Pharmaceutical and nutraceutical compositions having combinations of amino acids and their use in diseases characterized by lipid accumulation in tissues - Patents.com

Abstract

The present invention discloses a pharmaceutical or dietary supplement composition that contains a specific combination of compounds including histidine, serine, cysteine ​​and carnosine.The composition is proposed as an ingredient in food and has been proven to be effective in treating diseases characterized by the accumulation of fat in tissues, such as fatty liver disease, obesity and atherosclerosis.

Description

[Technical field]

[0001] This application claims the benefit of European Patent Application No. 22382390.7, filed April 25, 2022.

[0002] The present invention relates to the field of dietary supplements and pharmaceutical compositions for reducing the risks associated with adipose tissue deposition or for treating diseases associated with adipose tissue deposition. Thus, the present invention relates to the fields of nutrition and medicine. [Background technology]

[0003] Due to the increasing consumption and worldwide spread of the diet known as the "Western diet", which includes processed foods and foods rich in fat, diseases or disorders associated with the deposition of adipose tissue in several parts of the body, including visceral, vascular and subcutaneous tissues, are a trend that involves not only high costs for the health care system but also the most important health problems in subjects. Examples of these disorders are obesity, atherosclerosis and fatty liver disease, which are accompanied by associated visceral fat accumulation.

[0004] In other words, obesity is the accumulation of excess body fat to a degree that can have adverse health effects.

[0005] Atherosclerosis is a pattern of arteriosclerosis, a disease in which the walls of arteries develop abnormalities called lesions. These lesions can lead to narrowing due to the buildup of atheromatous plaque. If severe, this can lead to coronary artery disease, stroke, peripheral artery disease, or kidney problems, depending on which arteries are affected.

[0006] Fatty liver disease (FLD), also commonly known as hepatic steatosis (HS), is a condition in which excess fat accumulates in the liver. Fatty liver disease is represented by nonalcoholic fatty liver disease (NAFLD) and alcoholic liver disease. Westernized changes in diet and lifestyle can lead to the progression of several important risk factors associated with the development of NAFLD, such as obesity, leptin or insulin resistance (IR), dyslipidemia and metabolic syndrome. NAFLD is one of the highly prevalent human diseases affecting almost 1 / 3 of the world's population today. The disease is characterized by excessive accumulation of fat and low-grade inflammation in the liver. This abnormal lipid metabolism can affect lipid oxidation, which in more advanced stages disrupts redox homeostasis. Thus, the accumulation of reactive species of oxygen (ROS) and thus increased lipid peroxidation can induce hepatocellular injury and fibrosis. NAFLD is classified into different grades according to its severity and ability to restore its health: nonalcoholic fatty liver (NAFL) with simple steatosis; nonalcoholic steatohepatitis (NASH) with advanced steatosis and inflammation. Furthermore, NASH may develop fibrosis in advanced stages and eventually progress to irreversible stages such as cirrhosis or hepatocellular carcinoma. Treatment of NAFLD is generally by dietary modification to reduce weight and promoting exercise, unless severe progression necessitates liver transplantation.

[0007] Specific cocktails of compounds have been tested in vivo as dietary supplements to promote fat oxidation and synthesis of glutathione (GSH). GSH is required to prevent the accumulation of defective products of fatty acid oxidation. One example is the paper "Personal model-assisted identification of NAD+ and glutathione metabolism as intervention target in NAFLD”, Mol Syst Biol-2017,vol.no.13:916. These authors tested in mice fed a Western diet with high levels of fat and a cocktail consisting of sucrose, serine, NAC (N-acetyl-L-cysteine) and nicotinamide riboside (NR). Serine was included in the cocktail because it can be easily converted to glycine, and NAC because cysteine ​​could be a limiting metabolite after supplementation of glycine in the synthesis of GSH. The aim of this assay was to provide a proof of concept of a possible therapeutic strategy based on the promotion of the synthesis of GSH and NAD+, derived from an analysis by personalized genome-scale metabolic modeling of a human cohort with several degrees of hepatic steatosis. The rationale behind was to reduce oxidative stress in lipid metabolism. From the GEM analysis of the human cohort, the authors also concluded that the plasma levels of serine and glycine were negatively correlated with HS, and no significant correlation existed between HS and plasma levels of cysteine ​​and glutamine.

[0008] Another publication proposing dietary supplementation to reduce high-fat diet-induced hepatic steatosis is from Mong et al., “Histidine and carnosine alleviated hepatic steatosis in mice consumed highly saturated fat diet”, European Journal of Pharmacology 653(2011)82-88. The authors observed that mice fed these supplements had reduced body weight, epididymal fat, and hepatic triglyceride and cholesterol levels, and after observing a concomitant improvement in insulin sensitivity and attenuation of hyperinsulinemia, they propose using histidine and the dipeptide carnosine to reduce fatty liver.

[0009] Among other things, some of the roles attributed to histidine are its role as an anti-inflammatory and antioxidant agent, thus acting as a protective agent for the liver. Firstly, this anti-inflammatory property of histidine supplementation is related to the regulation of NF-kB and PPARα-induced pathways, which also reduce plasma and hepatic levels of TNFα, IL6 and C-reactive protein. In addition, histidine has an anti-glycation effect, which is associated with reduced inflammasome activation. On the other hand, the antioxidant properties of histidine treatment are due to this amino acid being a free radical scavenger and having the ability to bind divalent metal ions.

[0010] Based on this latter ability as a free radical scavenger, some authors have proposed to use newly synthesized dimeric histidine (H-bihistidine) to treat non-alcoholic liver injury (NLI) and other free radical-induced diseases (see Zhao, Z.; Fu, C.; Zhang, Y.; Fu, A. Dimeric Histidine as a Novel Free Radical Scavenger Alleviates Non-Alcoholic Liver Injury. Antioxidants 2021, 10, 1529. https: / / doi.org / 10.3390 / antiox10101529). Although this is a good approach, the complexity of using synthetic compounds is always a drawback. [Prior art documents] [Non-patent literature]

[0011] [Non-Patent Document 1] Mardinoglu et al., “Personal model-assisted identification of NAD+ and glutathione metabolism as intervention target in NAFLD”, Mol Syst Biol-2017, vol.no.13:916 [Non-Patent Document 2] Mong et al., “Histidine and carnosine allikated hepatic steatosis in mice consumed high saturated fat diet”, European Journal of Pharmacology 653(2011)82-88 [Non-Patent Document 3] Zhao, Z.;Fu,C.;Zhang,Y.;Fu,A.Dimeric Histidine as a Novel Free Radical Scavenger Alleviates Non-Alcoholic Liver Injury.Antioxidants 2021,10,1529.https: / / doi.org / 10.3390 / antiox10101529 Summary of the Invention [Problem to be solved by the invention]

[0012] Therefore, there remains a need for alternative approaches to address all these diseases or disorders associated with fat deposition in tissues. [Means for solving the problem]

[0013] The inventors have surprisingly found that a specific combination of three or more of histidine, cysteine, serine and carnosine can reduce the liver damage and liver characteristics of NAFLD, with damage seen macroscopically in terms of liver weight, whitish fatty liver, and at the biochemical level by analyzing the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The combination of compounds also reduced liver lipid content and fatty liver. Dietary supplementation with the combination of compounds reduced liver inflammation associated with NAFLD, increased antioxidant defense and antitumor activity, and reduced resistance to insulin. In short, the combination of three or more of these compounds allowed to reverse the symptoms of NAFLD in animal models of the disease.

[0014] Furthermore, the inventors found that the compound combination also prevents fat mass gain associated with menopause in ovariectomized (OVX) rats, reduces the loss of lean mass in OVX rats, and contributes to a healthier profile of body composition.These compounds also improve insulin resistance in this animal model.Thus, the authors found that the compound combination contributes to the restoration of metabolic changes associated with menopause in ovariectomized (OVX) rats.

[0015] Without being bound by any theory, the inventors propose that the combination of these compounds as ingredients in nutraceutical compositions, food supplements or as active ingredients in pharmaceutical compositions ultimately leads to an increase in plasma histidine levels, which in the end allows for a reduction in fat accumulation due to the complexity of factors. This envisages a new approach related to the proposed use of histidine as a radical scavenger or as an anti-inflammatory agent in diseases involving fat accumulation, as previously disclosed. Moreover, this approach is surprising, since histidine was only proposed to have a moderate effect on all these diseases, and the treatments in the prior art were more focused on promoting fat oxidation and the synthesis of glutathione (GSH). Moreover, the proposed approach, derived from the pure observations made by the inventors, allows to safely address these disorders related to fat accumulation in some body tissues in subjects whose control of fat oxidation capacity is not impaired, but who also suffer from clinical symptoms due to other mechanisms that ultimately reduce histidine circulating levels.

[0016] This rationale may also underlie the reduction of visceral fat in obese subjects, and the reduction of LDL-cholesterol in models of atherosclerosis.

[0017] Thus, a first aspect of the present invention is a pharmaceutical or nutraceutical composition comprising a therapeutically or nutraceutically effective amount of at least three of histidine, or a pharma- tically or nutraceutically acceptable salt or a solvate of any of them, cysteine, or a pharma-tically or nutraceutically acceptable salt or a solvate of any of them, serine, or a pharma-tically or nutraceutically acceptable salt or a solvate of any of them, together with one or more pharma-tically or nutraceutically acceptable excipients or carriers; and / or a therapeutically or nutraceutically effective amount of a peptide, the peptide comprising a sequence of amino acids comprising at least three of histidine, cysteine, serine and carnosine, in particular the pharmaceutical or nutraceutical composition comprising, together with one or more pharma-tically or nutraceutically acceptable excipients or carriers: (i) a therapeutically or nutraceutical effective amount of cysteine, or a pharma- ceutically or nutraceutically acceptable salt, or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of at least two of histidine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either thereof, serine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either thereof, and carnosine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either thereof; and / or a therapeutically or nutraceutically effective amount of a peptide comprising a sequence of amino acids that includes cysteine ​​and at least two of histidine, serine, and carnosine; or (ii) a therapeutically or nutraceutical effective amount of N-acetylcysteine ​​(NAC) or a pharma- ceutically or nutraceutically acceptable salt or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of at least two of histidine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, serine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, and carnosine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof; and / or a therapeutically or nutraceutically effective amount of a peptide comprising a sequence of amino acids that includes NAC and at least two of histidine, serine, and carnosine.

[0018] The pharmaceutical or nutraceutical composition may be used as an ingredient in general purpose foods such as beverages (i.e. soda), dairy products (i.e. yogurt), bakery and confectionery products, etc., along with being a common ingredient in these types of foods. Thus, a second aspect of the invention is a food product in solid or liquid form comprising the pharmaceutical or nutraceutical composition defined in the first aspect.

[0019] As illustrated in the examples and as shown above, the composition or foodstuff comprising at least three of the listed compounds has some beneficial effects in the animal model of NAFLD and the animal model of menopause.Therefore, another aspect of the present invention is the composition defined in the first aspect or the foodstuff of the second aspect for use in treatment (i.e. as a drug). [Brief description of the drawings]

[0020] [Figure 1]FIG. 1A shows the effect of treatment on liver weight. FIG. 1B shows the effect of treatment on representative gross appearance of the liver. FIG. 1C shows the effect of treatment on serum ALT (alanine aminotransferase). FIG. 1D shows the effect of treatment on serum AST (aspartate aminotransferase). Data are mean±SEM; n=16 animals / group. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 vs. control mice; #p<0.05, ##p<0.01, ###p<0.001 vs. HFHFr mice. [Diagram 2] Figure 2A shows the effect of treatment on total liver lipid content. Figure 2B shows the effect of treatment on total liver triglyceride (TG) content. Figure 2C shows the effect of treatment on total liver cholesterol content. Data are mean ± SEM; n = 16 animals / group. **p<0.01, ***p<0.001, ****p<0.0001 vs. control mice; #p<0.05, ####p<0.0001 vs. HFHFr mice. [Diagram 3] Figure 3A shows the histopathology and image analysis of the liver determined by H&E (hematoxylin-eosin staining) and the amplification of selected areas shown in the lower panel. Figure 3B shows the lipid droplet count. Figure 3C shows the lipid droplet surface field. Figure 3D shows the NAFLD / NASH score table. st., steatosis. Bar = 100 μm. Data are mean ± SEM; n = 6 animals / group. **p < 0.01, ****p < 0.0001 vs. control mice (C); ###p < 0.001 vs. HFHFr mice; ####p < 0.0001 vs. HFHFr mice. AA mice are treated mice. [Figure 4]Figure 4 shows the effect of treatment on hepatic insulin resistance. Figure 4A shows a representative Western blot analysis with Akt activation (pAktS473), total Akt protein levels (T-Akt), housekeeping β-actin levels and protein loading by Ponceau S membrane staining. Figure 4B shows densitometric analysis of phosphorylated Akt to total Akt ratio. Data are mean ± SEM; n=5–6 animals / group. *p<0.05 vs. control mice. Figure 4C shows analysis of hepatic expression of key genes after quaternary treatment in fatty acid β-oxidation (carnitine palmitoyltransferase, Cpt1a), inflammation (chemokine (CC motif) ligand 2, CCL2), lipolysis (hormone sensitive, HSL; and lipin, Plin) and adipogenesis (fatty acid synthase, FASN). Data are expressed as mean ± SEM; n=10–12 per condition. *p<0.05. [Diagram 5] Figure 5 shows the effect of treatment on circulating cholesterol levels. In Figure 5A, circulating HDL levels are shown. In Figure 5B, circulating low density lipoprotein (LDL) levels are shown. In Figure 5C, LDL / HDL ratios are shown. Data are mean ± SEM. n=16 animals / group. *p<0.05 **p<0.01, ***p<0.001, ****p<0.0001. [Figure 6]FIG. 6A shows the effect of treatment on body weight change after treatment. FIG. 6B shows the effect of treatment on the weight of the major visceral depots, epididymal white adipose tissue (EWAT), retroperitoneal white adipose tissue (RWAT) and mesenteric white adipose tissue (MWAT) after treatment. FIG. 6C shows the growth curves. Circles represent "lean", squares represent "obese" and triangles represent "obese+4AA". FIG. 6D shows visceral adipose tissue pathology and image analysis determined by H&E (hematoxylin-eosin staining). FIG. 6E shows BAT weight. FIG. 6F shows BAT histopathology and image analysis. FIG. 6G shows gene expression analysis of the indicated genes in BAT. FIG. 6H shows representative western blot analysis and quantification of the indicated proteins in BAT. FIG. 6I shows thermography analysis and quantification of mice. In Figures 6D-6I, the left column represents "lean", the center column represents "obese", and the right column represents "obese+4AA". Data are means ± SEM. n=16 animals / group. *p<0.05 **p<0.01, ****p<0.0001. [Figure 7] Figure 7 shows the effect of 24 h treatment with the indicated amino acids alone or in combination on Cpt1a, Ccl2, Hsl, Plin and Fasn mRNA expression in human hepatocytes (HepG2) with the following concentrations: histidine 10 mM, carnosine 5 mM, cysteine ​​5 mM and serine 5 mM. These concentrations were pooled in different combinations. Data are expressed as mean ± SEM; n = 3-4 per condition. *p<0.05, **p<0.01, ***p<0.001 vs vehicle. [Figure 8]Figure 8A shows the effect of ovariectomy, 17β-E2 injection and quaternary histidine + cysteine ​​+ serine + carnosine (4AA) treatment on changes in body weight in sham-operated (SH) and ovariectomized (OVX) female Sprague-Dawley rats after 57 days of treatment. Figure 8B shows the effect of ovariectomy, 17β-E2 injection and quaternary histidine + cysteine ​​+ serine + carnosine (4AA) treatment on changes in fat mass in sham-operated (SH) and ovariectomized (OVX) female Sprague-Dawley rats after 57 days of treatment. FIG. 8C shows the effect of ovariectomy, 17β-E2 injection and quaternary histidine+cysteine+serine+carnosine (4AA) treatment on changes in lean mass in sham-operated (SH) and ovariectomized (OVX) female Sprague-Dawley rats after 57 days of treatment. Body weight was recorded every 7 days, and fat mass and lean mass were recorded every 28 days. FIG. 8D shows weight gain in grams calculated as a percentage of final body weight accumulated every week throughout the study. Increases in fat mass and lean mass were calculated as the percentage difference between fat mass at the end of the experiment and fat mass at baseline. Data are shown as mean ± SEM (n=10). In each panel, different superscript lowercase letters (a, b, c, d) indicate mean values ​​that are significantly different between groups (one-way ANOVA and Duncan's post-hoc test, Welch's test and Games-Howell post-hoc test or Kruskal-Wallis test and Mann-Whitney U post-hoc test, p<0.05). I: effect of intervention, t: effect of time, Ixt: interaction between type of intervention and time (RM-ANOVA, p<0.05). SH: sham-operated rats, OVX: ovariectomized rats, OVX-E2: OVX rats treated with 17β-estradiol, OVX-4AA: OVX rats supplemented with the quaternary histidine + cysteine ​​+ serine + carnosine (4AA). In Figures 8A-8C, circles represent "SH", triangles represent "OVX", diamonds represent "OVX-E2" and squares represent "OVX-4AA". [Figure 9]Figure 9A shows the effect of ovariectomy, 17β-E2 injection and quaternary histidine + cysteine ​​+ serine + carnosine (4AA) treatment on RWAT depot weight in sham-operated (SH) and ovariectomized (OVX) rats after 57 days of intervention. Figure 9B shows the effect of ovariectomy, 17β-E2 injection and quaternary histidine + cysteine ​​+ serine + carnosine (4AA) treatment on MWAT depot weight in sham-operated (SH) and ovariectomized (OVX) rats after 57 days of intervention. Figure 9C shows the effect of ovariectomy, 17β-E2 injection and quaternary histidine + cysteine ​​+ serine + carnosine (4AA) treatment on IWAT depot weight in sham-operated (SH) and ovariectomized (OVX) rats after 57 days of intervention. Figure 9D shows the effect of ovariectomy, 17β-E2 injection and quaternary histidine + cysteine ​​+ serine + carnosine (4AA) treatment on body mass index in sham-operated (SH) and ovariectomized (OVX) rats after 57 days of intervention. Figure 9E shows the effect of ovariectomy, 17β-E2 injection and quaternary histidine + cysteine ​​+ serine + carnosine (4AA) treatment on circulating levels of leptin in sham-operated (SH) and ovariectomized (OVX) rats after 57 days of intervention. Data are shown as mean ± SEM (n = 9-10). Body mass index was calculated as the sum of IWAT, MWAT and RWAT depot weights (grams) and expressed as a percentage of body weight. In each figure, different superscript letters (a, b, c) indicate mean values ​​that are significantly different between groups (one-way ANOVA and Duncan's post hoc test, p < 0.05). I: Effect of intervention. SH: sham-operated rats, OVX: ovariectomized rats, OVX-E2: OVX rats treated with 17β-estradiol, OVX-4AA: OVX rats supplemented with four histidine-related amino acids (4AA). MWAT: mesenteric white adipose tissue; RWAT: retroperitoneal white adipose tissue; IWAT: inguinal white adipose tissue. [Figure 10]Figure 10a shows plasma levels of insulin at the end of the study in sham-operated (SH) and ovariectomized (OVX) rats after 57 days of intervention. Figure 10b shows plasma levels of glucose at the end of the study in sham-operated (SH) and ovariectomized (OVX) rats after 57 days of intervention. Figure 10c shows plasma levels of HOMA-IR at the end of the study in sham-operated (SH) and ovariectomized (OVX) rats after 57 days of intervention. Data are shown as mean ± SEM (n = 9-10). Superscript lowercase letters (a, b, c) indicate mean values ​​that are significantly different between groups (one-way ANOVA and Duncan's post hoc test, p < 0.05). I: Effect of intervention. SH: sham-operated rats, OVX: ovariectomized rats, OVX-E2: OVX rats treated with 17β-estradiol, OVX-4AA: OVX rats supplemented with quaternary histidine + cysteine ​​+ serine + carnosine (4AA). HOMA-IR: homeostasis model assessment estimated insulin resistance. [Figure 11]Figure 11 shows the association of plasma histidine and degree of steatosis with gut microbiota and Hut genes in humans. Figure 11a shows a volcano plot of different bacterial families associated with circulating histidine levels in a discovery cohort (n=73) identified using an analysis of microbiota with bias correction controlling for age, BMI, sex and country (ANCOM-BC). Figure 11b shows a volcano plot of different bacterial families associated with degree of fatty liver (liver biopsy) in a discovery cohort (n=73) identified using an analysis of microbiota with bias correction controlling for age, BMI, sex and country (ANCOM-BC). The log2 (fold change) associated with unit change in plasma histidine levels and -log10 (p-value) adjusted for multiple testing are plotted for each taxon. Significantly different taxa are colored according to phylum. Figure 11c shows histidine utilization pathways. The first three pathways appear to be universal. Formiminoglutamate has two different degradation pathways depending on the genus: hydrolysis to formamide and glutamate or hydrolysis to formylglutamate followed by hydrolysis to formate and glutamate. Figure 11d shows a violin plot of centered log-transformed microbial genes hutH, hutU, hutI and hutG involved in histidine utilization in subjects with lower or higher steatosis degree than 33%, respectively. Figure 11e shows qPCR of microbial gene expression involved in histidine utilization (hutH and hutG). Data are mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 vs. NAFLD mice. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] All terms used herein in this application are to be understood with their ordinary meaning known in the art unless otherwise specified. Other more specific definitions of certain terms used in this application are as follows, and are intended to be applied uniformly throughout the specification and claims, unless a definition expressly set forth provides a broader definition.

[0022] As used herein, the indefinite article "a" is synonymous with "at least one" or "one or more." Unless otherwise indicated, definite articles such as "the" and "the" as used herein also include the plural form of the noun.

[0023] For purposes of the present invention, any range given includes both the lower and upper endpoints of the range.

[0024] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, etc., and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or organic acids such as acetic acid, trifluoroacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by using other methods used in the art, such as ion exchange. Other pharma- ceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, and 2-hydroxy-ethanesulfonate. Salts derived from appropriate bases include alkali metal, alkaline earth metal, and ammonium salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharma-ceutically acceptable salts include non-toxic ammonium, quaternary ammonium and amine cations formed, where appropriate, using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkylsulfonates and arylsulfonates.

[0025] The term "solvate" refers to a molecular complex that comprises the compound of interest (i.e., L-histidine, L-cysteine, L-serine and L-carnosine or its pharma- ceutically acceptable salt) and one or more solvent molecules (e.g., water) in stoichiometric or non-stoichiometric amounts that are bound by non-covalent intermolecular forces.When one or more solvent molecules that form part of the molecular complex are water, the solvate is a hydrate.Methods of solvation are generally known in the art.

[0026] The term "medicine" also encompasses the notion of "veterinary compositions". They therefore relate to compositions that are therapeutically effective when administered by any desired or applicable route to any animal, including humans.

[0027] The expression "therapeutically effective amount" as used herein refers to an amount of compound that is sufficient when administered to prevent or alleviate to some extent one or more symptoms of the disease being treated.The specific dose of compound administered according to the present invention will of course be determined by the specific circumstances surrounding the case, including the compound being administered, the route of administration, the specific condition being treated, and similar considerations.

[0028] The expression "pharmaceutical or nutraceutical acceptable excipient or carrier" refers to a pharmaceutically acceptable material, composition, or vehicle. Each component must be pharmaceutically acceptable in the sense of being compatible with the other components of the pharmaceutical composition. It must also be suitable for use in contact with human and animal tissues or organs without undue toxicity, irritation, allergic reaction, immunogenicity or other problems or complications, commensurate with a reasonable benefit / risk ratio. Examples of suitable acceptable excipients are solvents, dispersion media, diluents, or other liquid vehicles, dispersing or suspending aids, surfactants, isotonicity agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, and the like. Use of any conventional excipient medium is contemplated within the scope of the present invention, except insofar as it is incompatible with the substance or its derivatives, such as by producing some undesirable biological effect or otherwise interacting in a deleterious manner with any one or more other components of the pharmaceutical composition.

[0029] The expression "nutraceutical effective amount" as used herein refers to the amount of compound that is sufficient to provide physiological benefits to subjects in a certain metabolic aspect when administered.The specific dose of compound administered according to the present invention is naturally determined by the specific circumstances surrounding the case, including the compound administered, the route of administration, the specific condition being treated, and similar considerations.

[0030] A "nutraceutical composition", also referred to herein as a "nutraceutical composition" or "dietary supplement", is a composition that provides a physiological benefit to a subject in a certain metabolic aspect. They cannot be labeled with drug claims, but can have health and nutritional claims, according to health agency regulations. This category includes several compositions that vary depending on national regulations. In general, it is a product taken orally, containing "dietary ingredients" intended to complement the diet. The "dietary ingredients" in these products can include substances such as vitamins, minerals, herbs or other botanicals, amino acids, and enzymes, organ tissues, glands and metabolites. Dietary supplements can also be extracts or concentrates and can be found in many forms, such as tablets, capsules, softgels, gelcaps, liquids or powders.

[0031] "Functional foods," also referred to herein as "foods," are foods that have been enriched or fortified during processing and then marketed as providing some benefit to the consumer.

[0032] As used herein, the term "NAC" or "N-acetylcysteine" or "acetylcysteine" refers to the acetylated form of the amino acid L-cysteine, and has the IUPAC name (2R)-2-acetamido-3-sulfanylpropanoic acid. As will be appreciated by those skilled in the art, NAC is a well-established cysteine ​​prodrug, and therefore cysteine ​​and NAC can be used interchangeably in the compositions of the present invention.

[0033] As mentioned above, a first aspect of the present invention is a pharmaceutical or nutraceutical composition comprising a therapeutically or nutraceutically effective amount of at least three of histidine or a pharma- tically or nutraceutically acceptable salt or a solvate of any of them, cysteine ​​or a pharma- tically or nutraceutically acceptable salt or a solvate of any of them, serine or a pharma-tically or nutraceutically acceptable salt or a solvate of any of them, together with one or more pharma- tically or nutraceutically acceptable excipients or carriers; and / or a therapeutically or nutraceutically effective amount of a peptide comprising a sequence of amino acids comprising at least three selected from the group of compounds consisting of histidine, cysteine, serine and carnosine, in particular the pharmaceutical or nutraceutical composition comprising, together with one or more pharma- tically or nutraceutically acceptable excipients or carriers: (i) a therapeutically or nutraceutical effective amount of cysteine, or a pharma- ceutically or nutraceutically acceptable salt, or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of at least two of histidine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either thereof, serine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either thereof, and carnosine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either thereof; and / or a therapeutically or nutraceutically effective amount of a peptide comprising a sequence of amino acids that includes cysteine ​​and at least two of histidine, serine, and carnosine; or (ii) a therapeutically or nutraceutical effective amount of N-acetylcysteine ​​(NAC) or a pharma- ceutically or nutraceutically acceptable salt or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of at least two of histidine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, serine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, and carnosine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof; and / or a therapeutically or nutraceutically effective amount of a peptide comprising a sequence of amino acids that includes NAC and at least two of histidine, serine, and carnosine.

[0034] In certain embodiments, the pharmaceutical or nutraceutical composition comprises a therapeutically or nutraceutically effective amount of at least three of histidine, or a pharma- ceutically or nutraceutically acceptable salt, or a solvate of any of them; cysteine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of any of them; serine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of any of them; and carnosine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of any of them, together with one or more pharma-ceutically or nutraceutically acceptable excipients or carriers.

[0035] In certain embodiments, the pharmaceutical or nutraceutical composition comprises, together with one or more pharma- ceutical or nutraceutical acceptable excipients or carriers, the following: (i) a therapeutically or nutraceutical effective amount of cysteine, or a pharma- ceutically or nutraceutically acceptable salt, or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of at least two of histidine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either thereof, serine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either thereof, and carnosine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either thereof; and / or a therapeutically or nutraceutically effective amount of a peptide comprising a sequence of amino acids that includes cysteine ​​and at least two of histidine, serine, and carnosine; or (ii) a therapeutically or nutraceutical effective amount of N-acetylcysteine ​​(NAC) or a pharma- ceutically or nutraceutically acceptable salt or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of at least two of histidine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, serine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, and carnosine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof; and / or a therapeutically or nutraceutically effective amount of a peptide comprising a sequence of amino acids that includes NAC and at least two of histidine, serine, and carnosine.

[0036] In another specific embodiment, the pharmaceutical or nutraceutical composition comprises, together with one or more pharma- ceutical or nutraceutical acceptable excipients or carriers: (i) a therapeutically or nutraceutically effective amount of cysteine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of at least two of histidine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, and carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof; or (ii) a therapeutically or nutraceutical effective amount of N-acetylcysteine ​​(NAC) or a pharma- ceutically or nutraceutically acceptable salt or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of at least two of histidine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, serine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, and carnosine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof.

[0037] In another specific embodiment, the pharmaceutical or nutraceutical composition comprises, together with one or more pharma- ceutical or nutraceutical acceptable excipients or carriers: (i) a therapeutically or nutraceutical effective amount of cysteine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, a therapeutically or nutraceutically effective amount of histidine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, a therapeutically or nutraceutically effective amount of serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, a therapeutically or nutraceutically effective amount of carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof; and / or a therapeutically or nutraceutically effective amount of a peptide comprising a sequence of amino acids including cysteine, histidine, serine and carnosine; or (ii) a therapeutically or nutraceutical effective amount of NAC, or a pharma- ceutically or nutraceutically acceptable salt, or a solvate of either thereof, a therapeutically or nutraceutically effective amount of histidine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either thereof, a therapeutically or nutraceutically effective amount of serine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either thereof, a therapeutically or nutraceutically effective amount of carnosine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either thereof; and / or a therapeutically or nutraceutically effective amount of a peptide comprising a sequence of amino acids including NAC, histidine, serine, and carnosine.

[0038] In another specific embodiment, the pharmaceutical or nutraceutical composition comprises, together with one or more pharma- ceutical or nutraceutical acceptable excipients or carriers: (i) a therapeutically or nutraceutical effective amount of cysteine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, a therapeutically or nutraceutically effective amount of histidine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, a therapeutically or nutraceutically effective amount of serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof; or (ii) a therapeutically or nutraceutical effective amount of NAC, or a pharma- ceutically or nutraceutically acceptable salt, or a solvate of either of them; a therapeutically or nutraceutically effective amount of histidine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either of them; a therapeutically or nutraceutically effective amount of serine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either of them; and a therapeutically or nutraceutically effective amount of carnosine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of either of them.

[0039] In a more specific embodiment, the pharmaceutical or nutraceutical composition comprises a therapeutically or nutraceutically effective amount of N-acetylcysteine ​​(NAC) or a pharma- ceutically or nutraceutically acceptable salt or a solvate of either thereof, and therapeutically or nutraceutically effective amounts of at least two of histidine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, serine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, and carnosine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, together with one or more pharma-ceutically or nutraceutically acceptable excipients or carriers.

[0040] In a more specific embodiment, the pharmaceutical or nutraceutical composition comprises a therapeutically or nutraceutically effective amount of NAC or a pharma- ceutically or nutraceutically acceptable salt or a solvate of either thereof, a therapeutically or nutraceutically effective amount of histidine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, a therapeutically or nutraceutically effective amount of serine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of carnosine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, together with one or more pharma-ceutically or nutraceutically acceptable excipients or carriers.

[0041] In certain embodiments, the pharmaceutical or nutraceutical composition comprises a therapeutically effective amount of cysteine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 2-550 mg per Kg of the subject; a therapeutically or nutraceutically effective amount of histidine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 7.5 mg-250 mg per Kg of the subject; a therapeutically effective amount of serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 1.3-250 mg per Kg of the subject; and a therapeutically effective amount of carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 4.5-200 mg per Kg of the subject.

[0042] In certain embodiments, the pharmaceutical or nutraceutical composition comprises a therapeutically or nutraceutical effective amount of histidine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 10 mg to 250 mg per Kg of the subject; a therapeutically or nutraceutically effective amount of cysteine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 30 to 550 mg per Kg of the subject; a therapeutically or nutraceutically effective amount of serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 10 to 250 mg per Kg of the subject; and a therapeutically or nutraceutically effective amount of carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 10 to 200 mg per Kg of the subject.

[0043] In another specific embodiment, the pharmaceutical or nutraceutical composition is a therapeutically effective amount of NAC or a pharma- ceutically or nutraceutically acceptable salt or a solvate of either thereof, providing a dose of 2-550 mg per Kg of the subject; a therapeutically or nutraceutically effective amount of histidine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, providing a dose of 7.5 mg to 250 mg per Kg of the subject; a therapeutically effective amount of serine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, providing a dose of 1.3 to 250 mg per Kg of the subject; and a therapeutically effective amount of carnosine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, providing a dose of 4.5 to 200 mg per Kg of the subject.

[0044] In another specific embodiment, the pharmaceutical or nutraceutical composition is a therapeutically effective amount of NAC or a pharma- ceutically or nutraceutically acceptable salt or a solvate of either thereof, providing a dose of 30-550 mg per Kg of the subject; a therapeutically or nutraceutically effective amount of histidine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, providing a dose of 10 mg-250 mg per Kg of the subject; a therapeutically effective amount of serine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, providing a dose of 10-250 mg per Kg of the subject; and a therapeutically effective amount of carnosine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, providing a dose of 10-200 mg per Kg of the subject.

[0045] The therapeutic or nutraceutical effective amount shown is the amount with respect to the weight of animals, particularly mammals, including mice and humans.In practice, the amount and dosage are generally determined by those skilled in the art by human equivalent dose (HED) calculations, which allows those skilled in the art to find the value for humans (Guidance for Industry Estimating Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers, U.S. Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER), July 2005 Pharmacology and Toxicology).

[0046] For example, a dose of 210 mg / Kg of histidine, serine and carnosine in mice assumes a HED of 17.027 mg / Kg (median human body weight of 70 kg). Similarly, a dose of 490 mg / Kg assumes a HED of 39.730 mg / Kg. The amount per Kg of subject is the amount used to calculate the daily dose (i.e. amount) in certain embodiments of the invention. Specific daily doses in humans are 660 mg / day to 1500 mg / day for histidine, serine and carnosine (or any salt or solvate thereof) and 1700 mg / day to 3000 mg / day for cysteine ​​(or any salt or solvate thereof).

[0047] In certain embodiments, a therapeutically or nutraceutical effective amount of any one of histidine, serine and carnosine or a pharma-ceutically acceptable salt or solvate thereof provides a dose of 12-230 mg / Kg, or even more specifically, 17-210 mg / Kg.

[0048] In certain embodiments, a therapeutically or nutraceutical effective amount of histidine, serine and carnosine or a pharma- ceutically acceptable salt or solvate thereof provides a dose of 12-25 mg / Kg in humans, or an amount selected from 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 and 25 mg / Kg. In mice, this is 200 mg / Kg to 230 mg / Kg, more specifically 210 mg / Kg.

[0049] In a particular embodiment, the therapeutically or nutraceutical effective amount of histidine, serine and carnosine or a pharma- ceutically acceptable salt or solvate thereof is one that provides a dose of 0.5-25, 1-25, 1.3-25, 1.5-25, 2.5-25, 4-25, 5-25, 6-25, 7.5-25, 8-25, 9-25, 10-25, 12-25, 15-25, 17-25, or 20-25 mg / kg in humans, particularly 15-25 mg / kg in humans. In another particular embodiment, the therapeutically or nutraceutical effective amount of histidine, serine and carnosine or a pharma- ceutically acceptable salt or solvate thereof is one that provides a dose of 10-25, 12-22, 15-20, or 16-18 mg / kg in humans, particularly 15-20 mg / kg in humans.

[0050] In another more specific embodiment, optionally in combination with any of the above or below embodiments, the therapeutically or nutraceutical effective amount of cysteine ​​or a pharma- ceutically or nutraceutically acceptable salt or solvate thereof provides a dose of 35 mg / Kg to 500 mg / Kg. In another more specific embodiment, optionally in combination with any of the above or below embodiments, the therapeutically or nutraceutical effective amount of cysteine ​​or a pharma- ceutically or nutraceutically acceptable salt or solvate thereof provides a dose of 35 to 50 mg / Kg in humans, or an amount selected from 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 36, 37, 38, 39, 39.5, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50 mg / Kg. In mice, this is between 480mg / Kg and 500mg / Kg, more specifically 490mg / Kg.

[0051] In another particular embodiment, optionally in combination with any of the above or below embodiments, the therapeutically or nutraceutical effective amount of cysteine ​​or a pharma- ceutically or nutraceutically acceptable salt or solvate thereof provides a dose of 1-10, 1.5-7.5, or 2-5 mg / kg in a human, particularly 2-5 mg / kg in a human.

[0052] In another specific embodiment, optionally in combination with any of the above or below embodiments, the therapeutically or nutraceutical effective amount of cysteine ​​or a pharma- ceutically or nutraceutically acceptable salt or solvate thereof is that which provides a dose in humans of 20-55, 25-50, 30-45, or 35-40, particularly 35-40 mg / kg.

[0053] In another more specific embodiment, optionally in combination with any of the above or below embodiments, the therapeutically or nutraceutical effective amount of NAC or a pharma- ceutically or nutraceutically acceptable salt or solvate thereof provides a dose of 35 mg / Kg to 500 mg / Kg. In another more specific embodiment, optionally in combination with any of the above or below embodiments, the therapeutically or nutraceutical effective amount of NAC or a pharma- ceutically or nutraceutically acceptable salt or solvate thereof provides a dose of 35 to 50 mg / Kg in humans, or an amount selected from 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 36, 37, 38, 39, 39.5, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50 mg / Kg. In mice, this is 480 mg / Kg to 500 mg / Kg, more specifically 490 mg / Kg.

[0054] In another specific embodiment, optionally in combination with any of the above or below embodiments, the therapeutically or nutraceutical effective amount of NAC or a pharma- ceutically or nutraceutically acceptable salt or solvate thereof provides a dose of 1-10, 1.5-7.5, or 2-5 mg / kg in a human, particularly 2-5 mg / kg in a human.

[0055] In another specific embodiment, optionally in combination with any of the above or below embodiments, the therapeutically or nutraceutical effective amount of cysteine ​​or a pharma- ceutically or nutraceutically acceptable salt or solvate thereof is that which provides a dose in humans of 20-55, 25-50, 30-45, or 35-40, particularly 35-40 mg / kg.

[0056] In more particular embodiments, the pharmaceutical or nutraceutical composition comprises a therapeutically effective amount of cysteine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 1-7.5 mg per Kg of the subject, the subject being preferably a human; a therapeutically or nutraceutically effective amount of histidine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 5 mg-25 mg per Kg of the subject, the subject being preferably a human; a therapeutically effective amount of serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 1-5 mg per Kg of the subject, the subject being preferably a human; and a therapeutically effective amount of carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 2-15 mg per Kg of the subject, the subject being preferably a human.

[0057] In even more particular embodiments, the pharmaceutical or nutraceutical composition comprises a therapeutically effective amount of cysteine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, provided in a dose of 2-5 mg per Kg of the subject, the subject being preferably a human; a therapeutically or nutraceutically effective amount of histidine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, provided in a dose of 7.5-20 mg per Kg of the subject, the subject being preferably a human; a therapeutically effective amount of serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, provided in a dose of 1.3-3.3 mg per Kg of the subject, the subject being preferably a human; and a therapeutically effective amount of carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, provided in a dose of 4.5-10 mg per Kg of the subject, the subject being preferably a human.

[0058] In more specific embodiments, the pharmaceutical or nutraceutical composition comprises a therapeutically effective amount of NAC or a pharma- ceutically or nutraceutically acceptable salt or a solvate of either thereof, the therapeutically or nutraceutically effective amount being provided at a dose of 1-7.5 mg per Kg of the subject, the subject being preferably a human; a therapeutically or nutraceutically effective amount of histidine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, the subject being preferably a human; a therapeutically effective amount of serine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, the subject being preferably a human; and a therapeutically effective amount of carnosine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, the subject being preferably a human;

[0059] In still further specific embodiments, the pharmaceutical or nutraceutical composition comprises a therapeutically effective amount of NAC or a pharma- ceutically or nutraceutically acceptable salt or a solvate of either thereof, the therapeutically or nutraceutically effective amount being provided at a dose of 2-5 mg per Kg of the subject, the subject being preferably a human; a therapeutically or nutraceutically effective amount of histidine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, the subject being preferably a human; a therapeutically effective amount of serine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, the subject being preferably a human; a therapeutically effective amount of carnosine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, the subject being preferably a human; a therapeutically effective amount of carnosine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, the subject being preferably a human;

[0060] In more particular embodiments, the pharmaceutical or nutraceutical composition comprises a therapeutically effective amount of cysteine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 35-40 mg / kg of subject, the subject being preferably a human; a therapeutically or nutraceutically effective amount of histidine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 15-20 mg / kg of subject, the subject being preferably a human; a therapeutically effective amount of serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 15-20 mg / kg of subject, the subject being preferably a human; and a therapeutically effective amount of carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, providing a dose of 15-20 mg / kg of subject, the subject being preferably a human.

[0061] In more particular embodiments, the pharmaceutical or nutraceutical composition comprises a therapeutically effective amount of NAC or a pharma- ceutically or nutraceutically acceptable salt thereof or a solvate of either thereof, providing a dose of 35-40 mg / kg of subject, the subject being preferably a human; a therapeutically or nutraceutically effective amount of histidine or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of either thereof, providing a dose of 15-20 mg / kg of subject, the subject being preferably a human; a therapeutically effective amount of serine or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of either thereof, providing a dose of 15-20 mg / kg of subject, the subject being preferably a human; and a therapeutically effective amount of carnosine or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of either thereof, providing a dose of 15-20 mg / kg of subject, the subject being preferably a human.

[0062] In another particular embodiment of the first aspect, the composition comprises a therapeutically or nutraceutical effective amount of at least three of L-histidine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; L-cysteine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; L-serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; and L-carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them.

[0063] Indeed, the amino acids in the composition can be either D or L enantiomers, but in certain embodiments are L-amino acids.Similarly, the dipeptide carnosine can be in the form of L-carnosine or D-carnosine, but in certain embodiments is L-carnosine.In one embodiment, the composition of the present invention comprises any combination of any of the compounds in L or D form.In another embodiment, the composition of the present invention comprises any combination of any of the amino acids histidine, serine, carnosine and / or cysteine ​​in L or D form.

[0064] In certain embodiments of the first aspect, the composition comprises: (i) a therapeutically or nutraceutical effective amount of L-cysteine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of at least two of L-histidine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, L-serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, and L-carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof; or (ii) a therapeutically or nutraceutical effective amount of N-acetylcysteine ​​(NAC) or a pharma- ceutically or nutraceutically acceptable salt or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of at least two of L-histidine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, L-serine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, and L-carnosine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof.

[0065] In another particular embodiment of the first aspect, the composition comprises: (i) a therapeutically or nutraceutical effective amount of L-cysteine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, a therapeutically or nutraceutically effective amount of L-histidine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, a therapeutically or nutraceutically effective amount of L-serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of L-carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof; or (ii) a therapeutically or nutraceutical effective amount of NAC, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of either of them; a therapeutically or nutraceutically effective amount of L-histidine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either of them; a therapeutically or nutraceutically effective amount of L-serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either of them; and a therapeutically or nutraceutically effective amount of L-carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either of them.

[0066] In another particular embodiment of the pharmaceutical composition according to the first aspect, the pharma- ceutically or nutraceutical acceptable salts of histidine and cysteine ​​are hydrochlorides, more particularly the hydrochlorides of L-histidine and L-cysteine.Even more particularly, the pharma-ceutically acceptable salt of L-histidine is L-histidine monohydrochloride monohydrate and the pharma-ceutically acceptable salt of L-cysteine ​​is L-cysteine ​​hydrochloride.

[0067] In another particular embodiment of the pharmaceutical composition according to the first aspect, the pharma- ceutically or nutraceutical acceptable salt of histidine is a hydrochloride, more particularly the hydrochloride of L-histidine.Even more particularly, the pharma-ceutically acceptable salt of L-histidine is L-histidine monohydrochloride monohydrate.

[0068] In another particular embodiment of the pharmaceutical composition according to the first aspect, the pharma- ceutically or nutraceutical acceptable salt of cysteine ​​is a hydrochloride, more particularly the hydrochloride of L-cysteine. Even more particularly, the pharma-ceutically acceptable salt of L-cysteine ​​is L-cysteine ​​hydrochloride.

[0069] In more specific embodiments of the pharmaceutical and dietary supplements of the first aspect, they comprise at least three of the indicated components, at least one of which is histidine, more specifically L-histidine, or any of their salts or solvates.Indeed, as shown in the following examples, the ternary combinations containing histidine provide good effects in terms of improving the rate of fatty acid β-oxidation by the enzyme carnitine palmitoyltransferase I. These ternary compositions containing at least histidine are in fact a cocktail of compounds that ultimately allow the elevation of histidine plasma levels in subjects.Serine and cysteine ​​are metabolized by several pathways to give histidine, and carnosine is the source of histidine, being a dipeptide composed of the amino acids β-alanine and histidine.NAC is the precursor of L-cysteine, and is converted to L-cysteine ​​after ingestion.

[0070] In another particular embodiment of the pharmaceutical or nutraceutical composition according to the first aspect, the pharmaceutical or nutraceutical composition comprises a therapeutically or nutraceutical effective amount of: (a) histidine, more specifically L-histidine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; cysteine, more specifically L-cysteine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them, and serine, more specifically L-serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; or (b) histidine, more specifically L-histidine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; cysteine, more specifically L-cysteine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them, and carnosine, more specifically L-carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; or (c) cysteine, more specifically L-cysteine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; serine, more specifically L-serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them, and carnosine, more specifically L-carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; or (d) histidine, more specifically L-histidine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; serine, more specifically L-serine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them, and carnosine, more specifically L-carnosine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them.

[0071] In another particular embodiment of the pharmaceutical or nutraceutical composition according to the first aspect, the composition comprises a therapeutically or nutraceutical effective amount of: (a) cysteine, more specifically L-cysteine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; histidine, more specifically L-histidine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them, and serine, more specifically L-serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; or (b) cysteine, more specifically L-cysteine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; histidine, more specifically L-histidine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them, and carnosine, more specifically L-carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; or (c) cysteine, more specifically L-cysteine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them; serine, more specifically L-serine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them, and carnosine, more specifically L-carnosine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of any of them.

[0072] In another particular embodiment of the pharmaceutical or nutraceutical composition according to the first aspect, the composition comprises a therapeutically or nutraceutical effective amount of: (a) NAC or a pharma- ceutically or nutraceutically acceptable salt thereof or a solvate of any of them; histidine, more particularly L-histidine, or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of any of them, and serine, more particularly L-serine, or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of any of them; or (b) NAC or a pharma- ceutically or nutraceutically acceptable salt thereof or a solvate of any of them; histidine, more particularly L-histidine, or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of any of them, and carnosine, more particularly L-carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of any of them; or (c) NAC or a pharma- ceutically or nutraceutically acceptable salt thereof or a solvate of any of them; serine, more specifically L-serine, or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of any of them, and carnosine, more specifically L-carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of any of them.

[0073] In a more particular embodiment of the first aspect of the invention, the composition comprises a therapeutically or nutraceutical effective amount of histidine, or a pharma- ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, a therapeutically or nutraceutically effective amount of cysteine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, a therapeutically or nutraceutically effective amount of serine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of carnosine, or a pharma-ceutically or nutraceutically acceptable salt thereof, or a solvate of either thereof, together with one or more pharma-ceutically or nutraceutically acceptable excipients or carriers.

[0074] More specifically, the compositions comprise a therapeutically or nutraceutical effective amount of L-histidine, or a pharma- ceutically acceptable salt thereof, or a solvate of either thereof, a therapeutically or nutraceutical effective amount of L-cysteine, or a pharma- ceutically acceptable salt thereof, or a solvate of either thereof, a therapeutically or nutraceutical effective amount of L-serine, or a pharma- ceutically acceptable salt thereof, or a solvate of either thereof, and a therapeutically or nutraceutical effective amount of L-carnosine, or a pharma- ceutically acceptable salt thereof, or a solvate of either thereof, together with one or more pharma- ceutically acceptable excipients or carriers.

[0075] In another more specific embodiment, the composition comprises a therapeutically or nutraceutical effective amount of L-histidine, or a pharma- ceutically acceptable salt thereof, or a solvate of either thereof, a therapeutically or nutraceutical effective amount of NAC, or a pharma-ceutically acceptable salt thereof, or a solvate of either thereof, a therapeutically or nutraceutical effective amount of L-serine, or a pharma-ceutically acceptable salt thereof, or a solvate of either thereof, and a therapeutically or nutraceutical effective amount of L-carnosine, or a pharma-ceutically acceptable salt thereof, or a solvate of either thereof, together with one or more pharma-ceutically acceptable excipients or carriers.

[0076] In an even more particular embodiment, the composition comprises: -A therapeutically or nutraceutical effective amount of L-histidine monohydrochloride monohydrate; - A therapeutically or nutraceutical effective amount of L-cysteine ​​hydrochloride; a therapeutically or nutraceutical effective amount of L-serine; and - A therapeutically or nutraceutical effective amount of L-carnosine together with one or more pharma- ceutically or nutraceutical acceptable excipients or carriers.

[0077] In another even more particular embodiment, the composition comprises: -A therapeutically or nutraceutical effective amount of L-histidine monohydrochloride monohydrate; - A therapeutically or nutraceutical effective amount of NAC; a therapeutically or nutraceutical effective amount of L-serine; and - A therapeutically or nutraceutical effective amount of L-carnosine together with one or more pharma- ceutically or nutraceutical acceptable excipients or carriers.

[0078] Specific therapeutic or nutraceutical amounts are disclosed in the previous paragraph.

[0079] In certain embodiments, a composition for human consumption comprises: - a therapeutically or nutraceutical effective amount of L-histidine monohydrochloride monohydrate providing a dose of 17 mg / Kg in humans; - a therapeutically or nutraceutical effective amount of L-cysteine ​​hydrochloride to provide a dose of 39.7 mg / Kg in humans; a therapeutically or nutraceutical effective amount of L-serine to provide a dose of 17 mg / Kg in humans; and - A therapeutically or nutraceutical effective amount of L-carnosine to provide a dose of 17mg / Kg in humans together with one or more pharma- ceutically acceptable excipients or carriers.

[0080] In another particular embodiment, the composition for human consumption comprises: - a therapeutically or nutraceutical effective amount of L-histidine monohydrochloride monohydrate providing a dose of 17 mg / Kg in humans; - a therapeutically or nutraceutical effective amount of NAC providing a dose of 39.7mg / Kg in humans; a therapeutically or nutraceutical effective amount of L-serine to provide a dose of 17 mg / Kg in humans; and - A therapeutically or nutraceutical effective amount of L-carnosine to provide a dose of 17mg / Kg in humans together with one or more pharma- ceutically acceptable excipients or carriers.

[0081] In another particular embodiment, the composition for human consumption comprises: - a therapeutically or nutraceutical effective amount of L-histidine monohydrochloride monohydrate to provide a dose of 7.5-20 mg / Kg in humans; - a therapeutically or nutraceutical effective amount of L-cysteine ​​hydrochloride to provide a dose of 2-5 mg / Kg in humans; a therapeutically or nutraceutical effective amount of L-serine to provide a dose of 1.3 to 3.3 mg / Kg in humans; and - A therapeutically or nutraceutical effective amount of L-carnosine to provide a dose of 4.5-10 mg / Kg in humans together with one or more pharma- ceutically acceptable excipients or carriers.

[0082] In another particular embodiment, the composition for human consumption comprises: - a therapeutically or nutraceutical effective amount of L-histidine monohydrochloride monohydrate to provide a dose of 7.5-20 mg / Kg in humans; - a therapeutically or nutraceutical effective amount of NAC to give a dose of 2-5mg / Kg in humans; a therapeutically or nutraceutical effective amount of L-serine to provide a dose of 1.3 to 3.3 mg / Kg in humans; and - A therapeutically or nutraceutical effective amount of L-carnosine to provide a dose of 4.5-10 mg / Kg in humans together with one or more pharma- ceutically acceptable excipients or carriers.

[0083] In another particular embodiment of the first aspect, the composition comprises a therapeutically or nutraceutical effective amount of a peptide, the peptide comprising a sequence of amino acids that includes at least three of histidine, cysteine, serine, and carnosine, in either the L or D configuration and combinations thereof (i.e., one compound is in the D isomeric form and another compound is in the L isomeric form).

[0084] In another particular embodiment of the first aspect, the composition comprises a therapeutically or nutraceutical effective amount of a peptide, the peptide comprising a sequence of amino acids comprising cysteine ​​and at least two of histidine, serine and carnosine in either the L- or D-configuration and combinations thereof (i.e., one compound is in the D-isomer form and another compound is in the L-isomer form). In another particular embodiment of the first aspect, the composition comprises a therapeutically or nutraceutical effective amount of a peptide, the peptide comprising a sequence of amino acids comprising cysteine, histidine, serine and carnosine in either the L- or D-configuration and combinations thereof (i.e., one compound is in the D-isomer form and another compound is in the L-isomer form).

[0085] In another particular embodiment of the first aspect, the composition comprises a therapeutically or nutraceutical effective amount of a peptide, the peptide comprising a sequence of amino acids including NAC and at least two of histidine, serine and carnosine, at least two of histidine, serine and / or carnosine being either in the L or D configuration and combinations thereof (i.e., one compound is in the D isomeric form and another compound is in the L isomeric form). In another particular embodiment of the first aspect, the composition comprises a therapeutically or nutraceutical effective amount of a peptide, the peptide comprising a sequence of amino acids including NAC, histidine, serine and carnosine, at least two of histidine, serine and / or carnosine being either in the L or D configuration and combinations thereof (i.e., one compound is in the D isomeric form and another compound is in the L isomeric form).

[0086] These peptides having a sequence that includes at least three of the indicated compounds, in another particular embodiment, have a length selected from 3 to 50 amino acids, more particularly 3 to 10 amino acids, and even more particularly 3, 4, 5, 6, 7, 8, 9, and 10 amino acids.

[0087] In a more particular embodiment, the peptide is selected from a tripeptide, a tetrapeptide and a pentapeptide.

[0088] In a preferred embodiment, the peptide in the composition comprises at least one histidine residue.In another preferred embodiment, the peptide in the composition comprises at least one cysteine ​​or NAC residue.In another preferred embodiment, the peptide in the composition comprises at least one histidine residue and at least one cysteine ​​residue, more preferably at least one histidine residue and at least one NAC residue.

[0089] In another particular embodiment, the tetrapeptide comprises at least three selected from histidine, cysteine, serine and carnosine, and any additional amino acids known to those skilled in the art, including all naturally occurring amino acids, in particular any amino acid selected from the 21 proteinogenic α-amino acids found in eukaryotic organisms.

[0090] In another particular embodiment, the tetrapeptide comprises cysteine ​​and at least two selected from histidine, serine and carnosine, which are any additional amino acids known to those skilled in the art, including any amino acid selected from all naturally occurring amino acids, particularly the 21 proteinogenic α-amino acids found in eukaryotes.In another particular embodiment, the tetrapeptide comprises NAC and at least two selected from histidine, serine and carnosine, which are any additional amino acids known to those skilled in the art, including any amino acid selected from all naturally occurring amino acids, particularly the 21 proteinogenic α-amino acids found in eukaryotes.

[0091] Amino acids within the meaning of the present invention include alanine (A), arginine (R), asparagine (N), aspartic acid (aspartate, D), cysteine ​​(C), glutamine (Q), glutamic acid (E), glycine (G), histidine (H), isoleucine (I), leucine (L), lysine (K), methionine (M), phenylalanine (F), proline (P), pyrrolysine (O), serine (S), selenocysteine ​​(U), threonine (T), tryptophan (W), tyrosine (Y) and valine (V). The modified amino acids were 2-aminoadipic acid (Aad), 3-aminoadipic acid (bAad), β-alanine (bAla), 2-aminobutyric acid (Abu), 6-aminocaproic acid (Acp), 2-aminoheptanoic acid (Ahe), 2-aminoisobutyric acid (Aib), 2-aminopimelic acid (Apm), 2,4-diaminobutyric acid (Dbu), desmosine, 2,2'-diaminopimelic acid (Dpm), 2,3-diaminopropionic acid (Dpr), N-ethylglycine (EtGly), and 1,2'-diaminopimelic acid (Dpm). ), N-ethylasparagine (EtAsn), hydroxylysine (Hyl), allo-hydroxylysine (aHyl), 3-hydroxyproline (3Hyp), 4-hydroxyproline (4Hyp), isodesmosine (Ide), allo-isoleucine (alle), N-methylisoleucine (MeIle), 6-N-methyllysine (MeLys), N-methylvaline (Meval), norvaline (Nva), norleucine (Nle) and ornithine (Orn). In some embodiments, one amino acid may be replaced with another while maintaining the hydrophobicity, aromaticity and basicity at each selected position.

[0092] In another particular embodiment, the peptide comprises histidine, cysteine, serine and carnosine and is at least a pentapeptide, preferably a pentapeptide.In another particular embodiment, the peptide comprises histidine, NAC, serine and carnosine and is at least a pentapeptide, preferably a pentapeptide.

[0093] In all these possible peptides of the composition, the amino acid residues may be in any order in the sequence read from the N-terminus to the C-terminus.

[0094] When the composition is administered, the peptides contained therein are naturally hydrolyzed by common cellular and body processes (ie, hydrolases, proteases, etc.) and single units are delivered.

[0095] Dosage adjustments for these peptides may vary, but in certain embodiments are adjusted to provide the indicated doses above for the single compounds disclosed above.

[0096] The preparation of all these peptides follows common practices known to those skilled in the art.

[0097] All pharmaceutical or nutraceutical compositions of the present invention will contain common excipients and / or carriers.

[0098] Examples of suitable pharma- ceutically or nutraceutical acceptable excipients are solvents, dispersion media, diluents, or other liquid vehicles, dispersing or suspending aids, surfactants, isotonicity agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, etc. Except insofar as any conventional excipient medium is incompatible with the substance or its derivatives, such as by producing some undesirable biological effect, or otherwise interacting in a deleterious manner with any one or more other components of the pharmaceutical composition, its use is contemplated within the scope of the present invention.

[0099] The relative amounts of the active ingredient, pharma- ceutical or nutraceutical acceptable excipient, and / or any additional ingredients in a pharmaceutical or nutraceutical composition of the present invention will vary depending on the identity, size, and / or condition of the subject being treated, as well as on the route by which the composition is administered.

[0100] Pharmaceutically or nutraceutical acceptable excipients used in the manufacture of pharmaceutical compositions include, but are not limited to, inert diluents, dispersing and / or granulating agents, surfactants and / or emulsifying agents, disintegrating agents, binders, preservatives, buffers, lubricants, and / or oils. Excipients such as colorants, coating agents, sweeteners, and flavoring agents can be present in the composition according to the judgment of the formulator.

[0101] The pharmaceutical or nutraceutical composition containing at least three combinations of histidine, carnosine, serine and cysteine ​​can be, for example, solid or liquid, and can be administered by any suitable route, for example, oral, parenteral, rectal, topical, intranasal, ocular, intraperitoneal or sublingual, and therefore includes the pharmaceutically or nutraceutical acceptable excipients required for formulation of desired dosage form.More specifically, the pharmaceutical or nutraceutical composition of the first aspect of the present invention can be, for example, solid or liquid, and can be administered by any suitable route, for example, oral, parenteral, rectal, topical, intranasal, ocular, intraperitoneal or sublingual, and therefore includes the pharmaceutically or nutraceutical acceptable excipients required for formulation of desired dosage form.

[0102] Exemplary diluents include, but are not limited to, calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate, lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and combinations thereof.

[0103] Exemplary granulating and / or dispersing agents include, but are not limited to, potato starch, corn starch, tapioca starch, sodium starch glycolate, clay, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose and wood products, natural sponge, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked polyvinylpyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethylcellulose, cross-linked sodium carboxymethylcellulose (croscarmellose), methylcellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethylcellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and combinations thereof.

[0104] Exemplary binders include, but are not limited to, starches (e.g., corn starch and starch paste); gelatin; sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol); natural and synthetic gums (e.g., acacia, sodium alginate, extract of Irish moss, panwar gum, ghatti gum, mucilage of isapol husk, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, microcrystalline cellulose, cellulose acetate, polyvinylpyrrolidone), magnesium aluminum silicate (Veegum), and larch arabinogalactan); alginates; polyethylene oxide; polyethylene glycol; inorganic calcium salts; silicic acid; polymethacrylates; waxes; water; alcohol; and combinations thereof.

[0105] Exemplary preservatives may include antioxidants, chelating agents, antibacterial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives, and other preservatives.Exemplary antioxidants include, but are not limited to, alpha tocopherol, ascorbic acid, ascorbyl palmitate, ascorbyl stearate, ascorbyl oleate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite.Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA), citric acid monohydrate, disodium edetate, dipotassium edetate, edetic acid, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid, and trisodium edetate.

[0106] Exemplary buffering agents include, but are not limited to, citrate buffer, acetate buffer, phosphate buffer, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixture, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixture, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic sodium phosphate, sodium phosphate mixture, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, and combinations thereof.

[0107] Exemplary lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behenate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and combinations thereof.

[0108] The preparation of all these pharmaceutical or nutraceutical compositions follows general practices known to those skilled in the art.

[0109] As mentioned above, another aspect of the present invention is a food product in solid or liquid form that comprises the pharmaceutical or nutraceutical composition defined in the first aspect.In a more particular embodiment, the food product is a beverage.These food products or nutraceutical products are in fact functional foods or supplements that provide health benefits even in the absence of pathology.Therefore, they are considered to be compositions that can prevent or reduce the risk of suffering from diseases, such as diseases related to fat deposition in tissues.

[0110] When the food or composition as disclosed in the first or second aspect and their embodiments is administered to mice with NAFLD, the animals are effectively treated and fat accumulated in the liver is reduced. Similarly, body weight is reduced due to fat mass. Furthermore, in an assay for the analysis of risk of atherosclerosis, it was observed that the risk is reduced when animals are fed the composition of the present invention. Furthermore, when the composition of the present invention is administered to ovariectomized rats that show metabolic changes associated with menopause, the values ​​of the corresponding metabolic parameters approach those of control rats. Administration of the composition of the present invention to NAFLD-induced mice reduces the expression of bacterial histidine utilization genes found to be increased in patients with NAFLD, thus alleviating the dysregulation or dysbiosis of the microbiota.

[0111] Thus, in a third aspect, this pharmaceutical or nutraceutical composition, or food, is proposed for therapeutic use.

[0112] In a particular embodiment of the third aspect of the invention, the composition of the first aspect or the food product of the second aspect is for use in the prevention and / or treatment of a disease selected from the group consisting of fatty liver disease, obesity, overweight, atherosclerosis, dysbiosis and combinations thereof.

[0113] This can also be formulated as the use of the composition or food of the present invention for the manufacture of a medicament for the prevention and / or treatment of a disorder or disease selected from the group consisting of fatty liver disease, obesity, overweight, atherosclerosis, dysbiosis and combinations thereof. The present invention also relates to a method for the prevention and / or treatment of a disorder or disease selected from the group consisting of fatty liver disease, obesity, overweight, atherosclerosis, dysbiosis and combinations thereof, comprising administering to a subject in need thereof, including a human, a therapeutically or nutraceutical effective amount of the composition or food, together with a pharma- ceutical or nutraceutical acceptable excipient and / or carrier.

[0114] The term "obesity" as used herein refers to abnormal or excessive fat accumulation in a subject, particularly a mammal, more particularly a human, that presents a risk to health. The BMI associated with obesity in humans, particularly adults, is 30 or higher. Obesity is often subdivided into three categories. Class 1 Obesity: BMI 30 to 35 Class 2 Obesity: BMI 35 to under 40 Class 3 Obesity: BMI above 40. Class 3 obesity is sometimes classified as "severe" obesity.

[0115] Child and teen BMI categories are often based on gender- and age-specific BMI percentiles, while adult BMI categories are based on BMI alone. Child and teen obesity is subdivided into two categories: Class 2 obesity: BMI ≥ 120%~< 140% of the 95th percentile or BMI ≥ 35~< 40kg / m2 Class 3 obesity: BMI ≥ 140% of the 95th percentile or BMI ≥ 40kg / m2

[0116] The term "body mass index" or "BMI" as used herein refers to a subject's weight in kilograms (or pounds) divided by the square of their height in meters (or feet).

[0117] In a particular embodiment of the third aspect of the present invention, the composition or food is for use in the prevention and / or treatment of obesity in adults. In another particular embodiment of the third aspect of the present invention, the composition or food is for use in the prevention and / or treatment of obesity in non-adults. As known to those skilled in the art, a non-adult is a human under 18 years of age, and an adult is a human over 18 years of age.

[0118] In a particular embodiment of the third aspect of the invention, the composition or food product is for use in the prevention and / or treatment of class 1 obesity, class 2 obesity and / or class 3 obesity, especially in adults. In a particular embodiment, it is for use in the prevention and / or treatment of class 1 obesity, especially in adults. In another embodiment, it is for use in the prevention and / or treatment of class 2 obesity, especially in adults. In another embodiment, it is for use in the prevention and / or treatment of class 3 obesity, especially in adults. In another embodiment, it is for use in the prevention and / or treatment of class 1 or class 2 obesity, especially in adults. In another embodiment, it is for use in the prevention and / or treatment of class 2 or class 3 obesity, especially in adults.

[0119] In a particular embodiment of the third aspect of the invention, the composition or food product is for use in the prevention and / or treatment of class 2 and / or class 3 obesity, particularly in non-adults. In a particular embodiment, the composition or food product is for use in the prevention and / or treatment of class 2 obesity, particularly in non-adults. In another embodiment, the composition or food product is for use in the prevention and / or treatment of class 3 obesity, particularly in non-adults. In a particular embodiment of the third aspect of the invention, the composition or food product is for use in the prevention and / or treatment of class 2 and class 3 obesity, particularly in non-adults. In a particular embodiment of the third aspect of the invention, the composition or food product is for use in the prevention and / or treatment of class 2 or class 3 obesity, particularly in non-adults.

[0120] The term "overweight" as used herein refers to abnormal or excessive adiposity in a subject, particularly a mammal, more particularly a human, who presents a risk to health associated with a BMI as defined above, in humans, particularly below the 25th to 30th percentile for adults, and the 85th to 95th percentile for children and teenagers.

[0121] In a particular embodiment of the third aspect of the invention, the composition or food product is for use in the prevention and / or treatment of overweight, particularly in adults. In another particular embodiment of the third aspect of the invention, the composition or food product is for use in the prevention and / or treatment of overweight in non-adults.

[0122] The term "dysbiosis" refers to any condition in which the normal diversity or function of the body's microbiota, particularly the intestinal microbiota, is disrupted.

[0123] In a more particular embodiment, the composition of the first aspect or the food product of the second aspect is for use in the prevention and / or treatment of fatty liver disease selected from non-alcoholic fatty liver disease and alcoholic fatty liver disease.

[0124] In a more particular embodiment, it is for use in the prevention and / or treatment of non-alcoholic fatty liver disease selected from simple steatosis and non-alcoholic fatty liver (NAFL), including non-alcoholic steatohepatitis (NASH).

[0125] In another particular embodiment of the third aspect of the invention, the composition or food product is for use in the prevention and / or treatment of menopause-related symptoms, particularly metabolic changes associated with menopause, in a female mammal, particularly a woman.

[0126] This may also be formulated as the use of the composition or food of the present invention for the manufacture of a medicament for the prevention and / or treatment of menopause-related symptoms, particularly metabolic changes associated with menopause in female mammals, especially women. The present invention also relates to a method for the prevention and / or treatment of menopause-related symptoms, particularly metabolic changes associated with menopause, comprising administering a therapeutically or nutraceutical effective amount of the composition or food, together with a pharma- ceutical or nutraceutical acceptable excipient and / or carrier, to a female mammal, including a woman, in need thereof.

[0127] In another particular embodiment of the third aspect of the invention, the composition or food product is for use in a female mammal, particularly a woman, for the prevention and / or treatment of menopause-related symptoms, particularly metabolic changes associated with menopause, and for the prevention and / or treatment of a disease selected from the group consisting of fatty liver disease, obesity, overweight, atherosclerosis, dysbiosis, and combinations thereof.

[0128] This can also be formulated as the use of the composition or food of the present invention for the manufacture of a medicament for the prevention and / or treatment of menopause-related symptoms, particularly metabolic changes associated with menopause, and for the prevention and / or treatment of diseases selected from the group consisting of fatty liver disease, obesity, overweight, atherosclerosis, dysbiosis, and combinations thereof, in female mammals, particularly women. The present invention also relates to a method for the prevention and / or treatment of menopause-related symptoms, particularly metabolic changes associated with menopause, and for the prevention and / or treatment of diseases selected from the group consisting of fatty liver disease, obesity, overweight, atherosclerosis, dysbiosis, and combinations thereof, comprising administering a therapeutically or nutraceutical effective amount of the composition or food, together with a pharma- ceutical or nutraceutical acceptable excipient and / or carrier, to a female mammal in need thereof, including a woman.

[0129] The term "menopause-related symptoms" as used herein refers to metabolic, body weight, cardiovascular and musculoskeletal changes, urogenital and skin atrophy, sexual dysfunction, and central nervous system-related disorders that generally occur as a result of menopause in female mammals, particularly women. The term "menopause-related metabolic changes" as used herein refers to changes in parameters related to metabolism that generally occur as a result of menopause in female mammals, particularly women. Non-limiting examples of parameters include body weight, obesity, and insulin resistance. More specifically, parameters include body weight, fat mass, lean mass, fat stores, adiposity index, and insulin resistance.

[0130] In certain embodiments, the metabolic changes associated with menopause are selected from the list consisting of weight gain, increased adiposity, and increased insulin resistance in a female mammal, particularly a woman, during the menopausal transition, during menopause or after menopause, as compared to premenopause, particularly before the onset of the menopausal transition.

[0131] In another particular embodiment, the metabolic changes associated with menopause are selected from the list consisting of weight gain, fat mass gain, lean mass loss, increased adiposity, increased body mass index, increased insulin resistance in a female mammal, particularly a woman, during the menopausal transition, menopause or after menopause, as compared to premenopause, particularly before the onset of the menopausal transition.

[0132] In certain embodiments, "pre-menopause" refers to one, two, three, four, preferably one year before the onset of menopause. In another particular embodiment, "before the onset of the menopausal transition" refers to one, two, three, four, preferably one year before the onset of the menopausal transition.

[0133] The term "menopause" as used herein refers to a condition in female mammals, preferably women, characterized by the ovaries no longer ovulating or producing significant amounts of sex hormones. Clinically, natural menopause is defined as one year of amenorrhea following the last menstrual period (FMP) without pathological or physiological causes.

[0134] The term "menopausal transition" or "perimenopause" refers to a defined period beginning with the onset of irregular menstrual cycles until the last menstrual period and characterized by fluctuations in reproductive hormones.

[0135] As used herein, the term "postmenopausal" refers to the period after menopause has occurred, ie, beginning 12 months after the last menstrual period (FMP).

[0136] In another particular aspect, the present invention relates to a non-therapeutic method for reducing weight in an overweight subject, comprising administering to the subject a composition according to the first aspect of the present invention or a food according to the second aspect of the present invention, the subject being particularly a mammal, and even more particularly a human. In a particular embodiment, the dosage of administration is the dosage disclosed in the first aspect of the present invention for the composition of the present invention. In another particular embodiment, the route of administration is the one disclosed in the first aspect of the present invention for the composition of the present invention. In a particular embodiment, the human is an adult. In another particular embodiment, the human is a non-adult.

[0137] In certain embodiments, reducing overweight in an overweight subject comprises reducing the subject's BMI by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, particularly at least 5%. In another particular embodiment, reducing overweight in an overweight subject comprises reducing the subject's BMI by at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7 points, particularly at least 1 point.

[0138] As shown in the Examples, a diet rich in fats and sugars when supplemented with a specific combination of the four histidines, serine, cysteine ​​and carnosine, allowed for the reduction of fat accumulation in the liver and other body tissues.

[0139] Thus, also disclosed herein for the first time is a combination comprising a therapeutically or nutraceutical effective amount of histidine or its pharma- ceutically or nutraceutical acceptable salt or any of its solvates, a therapeutically or nutraceutical effective amount of cysteine ​​or its pharma- ceutically or nutraceutical acceptable salt or any of its solvates, a therapeutically or nutraceutical effective amount of serine or its pharma- ceutically or nutraceutical acceptable salt or any of its solvates, and a therapeutically or nutraceutical effective amount of carnosine or its pharma- ceutically or nutraceutical acceptable salt or any of its solvates, together with one or more pharma- ceutical or nutraceutical acceptable excipients or carriers.This combination has not been disclosed before and is proposed as a dietary supplement for subjects at risk of suffering from one or more of fatty liver disease, obesity and atherosclerosis.Any of the above-mentioned embodiments shown for the first and second aspects, in relation to the amount (dosage) of any of the compounds and in relation to the isomers of the compounds, also apply to the indicated combination of the four components.

[0140] Any of the above embodiments indicated for the first and second aspects with respect to any amount (dosage) of the compound, as well as with respect to the isomers of the compound, also apply to the indicated combinations of the four components in which cysteine ​​is replaced by NAC.

[0141] Throughout the description and claims, the word "comprises" and variations of this word are not intended to exclude other technical features, additives, ingredients or steps. Furthermore, the word "comprises" encompasses the case of "consisting of". Additional objects, advantages and features of the present invention will become apparent to those skilled in the art upon consideration of the description or may be learned by practice of the present invention. The following examples are provided by way of illustration and are not intended to limit the present invention. Furthermore, the present invention encompasses all possible combinations of the specific and preferred embodiments described herein. EXAMPLES

[0142] 1. Assays in human cohorts. 1.1 Circulating histidine levels in different cohorts according to the degree of steatosis. To obtain detailed information on fatty liver in patients with NAFLD, metabolic profiling of urine and plasma by 1H-NMR spectroscopy was performed. Metabolome-wide association studies (MWAS) yielded 124 metabolite signals in urine and 80 metabolite signals in plasma that correlated with hepatic steatosis and related clinical traits. Among several hepatic steatosis metabolites strongly correlated with NAFLD grade, a notable negative correlation with histidine in plasma and a significant decrease in urine was observed. Plasma histidine was negatively correlated with fatty liver for 102 and 56 patients in the discovery study, respectively, which was also confirmed in two validation cohort studies with 313 and 283 patients. This cohort study of 283 patients also showed a decrease in histidine levels in obese patients without steatosis compared to lean patients without steatosis. Circulating histidine levels decreased as the degree of steatosis increased, confirming the negative correlation between histidine levels and fatty liver.

[0143] Example 1. A composition comprising histidine, serine, cysteine ​​and carnosine as supplements for use in ameliorating the clinical symptoms of hepatic steatosis. material and method Animal models and diets Forty-eight C57BL / 6J male mice (Envigo, Sant Feliu de Codines, Barcelona, ​​Spain), 6 weeks old at the start of the experiment, were used. Animals were housed in groups (4 mice per cage) under controlled conditions of temperature (22 ± 2 °C) and humidity (55 ± 10%) with food and water ad libitum and a 12-h light / dark cycle. Mice were left to acclimate to the animal facility for 1 week. After the acclimatization period, animals were randomly divided into two experimental groups with different diets. Sixteen control mice were kept on a standard diet (D12328, Research Diets, New Brunswick, NJ), and 32 animals (NAFLD group) were fed a HFHFr diet (HFHC:D12331, research diet) supplemented with 23.1 g / L fructose and 18.9 g / L sucrose in drinking water. Mice were maintained on these diets for 20 weeks under ad libitum conditions. These specific doses were determined based on previous studies and on calculations of dose conversion from human to animals. All experimental protocols were approved by the Animal Ethics Committee of the Technological Unit of Nutrition and Health of Eurecat (Reus, Spain) and the Generalitat de Catalunya approved all procedures (10281). The experimental protocols followed the “Principles of Laboratory Care” guidelines and were carried out in accordance with the EC Council Directive (86 / 609 / EEC).

[0144] During the last 4 weeks of the experiment (from week 16 to week 20), NAFLD mice were randomly divided into two groups: 16 mice were kept under the same feeding conditions as described above (HFHFr group) and 16 mice were subjected to histidine-related amino acid treatment (AA group). AA is a mixture of the following compounds: 210 mg / kg L-histidine monohydrochloride monohydrate (Merck, GmbH Germany), 490 mg / kg L-cysteine ​​hydrochloride (Merck, GmbH Germany), 210 mg / kg L-serine (Merck, GmbH Germany) and 210 mg / kg L-carnosine 98% (Acros Organics, Geel, Belgium). Histidine, cysteine, serine and carnosine were diluted in drinking water (vehicle) containing 23.1 g / L fructose and 18.9 g / L sucrose. Fresh solutions were prepared fresh three times a week, prepared from stock powders and protected from light. Prior to euthanasia, 10 animals per group were randomly selected to undergo insulin challenge. They were injected intraperitoneally with 1 mU / g insulin (n=5 per group) or saline (n=5 per group) and sacrificed 15 min later.

[0145] Serum was obtained by centrifugation and stored at -80°C for further analysis. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were quantified by enzymatic colorimetric assays (QCA, Barcelona, ​​Spain). Fasting insulinemia and glycemia were measured using a mouse insulin ELISA kit (Mercodia, Uppsala, Sweden) and a glucose liquid kit (QCA, Barcelona, ​​Spain), respectively. Livers were rapidly harvested, weighed, and divided into two sections - the left hepatic medial lobe was kept in formalin and the remaining tissue was frozen in liquid nitrogen and stored at -80°C until further analysis.

[0146] Quantification of liver fat Hepatic lipids were extracted and quantified. Briefly, total lipids were extracted from 80–100 mg liver slices by adding 1 mL of hexane / isopropanol (3:2, v / v) and degassing with nitrogen gas. They were then left overnight under orbital stirring, protected from light at room temperature. After extraction with 0.3 mL of Na2SO4 (0.47 M), the organic layer was separated and dried with nitrogen gas. Total lipids were quantified gravimetrically before emulsification. Triglycerides and total cholesterol were measured using a commercially available enzymatic kit (QCA).

[0147] Histological evaluation Liver sections fixed in buffered formalin (4% formaldehyde, 4 g / L NaH2PO4, 6.5 g / L Na2HPO4; pH 6.8) were cut to a thickness of 3.5 μm and stained with hematoxylin & eosin (H&E) and trichrome stain. Liver images (magnification 40x) were taken with a microscope (ECLIPSE Ti; Nikon, Tokyo, Japan) connected to a digital site camera (DS-Ri1, Nikon) and analyzed using ImageJ NDPI software (National Institutes of Health, Bethesda, MD, USA; https: / / imagej.nih.gov / ij, version 1.52a). To avoid any bias in the analysis, this study had a double-blind design, preventing reviewers from knowing any data from the mice during histopathological analysis. A common NAFLD scoring system was established to diagnose mice with NAFLD / NASH. The key features of NAFLD and NASH were classified as follows: steatosis was assessed by analyzing macrodroplet (0–3) and microdroplet steatosis (0–3) separately, followed by hepatocyte hypertrophy (0–3) assessing abnormal cellular enlargement, finally resulting in a total score of 9 points for the steatosis status. Inflammation was scored by counting cell aggregates (inflammatory foci). The score 0–3 depends on the grade of the feature. This was classified as 0 (<5%), 1 (5–33%), 2 (34–66%) and 3 (>66%), and this scoring was used for each feature of steatosis, then a total steatosis score was calculated. 23Because only quantitative measurements were considered for the NAFLD score in rodents, ballooning was not included in the scoring system. It is important to emphasize that hypertrophy is not a sign of cell damage, but rather refers to the abnormal enlargement of cells without acknowledging the cause of this enlargement.

[0148] RNA extraction and quantitative polymerase chain reaction Homogenates from eight livers per group were used for total RNA extraction using TriPure reagent (Roche Diagnostic, Sant Cugat del Valles, Barcelona, ​​Spain) according to the manufacturer's instructions. RNA concentration and purity were determined using a nanophotometer (Implen GmbH, Munchen, Germany). RNA was converted to cDNA using the High-Capacity RNA-to-cDNA Kit (Applied Biosystems, Wilmington, DE, USA). cDNA was diluted 1:10 and then incubated with the commercial LightCycler 480 Sybr Green I Master in a Lightcycler® 480 II (Roche Diagnostics GmbH, Manheim, Germany). Table 1 shows the list of primers used, previously described in other studies and validated with Primer-Blast software (National Center for Biotechnology Information, Bethesda, MD, USA). As previously described, 36b4 was used as a housekeeping gene.

[0149] [Table 1]

[0150] Protein extraction and Western blot analysis Approximately 20 mg of liver was homogenized in 300 μL of lysis buffer (8 mmol / L NaH2PO4, 42 mmol / L Na2HPO4, 1% SDS, 0.1 mol / L NaCl, 0.1% NP40, 1 mmol / L NaF, 10 mmol / L sodium orthovanadate, 2 mmol / L PMSF, and 1% protease inhibitor cocktail 1 (Millipore Sigma, Darmstadt, Germany)) for 50 s with a Tyssuelyser LT (Qiagen, Hilden, Germany). Protein extracts were quantified by the standardized BCA method (Bio-Rad protein assay; BioRad, Hercules, CA, USA). Protein extracts (20–25 μg) were electrophoretically separated on 10% SDS-PAGE and electroblotted onto nitrocellulose membranes (Li-cor biosciences, NE, USA). Efficient protein transfer was monitored by Ponceau S staining. Membranes were then blocked (5% BSA) at room temperature and probed with specific primary antibodies (diluted 1:1000) in 1% BSA:total Akt (4685) (CST, Danvers, MA, USA), phospho-Akt (Ser473) (4060) (CST) and β-actin (Santa Cruz Biotechnology, Inc; Dallas, TX, USA) overnight at 4°C. Infrared fluorescent secondary antibodies anti-rabbit 680, anti-rabbit 800 and anti-mouse 680 (LI-COR Biosciences, Lincoln, NE, USA; 926-32211, 926-68071 and 926-68070, respectively) were then used for detection and quantification using ImageJ.

[0151] statistical analysis Statistical analysis was performed using GraphPad Prism 9 software (Graph-Pad Software, La Jolla, CA, USA). Data are presented as mean ± SEM. Data distribution was analyzed by Shapiro-Wilk normality test. Differences between two groups were determined using unpaired t-test (two-tailed, 95% confidence interval). One-way analysis of variance (ANOVA) was performed to examine differences between three groups. A p-value of less than 0.05 was considered statistically significant. Statistical analysis was performed using GraphPad Prism 9 software (Graph-Pad Software, La Jolla, CA, USA). Data are presented as mean ± SEM. Data distribution was analyzed by Shapiro-Wilk normality test. Differences between two groups were determined using unpaired t-test (two-tailed, 95% confidence interval). One-way analysis of variance (ANOVA) was performed to examine differences between three groups. A p-value of less than 0.05 was considered statistically significant.

[0152] result Specific amino acid combinations reduced liver damage and liver features of NAFLD After 4 weeks of specific amino acid treatment (AA) consisting of histidine, serine, cysteine ​​and carnosine, animals were sacrificed and liver weights were analyzed. The AA supplemented group showed a tendency to reduce liver weight compared to HFHFr animals, while HFHFr animals showed a significant increase in liver weight compared to the control group (Figure 1(A)). These differences observed between the AA and HFHFr groups in liver weight did not correlate with differences in solid or liquid intake (data not shown). Furthermore, macroscopic observations showed whitish fatty liver in HFHFr animals compared to the control group, whereas AA treatment partially restored the appearance of a healthy liver compared to the HFHFr group (Figure 1(B)). The AA group showed significantly reduced AST levels compared to HFHFr animals. However, ALT showed no significant change in the AA group with respect to the HFHFr group counterparts, while the HFHFr group counterparts showed a significant increase in both transaminases compared to the control group (Figure 1(C) and (D)).

[0153] AA supplementation reduced hepatic lipid content and hepatic steatosis After evaluation of the liver status, this study analyzed liver contents and characteristic features of fatty liver. A significant increase in total lipid content, total cholesterol and triglyceride hepatic levels was observed in the HFHFr group compared to the control group. Interestingly, AA-supplemented animals showed a significant decrease in hepatic triglyceride levels compared to the HFHFr group (Figure 2(B)), but this effect was not reflected in hepatic total cholesterol levels (Figure 2(C)). However, total liver lipid levels showed a slight decrease in the AA group compared to the HFHFr group (Figure 2(A)).

[0154] In agreement with the liver status assessment and biochemical analysis, liver histology analysis showed that the HFHFr group developed prominent hepatic steatosis with the presence of macrodroplet and microdroplet steatosis accompanied by increased hypertrophy, which caused nuclear displacement. Importantly, treatment with AA supplementation improved hepatic steatosis, indeed, microdroplet steatosis was particularly reduced, as was the surface occupied by macrodroplet steatosis, hypertrophy was reduced, and nuclear displacement was lowered (Figure 3(A)). As a result, a significant reduction in lipid droplet volume and lipid surface was observed (Figure 3(B) and (C)). The diagnosis of mice with NAFLD / NASH was made through the establishment of a common NAFLD scoring system, which confirmed the significant improvement of the various parameters analyzed in animals treated with AA supplementation compared to the HFHFr group (Figure 3(D)).

[0155] To evaluate whether this combination of specific amino acids modulated the levels of pro-inflammatory markers, the expression levels of several representative inflammatory genes were analyzed. In relation to the previous results of this study, F4 / 80, a marker of macrophage infiltration, was significantly downregulated in AA-supplemented animals compared to the HFHFr group, which showed a significant upregulated level of this gene compared to the control group. Similar results were observed at the TNFα mRNA level, which was significantly increased in HFHFr animals compared to the control group, whereas TNFα expression was significantly downregulated in the AA-supplemented group compared to their counterparts (data not shown).

[0156] The beneficial effects of AA supplementation did not involve lipogenesis or lipid transport pathways To identify which metabolic pathways may be involved in the improvement of NAFLD after AA supplementation, we performed an analysis of gene expression related to lipid metabolism, such as hepatic lipogenesis and lipid transport. The HFHFr group showed a significant increase in lipid transport-related genes Cd36 and Fabp4. The AA group showed no effect in these two genes related to lipid transport, maintaining high levels of expression of these genes compared to the control group. A significant change was observed in the mRNA levels of Scd1, a gene involved in de novo hepatic lipogenesis, when compared to the control and HFHFr groups. The other de novo hepatic lipogenic genes Acc1 and Fasn did not show significant differences between the control and HFHFr groups. After AA treatment, no differences were found in de novo hepatic lipogenic gene levels compared to the HFHFr group, except for Acc1, where the AA group showed a significant increase compared to the HFHFr group.

[0157] AA supplementation reduced liver inflammation associated with NAFLD To evaluate whether supplementation with AA modulated the levels of pro-inflammatory markers, the expression levels of several representative inflammatory genes were evaluated. As expected, F4 / 80 mRNA levels, a marker of macrophage infiltration, were significantly upregulated in the HFHFr group compared to control mice. In contrast, AA treatment significantly corrected this upregulation of F4 / 80 expression. Similar significant results were observed in the expression of Tnfα, where the increased levels in the HFHFr group were corrected in the AA group, and an interesting statistical trend was found in Il1a gene expression, where there was a significant increase in the HFHFr group compared to the control group, but AA failed to significantly reduce the levels of Il1a. No significant results were shown for both Il6 and IL10 levels. However, the expression of Cd11c showed a significant increase in the HFHFr and AA groups compared to the control group, but no differences were found between HFHFR and AA mice.

[0158] AA supplementation increased antioxidant defense and antitumor activity in NAFLD We performed an analysis of genes related to autophagy, immune response, antioxidant defense, antitumor activity and vesicular transport. Focusing on autophagy, no differences were found in both genes between control, HFHFr and AA mice. However, when it comes to genes related to antioxidant activity, such as CBS or CTH, significant results were found. Although no differences were found between controls and HFHFr, the expression of both genes was significantly increased in AA mice compared to the HFHFr group. In the case of antitumor activity-related genes, Mtus1 showed a significant increase in AA mice compared to the HFHFr group. Regarding genes related to immune response, no differences were found in IRF4 expression. However, in Cebpb expression, a significant increase was observed in the AA group compared to the HFHFr group. Finally, the gene Sec24b related to vesicular transport showed a significant increase in AA mice compared to HFHFr mice.

[0159] AA supplementation reduced hepatic insulin resistance associated with NAFLD Considering that impaired hepatic insulin signaling plays a key role in NAFLD development, to assess insulin resistance, animals were challenged with an intraperitoneal insulin bolus or saline to determine Akt activation (phosphorylation at serine 473). As expected, the control group showed significant Akt phosphorylation (pAktS473) in insulin-challenged mice compared to vehicle-injected mice. This fact was not observed in the HFHFr group, indicating hepatic insulin resistance in these animals. In contrast, the AA supplemented group showed a significant improvement in insulin signaling, with a significant increase in phosphorylation of Akt in insulin-challenged mice compared to vehicle-injected AA mice. The data are presented in Figure 4, where (A) shows a representative Western blot analysis for Akt activation (pAktS473), total Akt protein levels (T-Akt), housekeeping β-actin levels and protein loading by Ponceau S membrane staining. (B) shows a densitometric analysis of the ratio of phosphorylated Akt to total Akt.

[0160] Furthermore, analysis of hepatic expression of key genes after quaternary treatment for fatty acid β-oxidation (carnitine palmitoyltransferase, Cpt1a), inflammation (chemokine (C-C motif) ligand 2, CCL2), lipolysis (hormone sensitive, HSL; and lipin, PLIN) as well as adipogenesis (fatty acid synthase, FASN) showed a trend towards an increase in Cpt1a, a significant decrease in hepatic inflammation (Ccl2) and an increase in lipolysis (higher expression of the lipase Hsl and lower levels of the surrounding lipid droplet protein Plin) (Figure 4(C)). In contrast, no changes were observed in adipogenesis (Fasn). Data are represented in Figure 4 as mean ± SEM. n = 10–12 per condition. *p < 0.05.

[0161] AA supplementation increases hepatic amino acid levels in NAFLD To analyze the effect of amino acid-based treatments on the liver, an evaluation by LC / MS was performed in serum with the aim of quantifying the amino acids involved in the treatment. In the case of histidine and serine, separately, no differences were observed between the groups. Taking the levels of amino acids together, a tendency to increase the concentration of these amino acids was observed, but not significant. This may occur due to tissue bioaccumulation of amino acids. Therefore, to prove this hypothesis, an analysis of these amino acids was performed in liver homogenates of the different groups by LC / MS. The results obtained showed a significant decrease in histidine and serine levels in the HFHFr group compared to the control group. Furthermore, a strong tendency to increase histidine and serine levels was observed in the AA group compared to the HFHFr group. Alanine, an amino acid that is part of carnosine and can be produced via carnosine synthetase, showed a significant decrease in the HFHFr group compared to the control group. Furthermore, a significant increase in AA mice compared to HFHFr mice. Glycine is an amino acid linked to the metabolism of serine and GSH, and its increased levels may be due to serine administration. Glycine levels were decreased in the HFHFr group compared with the control group, and glycine levels were restored in AA mice compared with those in HFHF mice.

[0162] Example 2. AA supplementation reduces the risk of atherosclerosis. Atherosclerosis is a complex disorder that exhibits many of the characteristics of a chronic inflammatory process. Atherosclerotic cardiovascular disease remains the leading cause of morbidity and mortality worldwide, and obesity increases the risk of atherosclerosis and mortality. Elucidating the mechanisms behind the differences between obese individuals with and without atherosclerosis could reveal therapeutic targets for treating the adverse cardiovascular consequences of obesity as an alternative or adjunct to weight loss programs that are known to have limited long-term success. One of the most compelling clinical challenges of our time is the increasing prevalence of obesity and its deleterious effects on the cardiovascular system. Obesity affects the pathophysiological processes involved in inflammation and the development of atherosclerosis. Obesity and overweight are accompanied by unfavorable blood lipid profile patterns. Dyslipidemia is a major risk factor for coronary artery disease. Among obese patients, the estimated prevalence of hypertriglyceridemia is two times higher than in nonobese individuals. Furthermore, the atherogenic combination of hypertriglyceridemia with high LDL and low HDL is more prevalent in obese and overweight patients.

[0163] Serum and blood analysis Serum fasting glucose, total cholesterol, and triglycerides were analyzed by enzymatic colorimetric assays (QCA, Barcelona, ​​Spain). Serum fasting insulinemia was analyzed using a mouse insulin ELISA kit (Mercodia, Uppsala, Sweden), and serum HDL and LDL levels were analyzed using EnzyChrom™ AF HDL and LDL / VLDL Assay Kits (BioAssay System, Hayward, CA, USA).

[0164] Sixteen control mice were fed a standard diet (D12328, Research Diets, New Brunswick, NJ, USA), and 32 animals (atheromatous group) were fed a HFHFr diet (HFHC:D12331, research diet) supplemented with 23.1 g / L fructose and 18.9 g / L sucrose in drinking water. Mice were fed these diets for 20 weeks under ad libitum feeding conditions. During the last 4 weeks of the experiment (from week 16 to week 20), atheromatous mice were randomly divided into two groups: 16 mice were fed under the same feeding conditions as above, and the remaining 16 mice were treated with the combination of histidine amino acids described above in Example 1.

[0165] The assay, with n=48 mice, also determined which were circulating levels of high density lipoprotein (HDL), low density lipoprotein (LDL) and the ratio of LDL / HDL. The data are shown in Figure 5, where the group receiving the supplement (histidine AA) showed higher levels of HDL compared to mice receiving the HFHFr diet (Figure 5(A)), a significant reduction in serum LDL compared to mice fed the HFHFr diet (Figure 5(B)), and an LDL / HDL ratio within the mean values ​​as the control (Figure 5(C)). The data confirm that the supplement allows for a reduction in levels of what is known as "bad cholesterol" (LDL) and an increase in levels of what is known as "good cholesterol" (HDL) reduces the risk of atherosclerosis.

[0166] Example 3. AA supplementation reduces visceral fat and obesity parameters. Adipose tissue is remarkably heterogeneous. Subcutaneous adipose tissue (scWAT) constitutes the majority of total fat storage, while visceral adipose (vWAT) tissue constitutes a smaller portion, yet excess vWAT is a more serious risk than excess scWAT, which has been reported to have a protective effect. In this regard, obesity is defined as an excess accumulation or abnormal distribution of body fat, which may also be complicated by associated diseases such as type 2 diabetes, fatty liver, cardiovascular disease, stroke, dyslipidemia, hypertension and certain types of cancer, among others. Although obesity is generally caused by excess energy consumption linked to energy expenditure, its etiology is complex and includes genetic, physiological, environmental, social, psychological, and economic factors, which interact to promote the development of obesity. This excess available energy promotes the expansion of adipose tissue depots mediated by the growth of adipocyte size (hypertrophy) and an increase in adipocyte number (hyperplasia).

[0167] In assays using the same number of mice as in Example 1, body weight, weight gain, among other parameters, were also determined to elucidate the ability of supplementation (AA) to reduce obesity risk and visceral fat.

[0168] material and method Histological analysis of EWAT and BAT EWAT and BAT sections fixed in buffered formalin (4% formaldehyde, 4 g / L NaH2PO4, 6.5 g / L Na2HPO4; pH 6.8) were cut to a thickness of 5 μm and stained with hematoxylin & eosin (H&E). BAT IWAT and EWAT images (4x magnification) were taken with a microscope (ECLIPSE Ti; Nikon, Tokyo, Japan) connected to a digital site camera (DS-Ri1, Nikon) and analyzed using ImageJ NDPI software (National Institutes of Health, Bethesda, MD, USA; https: / / imagej.nih.gov / ij, version 1.52a). To avoid any bias in the analysis, this study had a double-blind design, preventing reviewers from knowing any data from the mice during histopathological analysis. Adipocyte area quantification was analyzed by Adiposoft plugin to evaluate EWAT status between groups. Lipid droplet quantification in BAT was analyzed by the Droplet Finder plugin.

[0169] RNA extraction and quantitative polymerase chain reaction Homogenates from EWAT, IWAT and BAT were used for total RNA extraction using TriPure reagent (Roche Diagnostic, Sant Cugat del Valles, Barcelona, ​​Spain) according to the manufacturer's instructions. RNA concentration and purity were determined using a nanophotometer (Implen GmbH, Munchen, Germany). RNA was converted to cDNA using the High-Capacity RNA-to-cDNA Kit (Applied Biosystems, Wilmington, DE, USA). cDNA was diluted 1:10 and then incubated with the commercial LightCycler 480 Sybr Green I Master in a Lightcycler® 480 II (Roche Diagnostics GmbH, Manheim, Germany). Table 2 shows the list of primers used, previously described in other studies and validated with Primer-Blast software (National Center for Biotechnology Information, Bethesda, MD, USA). 36b4 was used as a housekeeping gene.

[0170] BAT temperature measurement The temperature surrounding the BAT was visualized using a high-resolution infrared camera (FLIR Systems) and analyzed with a specific software package (FLIR-Tools-Software, FLIR; Kent, UK) as previously described. For each image, the area surrounding the BAT was delimited and the mean temperature of the skin area was calculated as the average of three pictures / animal.

[0171] [Table 2]

[0172] result As seen in the data shown in Figure 6, the weight of mice fed the HFHFr diet supplemented with the quaternary combination of the present invention (histidine, cysteine, serine and carnosine) did not change initial weight and final weight after 4 weeks of treatment, in contrast to mice fed only the HFHFr diet, whose final weight was significantly higher than the initial weight (Figure 6(A)). Furthermore, obese animals treated with the quaternary combination of the present invention stopped gaining weight in the second week of treatment (Figure 6(C)). Furthermore, when measuring epididymal white adipose tissue (EWAT), retroperitoneal WAT (RWAT) and average weight adipose tissue, mice fed the HFHFr diet supplemented with the quaternary combination had reduced fat mass compared to mice not supplemented (Figure 6(B)). The reduction in hypertrophic visceral adipose tissue by the quaternary combination treatment was supported by a reduction in adipocyte size at the microscopic level, with an increase in the number of smaller adipocytes and a decrease in the number of larger adipocytes (Figure 6(D)). Moreover, the quaternary combination treatment also modulated brown adipose tissue (BAT) and reduced hypertrophy in this fat depot, with a reduction in brown adipocyte size (Figure 6(E)). These effects in BAT were accompanied by an increase in mRNA expression of key thermogenic genes (Ucp1, Fgf21, Pgc1a, Prdm16, Dio2 and Dio3), an insulin-sensitizing adipokine (Adipoq) and other genes related to fatty acid oxidation (Cpt1b) and glucose uptake (Glut1 and Glut4) (Figure 6(G)). The increase in mRNA levels of Ucp1 after treatment with the quaternary combination was corroborated at the protein level, indicating a higher capacity to burn fatty acids (Figure 6(H)). Indeed, this capacity was assessed by thermography, and higher temperatures were observed in the BAT region of animals treated with the quaternary combination (Figure 6(I)).

[0173] Example 4. In vitro assay In vitro assays were also performed using a hepatic cell line (HepG2) to test the effect of different combinations of supplemented amino acids on the hepatic expression of key genes for fatty acid β-oxidation (carnitine palmitoyltransferase, Cpt1a), inflammation (chemokine (CC motif) ligand 2, CCL2), lipolysis (hormone sensitive, HSL; and lipin, LPIN) and adipogenesis (fatty acid synthase, FASN). Gene expression was determined by qPCR as previously shown (see Materials and Methods in Example 1). Table 3 shows the list of primers used. The Cpt1a enzyme is located in the outer membrane and is a key enzyme in carnitine-dependent transport across the inner mitochondrial membrane. Its deficiency leads to a reduced rate of fatty acid β-oxidation.

[0174] [Table 3]

[0175] The data are presented in FIG. 7, where expression levels (mRNA) are shown for either supplemented medium. The supplementation vehicles tested were histidine (His), carnosine (Car), cysteine ​​(Cys), serine (Ser), histidine + carnosine (His+Car), histidine + serine (His+Ser), histidine + cysteine ​​(His+Cys), carnosine + cysteine ​​(Car+Cys), serine + carnosine (Ser+Car), serine + cysteine ​​(Ser+Cys), histidine + carnosine + serine (His+Car+Ser), histidine + cysteine ​​+ serine (His+Cys+Ser), histidine + carnosine + cysteine ​​(His+Car+Cys), serine + carnosine + cysteine ​​(Ser+Car+Cys), and the quaternary histidine + cysteine ​​+ serine + carnosine (4AA). The concentrations tested were: histidine 10 mM, carnosine 5 mM, cysteine ​​5 mM and serine 5 mM. These concentrations were stored in different combinations. Data are presented in FIG. 7 as mean ± SEM; n=3-4 per condition.

[0176] As shown in Figure 7, the best performance was with the supplements containing 4AA. The ternary combinations were also effective compared to the control group (all with histidine in the combination) and showed an increase in the effect compared to the expression of the cited enzymes caused by the single amino acids. It is also noteworthy that the ternary combinations containing histidine, serine and cysteine ​​show a significant increase. Taken together, these data make it possible to confirm that combinations of at least three of histidine, cysteine, serine, carnosine, and especially the ternary and quaternary combinations containing histidine, surprisingly, increased the levels of the analyzed enzymes, as proof of their beneficial role in diseases or disorders involving fat accumulation. It is also noteworthy that the quaternary combinations worked synergistically better, even in the presence of carnosine, a compound that showed a tendency to reduce its effect when added together with some of the other amino acids.

[0177] Example 5. AA supplementation ameliorates metabolic changes associated with menopause material and method Forty 24-week-old rats were ovariectomized according to the procedure described in Koebele, SV and Bimonte-Nelson, HA, 2016. An additional 10 rats of the same age were sham-operated. Two to three weeks after surgery, rats were randomly assigned to one of four experimental groups for 8 weeks: -SHAM, simulated surgery OVX: OVX rats administered placebo daily OVX-E2: OVX rats that received estrogen treatment (17β-estradiol) by subcutaneous injection every other week. OVX-E2 rats were injected subcutaneously every other week with 25 μg / kg body weight of 17β-estradiol (Sigma-Aldrich, St. Louis, MO) via a carrier solution in corn oil. -OVX+4AA: OVX rats supplemented daily with 4AA at doses of 105 mg / kg / biweekly for histidine, carnosine, and serine, and 245 mg / kg / biweekly for cysteine.

[0178] Body composition analysis: Lean and fat mass measurements (grams) were performed without anesthesia at the beginning and end of the study using an EchoMRI-700™ device (Echo Medical Systems, LLC, Houston, USA). Measurements were performed in triplicate at 8.00 am under ad libitum conditions and results were expressed as a percentage of body weight.

[0179] Body mass index: Body mass index was calculated as the sum of the weights of IWAT, MWAT and RWAT depot weights (in grams) and expressed as a percentage of body weight.

[0180] Serum leptin levels: Serum leptin levels were determined using a rat ELISA kit (EZRL-83K / Millipore, Barcelona, ​​Spain).

[0181] Glucose and insulin levels: Glucose and insulin levels were analyzed using a glucose enzymatic colorimetric kit (998282 / QCA, Barcelona, ​​Spain) and a rat insulin ELISA kit (10-1250-01 / Mercodia, Upssala, Sweden).

[0182] HOMA-IR analysis: Homeostasis model assessment estimated insulin resistance (HOMA-IR) was calculated using the following formula: (glucose_insulin) / 22.5, as previously described (Matthews et al., 1985).

[0183] result Previous studies conducted with ovariectomized (OVX) rats have shown that these animals exhibit weight gain, obesity and impaired insulin resistance compared to sham-operated rats (controls).

[0184] The results disclosed herein demonstrate that treatment with the quaternary histidine+cysteine+serine+carnosine (4AA) combination is effective in ameliorating the metabolic changes associated with menopause.

[0185] Example 6. Association of plasma histidine and adiposity with gut microbiota and the Hut gene in humans material and method Metagenomic analysis For metagenomics analysis, microbial families, genera or species with less than 10 in 10% of samples were excluded. Differential abundance analysis for taxa associated with circulating histidine levels, fatty liver or HAA treatment was performed using the Analysis of Composition of Microbiota with Bias Correction Method (ANCOM-BC). It accounts for bias due to differential sampling rates between specimens by adding a specimen-specific offset to a linear regression model estimated from the observed data. The log-scale linear regression model is similar to a log-ratio transformation to account for the compositional nature of metagenomics datasets, but the offset term acts as a bias correction. All models for humans were adjusted for age, sex, BMI and country. The Sequential Goodness of Fit implemented in the "SGoF" R package was used to adjust p-values ​​for multiple comparisons. It differs from the FDR method, which reduces statistical power as the number of hutG, hutH, hutI, and hutU, respectively.

[0186] Extraction of fecal genomic DNA and whole-genome shotgun sequencing (metagenomics) Fecal shotgun sequencing data were generated for 73 patients from the discovery cohort. Briefly, total DNA was extracted from frozen human stool using the QIAamp DNA Stool Mini kit (Qiagen), slightly modified by adding a bead-beating step. DNA quantification was performed using a Qubit 3.0 fluorometer (Thermo Fisher Scientific, USA) and 3 ng of extracted DNA for each sample was prepared using the Bioscientific PCR-free library kit according to the manufacturer's instructions and sequenced on a Hiseq 2500 (Illumina) with 2 × 150 paired-end chemistry. The resulting input fastq files were decompressed, filtered, and quality trimmed at the 3' end using the prinseq-lite-0.20.4 program, and overlapping pairs were joined using FLASH-1.2.11. The fastq files were then converted to fast files and human host reads were removed by mapping the reads against the reference human genome (GRCh38.p11, Dec 2013) using Bowtie 2, which has end-to-end and very sensitive options. Functional analysis was then performed by assembling non-host reads into contigs by MEGAHIT v1.1.2 and mapping those reads against the contigs using Bowtie 2. Reads that were not assembled were added to the contigs. Prodigal v2.6.342 was then used to predict coding regions. Functional annotation was performed using HMMER against the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, version 2016, to obtain functional subcategories of genes, pathways and annotations. Filtering of the best annotations and assignment of orf annotations to all reads were performed using R 3.1.0, which was also used to count aligned reads, add categories and their coverage, and finally build the abundance matrix. Taxonomic annotation was performed with Kaiju v1.6.2 on human free reads using greedy mode.Addition of phylogenetic information, taxon counting and generation of abundance matrices for all samples were performed using the package R.

[0187] Bacterial genomic DNA isolation, 16S rRNA sequencing and analysis (metagenomics) Bacterial genomic DNA was isolated from mouse fecal samples using the MBP DNA Soil extraction kit. Genomic DNA was normalized to 5ng / μL in EB (10mM Tris-HCl) and libraries were generated. Briefly, after the first PCR and cleanup using KAPA Pure Beads (Roche catalog no. 07983298001), a second PCR master mix was generated using P7 and P5 from the Nextera XT Index Kit v2 index primers (Illumina catalog no. FC-131-2001-2004). After the PCR reaction, libraries were quantified using the Quant-iT dsDNA Assay Kit, high sensitivity kit (catalog no. 10164582) and run on a FLUOstar Optima plate reader. Libraries were pooled and run on a High Sensitivity D1000 ScreenTape (Agilent catalog number 5067-5579) using an Agilent Tapestation 4200 to calculate the final library pool molar concentration. Pooling was performed on an Illumina MiSeq instrument using MiSeq® Reagent Kit v3 (600 cycles) (Illumina catalog FC-102-3003) following Illumina recommended denaturation and loading recommendations including 20% ​​PhiX spike-in (PhiX Control v3 Illumina catalog FC-110-3001). Raw data was analyzed locally on the MiSeq using MiSeq Reporter. For 16S sequence analysis, LotuS 1.36 used the pipeline in short amplicon mode with default quality filtering. Raw 16S rRNA gene reads were quality filtered to ensure a minimum length of 170 bp, no more than eight homonucleotides, no ambiguous bases, average quality ≥ 27 and accumulated error less than 0.5, with dereplication filter set to “6:1, 3:2”. Cleaned reads were clustered into amplicon sequencing variants (ASVs) using DADA2, and chimeric ASVs were removed using the DADA2 de novo chimera check.The remaining phiX reads were filtered by mapping ASVs against the phiX reference genome. ASV taxonomy was assigned using the LotuS2 LCA algorithm against the Silva 138.1 reference database. On average, 5500 ± 3108 reads could be assigned to each sample of cyanobacterial origin. Further data analysis was performed using the R statistical language version 3.00 (The R Foundation, https: / / www.r-project.org / ) using the rtk software or all data normalization. The 16S raw data can be found at https: / / 1drv.ms / u / s!ApVezdktX3aIlZVJXYWok9s2qhgfvg?e=VDyr2c.

[0188] Primer design for HutH and HutG microbiota functions To design degenerate primers that can amplify in conserved regions of HutH and HutG genes and cover all species used for primer design, several sequences from prokaryotic species were uploaded from the database to the ARDEP program. Based on the k-mer algorithm, the primer length is set as k and all sequences are divided into k-mers. The analysis platform allows to perform sequence length statistics, length filtering, and redundancy removal on the sequence database using tools located in the "Sequence Processing" section. After performing a quality control of the sequence database, ARDEP for primer design could be run in the "Primer Processing" section, resulting in a list of different possible primer sets. The "Target Classification Calculator" option was used to count the sequence numbers of taxonomic and functional groups covered by the primer sets. The best option was selected and basic primer characteristics (Tm, GC%, primer length) were considered.

[0189] result Microbiota-host interactions potentially contribute to the development of metabolic diseases, and molecular networks linking the gut microbiota to hepatic steatosis have recently been elucidated.

[0190] The gut microbiota plays an important role in the pathophysiology of non-alcoholic fatty liver disease (NAFLD) (Figure 11b) as well as other diseases and conditions such as obesity, IBD, visceral adipose tissue and unhealthy metabolic profiles.

[0191] The families Xanthomonadaceae, Rickettsiaceae, and Alteromonadales from the phylum Proteobacteria are increased in fatty liver. At the genus level, the genera Bilophila and Campylobacter have also been described to be increased in NAFLD.

[0192] Histidine can be utilized by many bacteria and is an important carbon, nitrogen, and / or energy source for Proteobacteria. The histidine utilization (Hut) pathway involves the removal of ammonia from histidine to produce urocanic acid, imidazolonepropionate, and formiminoglutamic acid (Figure 11c).

[0193] To evaluate the potential role of the gut microbiota in regulating plasma histidine levels, we performed a shotgun metagenomics analysis of 73 fecal samples of patients from the discovery cohort. We applied Analysis of Microbiota Composition with Bias Correction (ANCOM-BC) to explore the compositional nature of the metagenomic dataset and identified differentially abundant taxa associated with circulating histidine levels controlling for age, BMI, sex and country.

[0194] A strong negative association was identified between histidine levels and members of the phylum Proteobacteria. Families from the phylum Proteobacteria (Figure 11a) were also strongly associated with the degree of fatty liver. Circulating histidine levels were also strongly positively associated with Cyanobacteria as well as the orders Corynebacteriales (Phylum Actinobacteria) and Marinelabiliales (Phylum Bacteroidetes) (Figure 11a).

[0195] Circulating histidine was also positively associated with healthy gut bacterial indicators known to produce anti-inflammatory metabolites, such as species from the genera Faecalibacteirum, Bifidobacterium, and Odoribacter (Figure 11a).

[0196] Consistent with these results, patients with a higher degree of adiposity had increased levels of microbial Hut genes, which decreased after HAA supplementation ("HAA" as used herein corresponds to the composition used in the previous examples containing histidine, cysteine, serine and carnosine), indicating the effect of HAA administration on the associated dysbiosis.

[0197] Patients with a steatosis degree less than 33% and patients with hepatic steatosis greater than 33% were found to have a higher abundance of microbial genes involved in the first three histidine degradation steps, which appear to be universal and include the conversion of histidine to urocanic acid, the hydration of urocanic acid to imidazolepropionic acid, and the cleavage of the imidazole ring to give formiminoglutamic acid (Figure 11d).

[0198] Previous results have demonstrated that supplementation with histidine-related amino acids (HAAs) improved NAFLD and reduced liver inflammation in mice.

[0199] Herein, it is shown that HAA supplementation in NAFLD-induced mice reduces the expression of bacterial histidine utilization genes (hutH and hutG) that were found to be increased in NAFLD patients (Figure 11e).

[0200] Taken together, these results indicate that plasma histidine levels are negatively associated with dysregulated bacterial families in NAFLD. This microbiota dysregulation, known as dysbiosis, can be countered by HAA administration.

[0201] Further aspects / embodiments of the present invention can be found in the following clauses: 1. A pharmaceutical or nutraceutical composition comprising a therapeutically or nutraceutically effective amount of at least three of histidine, or a pharma- ceutically or nutraceutically acceptable salt, or a solvate of any of them, cysteine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of any of them, serine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of any of them, and carnosine, or a pharma-ceutically or nutraceutically acceptable salt, or a solvate of any of them, together with one or more pharma-ceutically or nutraceutically acceptable excipients or carriers; and / or a therapeutically or nutraceutically effective amount of a peptide comprising a sequence of amino acids that includes at least three of histidine, cysteine, serine, and carnosine.

[0202] 2. The composition of clause 1 comprising a therapeutically or nutraceutical effective amount of at least three of histidine, or a pharma- ceutically or nutraceutically acceptable salt or a solvate of either thereof, cysteine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, serine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, and carnosine, or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, together with one or more pharma-ceutically or nutraceutically acceptable excipients or carriers.

[0203] 3. The composition according to any one of clauses 1-2, wherein the therapeutically or nutraceutical effective amount of histidine or a pharma- ceutically or nutraceutically acceptable salt thereof or a solvate of either thereof provides a dose of 10 mg to 250 mg per Kg of the subject; the therapeutically effective amount of cysteine ​​or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of either thereof provides a dose of 30 to 550 mg per Kg of the subject; the therapeutically effective amount of serine or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of either thereof provides a dose of 10 to 250 mg per Kg of the subject; and the therapeutically effective amount of carnosine or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of either thereof provides a dose of 10 to 200 mg per Kg of the subject.

[0204] 4. The composition according to any one of clauses 1 to 3, comprising a therapeutically or nutraceutical effective amount of at least three of L-histidine or a pharma- ceutically or nutraceutically acceptable salt thereof or a solvate of either thereof, L-cysteine ​​or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of either thereof, L-serine or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of either thereof, and L-carnosine or a pharma-ceutically or nutraceutically acceptable salt thereof or a solvate of either thereof.

[0205] 5. The composition according to any one of clauses 1 to 4, wherein the pharma- ceutically or nutraceutical acceptable salts of histidine and cysteine ​​are the hydrochloride salts of L-histidine and L-cysteine.

[0206] 6. The composition according to any one of clauses 1 to 5, wherein the pharma- ceutically acceptable salt of histidine is L-histidine monohydrochloride monohydrate.

[0207] 7. The composition according to any one of clauses 1 to 6, comprising a therapeutically or nutraceutical effective amount of histidine or a pharma- ceutically or nutraceutically acceptable salt or a solvate of either thereof, a therapeutically or nutraceutically effective amount of cysteine ​​or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, a therapeutically or nutraceutically effective amount of serine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, and a therapeutically or nutraceutically effective amount of carnosine or a pharma-ceutically or nutraceutically acceptable salt or a solvate of either thereof, together with one or more pharma-ceutically or nutraceutically acceptable excipients or carriers.

[0208] 8.- A therapeutically or nutraceutical effective amount of L-histidine monohydrochloride monohydrate; - A therapeutically or nutraceutical effective amount of L-cysteine ​​hydrochloride; a therapeutically or nutraceutical effective amount of L-serine; and - A therapeutically or nutraceutical effective amount of L-carnosine The composition of claim 7, comprising, together with one or more pharma- ceutically or nutraceutical acceptable excipients or carriers.

[0209] 9. Food in solid or liquid form, comprising a pharmaceutical or dietary supplement composition as defined in any one of clauses 1 to 8.

[0210] 10. A food product as described in clause 9 which is a beverage.

[0211] 11. A pharmaceutical or nutraceutical composition as defined in any one of claims 1 to 8 or a food product as defined in any one of clauses 9 to 10 for use in therapy.

[0212] 12. A pharmaceutical or nutraceutical composition or food according to clause 11 for use in the prevention and / or treatment of a disease selected from the group consisting of fatty liver disease, obesity, atherosclerosis and combinations thereof.

[0213] 13. A pharmaceutical or dietary supplement composition or food according to any one of clauses 11 to 12 for use in the prevention and / or treatment of fatty liver disease selected from non-alcoholic fatty liver disease and alcoholic fatty liver disease.

[0214] 14. The pharmaceutical or dietary supplement composition or food for use according to clause 13, wherein the non-alcoholic fatty liver disease is selected from non-alcoholic fatty liver disease (NAFLD), including simple steatosis and non-alcoholic steatohepatitis (NASH).

[0215] References list “ -Koebele, SV & Bimonte-Nelson, HA “Modeling menopause: The utility of rodents in translational behavioral endocrinology research”.Maturitas 2016,87,5-17. -Mardinoglu et al., “Personal model-assisted identification of NAD+ and glutathione metabolism as intervention target in NAFLD”,Mol Syst Biol-2017,vol.no.13:916 -Mong et al., “Histidine and carnosine alleviated hepatic steatosis in mice consumed high saturated fat diet”, European Journal of Pharmacology 653(2011)82-88 -Zhao,Z.;Fu,C.;Zhang,Y.;Fu,A.Dimeric Histidine as a Novel Free Radical Scavenger Alleviates Non-Alcoholic Liver Injury.Antioxidants 2021,10,1529.https: / / doi.org / 10.3390 / antiox10101529

Claims

1. Together with one or more pharmaceutically or nutritionally acceptable excipients or carriers, (i) a therapeutically or nutritionally effective amount of cysteine ​​or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof, and a therapeutically or nutritionally effective amount of at least two of histidine or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof, serine or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof, and carnosine or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof; and / or a therapeutically or nutritionally effective amount of a peptide comprising a sequence of amino acids including cysteine ​​and at least two of histidine, serine and carnosine; or, (ii) A therapeutically or nutritionally effective amount of N-acetylcysteine ​​(NAC) or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof, and a therapeutically or nutritionally effective amount of at least two of histidine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof, serine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof, and carnosine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof; and / or a therapeutically or nutritionally effective amount of a peptide comprising NAC and a sequence of amino acids including at least two of histidine, serine, and carnosine. A pharmaceutical composition or nutritional supplement composition containing the above.

2. Together with one or more pharmaceutically or nutritionally acceptable excipients or carriers, (i) therapeutically or nutritionally effective amounts of cysteine ​​or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof, therapeutically or nutritionally effective amounts of histidine or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof, therapeutically or nutritionally effective amounts of serine or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof, and therapeutically or nutritionally effective amounts of carnosine or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof; and / or therapeutically or nutritionally effective amounts of peptides comprising a sequence of amino acids including cysteine, histidine, serine and carnosine; or, (ii) A therapeutically or nutritionally effective amount of NAC or a pharmaceutically or nutritionally acceptable salt or a solvate thereof; a therapeutically or nutritionally effective amount of histidine or a pharmaceutically or nutritionally acceptable salt or a solvate thereof; a therapeutically or nutritionally effective amount of serine or a pharmaceutically or nutritionally acceptable salt or a solvate thereof; and a therapeutically or nutritionally effective amount of carnosine or a pharmaceutically or nutritionally acceptable salt or a solvate thereof; and / or a therapeutically or nutritionally effective amount of a peptide comprising a sequence of amino acids including NAC, histidine, serine, and carnosine. The composition according to claim 1, comprising:

3. Together with one or more pharmaceutically or nutritionally acceptable excipients or carriers, (i) a therapeutically or nutritionally effective amount of cysteine ​​or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof, and a therapeutically or nutritionally effective amount of at least two of histidine or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof, serine or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof, and carnosine or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof; or, (ii) A therapeutically or nutritionally effective amount of N-acetylcysteine ​​(NAC) or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof, and a therapeutically or nutritionally effective amount of at least two of histidine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof, serine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof, and carnosine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof. The composition according to claim 1, comprising:

4. Together with one or more pharmaceutically or nutritionally acceptable excipients or carriers, (i) a therapeutically or nutritionally effective amount of cysteine ​​or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof; a therapeutically or nutritionally effective amount of histidine or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof; a therapeutically or nutritionally effective amount of serine or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof; and a therapeutically or nutritionally effective amount of carnosine or a pharmaceutically or nutritionally acceptable salt thereof or any solvate thereof; or, (ii) A therapeutically or nutritionally effective amount of NAC or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof; a therapeutically or nutritionally effective amount of histidine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof; a therapeutically or nutritionally effective amount of serine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof; and a therapeutically or nutritionally effective amount of carnosine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof. A composition according to any one of claims 1 to 3, comprising:

5. The composition according to any one of claims 1 or 3, comprising, together with one or more pharmaceutically or nutritionally acceptable excipients or carriers, a therapeutically or nutritionally effective amount of N-acetylcysteine ​​(NAC), a pharmaceutically or nutritionally acceptable salt thereof, or a solvate thereof, and a therapeutically or nutritionally effective amount of at least two of histidine, a pharmaceutically or nutritionally acceptable salt thereof, or a solvate thereof, serine, a pharmaceutically or nutritionally acceptable salt thereof, or a solvate thereof, and carnosine, a pharmaceutically or nutritionally acceptable salt thereof, or a solvate thereof.

6. A composition according to any one of claims 1 to 3, comprising, together with one or more pharmaceutically or nutritionally acceptable excipients or carriers, a therapeutically or nutritionally effective amount of NAC or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof, a therapeutically or nutritionally effective amount of histidine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof, a therapeutically or nutritionally effective amount of serine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof, and a therapeutically or nutritionally effective amount of carnosine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof.

7. The composition according to any one of claims 1 to 3, wherein the therapeutically effective amount of cysteine ​​or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof gives a dose of 2 to 550 mg per kg of the subject; the therapeutically or nutritionally acceptable amount of histidine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof gives a dose of 7.5 to 250 mg per kg of the subject; the therapeutically effective amount of serine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof gives a dose of 1.3 to 250 mg per kg of the subject; and the therapeutically effective amount of carnosine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof gives a dose of 4.5 to 200 mg per kg of the subject.

8. The composition according to claim 7, wherein the therapeutically effective amount of cysteine ​​or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof gives a dose of 30 to 550 mg per kg of the subject; the therapeutically or nutritionally acceptable amount of histidine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof gives a dose of 10 to 250 mg per kg of the subject; the therapeutically effective amount of serine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof gives a dose of 10 to 250 mg per kg of the subject; and the therapeutically effective amount of carnosine or a pharmaceutically or nutritionally acceptable salt thereof or a solvate thereof gives a dose of 10 to 200 mg per kg of the subject.

9. The composition according to any one of claims 1 to 3, wherein the therapeutically effective amount of NAC or a pharmaceutically or nutritionally acceptable salt or a solvate thereof gives a dose of 2 to 550 mg per kg of the subject; the therapeutically or nutritionally acceptable amount of histidine or a pharmaceutically or nutritionally acceptable salt or a solvate thereof gives a dose of 7.5 to 250 mg per kg of the subject; the therapeutically effective amount of serine or a pharmaceutically or nutritionally acceptable salt or a solvate thereof gives a dose of 1.3 to 250 mg per kg of the subject; and the therapeutically effective amount of carnosine or a pharmaceutically or nutritionally acceptable salt or a solvate thereof gives a dose of 4.5 to 200 mg per kg of the subject.

10. The composition according to claim 9, wherein the therapeutically effective amount of NAC or a pharmaceutically or nutritionally acceptable salt or a solvate thereof gives a dose of 30 to 550 mg per kg of the subject; the therapeutically or nutritionally effective amount of histidine or a pharmaceutically or nutritionally acceptable salt or a solvate thereof gives a dose of 10 to 250 mg per kg of the subject; the therapeutically effective amount of serine or a pharmaceutically or nutritionally acceptable salt or a solvate thereof gives a dose of 10 to 250 mg per kg of the subject; and the therapeutically effective amount of carnosine or a pharmaceutically or nutritionally acceptable salt or a solvate thereof gives a dose of 10 to 200 mg per kg of the subject.

11. The composition according to any one of claims 1 to 3, wherein the pharmaceutically or nutritionally acceptable salt of histidine is L-histidine hydrochloride.

12. The composition according to any one of claims 1 to 3, wherein the pharmaceutically acceptable salt of histidine is L-histidine monohydrochloride monohydrate.

13. The composition according to any one of claims 1 to 3, wherein the pharmaceutically or nutritionally acceptable salt of cysteine ​​is L-cysteine ​​hydrochloride.

14. - A therapeutically or nutritionally effective amount of L-histidine monohydrochloride monohydrate; - A therapeutically or nutritionally effective dose of NAC; - A therapeutically or nutritionally effective amount of L-serine; and - Therapeutic or nutritionally effective amounts of L-carnosine The composition according to any one of claims 1 to 3, comprising together with one or more pharmaceutically or nutritionally acceptable excipients or carriers.

15. - A therapeutically or nutritionally effective amount of L-histidine monohydrochloride monohydrate; - A therapeutically or nutritionally effective amount of L-cysteine ​​hydrochloride; - A therapeutically or nutritionally effective amount of L-serine; and - Therapeutic or nutritionally effective amounts of L-carnosine The composition according to any one of claims 1 to 3, comprising together with one or more pharmaceutically or nutritionally acceptable excipients or carriers.

16. A food in solid or liquid form comprising the pharmaceutical or nutritional supplement composition described in claim 1.

17. The food according to claim 16, which is a beverage.

18. A pharmaceutical composition or nutritional supplement composition according to any one of claims 1 to 3, or a food according to any one of claims 16 to 17, for use in treatment.

19. A pharmaceutical composition or nutritional supplement composition or food for use according to claim 18, for use in the prevention and / or treatment of diseases selected from the group consisting of fatty liver disease, obesity, overweight, atherosclerosis, dysbiosis, and combinations thereof.

20. A pharmaceutical composition or nutritional supplement composition or food for use according to claim 18, for use in the prevention and / or treatment of a fatty liver disease selected from non-alcoholic fatty liver disease and alcoholic fatty liver disease.

21. The pharmaceutical composition or nutritional supplement composition or food for use according to claim 20, wherein the non-alcoholic fatty liver disease is selected from non-alcoholic fatty liver (NAFLD), including simple steatosis and non-alcoholic steatohepatitis (NASH).

22. A pharmaceutical composition or nutritional supplement composition or food for use according to claim 18, for use in the prevention and / or treatment of menopausal symptoms in female mammals, particularly women.

23. The pharmaceutical composition or nutritional supplement composition or food for use according to claim 22, wherein the menopausal symptoms are metabolic changes associated with menopause in female mammals, particularly women.

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