymorphs or derivatives. More specifically, betaine may be in hygroscopic or non-hygroscopic form. In particular, betaine may be in the form of a monohydrate, an anhydrous form, glycine betaine, or pharmaceutically acceptable salts thereof, such as betaine hydrochloride.

In one embodiment, the pharmaceutical composition/formulation of the present invention includes betaine or pharmaceutically acceptable salts thereof, wherein the amount of betaine in the composition/formulation of the present invention is at least 0.5 wt.%, based on the weight of the composition. In another embodiment, the amount of betaine or pharmaceutically acceptable salts thereof in the composition/formulation of the present invention is less than or equal to 10 wt.%, based on the weight of the composition. In yet another embodiment, the composition/formulation of the present invention includes betaine or pharmaceutically acceptable salts thereof in a reasonably selected amount ranging from 0.5 to 10% by weight, based on the weight of the composition. In another embodiment, the amount of betaine or pharmaceutically acceptable salts thereof is from 0.5 to 5 wt.%, based on the weight of the composition. In yet another embodiment, the amount of betaine or pharmaceutically acceptable salts thereof is from 5 to 10% by weight, based on the weight of the composition.

In another embodiment, the pharmaceutical composition/formulation of the present invention includes betaine or pharmaceutically acceptable salts thereof, wherein the amount of betaine or pharmaceutically acceptable salts thereof in the composition/formulation of the present invention is at least 1 mg per dosage unit. In another embodiment, the composition/formulation of the present invention includes an amount of betaine or pharmaceutically acceptable salts thereof of less than or equal to 1000 mg per dosage unit. In yet another embodiment, the amount of betaine or pharmaceutically acceptable salts thereof that can be used in the pharmaceutical composition/formulation of the present invention is from 1 to 1000 mg per dosage unit. In a preferred embodiment, the amount of betaine or pharmaceutically acceptable salts thereof is from 1 to 1000 mg per dosage unit. In a preferred embodiment, the dosage of betaine or pharmaceutically acceptable salts thereof is from about 1 to 1000 mg per day.

In yet another embodiment, the pharmaceutical composition/formulation of the present invention includes a synergistic combination of inulin or pharmaceutically acceptable salts thereof ranging from 90 to 99% by weight, based on the weight of the composition, and betaine or pharmaceutically acceptable salts thereof ranging from 0.5 to 99% by weight of the composition. 10 wt.%, based on the weight of the composition.

The composition/formulation of the present invention uses betaine or pharmaceutically acceptable salts thereof in combination with inulin or pharmaceutically acceptable salts thereof to provide a synergistic effect and enhance the effect of inulin or pharmaceutically acceptable salts thereof in reducing the level of protein-bound uremic toxins in CKD. In another embodiment, the composition/formulation of the present invention uses inulin or pharmaceutically acceptable salts thereof in combination with betaine or pharmaceutically acceptable salts thereof in a reasonably optimized ratio (w/w) of 1:0.006 to 1:0.111 to synergistically reduce levels protein-bound uremic toxins in CKD.

In a preferred embodiment, the pharmaceutical composition/formulation of the present invention includes a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof with pharmaceutically acceptable excipients.

In yet another embodiment, the composition/formulation of the present invention further includes other active ingredients, such as resistant starch, omega-3 fatty acid, short chain fatty acid(s) (SCFA), or any combination thereof.

In yet another embodiment, the composition/formulation of the present invention includes a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof with additional active ingredients and pharmaceutically acceptable excipients. Additional active ingredients, such as resistant starch, omega-3 fatty acid, short-chain fatty acid(s) (SCFA), or any combination thereof, may further enhance the effect of the synergistic combination of inulin and betaine or their pharmaceutically acceptable salts.

In a preferred embodiment, the pharmaceutical composition/formulation of the present invention includes a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof with resistant starch. In another preferred embodiment, the pharmaceutical composition/formulation of the present invention includes a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof with resistant starch and pharmaceutically acceptable excipients.

In one embodiment, the amount of resistant starch that can be used in the pharmaceutical composition/formulation of the present invention is from 0.8 to 9 wt.%, based on the weight of the composition/formulation.

In another embodiment, the amount of resistant starch that can be used in the pharmaceutical composition/formulation of the present invention is from 0.5 to 20 g per dosage unit. In a preferred embodiment, the amount of resistant starch taken is from 0.5 to 20 g per day.

In a preferred embodiment, the pharmaceutical composition/formulation of the present invention includes a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof with an omega-3 fatty acid. In another preferred embodiment, the pharmaceutical composition/formulation of the present invention includes a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof with an omega-3 fatty acid and pharmaceutically acceptable excipients.

In one embodiment, the omega-3 fatty acid may be alphalinolenic acid (ALA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), or a combination thereof. In a preferred embodiment, the amount of omega-3 fatty acid that can be used in the pharmaceutical composition/formulation of the present invention is from 0.89 to 9% by weight, based on the weight of the composition/formulation.

In a preferred embodiment, the amount of omega-3 fatty acid that can be used in the pharmaceutical composition/formulation of the present invention is from 100 mg to 3 g per dosage unit. In a preferred embodiment, the amount of omega-3 fatty acid taken is from 100 mg to 3 g per day.

In a preferred embodiment, the pharmaceutical composition/formulation of the present invention includes a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof with SCFA or pharmaceutically acceptable salts thereof. In another preferred embodiment, the pharmaceutical composition/formulation of the present invention includes a synergistic combination of inulin and betaine or pharmaceutically acceptable salts thereof with SCFAs and pharmaceutically acceptable excipients.

In one embodiment, SCFAs are linear or branched C1-C5 monocarboxylic organic acids such as acetic, propionic, butyric and isovaleric acids. Pharmaceutically acceptable SCFA salts include, but are not limited to, sodium propionate, sodium butyrate, etc. In another embodiment, SCFA can be used in the pharmaceutical composition/formulation of the present invention in the range of from 0.8 to 8.2 wt.%, based on the weight of the composition/formulation.

In a preferred embodiment, the amount of SCFA that can be used in the pharmaceutical composition/formulation of the present invention is from 150 mg to 3 g per dosage unit. In a preferred embodiment, the amount of SCFA taken is from about 150 mg to 3 g per day.

In yet another embodiment, the pharmaceutical compositions/formulations of the present invention are prepared/provided in any suitable form for administration, such as solid and liquid dosage form. Solid dosage form includes oral dosage form such as powder, tablet, capsule, hard capsule filled with liquid or solid, soft capsule, pill, sachet, granule, etc. Liquid dosage form includes oral dosage form such as solution, suspension, emulsion, syrup, etc.

The term excipient or suitable excipient as used herein means a pharmacologically inactive component. Excipients that can be used in the preparation of the pharmaceutical composition/formulation of the present invention are generally safe and non-toxic. A reference to an excipient includes either one or more than one such excipient. Co-processed excipients are also included within the scope of the present invention. In yet another embodiment, the composition/formulation of the present invention contains pharmaceutically acceptable carriers/excipients/diluents or excipients to obtain the desired composition/formulation or dosage form. Pharmaceutically acceptable carriers/solvents/diluents or excipients in the context of this application means, without limitation, any adjuvants, carriers, sweeteners, flavor enhancers, diluents, preservatives, coloring agents, surfactants, wetting agents, dispersing agents , suspending agents, complexing agents, stabilizers, isotonic agents, solvents, emulsifiers, encapsulating agents, polymers, coating agents, waxes, encapsulating polymer delivery systems. Excipients may also include anti-blocking agents, antioxidants, binders, pH modifiers, solvents, coatings, pressing aids, disintegrants, emollients, fillers, film-forming agents, flavoring agents, glidants (flow enhancers), lubricants, preservatives, sorbents, anti-caking agents, nutritional supplements or water for hydration.

In a preferred embodiment, the pharmaceutically acceptable excipients are flavoring agents and sweeteners.

Sweeteners in the present invention include, but are not limited to, lactitol, maltitol, mannitol, sorbitol, sucrose, xylitol, acesulfame potassium, alitame, aspartame, compressed sugar, confectioners' sugar, dextrose, erythritol, fructose, glycerin, glycine, isomalt, liquid glucose , maltose, neohesperidin dihydrochalcone, neotame, saccharin, sodium saccharin, sodium cyclamate, sucralose, tagatose, thaumatin, trehalose, etc. The amount of sweetener in the pharmaceutical composition/formulation of the present invention is used in the range of from about 0.01 to 1 wt.%, based on the weight of the composition/formulation.

The flavor used may be orally acceptable natural or synthetic flavors, natural essences, extractable essences, essential oils, or a mixture thereof, but is not limited to adipic acid, n-butyl lactate, confectioner's sugar, denatonium benzoate, dibutyl sebacate, ethyl acetate, ethyl lactate, ethyl maltol, ethyl vanillin, ethyl cellulose , fumaric acid, leucine, malic acid, maltol, menthol, methionine, monosodium glutamate, phosphoric acid, propionic acid, sodium acetate, sodium lactate, sodium propionate, tartaric acid, thymol, triethyl citrate, vanillin, vanilla, pineapple, mixed fruit, banana , orange, geraniol, geranium essence, eucalyptus essential oil, almond oil, fruit flavors, honey or the like. The amount of flavoring agent in the pharmaceutical composition/formulation of the present invention is used in the range of 0.01 to 3 wt.%, based on the weight of the composition/formulation.

The diluent is selected from microcrystalline cellulose, lactose (anhydrous/monohydrate/spray dried), powdered cellulose, silicified microcrystalline cellulose, ammonium alginate, calcium carbonate, calcium lactate, dibasic calcium phosphate (anhydrous/dibasic dehydrate/tribasic), calcium silicate, calcium sulfate, cellulose acetate, compressed sugar, confectioner's sugar, corn starch, pregelatinized starch, dextrates, dextrin, dextrose, erythritol, ethylcellulose, fructose, fumaric acid, glyceryl palmitostearate, isomalt, kaolin, lactitol, magnesium carbonate, magnesium oxide, maltodextrin, maltose, mannitol , medium chain triglycerides, polydextrose, polymethacrylates, simethicone, sodium alginate, sodium chloride, sorbitol, sterilized corn, sucrose, sugar spheres, β-cyclodextrin sulfobutyl ether, talc, tragacanth, trehalose, xylitol and

etc. The amount of diluent in the pharmaceutical composition/formulation of the present invention is from 0 to 10% by weight, based on the weight of the composition/formulation.

The binder is selected from hypromellose, gum acacia, agar, alginic acid, calcium carbonate, calcium lactate, carbomers, sodium carboxymethylcellulose, carrageenan, cellulose acetate phthalate, ceratonia, chitosan, copovidone, corn starch, pregelatinized starch, cottonseed oil, dextrate, dextrin, dextrose , ethylcellulose, gelatin, glyceryl behenate, guar gum, hydrogenated vegetable oil type I, hydroxyethylcellulose, hydroxyethylmethylcellulose, hydroxypropylcellulose, lactose, liquid glucose, low-substituted hypromellose, magnesium aluminosilicate, maltodextrin, maltose, methylcellulose, microcrystalline cellulose, pectin, polox measure, polycarbophil, polydextrose, polyethylene oxide, polymethacrylates, povidone, sodium alginate, stearic acid, sucrose, sunflower oil, tricaprylin, vitamin E polyethylene glycol succinate, zein, etc. The amount of binder in the pharmaceutical composition/formulation of the present invention is from 0 to 10% by weight, based on the weight of the composition/formulation.

The baking powder is chosen from sodtrikarmallosis, crosparmallosis, carboxymethyl cellulose (sodium/calcium), sodium starch glycoles, alginic acid, alginate of calcium, powder cellulose, hitosan, colloidal silicon dioxide, sodium -sodium, glycine, and glycine, and glycine Cosemed hydroxypropyllulosis, magnesium aluminosilicate, methylcellulose, microcrystalline cellulose, polacrilin potassium, povidone, sodium alginate, pregelatinized starch, etc. The amount of disintegrant in the pharmaceutical composition/formulation of the present invention is from 0 to 10% by weight, based on the weight of the composition/formulation.

The lubricant is selected from magnesium stearate, zinc stearate, calcium stearate, glycerol monostearate, glycerol behenate, glycerol palmitostearate, hydrogenated castor oil, hydrogenated vegetable oil type I, light mineral oil, magnesium lauryl sulfate, medium chain triglycerides, mineral oil, myristic acid, palmitic acid , poloxamer, polyethylene glycol, sodium benzoate, sodium chloride, sodium lauryl sulfate, sodium stearyl fumarate, stearic acid, talc, potassium benzoate, etc. The amount of lubricant in the pharmaceutical composition/formulation of the present invention is from 0 to 5 wt.%, based on the weight of the composition/formulation.

The glidant is selected from colloidal silica, talc, tribasic calcium phosphate, powdered cellulose, magnesium oxide, magnesium silicate, magnesium trisilicate, and the like. The amount of glidant in the pharmaceutical composition/formulation of the present invention is from 0 to 5 wt.%, based on the weight of the composition/formulation.

The coating layer for the composition/formulation of the present invention is selected from a film coating, a seal coating or an enteric coating. The total amount of coating agent in the pharmaceutical composition/formulation of the present invention is from 0 to 10% by weight, based on the weight of the composition/formulation.

The sealant coating material is selected from Instamoistshield (hydroxypropyl methylcellulose, polyethylene glycol, talc, titanium dioxide, ethylcellulose), gelatin, copovidone, hydroxyethylcellulose, ethylcellulose, vanillin, hydroxypropylcellulose, guar gum, maleic acid, hypromellose, polymethacrylates, methylcellulose, and the like. The amount of sealant coating material in the pharmaceutical composition/formulation of the present invention is from 0 to 3.0% by weight, based on the weight of the composition/formulation.

The enteric coating material is selected from Instacoat EN HPMC P (hydroxypropyl methylcellulose phthalate, polyethylene glycol, titanium dioxide, red iron oxide), methacrylate copolymer, shellac, sodium alginate, acetyl tributyl citrate, carbomers, cellulose acetate phthalate, guar gum, hypromellose acetate succinate, phthal ata of hypromellose, polymethacrylates, polyvinyl acetate phthalate, potassium chloride, glycerin, Sureteric, tributyl citrate, triethyl citrate, triolein, white wax, zein, cellulose acetate phthalate with ethyl cellulose, chitosan, hydroxypropyl cellulose, etc. The amount of enteric coating material in the pharmaceutical composition/formulation of the present invention is from 0 to 10.0% by weight, based on the weight of the composition/formulation.

The film coating material is selected from Instacoat Universal (hydroxypropyl methylcellulose, polyethylene glycol, talc, titanium dioxide), guar gum, hypromellose, povidone, hydroxypropylcellulose, cellulose acetate, polydextrose, ethylcellulose, methylcellulose, gelatin, glycerin, maltodextrin, and the like. The amount of film coating material in the pharmaceutical composition/formulation of the present invention is from 0 to 3.0% by weight, based on the weight of the composition/formulation.

The solvent is selected from water, alcohol, isopropyl alcohol, propylene glycol, almond oil, benzyl alcohol, benzyl benzoate, butylene glycol, carbon dioxide, castor oil, corn oil, cottonseed oil, dibutyl phthalate, diethyl phthalate, dimethyl ether, albumin, dimethyl phthalate, dimethyl sulfoxide, dimethylacetamide, ethyl acetate, ethyl lactate, ethyl oleate,

- 8 043686 glycerin, glycofurol, isopropyl myristate, isopropyl palmitate, light mineral oil, medium chain triglycerides, methyl lactate, mineral oil, monoethanolamine, octyldodecanol, olive oil, peanut oil, polyethylene glycol, polyoxyl 35 castor oil, propylene carbonate, pyrrolidone, safflower oil, sesame oil, soybean oil, sunflower oil, triacetin, tricaprylin, triethanolamine, triethyl citrate, triolein, water-miscible solvents and the like. The amount of solvent in the pharmaceutical composition/formulation of the present invention is used in a sufficient amount.

The solubilizing agent is selected from polysorbate 80, sodium lauryl sulfate, anionic emulsifying wax, glycerin, nonionic emulsifying wax, glyceryl monooleate, phospholipids, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, polyoxyglycerides, sorbitan esters, triethyl citrate, Vitamin E polyethylene glycol succinate, microcrystalline cellulose, sodium carboxymethylcellulose, diethanolamine, ethylene glycol, palmitostearate, glycerol monostearate, hypromellose, hypromellose acetate succinate, lecithin, polyethylene alkyl ethers, aluminum oxide, poly(methyl vinyl ether/maleic anhydride), carbon calcium ata, crospovidone, cyclodextrins, fructose, hydroxypropyl betadex, oleyl alcohol, povidone, benzalkonium chloride, benzethonium chloride, benzyl alcohol, benzyl benzoate, cetylpyridinium chloride, meglumine, poloxamer, pyrrolidone, sodium bicarbonate, stearic acid, β-cyclodextrin sulfobutyl ether, tricaprylin, triolein, docuse that sodium, glycine, alcohol, self-emulsifying glyceryl monooleate, cationic benzethonium chloride, cetrimide, xanthan gum, lauric acid, myristyl alcohol, butylparaben, ethylparaben, methylparaben, propylparaben, sorbic acid and the like. The amount of solubilizing agent in the pharmaceutical composition of the present invention is from 0 to 5 wt.%, based on the weight of the composition.

The antioxidant is selected from propyl gallate, lecithin, vitamin E, tocopherol, sesamin, sesamol, sesamoline, alpha-tocopherol, ascorbic acid, ascorbyl palmitate, fumaric acid, malic acid, sodium metabisulfite, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), etc. P. The amount of antioxidant in the pharmaceutical composition of the present invention is from 0 to 1 wt.%, based on the weight of the composition.

The preservative is selected from diazolidinyl urea, iodopropnyl butyl carbamate, vitamin E (alpha tocopherol) and its derivatives including vitamin E acetate (alpha tocopherol acetate), vitamin C (ascorbic acid), butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), esters p-hydroxybenzoic acid, ethylparaben, propylparaben, butylparaben, etc. The amount of preservative in the pharmaceutical composition of the present invention is from 0 to 1 wt.%, based on the weight of the composition.

In another embodiment, the excipients are present in an amount sufficient to obtain a suitable composition/formulation or dosage form.

The active components that are present in the composition/formulation can be used in the most suitable physical state to prepare a suitable form for administration. The composition of the present invention is preferably for oral administration, and the preferred form is solid or liquid form. To obtain these solid forms, in particular powder or granules in sachet form, or to fill capsules or tablets, ingredients that are naturally in a liquid state can be suitably adapted to form solid forms. For example, since butyric acid is a liquid, a solid salt of the acid may be used, such as, for example, calcium butyrate, sodium butyrate, sodium propionate, or beta-hydroxybutyrate, or the acid itself may be applied to a solid substrate of inert material by known spray drying or adsorption techniques. . As solid substrates, excipients that are commonly used for the preparation of powders or granules can be used, such as, for example, pectin, monosaccharide and polysaccharide sugars, alginates, microcrystalline cellulose, gum acacia, corn starch, pregelatinized starch, alkyl derivatives or hydroxyalkyl derivatives of cellulose low, medium and high viscosity, monobasic and polybasic mineral salts, cyclodextrin, alkylcyclodextrin, hydroxyalkylcyclodextrin, pyrrolidones or derivatives, salts and/or esters of monocarboxylic organic acids, salts and/or esters of polycarboxylic organic acids, inorganic substrates such as colloidal silicon dioxide, talc and organic and inorganic ion exchange resins. To obtain a powder from a liquid, atomization is carried out by drying a suspension of liquid butyric acid and a solid substrate by spray drying, or the butyric acid is adsorbed on one of the above substrates.

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Some examples of compositions of the present invention are described below: ________ Composition/Formulation 1 - Synergistic Combination/Blend

No. Ingredients Quantity (% w/w) 1. Inulin or its pharmaceutically acceptable salts 90-99 2. Betaine or pharmaceutically acceptable salts thereof 0.5-10

Composition/Composition 2 - Synergistic combination/blend

No. Ingredients Quantity (% w/w) 1. Inulin or its pharmaceutically acceptable salts 90-99 2. Betaine or pharmaceutically acceptable salts thereof 0.5-10 3 Resistant starch 0.8-9

Composition/Composition 3 - Synergistic combination/blend

No. Ingredients Quantity (% w/w) 1. Inulin or its pharmaceutically acceptable salts 90-99 2. Betaine or pharmaceutically acceptable salts thereof 0.5-10 3 Omega-3 fatty acid 0.89-9

Composition/Composition 4 - Synergistic combination/blend

No. Ingredients Quantity (% w/w) 1. Inulin or its pharmaceutically acceptable salts 90-99 2. Betaine or pharmaceutically acceptable salts thereof 0.5-10 3 Short chain fatty acid (SCFA) or pharmaceutically acceptable salts thereof 0.8-8.2

Composition/Composition 5 - Synergistic combination/blend

No. Ingredients Quantity (% w/w) 1. Inulin or its pharmaceutically acceptable salts 90-99 2. Betaine or pharmaceutically acceptable salts thereof 0.5-10 O Resistant starch 0.8-9 4 Omega-3 fatty acid 0.89-9 5 Short chain fatty acid (SCFA) or pharmaceutically acceptable salts thereof 0.8-8.2

General method for preparing the compositions of the present invention

The present invention also provides a method for preparing the pharmaceutical composition/formulation of the present invention. Once the composition of the present invention has been prepared herein, one skilled in the art may further adapt the method presented below, with suitable modifications, to prepare the pharmaceutical compositions/formulations of the present invention.

General method of obtaining:

Manufacturing procedure: (dosage form in powder form)

1. Accurately weigh all ingredients into separate containers.

2. Separately sift pre-weighed ingredients through a #40 sieve.

3. Blend all sifted substances using a blender for about 30 minutes at a speed of about 20 rpm and store the mixture in a double-lined plastic container and seal the plastic bags tightly, and keep the silica packet between the plastic bags to protect from moisture.

4. Fill the mixture obtained in step 3 into a sachet or other suitable packaging material.

5. At all stages of manufacturing, the temperature is maintained at no more than 25°C and relative humidity 40%.

Manufacturing procedure: (dosage form in capsule form)

1. Accurately weigh all ingredients into separate containers.

2. Pass pre-weighed ingredients separately through a #30 sieve.

3. Blend the contents of step 2 in a blender on low speed.

4. Pass the pre-weighed lubricant through a #40 sieve. Transfer it to a blender and blend in the blender.

5. Fill and seal the mixture obtained in step 4 into HPMC capsule shells.

6. Transfer the filled capsules to the hopper of the polishing and visual inspection machine to remove any remaining powder adhering to the capsule shells.

7. At all stages of manufacturing, the temperature is maintained at no more than 25°C and relative humidity 40%.

- 10043686

Manufacturing procedure: (dosage form in tablet form)

1. Weigh the ingredients separately and sift them through a suitable sieve.

2. Mix the pre-weighed ingredients.

3. Prepare a thick mass by adding binder solution to the mixed ingredients and sift the thick mass to obtain granules.

4. Dry the granules obtained in step 3 until the moisture content (LOD) level is reduced to 1.3-1.7 w/w% to obtain semi-dried granules.

5. Sift the semi-dried granules obtained in step 4 through a suitable sieve.

6. Add lubricants or glidants to the semi-dried granules obtained in step 5 and compress the granules into a tablet.

7. The method further includes producing a seal coating and an enteric coating solution for enteric-coated tablets and a film coating solution for film-coated tablets.

8. At all stages of manufacturing, the temperature is maintained at no more than 25°C and a relative humidity of 40%.

Manufacturing procedure: (dosage form in the form of a solution)

1. Accurately weigh all ingredients into separate containers.

2. Separately sift pre-weighed ingredients through a #40 sieve.

3. Add all sifted ingredients to the mixer along with the solvent.

4. Mix well and continue stirring until a clear solution is obtained.

5. At all stages of manufacturing, the temperature is maintained at no more than 25°C and a relative humidity of 40%.

Examples

It should be understood that the above examples are merely illustrative of the present invention. Some modifications of the products and/or methods used are possible in order to achieve the objectives of the invention. Such modifications are considered to be within the scope of the claimed invention. The following examples are presented to illustrate the pharmaceutical compositions/formulations of the present invention. The Examples also provide and/or demonstrate the effectiveness or synergistic effect of the pharmaceutical composition/formulation of the present invention. These examples are not intended to include all aspects of the subject matter disclosed herein, but rather to illustrate representative methods, compositions, and results. These examples are not intended to exclude equivalents and variations of the present invention that will be apparent to one skilled in the art.

Example 1.__________________________________________________________________________

No. Ingredients % w/w 1 Inulin 88.52 2 Betaine 11.46 3 Flavor (orange) 0.01 4 Sweetener (xylitol) 0.01 Final mass of powder 100

Example 2.

Example 3.

No. Ingredients % w/w 1 Inulin 95.97 2 Betaine 2.88 3 Flavoring (vanilla) 0.96 4 Sweetener (sucralose) 0.19 Final mass of powder 100

Example 4.

No. Ingredients % w/w 1 Inulin 98.99 2 Betaine 0.99 3 Flavor (fruit mix) 0.01 4 Sweetener (sucrose) 0.01 Final mass of powder 100

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Example 5.

No. Ingredients % w/w 1 Inulin 99.67 2 Betaine oh, p 3 Flavor (banana) oh and 4 Sweetener (aspartame) oh and Final mass of powder 100

Example 6.

No. Ingredients % w/w 1 Inulin 90.01 2 Betaine 0.90 3 Resistant starch 9.00 4 Flavor (fruit mix) 0.09 Final mass of powder 100

Example 7.

No. Ingredients % w/w 1 Inulin propionate ester 94.07 2 Betaine hydrochloride 0.94 3 Resistant starch 4.70 4 Flavoring (vanilla) 0.09 5 Sweetener (aspartame) 0.19 Final mass of powder 100

Example 8.

No. Ingredients % w/w 1 Inulin 90.01 2 Betaine 0.90 3 Omega-3 fatty acid 9.00 4 Flavor (orange) 0.09 Final mass of powder 100

Example 9.

No. Ingredients % w/w 1 Inulin propionate ester 97.86 2 Betaine anhydrous 0.93 3 Omega-3 fatty acid 0.93 4 Flavor (fruit mix) 0.09 5 Sweetener (mannitol) 0.19 Final mass of powder 100

Example 10.

No. Ingredients % w/w 1 Inulin 90.66 2 Betaine 0.91 3 SCFA (sodium butyrate) 8.16 4 Flavor (banana) 0.09 5 Sweetener (sorbitol) 0.18 Final mass of powder 100

Example 11.

No. Ingredients % w/w 1 Inulin 98.89 2 Betaine 0.56 3 Flavor (fruit mix) 0.22 4 Sweetener (sucrose) 0.33 Final mass of powder 100

Example 12.

No. Ingredients % w/w 1 Oligofructose-enriched inulin 95.45 2 Betaine hydrochloride 4.09 3 Flavor (orange) 0.18 4 Sweetener (sucralose) 0.27 Final mass of powder 100

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Example 13.

No. Ingredients % w/w 1 Inulin 95.15 2 Betaine 1.90 3 Omega-3 fatty acid 1.90 4 Resistant starch 0.95 5 Flavor (banana) 0.10 Final mass of powder 100

Example 14.

No. Ingredients % w/w 1 Inulin 94.34 2 Betaine 1.80 3 Omega-3 fatty acid 2.70 4 SCFA (sodium propionate) 0.90 5 Flavoring (vanilla) 0.09 6 Sweetener (sucralose) 0.18 Final mass of powder 100

Example 15.

No. Ingredients % w/w 1 Inulin sulfate 93.75 2 Glycine Betaine 5.09 3 Omega-3 fatty acid 0.89 4 Flavor (fruit mix) 0.09 5 Sweetener (aspartame) 0.18 Final mass of powder 100

Example 16.

No. Ingredients % w/w 1 Inulin 93.94 2 Betaine 1.71 3 SCFA (sodium butyrate) 1.71 4 Resistant starch 2.56 5 Flavor (orange) 0.09 Final mass of powder 100

Example 17.

No. Ingredients % w/w 1 Inulin 96.40 2 Betaine 2.51 3 Resistant starch 0.84 4 Flavor (banana) 0.08 5 Sweetener (sorbitol) 0.17 Final mass of powder 100

Example 18.

No. Ingredients % w/w 1 Oligofructose-rich inulin 91.44 2 Glycine Betaine 7.48 3 SCFA (sodium butyrate) 0.83 4 Flavor (fruit mix) 0.08 5 Sweetener (xylitol) 0.17 Final mass of powder 100

Example 19.

No. Ingredients % w/w 1 Inulin 94.00 2 Betaine 1.00 3 Resistant starch 5.00 Final mass of powder 100

- 13 043686

Example 20.

No. Ingredients % w/w 1 Inulin 91.96 2 Betaine 2.68 3 Omega-3 fatty acid 5.36 Final mass of powder 100

Example 21.

No. Ingredients % w/w 1 Inulin 92.00 2 Betaine 3.20 3 SCFA (sodium butyrate) 4.80 Final mass of powder 100

Example 22.

No. Ingredients % w/w 1 Inulin propionate ester 92.88 2 Glycine Betaine 2.06 3 SCFA (sodium propionate) 5.06 Final mass of powder 100

Example 23.

No. Ingredients % w/w 1 Inulin propionate ester 94.74 2 Betaine hydrochloride 1.58 3 MCC pH 101 1.05 4 Croscarmellose sodium 1.05 5 Magnesium stearate 0.53 6 Talc 0.42 7 Zinc stearate 0.32 8 Colloidal silicon dioxide 0.32 Final capsule mass 100.00

Example 24.

Example

No. | Ingredients | % w/w Intragranular ingredients 1 Inulin sulfate 90.36 2 Betaine anhydrous 2.26 h MCC pH 101 1.66 4 Croscarmellose sodium 1.51 Binder solution 5 Hypromellose 1.51 6 Polysorbate 80 1.51 7 IPA how much is needed 8 Water how much is needed Extra granule ry ingredients 9 Magnesium stearate 0.45 10 Talc 0.15 AND Zinc stearate 0.30 12 Colloidal silicon dioxide 0.30 eleven Final capsule mass | 100.00 25. No. Intragranule 1 2 4 5 6 7 | Ingredients | [rich ingredients Inulin propionate ester Glycine Betaine Lactose monohydrate Sodium starch glycolate Binder solution PVP K-30 Polysorbate 80 % w/w 94.74 0.74 1.05 1.05 0.53 1.05 9 IPA how much is needed 10 Water how much is needed Extra granule ry ingredients AND Sodium starch glycolate 0.53 12 Magnesium stearate 0.16 13 Colloidal silicon dioxide 0.16 Final tablet weight 100

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Example 26.

No. Ingredients % w/w Dry mixing 1 Oligofructose-rich inulin 92.78 2 Betaine hydrochloride 1.03 3 MSSRN 102 1.24 4 Crospovidone 0.62 5 Croscarmellose sodium 0.41 6 Magnesium stearate 0.52 7 Colloidal silicon dioxide 0.31 Sealing Coat Ingredients 8 InstaMoistshield 0.62 9 Isopropyl alcohol how much is needed 10 Methylene dichloride how much is needed Enteric Coating Ingredients AND Instacoat ΕΝ HPMS R 2.47 12 Isopropyl alcohol how much is needed 13 Methylene dichloride how much is needed Final tablet weight enteric coated 100.00

Example 27.

No. Ingredients % w/w 1 Inulin sulfate 93.46 2 Betaine anhydrous 1.87 3 Flavoring (vanilla) 0.37 4 Sweetener (sucralose) 0.09 5 Glycerol 3.74 6 Preservative (methylparaben) 0.37 7 Antioxidant (lecithin) 0.09 8 Water how much is needed Final mass of solution 100

Example 28.

No. Ingredients % w/w 1 Oligofructose-enriched inulin 95.24 2 Glycine Betaine 2.86 3 Flavor (fruit mix) 0.19 4 Sweetener (aspartame) 0.05 5 Glycerol 1.43 6 Preservative (ethylparaben) 0.19 7 Antioxidant (fumaric acid) 0.05 8 Water how much is needed Final mass of solution 100

Example 29 Stability data from Example 2

No. 1 Duration Elementary Compliant ZM 40°С/75% RH Compliant ZM 25°С/60% RH Compliant 6M 40°C/75% RH Compliant 6M 25°C/60% RH Compliant Research Test Description Characteristic White free flowing powder 2 Analysis 2.1 Inulin 90% - 110% 101.50% 97.90% 98.10% 98.20% 97.60% 2.2 Betaine 90% - 110% 102.00% 99.80% 100.20% 98.50% 97.20%

Example 30 Stability data from Example 3

No. p/p Duration of the study Elementary ZM 40°С/75% RH ZM 25°С/60% RH 6M 40°C/75% RH 6M 25°C/60% RH Trial Characteristic 1 Description White free flowing powder Compliant Compliant Compliant Compliant Compliant 2 Analysis 2.1 Inulin 90% - 110% 99.60% 98.90% 98.60% 98.60% 97.90% 2.2 Betaine 90% - 110% 102.40% 100.20% 99.70% 99.30% 98.70%

Example 31 Stability data from Example 4

No. p/p Duration of the study Elementary ZM 40°С/75% RII ZM 25°С/60% RH 6M 40°C/75% RH 6M 25°C/60% RII Trial Characteristic 1 Description White free flowing powder Compliant Compliant Compliant Compliant Compliant 2 Analysis 2.1 Inulin 90% - 110% 98.70% 99.10% 98.00% 97.60% 97.90% 2.2 Betaine 90% - 110% 99.40% 97.90% 99.70% 97.20% 98.20%

- 15 043686

Example 32 Stability data from Example 6

No. Duration of the study Elementary ZM 40°С/75% RH ZM 25°С/60% RH 6M 40°C/75% RH 6M 25°C/60% RH Trial Characteristic 1 Description White free flowing powder Compliant Compliant Compliant Compliant Compliant 2 Analysis 2.1 Inulin 90% - 110% 99.80% 98.30% 97.40% 97.20% 98.20% 2.2 Betaine 90% - 110% 97.50% 98.50% 97.40% 98.10% 97.50% Example 33 Stability data from Example 8 No. Duration Elementary ZM 40°С/75% ZM 25°С/60% 6M 40°C/75% 6M 25°C/60% p/p Research Test Characteristic RH RH RH RH 1 Description White free flowing powder Compliant Compliant Compliant Compliant Compliant 2 Analysis 2.1 Inulin 90% - 110% 98.30% 97.60% 98.20% 97.20% 96.20% 2.2 Betaine Omega-3 90% - 110% 99.10% 98.90% 97.40% 98.10% 97.30% 2.3 fat 90% - 110% 97.30% 96.80% 97.40% 96.80% 96.70% acid Example 34 Stability data for Example 10 No. Duration Elementary ZM 40°C/75% ZM 25°С/60% 6M 40°C/75% 6M 25°C/60% p/p Research Test Characteristic RH RH RH RH 1 Description White free flowing powder Compliant Compliant Compliant Compliant Compliant _2____ Analysis 2.1 Inulin 90% - 110% 97.90% 97.20% 98.10% 97.30% 96.40% 2.2 Betaine SCFA 90% - 110% 97.20% 98.10% 98.20% 97.50% 96.80% 2.3 (sodium butyrate) 90% - 110% 95.40% 94.20% 95.40% 94.10% 94.50% Example 35 Example Stability Data eleven No. Duration Elementary ZM 40°С/75% ZM 25°С/60% 6M 40°C/75% 6M 25°C/60% p/p Research Test Characteristic RH RH RH RH 1 Description White free flowing powder Compliant Compliant Compliant Compliant Compliant _2____ Analysis 2.1 Inulin 90% - 110% 98.30% 97.60% 98.20% 97.20% 96.20% 2.2 Betaine 90% - 110% 97.50% 98.50% 97.40% 98.10% 97.50%

Example 36 Stability data for Example 13

No. Duration of the study Elementary ZM 40°С/75% RH ZM 25°С/60% RH 6M 40°C/75% RH 6M 25°C/60% RH Trial Characteristic 1 Description White free flowing powder Compliant Compliant Compliant Compliant Compliant 2 Analysis 2.1 Inulin 90% - 110% 96.90% 95.60% 96.40% 95.20% 96.30% 2.2 Betaine 90% - 110% 98.10% 97.80% 97.30% 96.90% 96.50% 2.3 Omega-3 fatty acid 90% - 110% 96.40% 95.70% 96.10% 95.10% 94.70%

Example 37 Stability data for Example 14

No. p/p Duration of the study Elementary ZM 40°С/75% RH ZM 25°С/60% RH 6M 40°C/75% RH 6M 25°C/60% RH Trial Characteristic 1 Description White free flowing powder Compliant Compliant Compliant Compliant Compliant 2 Analysis 2.1 Inulin 90% - 110% 95.60% 94.50% 94.70% 93.70% 94.30% 2.2 Betaine 90% - 110% 98.40% 97.50% 96.50% 96.40% 96.30% 2.3 Omega-3 fatty acid 90% - 110% 95.70% 94.60% 93.70% 94.60% 94.80% 2.4 SCFA (sodium propionate) 90% - 110% 94.20% 94.60% 94.10% 92.90% 93.10%

Example 38 Stability data for Example 16

No. Duration of the study Elementary ZM 40°С/75% RH ZM 25°С/60% RH 6M 40°C/75% RH 6M 25°C/60% RH Trial Characteristic 1 Description White free flowing powder Compliant Compliant Compliant Compliant Compliant 2 Analysis 2.1 Inulin 90% - 110% 95.60% 95.10% 96.00% 94.60% 94.70% 2.2 Betaine 90% - 110% 97.30% 96.30% 97.30% 96.40% 96.40% 2.3 SCFA (sodium butyrate) 90% - 110% 95.80% 94.60% 94.10% 92.90% 92.70%

- 16 043686

Example 39 Stability data for Example 17

No. p/p Duration of the study Elementary ZM 40°С/75% RH ZM 25°С/60% RH 6M 40°C/75% RH 6M 25°C/60% RH Trial Characteristic 1 Description White free flowing powder Compliant Compliant Compliant Compliant Compliant 2 Analysis 2.1 Inulin 90% - 110% 97.60% 97.10% 97.50% 97.30% 97.70% 2.2 Betaine 90% - 110% 97.30% 96.30% 97.30% 96.40% 96.20%

Example 40 Stability data for Example 19

No. p/p Duration of the study Elementary ZM 40°С/75% RH ZM 25°С/60% RH 6M 40°C/75% RH 6M 25°C/60% RH Trial Characteristic 1 2 2.1 2.2 At Description Analysis Inulin Betaine measures 41. Yes White free flowing powder 90% - 110% 90% - 110% stable Compliant 99.60% 97.30% example Compliant 98.90% 96.30% 20 Compliant 98.60% 97.30% Compliant 98.60% 96.40% Compliant 97.90% 96.40% No. p/p Duration of the study Elementary ZM 40°С/75% RII ZM 25°С/60% RH 6M 40°C/75% RII 6M 25°C/60% RH Trial Characteristic 1 Description White free flowing powder Compliant Compliant Compliant Compliant Compliant 2 Analysis 2.1 Inulin 90% - 110% 98.60% 98.10% 97.10% 97.60% 96.70% 2.2 Betaine 90% - 110% 98.30% 98.30% 97.80% 97.40% 97.40% 2.3 Omega-3 fatty acid 90% - 110% 96.80% 96.50% 96.10% 95.90% 95.70%

Example 42 Stability data for Example 21

No. Duration of the study Elementary ZM 40°C/75% RII ZM 25°С/60% RH 6M 40°C/75% RH 6M 25°C/60% RH Trial Characteristic 1 Description White free flowing powder Compliant Compliant Compliant Compliant Compliant 2 Analysis 2.1 Inulin 90% - 110% 97.60% 97.10% 98.10% 97.90% 97.70% 2.2 Betaine 90% - 110% 97.50% 97.30% 98.10% 97.90% 97.20% 2.3 SCFA (sodium butyrate) 90% - 110% 95.80% 94.60% 94.10% 94.50% 94.40%

*RH=relative humidity

Example 43 Animal Study

The effect of the compositions of the present invention and other test and control compositions was studied in animals. For this study, a comparative analysis of the effect of a synergistic combination or composition/formulation comprising inulin and other active ingredients against 5/6 nephrectomy-induced chronic kidney disease (CKD) in rats was carried out. The following tests were carried out:

Eighty-four (84) female rats (Wistar rats (Rattus norvegicus)), divided into 14 groups (6 in each group), were maintained in an animal house under a 12-hour light/dark cycle, with temperature and relative humidity ranging 19-25±2°С and 30-70%, respectively. To maintain appropriate conditions, temperature and relative humidity were recorded three times a day. All animals were allowed to acclimate for at least five days. Animals were kept under test conditions for at least 30 min once during the acclimatization period to reduce stress. Animals were weighed on the day of admission and abnormalities, if any, were observed daily. Detailed acclimatization records were also kept. Rats were housed 3 per cage in clean, sterilized polypropylene cages. Throughout the experiment, the animals were given standard certified rat food pellets and drinking water purified by reverse osmosis as needed.

To evaluate activity against chronic kidney disease, eighty-four (84) rats were screened and divided into fourteen (14) groups. For comparative analysis, the groups were divided as follows: normal control (group 1 (G1)), placebo control (group 2 (G2)), disease control (group 3 (G3)), treatment groups with individual components (group 4 (G4), group 5 (G5) and group 6 (G6), such as inulin, betaine and resistant starch, respectively), tested composition (group 7 (G7) - group 11 (G11) (combination of inulin with betaine at different dose levels), tested composition Group 12 (G12) - Group 13 (G13) (combination of inulin, betaine and resistant starch) and reference standard (Group 14 (G14)) (activated carbon) Table 1 provides detailed information on the different groups and treatments, carried out in a trial where G4-G14 are the treatment groups.The experiment was carried out for 3 months in two stages.

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Table 1

No. Group Group name Number of animals X (female rats) Dose (g/kg, po) 1 G1 Normal control 6 Normal control 2 G2 Placebo control 6 Placebo control 3 G3 Disease control 6 Disease control 4 G4 Inulin 6 Inulin (1.03 g/kg) 5 G5 Betaine 6 Betaine (0.03 g/kg) 6 G6 Resistant starch 6 Resistant starch (0.10 g/kg) 7 G7 Inulin combination (Inulin+Betaine) (Exp. 1) 6 Inulin (0.88 g/kg) + Betaine (0.11 g/kg) 8 G8 Inulin combination (Inulin+Betaine) (Project 2) 6 Inulin (0.93 g/kg) + Betaine (0.10 g/kg) 9 G9 Inulin combination (Inulin+Betaine) (Project 3) 6 Inulin (1.03 g/kg) + Betaine (0.03 g/kg) 10 G10 Inulin combination (Inulin+Betaine) (Project 4) 6 Inulin (1.03 g/kg) + Betaine (0.01 g/kg) AND G11 Inulin combination (Inulin+Betaine) (Project 5) 6 Inulin (0.93 g/kg) + Betaine (0.001 g/kg) 12 G12 Inulin combination (Inulin+Betaine+Resistant starch) (Project 6) 6 Inulin (1.03 g/kg) + Betaine (0.01 g/kg) + Resistant starch (0.10 g/kg) 13 G13 Inulin combination Inu l in + B etaine + Resistant starch (Project 7) 6 Inulin (1.03 g/kg) + Betaine (0.01 g/kg) + Resistant starch (0.05 g/kg) 14 G14 Standard for comparison 6 Activated carbon (0.62 g/kg)

Treatment protocol:

Study animals were monitored for 13 weeks and underwent a two-stage surgical procedure to ligate and remove the kidney.

Stage 1: A ventral incision is made along the midline of the abdomen to expose the animal's left kidney, and the organ is freed from surrounding tissue. A piece of suture material was placed and ligated around each pole of the kidney at its one-third position. One-third of the kidney at each pole was excised above the ligatures, and the abdominal incision was closed.

Stage 2: This procedure was performed 7 days after stage 1. Animals were placed in a prone position and an incision was made parallel to the midline. An entrance was made into the abdominal cavity and the right kidney was freed from surrounding tissue. The kidney was carefully removed from the incision, the adrenal gland was released and returned to the abdominal cavity. The renal blood vessels and ureter were ligated or cauterized. The kidney was then removed by making a transverse incision into the vessels and ureter just distal to the ligature or cauterized area. The skin incision was closed with wound clamps.

Treatment: All treatment groups received treatment daily for 13 weeks after stage 2 surgery. Blood collection and biochemical analysis were performed after completion of treatment for all treatment groups at the end of the 13th week of treatment.

Table 2 presents summarized results obtained upon administration of the Test Composition/Formulation against 5/6 nephrectomy-induced chronic kidney disease in rats.

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table 2

Group Group name IS PCS Average value (mg/l) % reduction Average value (mg/l) % reduction G1 Normal control 3.26 0 0.15 0 G2 Placebo control 3.16 0 0.22 0 G3 Disease control 7.33 0 1.13 0 G4 Inulin 5.55 24.29 0.84 25.66 G5 Betaine 7.01 4.37 1.09 3.54 G6 R.S. 6.5 11.33 1.01 10.62 G7 I+B (Ex. 1) 4.31 41.20 0.65 42.48 G8 I+B (Ex. 2) 3.83 47.75 0.55 51.33 G9 I+B (Ex. 3) 3.72 49.25 0.51 54.87 G10 I+B (Ex. 4) 3.78 48.43 0.52 53.98 G11 I+B (Ex. 5) 4.21 42.57 0.63 44.25 G12 I+B+RS (Ex. 6) 3.69 49.66 0.50 55.75 G13 I+B+RS (Project 7) 3.67 49.93 0.49 56.64 G14 Activated carbon 3.86 47.34 0.56 50.44

*All values shown above are average values

Interpretation and conclusions

Since there are many underlying causes of kidney disease, as discussed above in the description, there are important biochemical parameters such as Indoxyl Sulfate (IS) and Para-Cresyl Sulfate (PCS) for the diagnosis of CKD, where the level of these parameters is significantly increased.

The above study examined the % reduction in blood IS and PCS levels for the various compositions tested. For the normal control group (G1) and the placebo control group (G2), no significant difference in IS and PCS blood levels was observed. Compared with G1 and G2, a significant increase in IS and PCS levels in the blood was observed in the disease control group (G3). This ultimately confirms that the disease model was successfully induced with the CKD condition in all animals.

Analysis of important biomarkers

As can be seen from the data presented in table. 2, the following observation can be made:

Inulin (G4), when administered alone, showed a significant reduction in blood IS and PCS levels, i.e., 24.29 and 25.66%, respectively. But betaine (G5), when administered alone, did not show any significant effect. The reduction in blood IS and PCS levels was only 4.37 and 3.54%, respectively, for betaine, indicating that betaine itself does not play any role in reducing blood IS and PCS levels. As for resistant starch (G6), also when administered separately, no clinically significant reduction in IS and PCS levels in the blood was observed. The reduction in blood IS and PCS levels in the case of resistant starch was 11.33 and 10.62%, respectively. Separate administration of inulin, betaine and resistant starch shows that only inulin can be considered to have some significant effect on reducing IS and PCS levels in the blood.

Compared to above, when inulin and betaine were administered together or in combination (G7G11), a significant reduction in blood IS and PCS levels was observed, ranging from 41.20 to 49.25% and from 42.48 to 54.87% respectively. This shows that although betaine itself does not play any role in controlling IS and PCS levels, it synergistically enhances the effect of inulin, i.e. the combination of inulin and betaine provides a synergistic effect in reducing IS and PCS levels in the blood. It can also be observed that when the combination of inulin and betaine was administered in a judiciously selected amount in the range of 90-99% by weight based on the weight of the composition for inulin and 0.5-10% by weight based on the weight of the composition for betaine, observed significant synergistic effects for G8, G9 and G10.

It can also be observed that when resistant starch was added to the combination of inulin and betaine (G12 and G13), an even greater reduction in blood levels of IS and PCS was observed, indicating that even resistant starch further contributed to the synergistic effect of the composition of the present invention. The effect obtained with the combination of inulin and betaine is comparable and/or better than that of the reference group (G14), especially the combination of inulin and betaine when combined in judiciously selected amounts showed a higher effect compared to the reference activated carbon. Similarly, the combination of inulin, betaine and resistant starch showed a higher effect compared to the reference activated carbon.

Based on the result obtained, it can be concluded that the tested composition/formulation according to the present invention, i.e. the combination of inulin and betaine, or the combination of inulin, betaine and resistant starch, has synergistic activity compared with the individual components, and the treatment was effective in controlling kidney function by reducing levels of protein-bound uremic toxins such as indoxyl sulfate (IS) and para-cresyl sulfate (PCS).

Regarding mortality: during 13 weeks of treatment in all treatment groups (G4-G14) deaths

Claims (8)

ity was not observed. Conclusion Based on the experimental study conducted on animals, it can be concluded that the test composition/formulation of the present invention, i.e. the combination of inulin and betaine and the combination of inulin, betaine with additional active ingredient(s) such as resistant starch were more effective in reducing the levels of protein-bound uremic toxins in CKD and had a synergistic effect (G7-G13, especially G8-10 and G1213 ) compared to individual active ingredients. In particular, the effect when using a synergistic combination of inulin and betaine or inulin, betaine and additional active ingredients such as resistant starch was higher compared to the standard - activated carbon. Although the invention has been described and illustrated with reference to certain specific embodiments, those skilled in the art will appreciate that various adaptations, changes, modifications, substitutions, deletions or additions of procedures and protocols are possible without departing from the spirit and scope of the invention. For example, effective dosages other than the specific dosages described above may be useful as a consequence of changes in the response of a mammal being treated for any of the indications with the compounds of the invention described above. The specific pharmacological responses observed may vary based on and depending on the particular active compounds selected or the presence of pharmaceutical carriers, as well as the type of formulation and mode of administration used, and such expected variations or differences in results are contemplated by the objects and practice of the present invention. It is therefore intended that the invention should be defined by the scope of the claims that follow and that such claims should be interpreted as broadly as is reasonable. CLAIM 1. A pharmaceutical composition for reducing the level of protein-bound uremic toxins in chronic kidney disease (CKD), comprising a combination of: (a) inulin or pharmaceutically acceptable salts thereof; and (b) betaine or pharmaceutically acceptable salts thereof, wherein the amount of inulin or pharmaceutically acceptable salts thereof is from 90 to 99% by weight, based on the weight of the composition and wherein the amount of betaine or pharmaceutically acceptable salts thereof is salts range from 0.5 to 10 wt.%, based on the weight of the composition. 2. The pharmaceutical composition of claim 1, wherein the composition further comprises additional active ingredients selected from resistant starch, short-chain fatty acid(s) or pharmaceutically acceptable salts thereof, omega-3 fatty acid, or a combination thereof. 3. The pharmaceutical composition according to claim 2, wherein the amount of resistant starch is from 0.8 to 9 wt.% based on the weight of the composition. 4. The pharmaceutical composition according to claim 2, wherein the amount of omega-3 fatty acid is from 0.89 to 9 wt.% based on the weight of the composition. 5. The pharmaceutical composition according to claim 2, wherein the amount of short-chain fatty acid(s) or pharmaceutically acceptable salts thereof is from 0.8 to 8.2 wt.% based on the weight of the composition. 6. The pharmaceutical composition according to claim 1, further comprising pharmaceutically acceptable excipients. 7. The pharmaceutical composition according to claim 6, wherein the pharmaceutically acceptable excipients are selected from a diluent, a disintegrant, a binder, a glidant, a lubricant, a surfactant, a carrier, an antioxidant, a preservative, a flavoring agent, a sweetener, a solvent, a coating agent, a solubilizing agent, and any their combinations. 8. The pharmaceutical composition according to claim 1, where the composition is in the form of a tablet, capsule, sachet, pill, hard capsule filled with liquid or solid, soft capsule, powder, granule, suspension, solution, modified release dosage form, emulsion, syrup. -