Compositions comprising vitamins and minerals

Abstract

The present invention relates to pharmaceutical compositions comprising choline, betaine, methionine, folic acid, vitamin B12, vitamin B6 or a pharmaceutically acceptable form thereof and a pharmaceutically acceptable zinc salt. Said compositions have beneficial physiological effects such as reduced glucose and triglycerides in blood, lipolytic effects and improved abundance and diversity of the intestinal microbiota, such that they are useful in the treatment of at least one condition selected from the group consisting of obesity, dysbiosis, insulin resistance, steatotic liver disease, diabetes, metabolic syndrome, Crohn's disease, gastrointestinal disease and intestinal metabolic disorders.

Description

COMPOSITIONS INCLUDING VITAMINS AND MINERALS Field of Invention

[0001] The present invention relates to the field of the pharmaceutical industry, and more particularly relates to compositions comprising vitamins and minerals. Background of the Invention

[0002] Eating a balanced diet that includes foods rich in vitamins and minerals is essential to ensure an adequate intake of these nutrients and thus maintain good health. However, to address specific deficiencies or needs, supplementation can be achieved through pharmaceutical formulations or dietary supplements.

[0003] Dietary supplements are products that may include vitamins, minerals, amino acids, fatty acids, plant extracts, and other substances, whose primary purpose is to complement the regular diet and provide nutrients that may be lacking in the daily diet.

[0004] The health benefits derived from the intake of vitamins and minerals in food supplements have been found to be associated with different mechanisms, including the following: a) Modification of the intestinal microbiota: B vitamins play a role in the metabolism and growth of some bacteria, so the consumption of this type of vitamins may benefit the development of bacterial populations in the intestinal microbiota. b) Regeneration of the intestinal lining: Folate is involved in the regeneration of the intestinal lining. c) d) Modulation of the immune system: Vitamins and minerals improve and protect the integrity of epithelial barriers, mediate the migration of lymphocytes to the intestine, promote the survival of T cells in the small intestine, regulate the number and function of immune system cells (monocytes, neutrophils, lymphocytes), regulate the production of antimicrobial proteins and cytokines.d) e) Regulation of one-carbon metabolism: folate, vitamin B12, vitamin B6, betaine, choline, methionine, and zinc sulfate participate in the one-carbon metabolic pathway, in which a methyl group is transferred to the formation of the amino acid methionine, to which an adenosyl group is added by the action of the enzyme methionine-adenosyl transferase to form S-adenosyl methionine (SAM). The main function of SAM is to donate the methyl group through the enzymes DNA-methyltransferases and histone-methyltransferases; responsible for the methylation of DNA and histones respectively, so that they participate in the regulation of genetic expression, including the expression of genes involved in lipid catabolism.

[0005] Due to the above mechanisms, there is the possibility of developing dietary supplements that can help in conditions or diseases related to alterations in hepatic, carbohydrate and / or lipid metabolism, as well as gastrointestinal disorders such as obesity, diabetes, steatotic liver disease, dysbiosis, and Crohn's disease.

[0006] Lifestyle modification, through increased physical activity and dietary changes, remains the first line of treatment and prevention for these types of conditions; however, adherence to these strategies is low. Therefore, it is of great importance to have medications or supplements that prevent the development and / or progression of conditions related to alterations in liver, carbohydrate, and / or lipid metabolism, as well as gastrointestinal disorders.

[0007] Examples of technologies related to nutritional compositions or food supplements are listed below.

[0008] Patent US9539278B2 discloses an enteral nutritional or pharmaceutical composition comprising high amounts of aspartate in combination with vitamin B12 and / or biotin. The composition is intended to be used in the treatment and / or prevention of alterations in ketone and lactate metabolism, that is, elevated concentrations of ketone bodies, lactate and / or other organic acids and / or insufficient pH homeostasis in the blood of a mammal. In contrast, the present invention has the object of assisting in the treatment of at least one condition selected from the group consisting of obesity, dysbiosis, insulin resistance, steatotic liver disease, diabetes, metabolic syndrome, Crohn's disease, gastrointestinal disease and intestinal metabolic disorders by having an anti-lipogenic effect, reducing body weight, insulin resistance, and enhancing the intestinal microbiota.

[0009] Document US20210196663A1 describes a combination therapy comprising one or more omega-3 fatty acids and one or more B vitamins for treating cognitive impairment, such cognitive impairment disorders including, especially, but not exclusively, Mild Cognitive Impairment (MCI) and Alzheimer's disease. As indicated above, the food supplement of the present invention has a body weight reducing, anti-lipogenic, insulin resistance reducing and intestinal microbiota enhancing effect, making it an aid in the treatment of conditions related to alterations in hepatic, carbohydrate and / or lipid metabolism.

[0010] Patent US8545896B2 describes compositions comprising twelve-carbon chain fatty acids and / or twelve-carbon chain acylglycerols, vitamin D, iodine, vitamin B1, vitamin B6, vitamin B12, vitamin B2, vitamin B9, vitamin B3, vitamin E, vitamin A, vitamin C, iron, zinc, copper, magnesium, omega-3 fatty acids, and pharmaceutical excipients. The purpose of the compositions described in said document is to provide a nutritional supplement to patients with nutritional deficiencies; in contrast, the nutritional supplement of the present invention has the objective of decreasing lipid production, body weight, insulin resistance, and enriching the intestinal microbiota.

[0011] Document US20220079201 A1 discloses a pharmaceutical composition comprising vitamin D, iodine, vitamin B1, vitamin B6, vitamin B12, vitamin B2, vitamin B9, vitamin B3, vitamin E, vitamin A, vitamin C, iron, zinc, copper, magnesium, omega-3 fatty acids and excipients that is used to improve the nutritional status of patients under physiological stress, such as prenatal, pregnant or lactating patients. In contrast, the present invention focuses on reducing lipid production, body weight, insulin resistance, and enriching the intestinal microbiota.

[0012] Patent US6531171 B2 describes intermediate-moisture food products containing betaine that are particularly suitable for consumption by people seeking to maintain a low-carbohydrate diet. This document teaches that the inclusion of betaine in nutritional products allows for a reduction in the addition of ingredients that contribute to calorie content; on the contrary, the components of the supplement of the present invention have physiological effects, such as reducing lipid production, body weight, and insulin resistance, as well as enriching the intestinal microbiota.

[0013] Patent US10959936B2 discloses a composition that includes, at least, choline, betaine, methionine, folic acid and cyanocobalamin, as well as a zinc chelate compound, where the objective of the composition is the care of oral health through an antimicrobial, antioxidant and saliva production stimulating effect; in contrast, the present invention aims to assist in the treatment of conditions related to alterations in hepatic, carbohydrate and / or lipid metabolism.

[0014] Document CN110800880A describes a composition comprising, at least, choline, betaine, methionine, folic acid, cyanocobalamin and zinc sulfate, wherein said composition is used in the prevention and treatment of intestinal disorders. However, said composition is primarily intended for use in geese and, in addition, the essential components of the composition are Chinese medicinal herbs such as wormwood, Atractylodes macrocephala, Baigou, Chenpi, Fengfeng, Yinchen; on the contrary, the present invention focuses on use in humans and the essential components are vitamins and minerals.

[0015] As a result of the above, there has been an effort to develop pharmaceutical compositions or food supplements that are useful in the treatment of conditions related to alterations in hepatic, carbohydrate and / or lipid metabolism, developing a pharmaceutical composition that includes vitamins and minerals that have physiological effects such as the reduction of lipid production, body weight and insulin resistance, as well as the enrichment of the intestinal microbiota, among others. Objects of the invention

[0016] Taking into account the shortcomings of the prior art, it is an object of the present invention to provide a pharmaceutical composition comprising choline, betaine, methionine, folic acid, cyanocobalamin, pyhdoxin and zinc sulfate. Another object of the invention is to provide a pharmaceutical composition that can be used in the treatment of conditions related to alterations in hepatic, carbohydrate and / or lipid metabolism, as well as in gastrointestinal disorders by physiological effects that include lipolytic effects, improvement in the expression of genes involved in hepatic, carbohydrate and lipid metabolism, as well as in the enrichment of intestinal microbiota.

[0017] These and other objectives are achieved through compositions comprising vitamins and minerals. Brief Description of the Invention

[0018] For this purpose, a first aspect of the present invention relates to a pharmaceutical composition comprising choline, betaine, methionine, folic acid, pyhdoxine, vitamin B12 or a pharmaceutically acceptable form thereof and a pharmaceutically acceptable salt of zinc.

[0019] Another aspect of the invention relates to the use of the pharmaceutical composition described above for the treatment of at least one condition selected from the group consisting of obesity, insulin resistance, steatotic liver disease, diabetes, metabolic syndrome, Crohn's disease, gastrointestinal disease, intestinal metabolic disorders and intestinal dysbiosis. Brief description of the figures

[0020] The novel aspects considered characteristic of the present invention will be set forth with particularity in the appended claims. However, certain embodiments, features, objects, and advantages thereof will be better understood from the detailed description when read in conjunction with the accompanying drawings, in which: Figs.1 A-1G

[0021] [Figs. 1A-1G] FIGs. 1A-1G show the results of the effect of a composition according to an embodiment of the present invention on body weight and fat. FIG. 1A shows photographs of animals from the different treatments, as well as the extracted livers. FIG. 1B is a graph of the weight of the animals through the 18 weeks of the study. FIGs. 1C-1G correspond to graphs of the weight at sacrifice (FIG. 1C), liver weight (FIG. 1D), body weight / liver weight ratio (FIG. 1E), visceral fat weight (FIG. 1F) and epidemic fat weight (FIG. 1G). The data are presented as the mean per group ± standard deviation. *P<0.05, **P<0.01 , ***P<0.001. Figs. 2A-2L

[0022] [Figs.2A-2L] FIGs. 2A-2L illustrate the effects of a composition according to one embodiment of the present invention on biomarkers of hepatic, lipid, and glucose metabolism, in particular, blood glucose levels at sacrifice (FIG. 2A), insulin tolerance test (FIG. 2B), blood level of resistin (FIG. 2C), adiponectin (FIG. 2D), triglycerides (FIG. 2E), cholesterol (FIG. 2F), PAI-1 (FIG. 2G), leptin (FIG. 2H), AST (FIG. 2I), ALT (FIG. 2J), and insulin (FIG. 2K). FIG. 2L corresponds to the percentage of total DNA methylation. The data are presented as the mean per group ± standard deviation. *P<0.05, **P<0.01, ***P<0.001. Figs. 3A-3D

[0023] [Figs.3A-3D] FIGs. 3A-3D show the results of the effects of a composition according to an embodiment of the present invention on steatotic liver disease. FIGs. 3A-3C correspond to representative photographs of the livers of the study animals, that is, control group (FIG. 3A), treatment group with composition of the invention (FIG. 3B) and high-fat diet group (FIG. 3C). FIG. 3D is a graph of the percentage of liver with the presence of steatosis. The data are presented as the mean per group ± standard deviation. *P<0.05, **P<0.01 , ***P<0.001 . Figs. 4A-4D

[0024] [Figs.4A-4D] FIGs. 4A-4D illustrate the effects of a composition according to an embodiment of the present invention on adiposity. FIGs. 4A-4C correspond to representative photographs of the livers of the study animals, that is, control group (FIG. 4A), treatment group with composition of the invention (FIG. 4B) and high fat diet group (FIG. 4C). FIG. 4D is a graph of the adipocyte area. The data are presented as the mean per group ± standard deviation. *P<0.05, **P<0.01 , ***P<0.001 . Figs. 5A-5H

[0025] [Figs.5A-5H] FIGs. 5A-5H correspond to the gut microbiota analysis. FIGs. 5A-5C show the Beta Diversity results: observed ASV (FIG. 4A), Shannon (FIG. 4B) and Faith (FIG. 5C) indices. FIG. 5D is a three-dimensional scatterplot of beta diversity generated by principal coordinates analysis (PCoA). The relative abundance of bacterial phyla is shown in FIG. 5E. FIGs. 5F and 5G illustrate the abundance of Firmicutes and Bacteroidetes, while FIG. 5H shows the Firmicutes / Bacteroidetes (F / B) ratio. Data represent means ± SEM. *P<0.05. Fig.6

[0026] [Fig. 6] FIG. 6 is a heat map of the expression of various genes involved in hepatic lipid and carbohydrate metabolism. Detailed description of the invention

[0027] It should be understood that the present invention is not limited to the methodologies, protocols, and reagents provided in the detailed description, as these may vary. It should also be understood that the terminology used herein is merely for the purpose of describing particular embodiments and is not intended to limit the scope of the present disclosure, which will be limited only by the appended claims. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those commonly understood by one skilled in the art.

[0028] As used herein, the term "food supplement" refers to a product based on herbs, plant extracts, traditional foods, dehydrated or fruit concentrates, with or without added vitamins or minerals, which may be presented in pharmaceutical form and whose purpose is to increase the total dietary intake, complement it or supply some of its components.

[0029] The term "pharmaceutically acceptable form" refers to the presentation of a chemical compound or active ingredient in a form suitable for use. The pharmaceutically acceptable form of a compound includes, but is not limited to, pharmaceutically acceptable salts, hydrates, solvates, isomers, prodrugs, and isotopically labeled derivatives of the compounds.

[0030] “Pharmaceutically acceptable salt” means those salts that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of subjects without undue toxicity, irritation, allergic response, and the like, and are commensurate with a reasonable benefit / risk ratio.

[0031] “Recommended Daily Intake (RDI)” as used herein refers to the minimum dose that should be consumed to satisfy the nutritional needs of the majority of the healthy population.

[0032] As used herein, "treatment" refers to the application of medical, therapeutic measures or any intervention designed to address a disease, condition or disorder with the goal of alleviating symptoms, improving health or preventing disease progression.

[0033] The administration of compositions comprising vitamins and minerals has been found to have beneficial effects on health, particularly lipolytic effects, improvements in biomarkers and expression of genes associated with alterations in hepatic, carbohydrate and lipid metabolism, as well as improvements in the abundance and diversity of the intestinal microbiota.

[0034] Thus, in one aspect of the invention, a pharmaceutical composition is described comprising choline, betaine, methionine, folic acid, vitamin B6 and vitamin B12 or a pharmaceutically acceptable form thereof and a pharmaceutically acceptable salt of zinc.

[0035] In particular, doses higher than the recommended daily intake (RDI) have been found to have the following physiological effects: • Decreased blood levels of biomarkers associated with alterations in liver, carbohydrate, and lipid metabolism, including: o Glucose: Elevated blood glucose concentrations are associated with certain medical conditions such as diabetes, hormonal disorders, liver disease, or severe infections. o Triglycerides: Elevated blood triglyceride levels may be indicative of metabolic disorders and may also increase the risk of developing heart disease and stroke. o Plasminogen activator inhibitor-1 (PAI-1): Elevated PAI-1 levels have been associated with cancer, obesity, and metabolic syndrome; they have also been linked to the development of thrombosis. o Leptin: Serum leptin levels are proportional to the amount of body fat, which is why they are associated with obesity.o Aspartate aminotransferase (AST): Elevated serum levels reflect damage to the liver, lung, myocardium, skeletal muscle, kidney, and brain. o Alanine aminotransferase (ALT): It is primarily located in liver cells, so an increase in its blood concentration could suggest the destruction of hepatocytes. o Insulin: Elevated levels of insulin in the blood may be associated with various medical conditions, including insulin resistance, metabolic syndrome, diabetes, and liver disease. • Reduction of insulin resistance: Insulin resistance is a condition in which cells do not respond adequately to insulin, resulting in impaired glucose metabolism. Therefore, reducing insulin resistance is an expected beneficial effect in the treatment of insulin resistance, metabolic syndrome, diabetes, and liver disease. • Decreased visceral and epididymal fat: The accumulation of visceral fat and, in rodents, epididymal fat is related to obesity, as well as to the development of insulin resistance, type 2 diabetes, and metabolic syndrome. • Increased adiponectin: Adiponectin participates in the metabolism of glucose and fatty acids; its blood level is inversely proportional to the percentage of body fat. • Increased DNA methylation: DNA methylation regulates gene expression by preventing the binding of transcription factors or by creating a less accessible chromatin structure (heterochromatin). Differential methylation of specific genes has been correlated with metabolic alterations present in the development of steatotic liver disease. • Body weight reduction. • Decreased steatosis: Steatosis is the accumulation of fat vacuoles in the cytoplasm of hepatocytes • Increased expression of peroxisome proliferator-activated receptor alpha (PPARa): PPARa is involved in the regulation of carbohydrate, lipid and protein metabolism, prevents lipid accumulation in the liver. • Decrease in the expression of genes involved in the synthesis of fatty acids. • Modification of the abundance and diversity of the intestinal microbiota: Surprisingly, it was found that the dietary supplement of the present invention promotes the restoration of an adequate intestinal microbiota, thereby maintaining adequate levels of short-chain fatty acids produced by intestinal bacteria. Furthermore, it contributes to the enrichment of beneficial bacterial genera such as Acinetobacter, Anaeroplasma, Pseudomonas, Stenotrophomonas, among others.

[0036] Accordingly, in a non-limiting embodiment, the pharmaceutical composition of the present invention comprises: a) 125 to 1650 pg of choline; b) 119 to 500 mg of betaine; c) 38 to 8850 mg of methionine; d) 76 to 2250 pg of folic acid; e) 0.3 to 10.8 pg of vitamin B12 or a pharmaceutically acceptable form thereof; f) 0.8 to 4.8 pg of vitamin B6 or a pharmaceutically acceptable form thereof; g) 3.8 to 48 mg of a pharmaceutically acceptable salt of zinc.

[0037] Considering that the recommended daily intake (RDI) varies depending on the age or physiological state of individuals, Table 1 shows particular embodiments of the pharmaceutical composition of the present invention for different groups of individuals depending on their age and / or physiological state. Table 1. Modalities of pharmaceutical composition depending on the age and physiological state of the individuals.

[0038] As indicated above, a pharmaceutically acceptable form refers to the presentation of a chemical compound or active ingredient in a form that is suitable for use; in this regard, the pharmaceutically acceptable form of vitamin B12 may be selected from the group consisting of cyanocobalamin, methylcobalamin, deoxyadenosylcobalamin, and hydroxycobalamin.

[0039] With respect to the pharmaceutically acceptable salt of pyridoxine, in a non-limiting embodiment, the pyridoxine salt is selected from the group consisting of pyridoxine aceglumate, pyridoxine alpha-ketoglutarate, pyridoxine dipalmitate, pyridoxine phosphate, pyridoxine phosphosehenate, pyridoxine hydrochloride, pyridoxine palmitate or pyridoxine pyrrolidonecarboxylate.

[0040] With respect to the pharmaceutically acceptable salt of zinc, in a non-limiting embodiment, the zinc salt is selected from the group consisting of zinc oxide and zinc sulfate.

[0041] In another non-limiting embodiment, the pharmaceutical composition of the present invention may additionally comprise at least one ingredient selected from the group consisting of prebiotics, probiotics, and pharmaceutically acceptable excipients.

[0042] Regarding prebiotics, these are non-digestible substances that promote the growth and activity of beneficial microorganisms in the intestine. In a non-limiting embodiment, the prebiotic is selected from the group consisting of lactose, inulin, resistant starch, galacto-oligosaccharides (GOS), fructooligosaccharides (FOS), xylooligosaccharides (XOS), isomalto-oligosaccharide (IMO), oligosaccharides, such as those from soy (SOS), transgalactooligosaccharides (TOS), polydextrose, lactulose, and at least one extract of at least one plant containing said prebiotics, or combinations thereof. In a preferred embodiment, the extract of at least one plant is obtained from a plant selected from the group consisting of dandelion, green tea, Jerusalem artichoke, leek, onion, chicory root, asparagus, banana, barley, agave, wheat and garlic; or combinations thereof.

[0043] A probiotic is a microorganism that, when administered in adequate amounts, confers a benefit to the health of the host, in this sense, the genera that are commonly used as probiotics are Lactobacillus, Bifidobacterium, Saccharomyces, Streptococcus, Enterococcus, Escherichia and Bacillus, so that, in a non-limiting modality, the probiotic belongs to a genus that is selected from the group consisting of Lactobacillus, Bifidobacterium, Saccharomyces, Streptococcus, Enterococcus, Escherichia and Bacillus. In a preferred embodiment, the probiotic is selected from the group consisting of Lactobacillus acidophilus, L. casei, L. johnsonü, L. lactis, L. plantarum, L. reuteh, L. rhamnosus, L. salivarius, Bifidobacterium animalis, B. breve, B. infantis, B. lactis, B. longum, Saccharomyces cerevisiae, S. boulardii, Escherichia coli, Bacillus clausii, B. subtilis, B. coagulans, B. licheniformis, B. polyfermenticcus and B. pumilus.

[0044] On the other hand, in order to improve stability, bioavailability of the components, appearance and facilitate the consumption of the pharmaceutical composition of the present invention, pharmaceutically acceptable excipients may be included, which in a non-limiting embodiment are selected from the group consisting of: lactose, dextrins, glucose, sucrose, sorbitol, silicates, calcium salts, magnesium salts, potassium chloride, sodium chloride, starches, sugars, sugar alcohols, cellulose derivatives, alginates and derivatives, crospovidone, colloidal anhydrous silicon and other silica compounds, Stearic acid and its salts, cellulose acetate phthalate, colorants, acacia, alginic acid, aluminum acetate, benzyl alcohol, butyl paraben, butyl hydroxytoluene, citric acid, calcium carbonate, waxes, carboxymethylcellulose and derivatives, calcium stearate, ethylenediaminetetraacetic acid (EDTA), copolyvidone, hydrogenated castor oil, calcium hydrogen phosphate dihydrate, cetylpyridone chloride, dimethi, sodium erythrosine, gelatin, glyceryl monolate, glycerin, glycine, glyceryl monostearate, hydroxymethylcellulose, hypromellose, ferric oxide, iron oxide, mannitol, mineral oil, vegetable oil, methacrylic acid copolymer, povidone, polyethylene glycol, polysorbate 80, polaxamers, potassium bicarbonate, phosphoric acid, sodium benzoate, sodium metabisulfite, acid succinic, sodium propionate, titanium dioxide, talc, triacetin, triethyl citrate, gums.

[0045] The pharmaceutical composition of the present invention can be presented in various pharmaceutical forms, which are the physical arrangement of its components to form a product and facilitate its dosage and administration. In a non-limiting embodiment, the pharmaceutical composition of the present invention has a pharmaceutical form selected from the group consisting of powder, capsule, emulsion, suspension, syrup, and solution.

[0046] Additionally, one or more components of the pharmaceutical composition of the present invention may be formulated as nanoparticles, such that they may be included in the various applicable pharmaceutical forms.

[0047] Preferably, the composition of the present invention is designed to be administered in at least one dose per day orally.

[0048] In one embodiment, the pharmaceutical composition of the present invention can be added to a food, for its fortification.

[0049] In another embodiment, the pharmaceutical composition of the present invention is a dietary supplement.

[0050] Considering the physiological effects obtained by administering the pharmaceutical composition of the present invention, another aspect relates to the treatment of at least one condition selected from the group consisting of obesity, insulin resistance, steatotic liver disease, diabetes, metabolic syndrome, Crohn's disease, gastrointestinal disease, Intestinal metabolic disorders and intestinal dysbiosis. In a preferred embodiment, the pharmaceutical composition of the present invention can be used as an aid in the treatment of the aforementioned conditions, that is, it contributes in a complementary manner to the treatment process.

[0051] Steatotic liver disease (also known as fatty liver disease or SLD) is the most common form of steatosis, or fat accumulation. It is not always accompanied by liver injury due to the liver's high functional capacity. Steatosis includes diseases that present with the symptom of fat accumulation in the liver, either due to alcohol consumption or metabolic abnormalities present in patients with overweight, obesity, diabetes mellitus, or other metabolic disorders.

[0052] The gut microbiota is a diverse community of microorganisms residing in the gastrointestinal tract, primarily in the large intestine. It plays an important role in metabolic homeostasis, participating in digestion, energy regulation, short-chain fatty acid (SCFA) production, vitamin synthesis, protection against pathogens, and modulation of the host immune system. Alterations in the gut microbiota (gut dysbiosis) have been associated with diseases such as obesity, type 2 diabetes mellitus, and steatotic liver disease.

[0053] The present invention will be better understood from the following examples, which are presented solely for illustrative purposes to allow a full understanding of the preferred embodiments of the present invention, without implying that there are no other non-illustrated embodiments that can be put into practice based on the detailed description given above. Examples

[0054] Example 1. Effects of compositions comprising vitamins and minerals in an animal model

[0055] Twenty-four C57BL / 6J mice, aged 7 to 8 weeks, were randomly assigned to three groups of eight mice each. Each group received a different diet, as detailed below: • ND Diet: This is a normocaloric diet, providing a calorie intake considered normal or adequate to maintain body weight and meet individual energy needs. This diet was used as a control. • HF Diet: The high-fat diet, where 60% of calories come from fat. The goal of this group was to promote metabolic alterations that result in the development of steatotic liver disease. • HFMS diet: Patients were provided a HF (high-fat) diet; from week 8 to week 18, the composition described in Table 1, referred to as the “MetMix Supplement,” was also administered daily. This group corresponds to the treatment group. Table 2. Composition of the “MetMix Supplement”

[0056] Weight, food intake, and daily water intake were monitored per mouse during the 18 weeks to calculate total caloric intake. All mice were sacrificed at week 18. Blood, liver, and epididymal and visceral adipose tissue were collected. The blood was centrifuged to obtain serum.

[0057] Food intake was measured three times per week between 9 and 10 am, and energy was calculated as kilocalories (kcal) from food and drink. Body weight was recorded each week during the study. Blood glucose levels were measured with a glucometer from blood obtained from the tail vein before sacrifice. At the end of treatment, 48 h before sacrifice, mice were fasted for 4 h to perform the insulin tolerance test (ITT) after an intraperitoneal injection of short-acting human recombinant insulin at a standardized dose of 0.025 U / mouse. Glucose levels were measured at 0, 30, 60, and 90 minutes after insulin administration. Finally, the area under the curve (AUC) was calculated.

[0058] To evaluate insulin resistance, as well as levels of adiponectin, PAI-I, leptin and insulin, blood was collected from the ocular vein to obtain serum and commercial kits were used.

[0059] The 16S ribosomal RNA subunit is considered the universal target for bacterial identification, due to its characteristics and properties, which allow for the taxonomic characterization of bacteria, allowing for the identification down to the genus and, in some cases, the species level. Consequently, the intestinal microbiota was evaluated by sequencing this subunit. a) Effect on body weight and fat

[0060] Referring to FIGS. 1A-1G, as expected, the group fed the high-fat diet (HF) increased their body weight, liver weight and size, body weight / liver weight ratio, as well as visceral and epididymal fat compared to the control (ND). At the beginning of the trial, the treatment group (HFMS) had a weight increase similar to the group that was only administered the high-fat diet; however, once the administration of the composition of the present invention began, the trend changed, observing a decrease in body weight, liver weight, body weight / liver weight ratio, as well as visceral and epididymal fat, thus confirming that the composition of the present invention has a lipolytic effect. b) Biomarkers of liver, lipid and glucose metabolism

[0061] As seen in FIGS. 2A-2L, animals fed the high-fat (HF) diet had increased serum levels of glucose, glycerols, PAI-1, ALT, insulin, and leptin compared to ND control animals. In contrast, The MetMix-treated group (HFMS) decreased glucose, triglycerides, cholesterol, PAI-1, AST, ALT, insulin, and leptin levels. Additionally, the treated group (HFMS) showed an increase in insulin sensitivity as measured by the insulin tolerance test (ITT), which is shown by the decreased area under the curve (AUC) value.

[0062] The above reflects that treatment with the composition of the present invention (MetMix) has a beneficial effect on insulin resistance and lipid metabolism, which allows for a reduction in body weight and is reflected in a lower accumulation of adipose tissue.

[0063] On the other hand, the tendency towards an increase in adiponectin in the group treated with the composition of the invention (HFMS) to levels similar to the control group (ND), suggests an improvement in insulin sensitivity shown by the treated animals, since adiponectin helps maintain glucose homeostasis in the body. c) Steatotic liver disease and adiposity

[0064] In FIG. 3C, a photomicrograph of the liver of mice fed a high-fat (HF) diet, the presence of steatosis is clearly seen; in contrast, no steatosis is seen in the liver of mice in the control group (FIG. 3A), while small lipid vacuoles are observed in the group administered the composition of the invention (FIG. 3B). The graph in FIG. 3D confirms that the composition of the invention reduces steatosis, since a significant difference was observed in the ratio of the area of ​​vacuoles to the total area.

[0065] Regarding adiposity, the size of the adipocytes in the control group (FIG. 4A) and in the group administered the composition of the invention is smaller compared to the adipocytes of mice administered a high-fat diet (FIG. 4C). The graph in FIG. 4D shows that the area of ​​the adipocytes was considerably reduced in animals treated with MetMix, with an increase in the frequency of adipocytes with a smaller area.

[0066] The above results reflect that the composition of the invention has an effect on fat storage, decreasing lipid accumulation and, therefore, reducing obesity in the same. d) Modification of intestinal microbiota

[0067] Referring to FIGs. 5A-5C, the phylogenetic alpha diversity is observed according to the analysis of observed characteristics and Shannon and Faith indices. FIG. 5D shows the beta diversity among the analyzed samples, showing a grouping of the samples. In FIG. 5E, a relative abundance of genera is observed between the control group (ND) and the group treated with the composition of the invention (HFMS), indicating similarity between them and differences compared to the HF group. FIGs. 5F and 5G show graphs of the abundance of the most abundant microbial groups of the intestinal microbiota, i.e., Firmucutes and Bacteroidetes, respectively, between the study groups. FIG. 5H shows a similar Firmicutes / Bacteroidetes ratio between the control groups (ND) and those supplemented with methyl group donors, clearly different from that present in animals with SLD.Administration of a composition of the present invention improved the Firmicutes / Bacteroidetes ratio. In this regard, it has been reported that the increase in the Firmicutes / Bacteroidetes ratio is related to an "obese-type" human microbiota, which is associated with excess weight and metabolic syndrome. e) Gene expression.

[0068] In FIG. 6 it can be seen that the group administered a composition according to the present invention (HFMS) presents similarities in expression levels, according to the coloring on the map, with the control group (ND), however, there is a deregulation and changes in expression in the group subjected to a high-fat diet (HF). It is highlighted that in the HFMS group there was an increase in the expression of PPARa, a transcription factor important in lipid metabolism, regulator of processes such as beta-oxidation and triglyceride degradation, and by its function, exerts a protective effect by preventing lipid accumulation in the liver. On the other hand, a decrease in the expression of genes involved in the synthesis of fatty acids such as Acaca and Fasn was observed. In contrast, in the HF group there is an increase in these genes despite the consumption of fat in the diet.

[0069] In accordance with the above, it can be observed that the compositions comprising vitamins and minerals of the present invention, have been devised for use in the treatment of at least one condition selected from the group consisting of obesity, dysbiosis, insulin resistance, steatotic liver disease, diabetes, metabolic syndrome, Crohn's disease, gastrointestinal disease, intestinal metabolic disorders and intestinal dysbiosis, and it will be apparent to any person skilled in the art that the embodiments of the compositions comprising vitamins and minerals as described above and illustrated in the accompanying drawings are merely illustrative but not limiting of the present invention, since numerous considerable changes in its details are possible without departing from the scope of the invention.

[0070] Therefore, the present invention should not be considered as restricted except as required by the prior art and by the scope of the appended claims.

Claims

Claims

1. ^ A pharmaceutical composition characterized in that it comprises choline, betaine, methionine, folic acid, vitamin B12, vitamin B6 or a pharmaceutically acceptable form thereof and a pharmaceutically acceptable salt of zinc.

2. The pharmaceutical composition according to claim 1, further characterized in that it comprises: a) 125 to 1650 pg of choline; b) 119 to 500 mg of betaine; c) 38 to 8850 mg of methionine; d) 76 to 2250 pg of folic acid; e) 0.3 to 10.8 pg of vitamin B12 or a pharmaceutically acceptable form thereof; f) 0.8 to 4.8 pg of vitamin B6 or a pharmaceutically acceptable form thereof; g) 3.8 to 48 mg of a pharmaceutically acceptable salt of zinc.

3. The pharmaceutical composition according to claim 2, further characterized in that it comprises: a) 125 to 1125 pg of choline; b) 119 to 381 mg of betaine; c) 114 to 5130 mg of methionine; d) 76 to 1 170 pg of folic acid; e) 0.3 to 6.6 pg of vitamin B12 or a pharmaceutically acceptable form thereof; f) 0.8 to 3.3 pg of vitamin B6 or a pharmaceutically acceptable form thereof; g) 3.8 to 41.7 mg of a pharmaceutically acceptable salt of zinc.

4. The pharmaceutical composition according to claim 2, further characterized in that it comprises: a) 400 to 1650 pg of choline; b) 119 to 381 mg of betaine; c) 2280 to 8550 mg of methionine; d) 460 to 1380 pg of folic acid; e) 2.4 to 10.8 pg of vitamin B12 or a pharmaceutically acceptable form thereof; f) 1 to 3.9 pg of vitamin B6 or a pharmaceutically acceptable form thereof; g) 11 to 45 mg of a pharmaceutically acceptable salt of zinc.

5. The pharmaceutical composition according to claim 2, further characterized in that it comprises: a) 450 to 1350 pg of choline; b) 119 to 381 mg of betaine; c) 2280 to 8550 mg of methionine; d) 750 to 2250 pg of folic acid; e) 2.6 to 7.8 pg of vitamin B12 or a pharmaceutically acceptable form thereof; f) 1.4 to 4.2 pg of vitamin B6 or a pharmaceutically acceptable form thereof; g) 14 to 42 mg of a pharmaceutically acceptable salt of zinc.

6. The pharmaceutical composition according to claim 2, further characterized in that it comprises: a) 550 to 1650 pg of choline; b) 119 to 381 mg of betaine; c) 2280 to 8550 mg of methionine; d) 650 to 1950 pg of folic acid; e) 2.8 to 8.4 ng of vitamin B12 or a pharmaceutically acceptable form thereof; f) 1.6 to 4.8 pg of vitamin B6 or a pharmaceutically acceptable form thereof; g) 16 to 48 mg of a pharmaceutically acceptable zinc salt.

7. The pharmaceutical composition according to any one of claims 1 to 6, further characterized in that the pharmaceutically acceptable form of vitamin B12 is selected from the group consisting of cyanocobalamin, methylcobalamin, deoxyadenosylcobalamin, and hydroxycobalamin.

8. The pharmaceutical composition according to any one of claims 1 to 7, further characterized in that the pharmaceutically acceptable form of vitamin B6 is selected from the group consisting of Pyridoxine aceglumate, Pyridoxine alpha-ketoglutarate, Pyridoxine dipalmitate, Pyridoxine phosphate, Pyridoxine phosphoserinate, Pyridoxine hydrochloride, Pyridoxine palmitate or Pyridoxine pyrrolidonecarboxylate.

9. The pharmaceutical composition according to any one of claims 1 to 8, further characterized in that the pharmaceutically acceptable salt of zinc is selected from the group consisting of zinc oxide and zinc sulfate.

10. The pharmaceutical composition according to any of claims 1 to 9, further characterized in that it additionally comprises at least one prebiotic.

11. The pharmaceutical composition according to claim 10, further characterized in that the prebiotic is selected from the group consisting of lactose, inulin, resistant starch, galactooligosaccharides (GOS), fructooligosaccharides (FOS), xylooligosaccharides (XOS), isomaltooligosaccharide (IMO), oligosaccharides, such as those from soy (SOS), transgalactooligosaccharides (TOS), polydextrose, lactulose, and at least one extract of at least one plant containing said prebiotics; or combinations thereof.

12. The pharmaceutical composition according to claim 11, further characterized in that the extract of at least one plant is obtained from a plant selected from the group consisting of dandelion, green tea, Jerusalem artichoke, onion, garlic, chicory root, asparagus, plantain, barley, agave, wheat, leek or combinations thereof.

13. The pharmaceutical composition according to any of claims 1 to 7, further characterized in that it additionally comprises at least one probiotic.

14. The pharmaceutical composition according to claim 13, characterized in that the probiotic is a microorganism of a genus selected from the group consisting of Lactobacillus, Bifidobacterium, Saccharomyces, Streptococcus, Enterococcus, Escherichia and Bacillus.

15. The pharmaceutical composition according to any of claims 1 to 14, further characterized in that it additionally comprises at least one pharmaceutically acceptable excipient.

16. The pharmaceutical composition according to claim 15, further characterized in that the pharmaceutically acceptable excipient is selected from the group consisting of: lactose, dextrins, glucose, sucrose, sorbitol, silicates, calcium salts, magnesium salts, potassium chloride, sodium chloride, starches, sugars, sugar alcohols, cellulose derivatives, alginates and derivatives, crospovidone, colloidal anhydrous silicon and other silica compounds, stearic acid and its salts, cellulose acetate phthalate, dyes, acacia, alginic acid, aluminum acetate, benzyl alcohol, butyl paraben, butyl hydroxytoluene, citric acid, calcium carbonate, waxes, carboxymethylcellulose and derivatives, calcium stearate, ethylenediaminetetraacetic acid (EDTA), copovidone, hydrogenated castor oil, calcium hydrogen phosphate dihydrate, chloride cetylpyridone, dimethi, sodium ethrosine, gelatin, glyceryl monolate, glycehna, glycine,glyceryl monostearate, hydroxymethylcellulose, hypromellose, ferric oxide, iron oxide, mannitol, mineral oil, vegetable oil, methacrylic acid copolymer, povidone, polyethylene glycol, polysorbate 80, polaxamers, potassium bicarbonate, phosphoric acid, sodium benzoate, sodium metabisulfite, succinic acid, sodium propionate, titanium dioxide, talc, triacetin, triethyl citrate, gums.

17. The pharmaceutical composition according to any one of claims 1 to 16, further characterized in that it has a pharmaceutical form selected from the group consisting of powder, capsule, tablets, emulsion, suspension, syrup and solution.

18. The pharmaceutical composition according to any one of claims 1 to 17, further characterized in that one or more components can be formulated into nanoparticles.

19. The pharmaceutical composition according to any of claims 1 to 18, further characterized in that it is added to a food.

20. The pharmaceutical composition according to any of claims 1 to 18, further characterized in that it is a food supplement.

21. A pharmaceutical composition as claimed in any one of claims 1 to 20 for use as an aid in the treatment of at least one condition selected from the group consisting of obesity, insulin resistance, steatotic liver disease, diabetes, metabolic syndrome, Crohn's disease, gastrointestinal disease, intestinal metabolic disorders and intestinal dysbiosis.

22. The pharmaceutical composition for use according to claim 21, wherein the supplement induces the enrichment of at least one beneficial bacterial genus.

23. The pharmaceutical composition for use according to claim 22, wherein the beneficial bacterial genus is selected from the group consisting of Acinetobacter, Anaeroplasma, Pseudomonas and Stenotrophomonas.

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