Pharmaceutical and health food compositions containing combinations of amino acids and their use in diseases characterized by lipid accumulation in tissues

Abstract

Disclosed are pharmaceutical compositions or dietary supplement compositions comprising a specific combination of compounds comprising histidine, serine, cysteine, and carnosine. The composition is proposed as an ingredient of a food product, and it has been proved to be effective in the treatment of diseases characterized by fat accumulation in tissue, such as fatty liver disease, obesity and atherosclerosis.

Description

[0001] This application claims priority from European patent application 22382390.7 filed on April 25, 2022. Technical Field

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

[0003] Due to the growth and worldwide expansion of the consumption of a diet known as the Western diet, which includes processed foods and foods rich in fat, diseases or conditions related to the deposition of adipose tissue in several body parts, including visceral, vascular and subcutaneous tissue, are becoming a trend, which involves not only high costs to the health system, but also, most importantly, health problems in the subject. Examples of these conditions are obesity with associated visceral fat accumulation, atherosclerosis and fatty liver disease.

[0004] Simply put, obesity is a condition in which excess body fat accumulates to a point where it can have a negative impact on health.

[0005] Atherosclerosis is a pattern of disease arteriosclerosis in which abnormalities called lesions develop in the artery walls. These lesions may cause the arteries to narrow due to a buildup of atherosclerotic plaque. When severe, it 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 referred to as hepatic steatosis (HS), is a condition in which excess fat accumulates in the liver. Fatty liver disease is divided into non-alcoholic fatty liver disease (NAFLD) and alcoholic liver disease. Westernized changes in diet and lifestyle can lead to the development of key risk factors such as obesity, leptin or insulin resistance (IR), dyslipidemia and metabolic syndrome, which are associated with the development of NAFLD. NAFLD is one of the most prevalent human diseases today, affecting nearly one-third of the world's population. The disease is characterized by excessive fat accumulation and low-grade inflammation in the liver. This abnormal lipid metabolism may affect lipid oxidation and disturb the more advanced redox homeostasis. As a result, the accumulation of reactive oxygen species (ROS) and the resulting increase in lipid peroxidation may lead to liver cell damage and fibrosis. NAFLD is divided into different grades according to the severity and the ability to return to its healthy state: non-alcoholic fatty liver (NAFL) with simple steatosis; non-alcoholic steatohepatitis (NASH) with advanced steatosis and inflammation. Furthermore, NASH may eventually progress to fibrosis in advanced stages and eventually to irreversible stages such as cirrhosis or liver cancer. Treatment of NAFLD is usually through weight loss through changes in diet and promotion of exercise, unless severe progression occurs and liver transplantation is necessary.

[0007] Mixtures of certain compounds as dietary supplements have been tested in vivo to promote fat oxidation and glutathione (GSH) synthesis. GSH is required to prevent the accumulation of incomplete products of fatty acid oxidation. An example is the literature of Mardinoglu et al., "Personal model-assisted identification of NAD+ andglutathione metabolism as intervention target in NAFLD", Mol Syst Biol-2017, vol. no. 13:916. These authors tested a mixture of serine, NAC (N-acetyl-L-cysteine) and nicotinamide riboside (NR) in mice fed with a Western diet containing high levels of fat and sucrose. Serine is included in the mixture because it can be easily converted to glycine, and NAC is included because cysteine ​​may be a limiting metabolite after glycine supplementation in GSH synthesis. The purpose of this trial is to provide proof of concept for a possible therapeutic strategy based on promoting GSH and NAD+ synthesis, and to draw results from an analysis of personalized genome-scale metabolic modeling of a human cohort with varying degrees of hepatic steatosis. The rationale behind this is to reduce oxidative stress in lipid metabolism. From the GEM analysis of the human cohort, the authors also concluded that plasma serine and glycine levels were negatively correlated with HS, while there was no significant correlation between HS and plasma cysteine ​​and glutamine levels.

[0008] Another publication proposing dietary supplementation to alleviate high-fat diet-induced hepatic steatosis is Mong et al., “Histidine and carnosine alleviated hepatic steatosis in mice consumed highsaturated fat diet”, European Journal of Pharmacology 653 (2011) 82–88. The authors proposed the use of histidine and carnosine to alleviate hepatic steatosis after observing that mice fed with histidine and carnosine supplementation had reduced body weight, epididymal fat, and liver triglyceride and cholesterol levels, as well as improved insulin sensitivity and attenuated hyperinsulinemia.

[0009] Some of the effects attributed to histidine are, among others, 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 associated with the modulation of NF-kB and PPARα-induced pathways, which reduces the concentrations of TNFα, IL6, and C-reactive protein in plasma and levels in the liver. In addition, histidine has an anti-glycation effect associated with reduced inflammasome activation. On the other hand, the antioxidant properties of histidine treatment are due to the fact that this amino acid is a free radical scavenger, which has the ability to bind divalent metal ions.

[0010] Based on the ability of the latter to act as a free radical scavenger, some authors have proposed the use of 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 ScavengerAlleviates Non-Alcoholic Liver Injury. Antioxidants 2021, 10, 1529. https: / / doi.org / 10.3390 / antiox10101529). Although this is an excellent approach, the complexity of using synthetic compounds is always a disadvantage.

[0011] Therefore, there is still a need for alternative approaches to deal with all these diseases or conditions associated with fat deposition in tissues. Summary of the invention

[0012] The inventors unexpectedly found that a specific combination of three or more of histidine, cysteine, serine and carnosine can reduce liver damage and liver characteristics of NAFLD, which is manifested macroscopically as liver weight, liver whitening and fatty liver; at the biochemical level, by analyzing alanine aminotransferase (ALT) levels and aspartate aminotransferase (AST) levels. The combination of these compounds also reduces liver lipid content and hepatic steatosis. Supplementing the combination of the compounds in the diet reduces liver inflammation associated with NAFLD, increases antioxidant defense and anti-tumor activity, and reduces insulin resistance. In short, in animal models suffering from the disease, a combination of three or more of these compounds can reverse NAFLD symptoms.

[0013] In addition, the inventors have found that the combination of compounds can also counteract the accumulation of fat mass associated with menopause in ovariectomized (OVX) rats and reduce the loss of lean body mass in OVX rats, contributing to a healthier body composition. In this animal model, these compounds can also improve insulin resistance. Therefore, the authors found that the combination of compounds helps to reverse the metabolic changes associated with menopause in ovariectomized (OVX) rats.

[0014] Without being bound by any theory, the inventors propose that the combination of these compounds as an ingredient in a health food composition, a food supplement or as an active ingredient in a pharmaceutical composition ultimately leads to an increase in plasma histidine levels, which ultimately and due to the complexity of various factors can reduce fat accumulation. This assumes a new method related to the previously disclosed use of histidine as a free radical scavenger or as an anti-inflammatory agent in diseases involving fat accumulation. The method is unexpected because, in addition, histidine has only been proposed as having a modest effect on all these diseases, while the treatments in the prior art are more focused on promoting the oxidation of fat and the synthesis of glutathione (GSH). In addition, the proposed method, which stems from the real observations of the inventors, allows to safely deal with diseases related to the accumulation of fat in several body tissues in these subjects, in which the control of fat oxidation capacity is not impaired, but these subjects also suffer from clinical symptoms due to other mechanisms that ultimately reduce the circulating levels of histidine.

[0015] This mechanism may also be responsible for the reduction of visceral fat in obese subjects and the reduction of LDL-cholesterol in models of atherosclerosis.

[0016] Therefore, the first aspect of the present invention is a pharmaceutical composition or dietary supplement composition, which comprises one or more than one pharmaceutically or nutraceutically acceptable excipients or carriers, and the following substances: a therapeutically or nutraceutically effective amount of at least three of the following substances: histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two; cysteine ​​or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, and carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two; and / or a therapeutically or nutraceutically effective amount of a peptide, the peptide comprising an amino acid sequence containing at least three of histidine, cysteine, serine and carnosine. Specifically, the pharmaceutical composition or dietary supplement composition comprises one or more than one pharmaceutically or nutraceutically acceptable excipients or carriers, and the following substances:

[0017] (i) a therapeutically or health food effective amount of cysteine ​​or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of at least two of the following substances: histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two; and / or a therapeutically or health food effective amount of a peptide comprising an amino acid sequence containing cysteine ​​and at least two of histidine, serine and carnosine; or, alternatively,

[0018] (ii) a therapeutically or health food effective amount of N-acetylcysteine ​​(NAC) or a pharmaceutically or health food acceptable salt or a solvate of either of the two, and a therapeutically or health food effective amount of at least two of the following substances: histidine or a pharmaceutically or health food acceptable salt or a solvate of either of the two, serine or a pharmaceutically or health food acceptable salt or a solvate of either of the two, and carnosine or a pharmaceutically or health food acceptable salt or a solvate of either of the two; and / or a therapeutically or health food effective amount of a peptide comprising an amino acid sequence containing NAC and at least two of histidine, serine and carnosine.

[0019] The pharmaceutical composition or dietary supplement composition can be used as an ingredient of commonly used foods together with common ingredients of such foods, such as beverages (i.e., soda), dairy products (i.e., yogurt), baked goods, etc. Therefore, the second aspect of the present invention is a food in solid or liquid form, which comprises the pharmaceutical composition or dietary supplement composition defined in the first aspect.

[0020] As will be illustrated in the examples, and as previously indicated, a composition or food comprising at least three of the listed compounds provides multiple beneficial effects in an animal model of NAFLD and an animal model of menopause. Therefore, another aspect of the invention is the use of a composition as defined in the first aspect or a food of the second aspect in therapy (i.e. as a medicine). BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1Shown are the effects of treatment on (A) liver weight; (B) representative macroscopic appearance of the liver; and (C) serum ALT (alanine aminotransferase); and (D) serum AST (aspartate aminotransferase). Data are mean ± SEM; n = 16 animals / group. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 compared to control mice; #p < 0.05, ##p < 0.01, ###p < 0.001 compared to HFHFr mice.

[0022] Figure 2 Shown are the effects of treatment on (A) total liver lipid content; (B) total liver triglyceride (TG) content; (C) total liver cholesterol content. Data are mean ± SEM; n = 16 animals / group. **p < 0.01, ***p < 0.001, ****p < 0.0001 compared with control mice; #p < 0.05, ####p < 0.0001 compared with HFHFr mice.

[0023] Figure 3 Shown are the histopathological analysis and image analysis of liver determined by (A) H&E (hematoxylin-eosin staining) and magnification of selected areas, with magnified areas shown in the lower panels; (B) lipid droplet counts; (C) lipid droplet surface area field; (D) NAFLD / NASH scoring table. st., steatosis. Bar = 100 µm. Data are mean ± SEM; n = 6 animals / group. **p < 0.01, ****p < 0.0001 compared with control mice (C); ###p < 0.001, ####p < 0.0001 compared with HFHFr mice. AA mice are treated mice.

[0024] Figure 4 The effects of treatment on hepatic insulin resistance are shown. (A) Representative Western blot analysis of Akt activation (pAktS473), total Akt protein levels (T-Akt), housekeeping β-actin levels, and protein loading using Ponceau S membrane staining; (B) Densitometry analysis of the ratio of phosphorylated Akt to total Akt. Data are mean ± SEM. n = 5 to 6 animals / group. *p < 0.05 compared with control mice. (C) Analysis of hepatic expression of key genes for fatty acid β-oxidation (carnitine palmitoyltransferase, Cpt1a), inflammation (chemokine (CC motif) ligand 2, CCL2), lipolysis (hormone-sensitive lipase, HSL; and lipid droplet coating protein, Plin) and lipogenesis (fatty acid synthase, FASN) after quadruple treatment. Data are expressed as mean ± SEM; n = 10 to 12 per condition. *p < 0.05.

[0025] Figure 5 Effects of treatment on circulating cholesterol levels are shown. Circulating (HDL) levels are depicted in (A), low-density lipoprotein (LDL) levels are depicted in (B), and LDL / HDL ratios are depicted in (C). Data are mean ± SEM. n = 16 animals / group. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0026] Figure 6 The effects of treatment on post-treatment weight change (A) and on the weight of major visceral depots after treatment (B) are shown; epididymal white adipose tissue (EWAT); retroperitoneal white adipose tissue (RWAT) and mesenteric white adipose tissue (MWAT). (C) Growth curve. Circles represent "thin", squares represent "obese", and triangles represent "obese + 4AA". (D) Histopathological analysis and image analysis of visceral adipose tissue determined by H&E (hematoxylin-eosin staining). (E) BAT weight. (F) Histopathological analysis and image analysis of BAT. (G) Gene expression analysis of genes indicated in BAT. (H) Representative protein blot analysis and quantification of proteins indicated in BAT. (I) Mouse thermal imaging analysis and quantification. In (D) to (I), the left column represents "thin", the middle column represents "obese", and the right column represents "obese + 4AA". Data are mean ± SEM. n = 16 animals / group. *p<0.05, **p<0.01, ****p<0.0001.

[0027] Figure 7 The effect of 24-hour treatment with the indicated amino acids, alone or in combination, on the expression of Cpt1a mRNA, Ccl2 mRNA, Hsl mRNA, Plin mRNA, and Fasn mRNA in human liver cells (HepG2) is shown: histidine 10 mmol / L, carnosine 5 mmol / L, cysteine ​​5 mmol / L, and serine 5 mmol / L. These concentrations were constant in the different combinations. Data are expressed as mean ± SEM; n = 3 to 4 per condition. *p < 0.05, **p < 0.01, ***p < 0.001 compared to vehicle.

[0028] Figure 8Shown is the effect of ovariectomy, 17β-E2 injection and tetrahistidine+cysteine+serine+carnosine (4AA) treatment on weight change (A), fat body mass change (B) and lean body mass change (C) of female Sprague-Dawley rats with sham operation (SH) and ovariectomy (OVX) after 57 days of treatment. Body weight was recorded once every 7 days, while fat body mass and lean body mass were recorded once every 28 days. Weight gain (D) in grams was calculated as the quotient of the final body weight accumulated weekly during the entire study. Fat body mass increase and lean body mass increase were calculated as the difference in percentage between fat body mass and baseline point at the end of the experiment. Data are given with mean ± SEM (n=10). In each panel, different superscript lowercase letters (a, b, c, d) indicate means that are significantly different between groups (one-way ANOVA with Duncan post hoc test, Welch test with Games-Howell post hoc test or Kruskal-Wallis test with Mann-Whitney U post hoc test, p<0.05). I: effect of intervention, t: effect of time, Ixt: interaction between intervention type 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 tetrahistidine+cysteine+serine+carnosine (4AA). In (A) to (C), circles represent "SH", triangles represent "OVX", diamonds represent "OVX-E2", and squares represent "OVX-4AA".

[0029] Fig. 9 Shown is the effect of ovariectomy, 17β-E2 injection and tetrahistidine+cysteine+serine+carnosine (4AA) treatment on RWAT (A) depot weight, MWAT (B) depot weight, IWAT (C) depot weight, obesity index (D) and circulating leptin level (E) of sham-operated (SH) and ovariectomized (OVX) rats after 57 days of intervention.Data are given with mean ± SEM (n=9 to 10).Obesity index is calculated as the sum (in grams) of IWAT depot weight, MWAT depot weight and RWAT depot weight, and is expressed as the percentage of body weight.In each figure, different superscript lowercase letters (a, b, c) indicate the significantly different mean values ​​(single factor ANOVA and Duncan post hoc test, p<0.05) between groups.I: the effect of intervention. SH: sham-operated rats, OVX: ovariectomized rats, OVX-E2: 17β-estradiol-treated OVX rats, OVX-4AA: OVX rats supplemented with tetrahistidine-related amino acids (4AA). MWAT: mesenteric white adipose tissue; RWAT: retroperitoneal white adipose tissue; IWAT: inguinal white adipose tissue.

[0030] Fig.10 Plasma insulin levels (a), plasma glucose levels (b) and plasma HOMA-IR levels (c) at the end of the study in sham-operated (SH) and ovariectomized (OVX) rats after 57 days of intervention are shown. Data are given as mean ± SEM (n = 9 to 10). Superscript lowercase letters (a, b, c) indicate significantly different means between groups (one-way ANOVA and Duncan 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 tetrahistidine + cysteine ​​+ serine + carnosine (4AA). HOMA-IR: insulin resistance estimated by homeostatic model assessment.

[0031] Fig.11 Figure 2. The relationship between plasma histidine and the degree of steatosis and the human gut microbiota and Hut genes is shown. a) Volcano plot of differential bacterial families associated with circulating histidine levels in the discovery cohort (n=73) using bias-corrected microbiome analysis (ANCOM-BC), controlling for age, BMI, sex, and country. b) Volcano plot of differential bacterial families associated with the degree of hepatic steatosis (liver biopsy) in the discovery cohort (n=73), controlling for age, BMI, sex, and country using bias-corrected microbiome analysis (ANCOM-BC). The log2 (fold change) associated with a unit change in plasma histidine levels and the -log10 (p-value) adjusted for multiple testing are plotted for each taxon. Significantly different taxa are colored according to phylum. c) Histidine utilization pathways. The first three pathways appear to be universal. Depending on the genus, there are two different degradation pathways for imidomethylglutamate: hydrolysis to formamide and glutamate, or hydrolysis to formylglutamate and subsequent hydrolysis to formate and glutamate. d) Violin plots of centered log-ratio transformed microbial genes involved in histidine utilization, hutH, hutU, hutI, and hutG, in subjects with less than 33% or more steatosis. e) qPCR of expression of microbial genes involved in histidine utilization (hutH and hutG). Data are mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 relative to NAFLD mice.

[0032] Detailed description of the invention

[0033] Unless otherwise specified, all terms used in this application should be understood as their ordinary meanings known in the art. Other more specific definitions of certain terms used in this application are described below and are intended to be consistently applied throughout the specification and claims, unless otherwise clearly described definitions provide a broader definition.

[0034] As used herein, the singular references "a" and "an" are synonymous with "at least one" or "one or more than one." Unless otherwise indicated, particular references such as "the" used herein also include the plural noun.

[0035] For purposes of the present invention, any range given includes the lower and upper endpoints of the range.

[0036] As used herein, the term "pharmaceutically acceptable salt" refers to salts that are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, etc., and are commensurate with a reasonable benefit / risk ratio, within the scope of sound medical judgment. Pharmaceutically acceptable salts are well known in the art. Examples of pharmaceutically acceptable salts, non-toxic acid addition salts, are amino salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with 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 pharmaceutically 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, 2-hydroxyethanesulfonate, lactobionate, lactate, dodecanoate, dodecylsulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Salts derived from suitable bases include alkali metal salts, alkaline earth metal salts and ammonium salts. Representative alkali metal salts or alkaline earth metal salts include salts of sodium, lithium, potassium, calcium, magnesium and the like. Where appropriate, other pharmaceutically acceptable salts include salts of non-toxic ammonium, quaternary ammonium and amine cations formed using counterions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates and aryl sulfonates.

[0037] The term "solvate" refers to a molecular complex comprising a compound of interest (i.e., L-histidine, L-cysteine, L-serine, and L-carnosine, or a pharmaceutically acceptable salt thereof) and a stoichiometric or non-stoichiometric amount of one or more solvent molecules (e.g., water) bound by non-covalent intermolecular forces. When the one or more solvent molecules forming part of the molecular complex is water, the solvate is a hydrate. Methods of solvation are generally known in the art.

[0038] The term "medicament" also includes the concept of a "veterinary composition". It therefore relates to a composition which is therapeutically effective when administered to any animal, including humans, by any desired or applicable route.

[0039] As used herein, the expression "therapeutically effective amount" refers to an amount of a compound that, when administered, is sufficient to prevent the development of one or more symptoms of the disease being treated, or to alleviate to some extent one or more symptoms of the disease being treated. The specific dosage of a compound administered in accordance with the present invention will, of course, be determined by the specific circumstances associated with the case, including the compound administered, the route of administration, the specific condition being treated, and similar considerations.

[0040] The expression "pharmaceutically or health food acceptable excipient or carrier" refers to a pharmaceutically acceptable material, composition or carrier. Each component must be pharmaceutically acceptable, i.e. compatible with the other ingredients of the pharmaceutical composition. It must also be suitable for contact with human and animal tissues or organs without excessive toxicity, irritation, allergic reaction, immunogenicity or other problems or other complications, and commensurate with a reasonable benefit / risk ratio. Examples of suitable acceptable excipients are solvents, dispersion media, diluents or other liquid carriers, dispersing aids or suspension aids, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, etc. Unless any conventional excipient medium is incompatible with the substance or its derivatives, such as by producing any undesirable biological effects or interacting with any other one or more than one component of the pharmaceutical composition in other harmful ways, its use should be considered within the scope of the present invention.

[0041] As used herein, the expression "nutraceutically effective amount" refers to an amount of a compound that is sufficient to provide a physiological benefit to a subject in some specific metabolic aspect when administered. The specific dosage of a compound administered according to the present invention will of course be determined by the specific circumstances associated with the case, including the compound administered, the route of administration, the specific condition being treated, and similar considerations.

[0042] "Dietary supplement compositions", also referred to herein as "health food compositions" or "food supplements", are compositions that provide a physiological benefit to a subject in some specific metabolic aspect. Although they cannot be labeled with drug claims, they may carry health claims and nutritional claims as regulated by health administration agencies. Under this category, several compositions are included, which vary according to regulations in different countries. Generally, it is an oral product containing "dietary ingredients" intended to supplement the diet. The "dietary ingredients" in these products may include: vitamins, minerals, herbs or other botanicals, amino acids, and substances such as enzymes, organ tissues, glands, and metabolites. Dietary supplements can also be extracts or concentrates, which can have a variety of forms, such as tablets, capsules, softgels, gelatin capsules, liquids or powders.

[0043] "Functional foods", also referred to herein as "foods", have their nutritional value increased or have been enriched during processing and are then marketed as providing a benefit to the consumer.

[0044] 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 of (2R)-2-acetylamino-3-mercaptopropionic acid. As will be appreciated by the skilled artisan, NAC is a well-established prodrug of cysteine ​​and thus cysteine ​​and NAC can be used interchangeably in the compositions of the present invention.

[0045] As indicated above, the first aspect of the present invention is a pharmaceutical composition or dietary supplement composition, which comprises one or more than one pharmaceutically or nutraceutically acceptable excipient or carrier, and the following substances: a therapeutically or nutraceutically effective amount of at least three of the following substances: histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two; cysteine ​​or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, and carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two; and / or a therapeutically or nutraceutically effective amount of a peptide, the peptide comprising an amino acid sequence containing at least three compounds selected from histidine, cysteine, serine and carnosine. Specifically, the pharmaceutical composition or dietary supplement composition comprises one or more than one pharmaceutically or nutraceutically acceptable excipient or carrier, and the following substances:

[0046] (i) a therapeutically or health food effective amount of cysteine ​​or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of at least two of the following substances: histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two; and / or a therapeutically or health food effective amount of a peptide comprising an amino acid sequence containing cysteine ​​and at least two of histidine, serine and carnosine; or, alternatively,

[0047] (ii) a therapeutically or health food effective amount of N-acetylcysteine ​​(NAC) or a pharmaceutically or health food acceptable salt or a solvate of either of the two, and a therapeutically or health food effective amount of at least two of the following substances: histidine or a pharmaceutically or health food acceptable salt or a solvate of either of the two, serine or a pharmaceutically or health food acceptable salt or a solvate of either of the two, and carnosine or a pharmaceutically or health food acceptable salt or a solvate of either of the two; and / or a therapeutically or health food effective amount of a peptide comprising an amino acid sequence containing NAC and at least two of histidine, serine and carnosine.

[0048] In a specific embodiment, the pharmaceutical composition or dietary supplement composition comprises one or more than one pharmaceutically or nutraceutically acceptable excipient or carrier, and the following substances: therapeutically or nutraceutically effective amounts of at least three of the following substances: histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either; cysteine ​​or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, and carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either.

[0049] In certain embodiments, the pharmaceutical composition or dietary supplement composition comprises one or more than one pharmaceutically or nutraceutically acceptable excipient or carrier, and the following substances:

[0050] (i) a therapeutically or health food effective amount of cysteine ​​or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of at least two of the following substances: histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two; and / or a therapeutically or health food effective amount of a peptide comprising an amino acid sequence containing cysteine ​​and at least two of histidine, serine and carnosine; or, alternatively,

[0051] (ii) a therapeutically or health food effective amount of N-acetylcysteine ​​(NAC) or a pharmaceutically or health food acceptable salt or a solvate of either of the two, and a therapeutically or health food effective amount of at least two of the following substances: histidine or a pharmaceutically or health food acceptable salt or a solvate of either of the two, serine or a pharmaceutically or health food acceptable salt or a solvate of either of the two, and carnosine or a pharmaceutically or health food acceptable salt or a solvate of either of the two; and / or a therapeutically or health food effective amount of a peptide comprising an amino acid sequence containing NAC and at least two of histidine, serine and carnosine.

[0052] In another specific embodiment, the pharmaceutical composition or dietary supplement composition comprises one or more than one pharmaceutically or nutraceutically acceptable excipient or carrier, and the following substances:

[0053] (i) a therapeutically or health food effective amount of cysteine ​​or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of at least two of the following substances: histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two; or, alternatively,

[0054] (ii) a therapeutically or healthily effective amount of N-acetylcysteine ​​(NAC) or a pharmaceutically or healthily acceptable salt thereof or a solvate of either of the two, and a therapeutically or healthily effective amount of at least two of the following substances: histidine or a pharmaceutically or healthily acceptable salt thereof or a solvate of either of the two, serine or a pharmaceutically or healthily acceptable salt thereof or a solvate of either of the two, and carnosine or a pharmaceutically or healthily acceptable salt thereof or a solvate of either of the two.

[0055] In another specific embodiment, the pharmaceutical composition or dietary supplement composition comprises one or more than one pharmaceutically or nutraceutically acceptable excipient or carrier, and the following substances:

[0056] (i) a therapeutically or health food effective amount of cysteine ​​or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two; and / or a therapeutically or health food effective amount of a peptide comprising an amino acid sequence containing cysteine, histidine, serine and carnosine; or, alternatively,

[0057] (ii) a therapeutically or nutraceutically effective amount of NAC or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, a therapeutically or nutraceutically effective amount of histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, a therapeutically or nutraceutically effective amount of serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, and a therapeutically or nutraceutically effective amount of carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two; and / or a therapeutically or nutraceutically effective amount of a peptide comprising an amino acid sequence containing NAC, histidine, serine and carnosine.

[0058] In another specific embodiment, the pharmaceutical composition or dietary supplement composition comprises one or more than one pharmaceutically or nutraceutically acceptable excipient or carrier, and the following substances:

[0059] (i) a therapeutically or health food effective amount of cysteine ​​or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two; or, alternatively,

[0060] (ii) a therapeutically or health food effective amount of NAC or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two.

[0061] In a more particular embodiment, the pharmaceutical composition or dietary supplement composition comprises one or more than one pharmaceutically or nutraceutically acceptable excipient or carrier, and the following substances: a therapeutically or nutraceutically effective amount of N-acetylcysteine ​​(NAC) or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either of the two, and a therapeutically or nutraceutically effective amount of at least two of the following substances: histidine or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either of the two, serine or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either of the two, and carnosine or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either of the two.

[0062] In a more particular embodiment, the pharmaceutical composition or dietary supplement composition comprises one or more than one pharmaceutically or nutraceutically acceptable excipient or carrier, and the following: a therapeutically or nutraceutically effective amount of NAC or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, a therapeutically or nutraceutically effective amount of histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, a therapeutically or nutraceutically effective amount of serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, and a therapeutically or nutraceutically effective amount of carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either.

[0063] In a particular embodiment, the pharmaceutical composition or dietary supplement composition is one wherein the therapeutically effective amount of cysteine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either, is a dosage of 2 mg per kg of subject to be administered to 550 mg per kg of subject; the therapeutically effective amount of histidine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either, is a dosage of 7.5 mg per kg of subject to be administered to 250 mg per kg of subject; the therapeutically effective amount of serine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either, is a dosage of 1.3 mg per kg of subject to be administered to 250 mg per kg of subject; and the therapeutically effective amount of carnosine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either, is a dosage of 4.5 mg per kg of subject to be administered to 200 mg per kg of subject.

[0064] In a particular embodiment, the pharmaceutical composition or dietary supplement composition is one wherein the therapeutically or nutraceutically effective amount of histidine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either of the two, is a dosage of 10 mg per kg of subject to be administered to 250 mg per kg of subject; the therapeutically or nutraceutically effective amount of cysteine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either of the two, is a dosage of 30 mg per kg of subject to be administered to 550 mg per kg of subject; the therapeutically or nutraceutically effective amount of serine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either of the two, is a dosage of 10 mg per kg of subject to be administered to 250 mg per kg of subject; and the therapeutically or nutraceutically effective amount of carnosine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either of the two, is a dosage of 10 mg per kg of subject to be administered to 200 mg per kg of subject.

[0065] In another specific embodiment, the pharmaceutical composition or dietary supplement composition is one wherein the therapeutically effective amount of NAC or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 2 mg per kg of subject to be administered to 550 mg per kg of subject; the therapeutically or nutraceutically effective amount of histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 7.5 mg per kg of subject to be administered to 250 mg per kg of subject; the therapeutically effective amount of serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 1.3 mg per kg of subject to be administered to 250 mg per kg of subject; and the therapeutically effective amount of carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 4.5 mg per kg of subject to be administered to 200 mg per kg of subject.

[0066] In another specific embodiment, the pharmaceutical composition or dietary supplement composition is one wherein the therapeutically effective amount of NAC, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either, is a dosage of 30 mg per kg of subject to be administered to 550 mg per kg of subject; the therapeutically or nutraceutically effective amount of histidine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either, is a dosage of 10 mg per kg of subject to be administered to 250 mg per kg of subject; the therapeutically effective amount of serine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either, is a dosage of 10 mg per kg of subject to be administered to 250 mg per kg of subject; and the therapeutically effective amount of carnosine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either, is a dosage of 10 mg per kg of subject to be administered to 200 mg per kg of subject.

[0067] The indicated therapeutic or health product effective amount refers to the amount related to the weight of the animal, particularly a mammal, including mice and humans. In practice, the amount and dosage are usually determined by a person skilled in the art by a human equivalent dose (HED) calculation, and the technician can find the value for humans (Guidance for Industry Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers, US Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER), July 2005 Pharmacology and Toxicology).

[0068] For example, the dose of histidine, serine and carnosine in mice is 210 mg / Kg, and the inferred HED is 17.027 mg / Kg (the median human body weight is 70 Kg). Similarly, the dose is 490 mg / Kg, and the inferred HED is 39.730 mg / Kg. In a particular embodiment of the present invention, the amount per Kg of object is the amount used to calculate the daily dose (i.e., amount). In humans, the specific daily dose of histidine, serine and carnosine (or any salt or solvate thereof) is 660 mg / day to 1500 mg / day, and the specific daily dose of cysteine ​​(or any salt or solvate thereof) is 1700 mg / day to 3000 mg / day.

[0069] In a particular embodiment, the therapeutically or nutraceutically effective amount of any of histidine, serine and carnosine or a pharmaceutically acceptable salt or solvate thereof is a dose of 12 mg / Kg to 230 mg / Kg, or even more particularly 17 mg / Kg to 210 mg / Kg.

[0070] In a particular embodiment, the therapeutic or health food effective amount of histidine, serine and carnosine or their pharmaceutically acceptable salts or solvates in humans is a dose of 12 mg / Kg to 25 mg / Kg, or selected from 0.5 mg / Kg, 1 mg / Kg, 1.5 mg / Kg, 2 mg / Kg, 2.5 mg / Kg, 3 mg / Kg, 3.5 mg / Kg, 4 mg / Kg, 4.5 mg / Kg, 5 mg / Kg, 5.5 mg / Kg, 6 mg / Kg. In some embodiments, the amount of the present invention is 200mg / Kg to 230mg / Kg, more particularly 210mg / Kg. In mice, it is 200mg / Kg to 230mg / Kg, more particularly 210mg / Kg.

[0071] In a particular embodiment, the therapeutic or health food effective amount of histidine, serine and carnosine or their pharmaceutically acceptable salts or solvates in the human body is 0.5 mg / Kg to 25 mg / Kg, 1 mg / Kg to 25 mg / Kg, 1.3 mg / Kg to 25 mg / Kg, 1.5 mg / Kg to 25 mg / Kg, 2.5 mg / Kg to 25 mg / Kg, 4 mg / Kg to 25 mg / Kg, 5 mg / Kg to 25 mg / Kg. , 6mg / Kg to 25mg / Kg, 7.5mg / Kg to 25mg / Kg, 8mg / Kg to 25mg / Kg, 9mg / Kg to 25mg / Kg, 10mg / Kg to 25mg / Kg, 12mg / Kg to 25mg / Kg, 15mg / Kg to 25mg / Kg, 17mg / Kg to 25mg / Kg or 20mg / Kg to 25mg / Kg, in particular a dosage of 15mg / Kg to 25mg / Kg. In another specific embodiment, the therapeutically or nutraceutically effective amount of histidine, serine and carnosine or their pharmaceutically acceptable salts or solvates in humans is a dose of 10 mg / Kg to 25 mg / kg, 12 mg / Kg to 22 mg / kg, 15 mg / Kg to 20 mg / Kg or 16 mg / Kg to 18 mg / kg, particularly 15 mg / kg to 20 mg / kg.

[0072] In another more specific embodiment, optionally in combination with any of the above or following embodiments, the therapeutic or health food effective amount of cysteine ​​or its pharmaceutically or health food acceptable salt or solvate is a dose of 35 mg / Kg to 500 mg / Kg. In another more specific embodiment, optionally in combination with any of the above or following embodiments, the therapeutic or health food effective amount of cysteine ​​or its pharmaceutically or health food acceptable salt or solvate in the human body is a dose of 35 mg / Kg to 50 mg / Kg, or selected from 0.5 mg / Kg, 1 mg / Kg, 2 mg / Kg, 3 mg / Kg, 4 mg / Kg, 5 mg / Kg, 6 mg / Kg, 7 mg / Kg, 8 mg / Kg, 9 mg / Kg, 10 mg / In some embodiments, the amount of the present invention is 480mg / Kg to 500mg / Kg, more particularly 490mg / Kg. In mice, it is 480mg / Kg to 500mg / Kg, more particularly 490mg / Kg.

[0073] In another specific embodiment, optionally in combination with any of the above or following embodiments, the therapeutically or nutraceutically effective amount of cysteine ​​or its pharmaceutically or nutraceutically acceptable salt or solvate in the human body is a dose of 1 mg / Kg to 10 mg / Kg, 1.5 mg / Kg to 7.5 mg / Kg or 2 mg / Kg to 5 mg / Kg, particularly 2 mg / Kg to 5 mg / Kg.

[0074] In another specific embodiment, optionally in combination with any of the above or following embodiments, the therapeutically or nutraceutically effective amount of cysteine ​​or its pharmaceutically or nutraceutically acceptable salt or solvate in the human body is a dose of 20 mg / Kg to 55 mg / Kg, 25 mg / Kg to 50 mg / Kg, 30 mg / Kg to 45 mg / Kg or 35 mg / Kg to 40 mg / Kg, particularly 35 mg / Kg to 40 mg / Kg.

[0075] In another more specific embodiment, optionally in combination with any of the above or following embodiments, the therapeutically or health-food effective amount of NAC or its pharmaceutically or health-food acceptable salt or solvate is administered at a dose of 35 mg / Kg to 500 mg / Kg. In another more specific embodiment, optionally in combination with any of the above or following embodiments, the therapeutically or health-food effective amount of NAC or its pharmaceutically or health-food acceptable salt or solvate in the human body is administered at a dose of 35 mg / Kg to 50 mg / Kg, or selected from 0.5 mg / Kg, 1 mg / Kg, 2 mg / Kg, 3 mg / Kg, 4 mg / Kg, 5 mg / Kg, 6 mg / Kg, 7 mg / Kg, 8 mg / Kg, 9 mg / Kg, 10 mg / Kg, In some embodiments, the amount of the present invention is 480mg / Kg to 500mg / Kg, more particularly 490mg / Kg. In mice, it is 480mg / Kg to 500mg / Kg, more particularly 490mg / Kg.

[0076] In another specific embodiment, optionally in combination with any of the above or below embodiments, the therapeutically or nutraceutically effective amount of NAC or a pharmaceutically or nutraceutically acceptable salt or solvate thereof in a human body is a dose of 1 mg / Kg to 10 mg / Kg, 1.5 mg / Kg to 7.5 mg / Kg or 2 mg / Kg to 5 mg / Kg, particularly 2 mg / Kg to 5 mg / Kg.

[0077] In another specific embodiment, optionally in combination with any of the above or following embodiments, the therapeutically or nutraceutically effective amount of cysteine ​​or its pharmaceutically or nutraceutically acceptable salt or solvate in the human body is a dose of 20 mg / Kg to 55 mg / Kg, 25 mg / Kg to 50 mg / Kg, 30 mg / Kg to 45 mg / Kg or 35 mg / Kg to 40 mg / Kg, particularly 35 mg / Kg to 40 mg / Kg.

[0078] In a more particular embodiment, the pharmaceutical composition or dietary supplement composition is one in which the therapeutically effective amount of cysteine ​​or its pharmaceutically or nutraceutically acceptable salt or solvate of either is a dosage of 1 mg per Kg of subject to be administered to 7.5 mg per Kg of subject, wherein the subject is preferably a human; the therapeutically or nutraceutically effective amount of histidine or its pharmaceutically or nutraceutically acceptable salt or solvate of either is a dosage of 5 mg per Kg of subject to be administered to 25 mg per Kg of subject, wherein the subject is preferably a human; the therapeutically effective amount of serine or its pharmaceutically or nutraceutically acceptable salt or solvate of either is a dosage of 1 mg per Kg of subject to be administered to 5 mg per Kg of subject, wherein the subject is preferably a human; and the therapeutically effective amount of carnosine or its pharmaceutically or nutraceutically acceptable salt or solvate of either is a dosage of 2 mg per Kg of subject to be administered to 15 mg per Kg of subject, wherein the subject is preferably a human.

[0079] In an even more particular embodiment, the pharmaceutical composition or dietary supplement composition is one in which the therapeutically effective amount of cysteine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either, is a dosage of 2 mg per Kg of subject to be administered to 5 mg per Kg of subject, wherein the subject is preferably a human; the therapeutically or nutraceutically effective amount of histidine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either, is a dosage of 7.5 mg per Kg of subject to be administered to 20 mg per Kg of subject, wherein the subject is preferably a human; the therapeutically effective amount of serine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either, is a dosage of 1.3 mg per Kg of subject to be administered to 3.3 mg per Kg of subject, wherein the subject is preferably a human; and the therapeutically effective amount of carnosine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either, is a dosage of 4.5 mg per Kg of subject to be administered to 10 mg per Kg of subject, wherein the subject is preferably a human.

[0080] In a more particular embodiment, the pharmaceutical composition or dietary supplement composition is one wherein the therapeutically effective amount of NAC or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 1 mg per kg of subject to be administered to 7.5 mg per kg of subject, wherein the subject is preferably a human; the therapeutically or nutraceutically effective amount of histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 5 mg per kg of subject to be administered to 25 mg per kg of subject, wherein the subject is preferably a human; the therapeutically effective amount of serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 1 mg per kg of subject to be administered to 5 mg per kg of subject, wherein the subject is preferably a human; and the therapeutically effective amount of carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 2 mg per kg of subject to be administered to 15 mg per kg of subject, wherein the subject is preferably a human.

[0081] In an even more particular embodiment, the pharmaceutical composition or dietary supplement composition is one wherein the therapeutically effective amount of NAC or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 2 mg per Kg of subject to be administered to 5 mg per Kg of subject, wherein the subject is preferably a human; the therapeutically or nutraceutically effective amount of histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 7.5 mg per Kg of subject to be administered to 20 mg per Kg of subject, wherein the subject is preferably a human; the therapeutically effective amount of serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 1.3 mg per Kg of subject to be administered to 3.3 mg per Kg of subject, wherein the subject is preferably a human; and the therapeutically effective amount of carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 4.5 mg per Kg of subject to be administered to 10 mg per Kg of subject, wherein the subject is preferably a human.

[0082] In a more particular embodiment, the pharmaceutical composition or dietary supplement composition is one in which the therapeutically effective amount of cysteine ​​or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 35 mg per Kg to 40 mg per Kg of subject administered, wherein the subject is preferably a human; the therapeutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 15 mg per Kg to 20 mg per Kg of subject administered, wherein the subject is preferably a human; the therapeutically effective amount of serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 15 mg per Kg to 20 mg per Kg of subject administered, wherein the subject is preferably a human; and the therapeutically effective amount of carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 15 mg per Kg to 20 mg per Kg of subject administered, wherein the subject is preferably a human.

[0083] In a more particular embodiment, the pharmaceutical composition or dietary supplement composition is one wherein the therapeutically effective amount of NAC or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 35 mg per Kg to 40 mg per Kg of subject administered, wherein the subject is preferably a human; the therapeutically or nutraceutically effective amount of histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 15 mg per Kg to 20 mg per Kg of subject administered, wherein the subject is preferably a human; the therapeutically effective amount of serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 15 mg per Kg to 20 mg per Kg of subject administered, wherein the subject is preferably a human; and the therapeutically effective amount of carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either is a dosage of 15 mg per Kg to 20 mg per Kg of subject administered, wherein the subject is preferably a human.

[0084] In another particular embodiment of the first aspect, the composition comprises therapeutically or nutraceutically effective amounts of at least three of the following substances: L-histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, L-cysteine ​​histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, L-serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, and L-carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either.

[0085] In fact, the amino acids in the composition can be either the D-enantiomer or the L-enantiomer, although in particular embodiments they are L-amino acids. Similarly, the dipeptide carnosine can be present in the form of L-carnosine or D-carnosine, although in particular embodiments it is L-carnosine. In one embodiment, the composition of the present invention comprises any combination of any L-form or D-form of the compound. In another embodiment, the composition of the present invention comprises any combination of any amino acid in the L-form or D-form of histidine, serine, carnosine and / or cysteine.

[0086] In a particular embodiment of the first aspect, the composition comprises:

[0087] (i) a therapeutically or health food effective amount of L-cysteine ​​or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of at least two of the following substances: L-histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, L-serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and L-carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two; or, alternatively,

[0088] (ii) a therapeutically or health food effective amount of N-acetylcysteine ​​(NAC) or a pharmaceutically or health food acceptable salt or a solvate of either of the two, and a therapeutically or health food effective amount of at least two of the following substances: L-histidine or a pharmaceutically or health food acceptable salt or a solvate of either of the two, L-serine or a pharmaceutically or health food acceptable salt or a solvate of either of the two, and L-carnosine or a pharmaceutically or health food acceptable salt or a solvate of either of the two.

[0089] In another particular embodiment of the first aspect, the composition comprises:

[0090] (i) a therapeutically or health food effective amount of L-cysteine ​​or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of L-histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of L-serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of L-carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two; or, alternatively,

[0091] (ii) a therapeutically or health food effective amount of NAC or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of L-histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of L-serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of L-carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two.

[0092] In another particular embodiment of the pharmaceutical composition according to the first aspect, the pharmaceutically or health food acceptable salts of histidine and cysteine ​​are hydrochlorides, more particularly the hydrochlorides of L-histidine and L-cysteine. Even more particularly, the pharmaceutically acceptable salt of L-histidine is L-histidine monohydrochloride monohydrate, and the pharmaceutically acceptable salt of L-cysteine ​​is L-cysteine ​​hydrochloride.

[0093] In another particular embodiment of the pharmaceutical composition according to the first aspect, the pharmaceutically or health food acceptable salt of histidine is a hydrochloride, more particularly the hydrochloride of L-histidine. Even more particularly, the pharmaceutically acceptable salt of L-histidine is L-histidine monohydrochloride monohydrate.

[0094] In another particular embodiment of the pharmaceutical composition according to the first aspect, the pharmaceutically or nutraceutically acceptable salt of cysteine ​​is a hydrochloride, more particularly the hydrochloride of L-cysteine. Even more particularly, the pharmaceutically acceptable salt of L-cysteine ​​is L-cysteine ​​hydrochloride.

[0095] In a more particular embodiment of the pharmaceutical composition and dietary supplement composition of the first aspect, it comprises at least three of the indicated ingredients, at least one of which is histidine, more particularly L-histidine or a salt or a solvate of either. In fact, as will be described in the examples below, triple combinations comprising histidine provide good results in increasing the rate of fatty acid β-oxidation by carnitine palmitoyltransferase I. These triple compositions containing at least histidine are actually mixtures of compounds that are ultimately able to increase histidine plasma levels in a subject. Serine and cysteine ​​are metabolized to histidine by various pathways, and carnosine is a source of histidine because it is a dipeptide composed of the amino acids β-alanine and histidine. NAC is a precursor of L-cysteine, which is converted to L-cysteine ​​after ingestion.

[0096] In another particular embodiment of the pharmaceutical composition or dietary supplement composition according to the first aspect, it comprises a therapeutically or nutraceutically effective amount of the following substances:

[0097] (a) histidine, more particularly L-histidine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either; cysteine, more particularly L-cysteine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either, and serine, more particularly L-serine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either; or alternatively,

[0098] (b) histidine, more particularly L-histidine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either; cysteine, more particularly L-cysteine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either, and carnosine, more particularly L-carnosine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either; or alternatively,

[0099] (c) cysteine, more particularly L-cysteine, or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, serine, more particularly L-serine, or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, and carnosine, more particularly L-carnosine, or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either; or alternatively,

[0100] (d) histidine, more particularly L-histidine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, serine, more particularly L-serine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and carnosine, more particularly L-carnosine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two.

[0101] In another particular embodiment of the pharmaceutical composition or dietary supplement composition according to the first aspect, the composition comprises a therapeutically or nutraceutically effective amount of the following substances:

[0102] (a) cysteine, more particularly L-cysteine, or a pharmaceutically or health food acceptable salt thereof, or a solvate of either; histidine, more particularly L-histidine, or a pharmaceutically or health food acceptable salt thereof, or a solvate of either; and serine, more particularly L-serine, or a pharmaceutically or health food acceptable salt thereof, or a solvate of either; or alternatively,

[0103] (b) cysteine, more particularly L-cysteine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either; histidine, more particularly L-histidine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either; and carnosine, more particularly L-carnosine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either; or alternatively,

[0104] (c) cysteine, more particularly L-cysteine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, serine, more particularly L-serine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and carnosine, more particularly L-carnosine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two.

[0105] In another particular embodiment of the pharmaceutical composition or dietary supplement composition according to the first aspect, the composition comprises a therapeutically or nutraceutically effective amount of the following substances:

[0106] (a) NAC or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either; histidine, more particularly L-histidine, or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either; and serine, more particularly L-serine, or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either; or alternatively,

[0107] (b) NAC or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either; histidine, more particularly L-histidine, or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either; and carnosine, more particularly L-carnosine, or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either; or alternatively,

[0108] (c) NAC or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, serine, more particularly L-serine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and carnosine, more particularly L-carnosine, or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two.

[0109] In a more particular embodiment of the first aspect of the present invention, the composition comprises a therapeutically or nutraceutically effective amount of histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two; a therapeutically or nutraceutically effective amount of cysteine ​​or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, a therapeutically or nutraceutically effective amount of serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, and a therapeutically or nutraceutically effective amount of carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, and one or more pharmaceutically or nutraceutically acceptable excipients or carriers.

[0110] More specifically, it comprises a therapeutically or health food effective amount of L-histidine or a pharmaceutically acceptable salt thereof or a solvate of either of the two; a therapeutically or health food effective amount of L-cysteine ​​or a pharmaceutically acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of L-serine or a pharmaceutically acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of L-carnosine or a pharmaceutically acceptable salt thereof or a solvate of either of the two, and one or more pharmaceutically acceptable excipients or carriers.

[0111] In another more specific embodiment, the composition comprises a therapeutically or nutraceutically effective amount of L-histidine or a pharmaceutically acceptable salt thereof or a solvate of either of the two; a therapeutically or nutraceutically effective amount of NAC or a pharmaceutically acceptable salt thereof or a solvate of either of the two, a therapeutically or nutraceutically effective amount of L-serine or a pharmaceutically acceptable salt thereof or a solvate of either of the two, and a therapeutically or nutraceutically effective amount of L-carnosine or a pharmaceutically acceptable salt thereof or a solvate of either of the two, and one or more pharmaceutically acceptable excipients or carriers.

[0112] In an even more particular embodiment, the composition comprises:

[0113] - a therapeutically or health food effective amount of L-histidine monohydrochloride monohydrate;

[0114] - a therapeutically or health food effective amount of L-cysteine ​​hydrochloride;

[0115] - a therapeutically or health food effective amount of L-serine; and

[0116] - L-carnosine in an effective amount for treatment or health food,

[0117] and one or more than one excipient or carrier acceptable in medicine or health food.

[0118] In another even more particular embodiment, the composition comprises:

[0119] - a therapeutically or health food effective amount of L-histidine monohydrochloride monohydrate;

[0120] - A therapeutically or nutraceutically effective amount of NAC;

[0121] - a therapeutically or health food effective amount of L-serine; and

[0122] - L-carnosine in an effective amount for treatment or health food,

[0123] and one or more than one excipient or carrier acceptable in medicine or health food.

[0124] Specific therapeutic or nutraceutical amounts have been disclosed in the preceding paragraphs.

[0125] In particular embodiments, the composition for human consumption comprises:

[0126] - a therapeutically effective amount of L-histidine monohydrochloride monohydrate in humans or in health food, administered at a dose of 17 mg / kg;

[0127] - a therapeutically or health food effective amount of L-cysteine ​​hydrochloride in humans, administered at a dose of 39.7 mg / kg;

[0128] - a therapeutically or health food effective amount of L-serine in humans, administered at a dose of 17 mg / Kg; and

[0129] - L-carnosine effective in human body in therapeutic or health food, administered in a dose of 17 mg / Kg;

[0130] and one or more than one pharmaceutically acceptable excipient or carrier.

[0131] In another particular embodiment, a composition for human consumption comprises:

[0132] - a therapeutically effective amount of L-histidine monohydrochloride monohydrate in humans or in health food, administered at a dose of 17 mg / kg;

[0133] - a therapeutically or health food effective amount of NAC in humans, administered at a dose of 39.7 mg / Kg;

[0134] - a therapeutically or health food effective amount of L-serine in humans, administered at a dose of 17 mg / Kg; and

[0135] - L-carnosine effective in human body in therapeutic or health food, administered in a dose of 17 mg / Kg;

[0136] and one or more than one pharmaceutically acceptable excipient or carrier.

[0137] In another particular embodiment, a composition for human consumption comprises:

[0138] - a therapeutically or health food effective amount of L-histidine monohydrochloride monohydrate in humans, administered at a dose of 7.5 mg / Kg to 20 mg / Kg;

[0139] - a therapeutically or health food effective amount of L-cysteine ​​hydrochloride in humans, administered at a dose of 2 mg / Kg to 5 mg / Kg;

[0140] - a therapeutically or health food effective amount of L-serine in humans, administered at a dose of 1.3 mg / Kg to 3.3 mg / Kg; and

[0141] - a therapeutically or health food effective amount of L-carnosine in the human body, administered at a dose of 4.5 mg / Kg to 10 mg / Kg;

[0142] and one or more than one pharmaceutically acceptable excipient or carrier.

[0143] In another particular embodiment, a composition for human consumption comprises:

[0144] - a therapeutically or health food effective amount of L-histidine monohydrochloride monohydrate in humans, administered at a dose of 7.5 mg / Kg to 20 mg / Kg;

[0145] - a therapeutically or nutraceutically effective amount of NAC in humans, administered at a dose of 2 mg / Kg to 5 mg / Kg;

[0146] - a therapeutically or health food effective amount of L-serine in humans, administered at a dose of 1.3 mg / Kg to 3.3 mg / Kg; and

[0147] - a therapeutically or health food effective amount of L-carnosine in the human body, administered at a dose of 4.5 mg / Kg to 10 mg / Kg;

[0148] and one or more than one pharmaceutically acceptable excipient or carrier.

[0149] In another particular embodiment of the first aspect, the composition comprises a therapeutically or nutraceutically effective amount of a peptide comprising an amino acid sequence containing at least three of histidine, cysteine, serine and carnosine in any of their L-configuration or D-configuration and combinations thereof (i.e. one compound is in the D-isomer form and the other is in the L-isomer form).

[0150] In another specific embodiment of the first aspect, the composition comprises a therapeutically or nutraceutically effective amount of a peptide comprising an amino acid sequence containing cysteine ​​in any of its L-configuration or D-configuration and combinations thereof (i.e. one compound is in the form of a D-isomer and the other is in the form of an L-isomer). In another specific embodiment of the first aspect, the composition comprises a therapeutically or nutraceutically effective amount of a peptide comprising an amino acid sequence containing cysteine, histidine, serine and carnosine in any of its L-configuration or D-configuration and combinations thereof (i.e. one compound is in the form of a D-isomer and the other is in the form of an L-isomer).

[0151] In another specific embodiment of the first aspect, the composition comprises a therapeutically or nutraceutically effective amount of a peptide comprising an amino acid sequence comprising NAC and at least two of histidine, serine and carnosine, wherein at least two of histidine, serine and / or carnosine are in any of their L-configurations or D-configurations and combinations thereof (i.e., one compound is in the form of a D-isomer and the other is in the form of an L-isomer). In another specific embodiment of the first aspect, the composition comprises a therapeutically or nutraceutically effective amount of a peptide comprising an amino acid sequence comprising NAC, histidine, serine and carnosine, wherein histidine, serine and carnosine are in any of their L-configurations or D-configurations and combinations thereof (i.e., one compound is in the form of a D-isomer and the other is in the form of an L-isomer).

[0152] In another particular embodiment, these peptides having a sequence containing at least three of the indicated compounds have a length of 3 to 50 amino acids, more particularly a length of 3 to 10 amino acids, even more particularly a length selected from 3, 4, 5, 6, 7, 8, 9 and 10 amino acids.

[0153] In a more particular embodiment, the peptide is selected from the group consisting of a tripeptide, a tetrapeptide and a pentapeptide.

[0154] In a preferred embodiment, the peptide in the composition is a peptide comprising at least one histidine residue. In another preferred embodiment, the peptide in the composition is a peptide comprising at least one cysteine ​​residue or NAC residue. In another preferred embodiment, the peptide in the composition is a peptide comprising at least one histidine residue and at least one cysteine ​​residue, more preferably at least one histidine residue and at least one NAC residue.

[0155] In another particular embodiment, the tetrapeptide comprises at least three amino acids selected from histidine, cysteine, serine and carnosine, and the other amino acids are any amino acids known to those skilled in the art, including all naturally occurring amino acids, in particular any amino acids selected from the 21 proteinogenic α-amino acids found in eukaryotic organisms.

[0156] In another particular embodiment, the tetrapeptide comprises cysteine ​​and at least two amino acids selected from histidine, serine and carnosine, the other amino acids being any amino acids known to those skilled in the art, including all naturally occurring amino acids, in particular any amino acids selected from the 21 proteinogenic α-amino acids found in eukaryotes. In another particular embodiment, the tetrapeptide comprises NAC and at least two amino acids selected from histidine, cysteine ​​and carnosine, the other amino acids being any amino acids known to those skilled in the art, including all naturally occurring amino acids, in particular any amino acids selected from the 21 proteinogenic α-amino acids found in eukaryotes.

[0157] Amino acids in the sense 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 include 2-aminoadipic acid (Aad), 3-aminoadipic acid (bAad), β-alanine (bAla), 2-aminobutyric acid (Abu), 6-aminohexanoic 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 (EtG ly), N-ethylasparagine (EtAsn), hydroxylysine (Hyl), allo-hydroxylysine (aHyl), 3-hydroxyproline (3Hyp), 4-hydroxyproline (4Hyp), isodesmosine (Ide), allo-isoleucine (alle), N-methylisoleucine (Melle), 6-N-methyllysine (MeLys), N-methylvaline (Meval), norvaline (Nva), norleucine (Nle), and ornithine (Orn). In various embodiments, one amino acid can be substituted by another while maintaining hydrophobicity, aromaticity, and basicity at each selected position.

[0158] In another specific embodiment, the peptide comprises histidine, cysteine, serine and carnosine, and is at least a pentapeptide, preferably a pentapeptide. In another specific embodiment, the peptide comprises histidine, NAC, serine and carnosine, and is at least a pentapeptide, preferably a pentapeptide.

[0159] In all of these possible peptides of the composition, the amino acid residues are arranged in any order in the sequence read from the N-terminus to the C-terminus.

[0160] Once the composition is administered, the peptides contained therein will be naturally hydrolyzed by common cellular and bodily processes (ie, hydrolases, proteases, etc.) and the separate portions will be delivered.

[0161] Dosage adjustments for these peptides will vary, but in particular embodiments will be adjusted to administer the above indicated doses of the individual compounds as previously disclosed.

[0162] The preparation of all these peptides is according to common practices known to those skilled in the art.

[0163] All pharmaceutical compositions or dietary supplement compositions of the present invention contain common excipients and / or carriers.

[0164] Examples of suitable pharmaceutically and nutraceutically acceptable excipients are solvents, dispersion media, diluents or other liquid carriers, dispersing aids or suspension aids, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, etc. Unless any conventional excipient medium is incompatible with the substance or its derivatives, for example by producing any undesirable biological effect or otherwise interacting with any other one or more components of the pharmaceutical composition in a deleterious manner, its use should be considered within the scope of the present invention.

[0165] The relative amounts of the active ingredients, pharmaceutically or nutraceutically acceptable excipients, and / or any other ingredients in the pharmaceutical or nutraceutical compositions of the present invention will vary depending on the identity, size and / or condition of the subject being treated and the route of administration of the composition.

[0166] The acceptable excipients for the manufacture of the pharmaceutical composition or the health food include, but are not limited to, inert diluents, dispersants and / or granulating agents, surfactants and / or emulsifiers, disintegrating agents, binding agents, preservatives, buffers, lubricants and / or oils. According to the judgement of the formulation technician, excipients such as coloring agents, film-forming agents, sweeteners and aromatics can be added to the composition.

[0167] The pharmaceutical composition or health food composition containing a combination of at least three of histidine, carnosine, serine and cysteine ​​is, for example, solid or liquid, and can be administered by any suitable route, such as oral route, parenteral route, rectal route, topical route, intranasal route, intraocular route, peritoneal route or sublingual route, for which it will contain the pharmaceutically or health food acceptable excipients required for the preparation of the desired dosage form. More particularly, the pharmaceutical composition or dietary supplement composition of the first aspect of the present invention is, for example, solid or liquid, and can be administered by any suitable route, such as oral route, parenteral route, rectal route, topical route, intranasal route, intraocular route, peritoneal route or sublingual route, for which it will contain the pharmaceutically or health food acceptable excipients required for the preparation of the desired dosage form.

[0168] Exemplary diluents include, but are not limited to, calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, dibasic calcium phosphate, sodium phosphate, lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and combinations thereof.

[0169] 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 sponges, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked polyvinyl pyrrolidone (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (croscarmellose), methyl cellulose, pregelatinized starch (starch 1500), microcrystalline starch, water-insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and combinations thereof.

[0170] 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, Irish moss extract, panwar gum, ghatti gum, mucilage of isapolhusks, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, microcrystalline cellulose, cellulose acetate, polyvinylpyrrolidone), magnesium aluminum silicate (Veegum), and larch arabogalactan); alginates; polyethylene oxides; polyethylene glycols; inorganic calcium salts; silicic acid; polymethacrylates; waxes; water; alcohols; and combinations thereof.

[0171] 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 pyrosulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium pyrosulfite and sodium sulfite. Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA), citric acid monohydrate, disodium ethylenediaminetetraacetate, dipotassium ethylenediaminetetraacetate, ethylenediaminetetraacetic acid, fumaric acid, malic acid, phosphoric acid, sodium ethylenediaminetetraacetate, tartaric acid and trisodium ethylenediaminetetraacetate.

[0172] Exemplary buffers include, but are not limited to, citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glucuronate, calcium glucoheptonate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propionic acid, calcium levulinate, valeric acid, dicalcium phosphate, phosphoric acid, tricalcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dipotassium phosphate, monopotassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, disodium phosphate, monosodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethanol, and combinations thereof.

[0173] Exemplary lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, silicon dioxide, 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.

[0174] The preparation of all these pharmaceutical compositions or health food compositions is according to conventional practices known to those skilled in the art.

[0175] As indicated above, another aspect of the invention is a food in solid or liquid form comprising a pharmaceutical composition or dietary supplement composition as defined in the first aspect. In a more particular embodiment, the food is a beverage. These foods or dietary supplements are in fact functional foods or supplements that provide health benefits even in the absence of pathological conditions. Therefore, they are considered to be compositions that can prevent or reduce the risk of diseases such as those associated with fat deposition in tissues.

[0176] When the food or composition disclosed in the first aspect or the second aspect and embodiments thereof is applied to mice suffering from NAFLD, the animals are effectively treated and the fat accumulated in the liver is reduced. Similarly, the body weight due to fat body weight is also reduced. In addition, in experiments analyzing the risk of atherosclerosis, it was observed that when animals received the composition of the present invention, the risk was reduced. In addition, when the composition of the present invention is applied to ovariectomized rats showing metabolic changes associated with menopause, the values ​​of the corresponding metabolic parameters become closer to those of control rats. The composition of the present invention is applied to mice induced with NAFLD, reducing the expression of bacterial histidine utilization genes found to be increased in NAFLD patients, thereby offsetting microbiome imbalance or dysbiosis. Therefore, as a third aspect, the pharmaceutical composition or dietary supplement composition or food is proposed for treatment.

[0177] In a particular embodiment of the third aspect of the present invention, the composition of the first aspect or the food of the second aspect is used to prevent and / or treat a disease selected from fatty liver disease, obesity, overweight, atherosclerosis, dysbiosis and a combination thereof.

[0178] This can also be expressed as the use of the composition or food of the present invention in the preparation of a medicament for preventing and / or treating a condition or disease selected from fatty liver disease, obesity, overweight, atherosclerosis, dysbiosis and a combination thereof. The present invention also relates to a method for preventing and / or treating a condition or disease selected from fatty liver disease, obesity, overweight, atherosclerosis, dysbiosis and a combination thereof, which comprises administering a therapeutically or health food effective amount of the composition or food, together with a pharmaceutically or health food acceptable excipient and / or carrier, to a subject including a human being in need thereof.

[0179] The term "obesity" as used herein refers to abnormal or excessive fat accumulation in a subject, particularly a mammal, more particularly a human, that poses a risk to health. A BMI associated with obesity in humans, particularly adults, is 30 or above. Obesity is generally subdivided into 3 categories:

[0180] Obesity level 1: BMI 30 to <35

[0181] Stage 2 obesity: BMI 35 to <40

[0182] Class 3 obesity: A BMI of 40 or higher. Class 3 obesity is sometimes classified as “severe” obesity.

[0183] BMI classification for children and adolescents is usually based on sex-specific and age-specific BMI percentiles, whereas BMI classification for adults is based on BMI alone. Obesity in children and adolescents is further subdivided into two categories:

[0184] Grade 2 obesity: BMI ≥120% to <140% of the 95th percentile, or BMI ≥35kg / m2 to <40kg / m2

[0185] Grade 3 obesity: BMI ≥ 140% of the 95th percentile, or BMI ≥ 40 kg / m2

[0186] As used herein, the term "body mass index" or "BMI" refers to a subject's weight in kilograms (or pounds) divided by the square of their height in meters (or feet).

[0187] In a particular embodiment of the third aspect of the present invention, the composition or food is used to prevent and / or treat obesity in adults. In another particular embodiment of the third aspect of the present invention, the composition or food is used to prevent and / or treat obesity in minors. As is well known to experts in the field, minors are people less than 18 years old, and adults are people 18 years old or older.

[0188] In a particular embodiment of the third aspect of the present invention, the composition or food is used to prevent and / or treat grade 1 obesity, grade 2 obesity and / or grade 3 obesity, in particular obesity in adults. In a particular embodiment, it is used to prevent and / or treat grade 1 obesity, in particular grade 1 obesity in adults. In another embodiment, it is used to prevent and / or treat grade 2 obesity, in particular grade 2 obesity in adults. In another embodiment, it is used to prevent and / or treat grade 3 obesity, in particular grade 3 obesity in adults. In another embodiment, it is used to prevent and / or treat grade 1 obesity or grade 2 obesity, in particular grade 1 obesity or grade 2 obesity in adults. In another embodiment, it is used to prevent and / or treat grade 2 obesity or grade 3 obesity, in particular grade 2 obesity or grade 3 obesity in adults.

[0189] In a special embodiment of the third aspect of the present invention, the composition or food is used to prevent and / or treat grade 2 obesity and / or grade 3 obesity, in particular obesity in minors. In a special embodiment, it is used to prevent and / or treat grade 2 obesity, in particular grade 2 obesity in minors. In another embodiment, it is used to prevent and / or treat grade 3 obesity, in particular grade 3 obesity in minors. In a special embodiment of the third aspect of the present invention, the composition or food is used to prevent and / or treat grade 2 obesity and grade 3 obesity, in particular grade 2 obesity and grade 3 obesity in minors. In a special embodiment of the third aspect of the present invention, the composition or food is used to prevent and / or treat grade 2 obesity or grade 3 obesity, in particular grade 2 obesity or grade 3 obesity in minors.

[0190] The term "overweight" as used herein refers to an abnormal or excessive accumulation of fat in a subject, particularly a mammal, more particularly a human being, which constitutes a risk to health and which in humans, particularly adults, is associated with a BMI of 25 to 30 as defined above, and for children and adolescents, is associated with a BMI of the 85th percentile to less than the 95th percentile.

[0191] In a particular embodiment of the third aspect of the present invention, the composition or food is used to prevent and / or treat overweight, particularly overweight in adults. In another particular embodiment of the third aspect of the present invention, the composition or food is used to prevent and / or treat overweight in minors.

[0192] The term "dysbiosis" refers to any condition in which the normal diversity or normal function of the body's microbiome, particularly the intestinal microbiome, is disrupted.

[0193] In a more particular embodiment, the composition of the first aspect or the food of the second aspect is used for preventing and / or treating fatty liver disease selected from non-alcoholic fatty liver disease and alcoholic fatty liver disease.

[0194] In a more particular embodiment, it is used for the prevention and / or treatment of non-alcoholic fatty liver disease selected from non-alcoholic fatty liver disease with simple steatosis (NAFL) and non-alcoholic steatohepatitis (NASH).

[0195] In another particular embodiment of the third aspect of the present invention, the composition or food is used for preventing and / or treating symptoms associated with menopause, especially metabolic changes associated with menopause, in female mammals, especially female humans.

[0196] This can also be expressed as the use of the composition or food of the present invention in the preparation of a medicament for preventing and / or treating symptoms associated with menopause, especially metabolic changes associated with menopause, in female mammals, especially female humans. The present invention also relates to a method for preventing and / or treating symptoms associated with menopause, especially metabolic changes associated with menopause, which comprises administering a therapeutically or health food effective amount of the composition or food, together with a pharmaceutically or health food acceptable excipient and / or carrier, to a female mammal including a female human in need thereof.

[0197] In another particular embodiment of the third aspect of the present invention, the composition or food is used to prevent and / or treat symptoms associated with menopause, especially metabolic changes associated with menopause, and to prevent and / or treat diseases selected from fatty liver disease, obesity, overweight, atherosclerosis, dysbiosis and combinations thereof in female mammals, especially female humans.

[0198] This can also be expressed as the use of the composition or food of the present invention in the preparation of a medicament for preventing and / or treating symptoms associated with menopause, especially metabolic changes associated with menopause, and preventing and / or treating diseases selected from fatty liver disease, obesity, overweight, atherosclerosis, dysbiosis and combinations thereof in female mammals, especially female humans. The present invention also relates to a method for preventing and / or treating symptoms associated with menopause, especially metabolic changes associated with menopause, and preventing and / or treating diseases selected from fatty liver disease, obesity, overweight, atherosclerosis, dysbiosis and combinations thereof, which comprises administering a therapeutically or health food effective amount of the composition or food, together with a pharmaceutically or health food acceptable excipient and / or carrier, to a female mammal, including a female human, in need thereof.

[0199] As used herein, the term "symptoms associated with menopause" refers to metabolic changes, changes in body weight, cardiovascular changes and musculoskeletal changes, urogenital atrophy and skin atrophy; sexual dysfunction and disorders associated with the central nervous system, which usually occur in female mammals, especially female humans, as a consequence of menopause. The term "metabolic changes associated with menopause" as used herein refers to changes in metabolic-related parameters that occur in female mammals, especially female humans, as a consequence of menopause. Non-limiting examples of the parameters include body weight, obesity and insulin resistance. More specifically, the parameters include body weight, fat body mass, lean body mass, fat accumulation, obesity index, insulin resistance.

[0200] In a particular embodiment, the metabolic changes associated with menopause are selected from the group consisting of increased weight, increased fat, increased insulin resistance in a female mammal, particularly a human female, during the menopausal transition, menopause or postmenopause compared to before menopause, particularly before starting the menopausal transition.

[0201] In another particular embodiment, the metabolic changes associated with menopause are selected from the group consisting of increased body weight, increased fat body mass, decreased lean body mass, increased fat accumulation, increased adiposity index, and increased insulin resistance in female mammals, particularly human females, during the menopausal transition, menopause or postmenopause, compared to before menopause, particularly before the onset of the menopausal transition.

[0202] In a specific embodiment, "premenopause" refers to 1 year, 2 years, 3 years, 4 years, preferably 1 year before the onset of menopause. In another specific embodiment, "before the onset of the menopausal transition" refers to 1 year, 2 years, 3 years, 4 years, preferably 1 year before the onset of the menopausal transition.

[0203] The term "menopause" as used herein refers to a state in female mammals, preferably female humans, characterized by the ovaries no longer releasing eggs or producing very little sex hormones. Clinically, natural menopause is defined as a 1-year amenorrhea period after the final menstrual period (FMP) without any pathological or physiological reasons.

[0204] The term "menopausal transition" or "perimenopause" refers to a defined period from the onset of irregular menstrual cycles until the last menstrual period, which is characterized by fluctuations in reproductive hormones.

[0205] As used herein, the term "postmenopause" refers to the period of time after the onset of menopause, ie, the period beginning 12 months after the final menstrual period (FMP).

[0206] In another particular aspect, the present invention relates to a non-therapeutic method for reducing the weight of an overweight subject, comprising administering the composition of the first aspect of the present invention or the food of the second aspect of the present invention to the subject, wherein the subject is particularly a mammal, even more particularly a human being. In a particular embodiment, the dosage administered is the dosage of the composition of the present invention as disclosed in the first aspect of the present invention. In another particular embodiment, the route of administration is those of the composition of the present invention as disclosed in the first aspect of the present invention. In a particular embodiment, the human being is an adult. In another particular embodiment, the human being is a minor.

[0207] In a particular embodiment, reducing the overweight of an overweight subject involves reducing the BMI of the subject by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, in particular at least 5%. In another particular embodiment, reducing the overweight of an overweight subject involves reducing the BMI of the subject by at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, in particular at least 1.

[0208] As will be demonstrated in the examples, a diet rich in fat and sugar, when supplemented with a specific combination of histidine, serine, cysteine ​​and carnosine, can reduce fat accumulation in the liver and other body tissues.

[0209] Therefore, the present invention also discloses for the first time a combination comprising: a therapeutically or health food effective amount of histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either; a therapeutically or health food effective amount of cysteine ​​or a pharmaceutically or health food acceptable salt thereof or a solvate of either, a therapeutically or health food effective amount of serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either, and a therapeutically or health food effective amount of carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either, and one or more pharmaceutically or health food acceptable excipients or carriers. This combination has never been disclosed and is now proposed as a dietary supplement for subjects at risk of one or more of fatty liver disease, obesity and atherosclerosis. The first and second aspects related to the amount (dosage) of any compound and to the isomers of the compound indicated in any of the aforementioned embodiments also apply to the combination of the four components indicated.

[0210] The first and second aspects indicated in any of the preceding embodiments relating to the amount (dosage) of any compound and to the isomers of the compound also apply to the indicated combination of four components, wherein cysteine ​​is substituted by NAC.

[0211] Throughout the specification and claims, the words "comprise" or "include" and their variations are not intended to exclude other technical features, additives, components or steps. In addition, the word "comprising" covers the situation of "consisting of..." Other objects, advantages and features of the present invention will become obvious to those skilled in the art after reviewing the specification, or can be learned by practicing the present invention. The examples below are provided by way of illustration and are not intended to limit the present invention. In addition, the present invention covers all possible combinations of the particular embodiments and preferred embodiments described herein. Example

[0212] 1.-Human cohort trials.

[0213] 1.1.- Circulating histidine levels in different cohorts depending on the degree of steatosis.

[0214] To obtain detailed information on hepatic steatosis in patients with NAFLD, metabolic profiling of urine and plasma was performed by 1H-NMR spectroscopy. Metabolome-wide association analysis (MWAS) found that 124 metabolite signals in urine and 80 metabolite signals in plasma were associated with hepatic steatosis and related clinical features. Among some metabolites of hepatic steatosis that were strongly associated with the grade of NAFLD, a significant negative correlation was found between plasma histidine and a significant decrease in urine histidine. In the discovery study, plasma histidine was negatively correlated with hepatic steatosis in 102 patients and 56 patients, respectively, which was also confirmed in two validation cohort studies of 313 patients and 283 patients, respectively. This cohort study of 283 patients also showed that obese patients without steatosis had decreased histidine levels compared with lean patients without steatosis. When the degree of steatosis increased, the circulating histidine level decreased, confirming the negative correlation between histidine level and hepatic steatosis.

[0215] Example 1. A composition containing histidine, serine, cysteine ​​and carnosine is used as a supplement to improve clinical symptoms of hepatic steatosis.

[0216] Materials and methods

[0217] Animal models and diet

[0218] At the beginning of the experiment, 48 6-week-old C57BL / 6J male mice (Envigo, Sant Feliu de Codines, Barcelona, ​​Spain) were used. The animals were housed in groups (4 mice per cage) under controlled temperature (22°C ± 2°C) and controlled humidity (55% ± 10%) conditions with a 12-h light-dark cycle and free access to food and water. The mice were allowed to acclimate to the animal facility for one week without interference. After the acclimatization period, the animals were randomly divided into two experimental groups with different diets. 16 control mice were maintained on a standard diet (D12328, Research Diets, New Brunswick, NJ), and 32 animals (NAFLD group) were fed with a HFHFr diet (HFHC: D12331, Research Diets) supplemented with 23.1 g / L fructose and 18.9 g / L sucrose in the drinking water. The mice were maintained on these diets under free conditions for 20 weeks. These specific doses were determined based on previous studies and dose conversion calculations from human doses to animal doses. 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 were guided by the “Principles of Laboratory Care” and were performed in accordance with the European Communities Council Directive (86 / 609 / EEC).

[0219] In the last 4 weeks of the experiment (weeks 16 to 20), NAFLD mice were randomly divided into two groups: 16 mice were kept under the same feeding conditions as before (HFHFr group), and 16 mice were exposed to histidine-related amino acid treatment (AA group). AA is a mixture of the following compounds: 210 mg / kg of L-histidine monohydrochloride monohydrate (Merck, Germany GmbH), 490 mg / kg of L-cysteine ​​hydrochloride (Merck, Germany GmbH), 210 mg / kg of L-serine (Merck, Germany GmbH) and 210 mg / kg of L-carnosine 98% (Acros Organics, Geel, Belgium). Histidine, cysteine, serine and carnosine were diluted with drinking water (vehicle) containing 23.1 g / L fructose and 18.9 g / L sucrose. Fresh solutions were freshly prepared three times a week, prepared with storage powders, and protected from light. Before euthanasia, 10 animals were randomly selected from each group for insulin stimulation. They were intraperitoneally injected with 1 mU / g insulin (n=5 / group) or saline (n=5 / group) and sacrificed 15 min later.

[0220] 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 assay (QCA, Barcelona, ​​Spain). Fasting blood insulin and blood glucose were measured using a mouse insulin ELISA kit (Mercodia, Uppsala, Sweden) and a glucose liquid kit (QCA, Barcelona, ​​Spain), respectively. The liver was quickly collected, weighed and divided into two parts - the left inner lobe of the liver was preserved in formalin, and the remaining tissue was frozen in liquid nitrogen and stored at -80°C until further analysis.

[0221] Liver fat quantification

[0222] Extraction and quantification of liver lipids. Briefly, total lipids were extracted from 80 mg to 100 mg liver sections by adding 1 mL of hexane / isopropanol (3:2, v / v) and degassing with nitrogen. They were then orbitally shaken overnight at room temperature in the dark. After extraction with 0.3 mL of Na2SO4 (0.47 M), the organic layer was separated and dried with nitrogen. Total lipids were quantified by weight analysis before emulsification. Triglycerides and total cholesterol were detected using a commercial enzyme kit (QCA).

[0223] Histological evaluation

[0224] Parts of the liver fixed in buffered formalin (4% formaldehyde, 4 g / L NaH2PO4, 6.5 g / L Na2HPO4; pH 6.8) were cut at a thickness of 3.5 μm and stained with hematoxylin-eosin (H&E) and trichrome staining. Liver images (magnification 40 times) were taken using a microscope (ECLIPSE Ti; Nikon, Tokyo, Japan) connected to a digital sighting 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, the study had a double-blind design to prevent the reviewers from knowing any data from the mice during the histopathological analysis. A universal NAFLD scoring system was established to diagnose mice with NAFLD / NASH. The main features of NAFLD and NASH are classified as follows: Steatosis is assessed by analyzing macrovesicular steatosis (0 to 3) and microvesicular steatosis (0 to 3) separately, followed by hepatocyte hypertrophy (0 to 3), which assesses abnormal enlargement of cells, giving a total of 9 points for the steatosis status. Inflammation is scored by counting cell aggregates (inflammatory foci). The scores range from 0 to 3 depending on the degree of the feature. It is divided into 0 (<5%), 1 (5% to 33%), 2 (34% to 66%) and 3 (>66%), and this score is used for each feature of steatosis and then added to the total steatosis score. 23 Ballooning was not included in the scoring system because the rodent NAFLD score considers only quantitative parameters. It is important to emphasize that hypertrophy is not a sign of cell damage but rather refers to an abnormal enlargement of cells without distinguishing the cause of this enlargement.

[0225] RNA extraction and quantitative polymerase chain reaction

[0226] Homogenates of 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 measured using a nanophotometer (Implen GmbH, Munich, Germany). RNA was converted to cDNA using a high-capacity RNA-to-cDNA kit (Applied Biosystems, Wilmington, DE, USA). The cDNA was expressed in 4% cDNA using a commercial LightCycler 480 Sybrgreen I master. ®Before incubation on 480 II (Roche Diagnostics GmbH, Mannheim, Germany), cDNA was diluted 1:10. Table 1 shows a list of primers used, which have been previously described in other studies and validated using Primer-Blast software (National Center for Biotechnology Information, Bethesda, MD, USA). As previously described, 36b4 was used as a housekeeping gene.

[0227] Table 1. Sequences of oligonucleotides used in RT-PCR.

[0228]

[0229] Protein extraction and western blot analysis

[0230] 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 orthovanadate, 2 mmol / L PMSF, and 1% protease inhibitor cocktail (MMilliporeSigma, Darmstadt, Germany)) for 50 s using 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 µg to 25 µg) were separated by electrophoresis on 10% SDS-PAGE and electrotransfected onto nitrocellulose membranes (Li-cor biosciences, NE, USA). Efficient protein transfer was monitored by Ponceau S staining. Next, the membrane was blocked (5% BSA) at room temperature and probed with specific primary antibodies (diluted 1:1000) in 1% BSA at 4°C overnight: total Akt (4685) (CST, Danvers, MA, USA), phosphorylated Akt (Ser473) (4060) (CST), and β-actin (Santa Cruz Biotechnology, Inc; Dallas, TX, USA). Thereafter, 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 used for detection and quantified using ImageJ.

[0231] Statistical analysis

[0232] Statistical analysis was performed using GraphPad Prism 9 software (Graph-Pad Soft-ware, La Jolla, CA, USA). Data are expressed as mean ± SEM. Data distribution was analyzed using the Shapiro–Wilk normality test. Unpaired t-test (two-tailed, 95% confidence interval) was used to determine the differences between the two groups. One-way analysis of variance (ANOVA) was used to test the differences between the three groups. P values ​​below 0.05 were considered statistically significant. Statistical analysis was performed using GraphPad Prism 9 software (Graph-Pad Soft-ware, La Jolla, CA, USA). Data are expressed as mean ± SEM. Data distribution was analyzed using the Shapiro–Wilk normality test. Unpaired t-test (two-tailed, 95% confidence interval) was used to determine the differences between the two groups. One-way analysis of variance (ANOVA) was used to test the differences between the three groups. P values ​​below 0.05 were considered statistically significant.

[0233] result

[0234] Specific amino acid combination alleviates liver damage and liver characteristics in NAFLD

[0235] After 4 weeks of treatment with specific amino acids (AA) consisting of histidine, serine, cysteine, and carnosine, the animals were sacrificed and liver weights were analyzed. Liver weights in the AA-supplemented group showed a trend toward reduction compared with HFHFr animals, which had a significant increase in liver weight compared with the control group ( Figure 1 (A)). The differences in liver weight found between the AA and HFHFr groups were not associated with differences in solid or liquid intake (data not shown). In addition, macroscopic observations were performed and showed that the HFHFr animals presented with pale liver and fatty liver compared with the control group, whereas AA treatment partially restored a healthy liver appearance compared with the HFHFr group ( Figure 1 (B)). The AA group showed significantly decreased AST levels compared to the HFHFr animals. However, the ALT in the AA group did not show any significant changes compared to the ALT in the HFHFr group, whereas the HFHFr group showed significant increases in both aminotransferases compared to the control group ( Figure 1 (C) and Figure 1 (D)).

[0236] AA supplementation reduces liver lipid content and hepatic steatosis

[0237] After evaluating the liver status, the present study analyzed the liver lipid content and the characteristics of hepatic steatosis. A significant increase in the levels of total lipid content, total cholesterol, and triglycerides in the liver was observed in the HFHFr group compared with the control group. Interestingly, AA-supplemented animals showed a significant decrease in hepatic triglyceride levels compared with the HFHFr group ( Figure 2 (B)), but this effect was not reflected in total liver cholesterol levels ( Figure 2 (C)). However, the total liver lipid levels in the AA group showed a slight decrease compared with the HFHFr group ( Figure 2 (A)).

[0238] In line with the liver status assessment and biochemical analysis, liver histological analysis showed that the HFHFr group had significant hepatic steatosis, manifested by the presence of macrovesicular steatosis and microvesicular steatosis, accompanied by increased hypertrophy, which triggered nuclear displacement. Of note, treatment with AA supplementation improved hepatic steatosis, in fact, microvesicular steatosis was particularly reduced, the surface area occupied by macrovesicular steatosis was also reduced, hypertrophy was reduced, and nuclear displacement was alleviated ( Figure 3 (A)). As a result, a significant decrease in lipid droplet amount and lipid surface area was observed ( Figure 3 (B) and Figure 3 (C)). NAFLD / NASH mice were diagnosed by establishing a universal NAFLD scoring system, which confirmed significant improvements in the different parameters analyzed in animals treated with AA supplementation compared to the HFHFr group ( Figure 3 (D)).

[0239] To evaluate whether this specific combination of amino acids modulates the levels of pro-inflammatory markers, the expression levels of some representative inflammatory genes were analyzed. In correlation with previous results of this study, the macrophage infiltration marker F4 / 80 was significantly downregulated in AA-supplemented animals compared to the HFHFr group, while the HFHFr group showed a significant upregulation of this gene level compared to the control group. Similar results were also observed for TNFα mRNA levels, which were significantly increased in HFHFr animals compared to the control group, although TNFα expression was significantly downregulated in the AA-supplemented group compared to their counterparts (data not shown).

[0240] The beneficial effects of AA supplementation do not involve lipogenesis or lipid transport pathways

[0241] To understand which metabolic pathways may be involved in the improvement of NAFLD after AA supplementation, analysis of gene expression related to lipid metabolism, such as hepatic lipogenesis, lipid transport, etc., was performed. The HFHFr group showed a significant increase in the genes Cd36 and Fabp4, which are related to lipid transport. In the AA group, no effect was observed on these two genes related to lipid transport, and these genes maintained high expression levels compared to the control group. Significant changes in the mRNA levels of Scd1, a gene related to de novo hepatic lipogenesis, were observed when compared to the control and HFHFr groups. Other de novo hepatic lipogenesis genes, Acc1 and Fasn, did not show significant differences between the control and HFHFr groups. After AA treatment, there was no difference in the levels of de novo hepatic lipogenesis genes compared to the HFHFr group, except for Acc1, in which the AA group showed a significant increase compared to the HFHFr group.

[0242] AA supplementation reduces liver inflammation associated with NAFLD

[0243] To evaluate whether AA supplementation modulates the levels of pro-inflammatory markers, the expression levels of some representative inflammatory genes were evaluated. As expected, the mRNA level of F4 / 80, a marker of macrophage infiltration, was 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 for 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 for the expression of II1a gene, which was significantly increased in the HFHFr group compared to the control group, but AA did not significantly reduce the levels of II1a. No significant results were shown for the levels of II6 and IL10. However, the expression of Cd11c showed a significant increase in the HFHFr and AA groups compared to the control group, although there was no difference between HFHFr mice and AA mice.

[0244] AA supplementation promotes antioxidant defense and antitumor activity in NAFLD

[0245] Genes related to autophagy, immune response, antioxidant defense, antitumor activity, and vesicle trafficking were analyzed. In terms of autophagy, no differences were found in either gene between control mice, HFHFr mice, and AA mice. However, when it came to genes related to antioxidant activity, such as CBS or CTH, significant results were found. No differences were found between control and HFHFr, but in AA mice, the expression of both genes was significantly increased compared to the HFHFr group. Regarding genes related to antitumor activity, the expression of Mtus1 in AA mice showed a significant increase compared to the HFHFr group. Regarding genes related to immune response, no difference was observed in IRF4 expression. However, a significant increase in Cebpb expression in the AA group was observed compared to the HFHFr group. Finally, Sec24b, a gene related to vesicle trafficking, showed a significant increase in AA mice compared to HFHFr mice.

[0246] AA supplementation alleviates hepatic insulin resistance associated with NAFLD

[0247] Considering that impaired hepatic insulin signaling plays an important role in the development of NAFLD, to assess insulin resistance, animals were stimulated with an intraperitoneal insulin bolus, and Akt activation (phosphorylation of serine 473) was determined, or with saline. As expected, the control group exhibited significant Akt phosphorylation (pAktS473) in insulin-stimulated mice compared to insulin-injected mice. The fact that this was not observed in the HFHFr group suggests the presence of hepatic insulin resistance in these animals. In contrast, the AA-supplemented group exhibited a significant promotion of insulin signaling in insulin-stimulated mice and a significant increase in Akt phosphorylation compared to vehicle-injected AA mice. Data are shown in Figure 2. Figure 4 Figure 2 shows representative Western blot analysis of Akt activation (pAktS473), total Akt protein levels (T-Akt), housekeeping β-actin levels, and protein loading using membrane staining with Ponceau S. (B) shows densitometry analysis of the ratio of phosphorylated Akt to total Akt.

[0248] Furthermore, analysis of the hepatic expression of key genes involved in fatty acid β-oxidation (carnitine palmitoyltransferase, Cpt1a), inflammation (chemokine (CC motif) ligand 2, CCL2), lipolysis (hormone-sensitive lipase, HSL; and lipid droplet envelope protein, PLIN), and lipogenesis (fatty acid synthase, FASN) after quadruple treatment showed a trend toward increased Cpt1a, a reduction in important hepatic inflammation (Ccl2), and an increase in lipolysis (higher expression of the lipase HSL and lower levels of the lipid droplet envelope protein PLIN) ( Figure 4 (C)). In contrast, no changes in adipogenesis (Fasn) were observed. Data are Figure 4 Figures represent mean ± SEM; n = 10 to 12 per condition. *p < 0.05.

[0249] AA supplementation increases amino acid levels in the liver of NAFLD

[0250] To analyze the effects of amino acid-based treatment on the liver, LC / MS evaluation of the serum was performed with the goal of quantifying the amino acids in the participants under treatment. In the case of histidine and serine alone, no differences were found between the groups. When the levels of the amino acids were combined, a trend of increase in the concentrations of these amino acids was observed, but it was not significant. This could be due to tissue bioaccumulation of amino acids. Therefore, liver homogenates from different groups were analyzed for these amino acids by LC / MS to prove this hypothesis. The results obtained showed that histidine and serine levels were significantly reduced in the HFHFr group compared to the control group. In addition, a significant trend of increase in 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 by carnosine synthetase, showed a significant decrease in the HFHFr group compared to the control group. In addition, it was significantly increased in AA mice compared to HFHFr mice. Glycine is an amino acid associated with serine and GSH metabolism, and the increase in its levels could be due to the administration of serine. Glycine levels were found to be reduced in the HFHFr group compared to the control group. Furthermore, glycine levels were restored in AA mice compared with those in HFHFr mice.

[0251] Example 2. AA supplementation reduces the risk of atherosclerosis

[0252] Atherosclerosis is a complex disorder that exhibits many features of a chronic inflammatory process. Atherosclerotic cardiovascular disease remains a major cause of morbidity and mortality worldwide, and obesity increases the risk of atherosclerosis and death. Elucidating the mechanisms underlying the differences between obese individuals with and without atherosclerosis could reveal therapeutic targets for treating the deleterious cardiovascular consequences of obesity as an alternative or adjunct to weight loss programs, whose long-term success is known to be limited. 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 influences inflammation and the pathophysiological processes involved in the development of atherosclerotic disease. Obesity and overweight are accompanied by an unfavorable lipid profile pattern. Dyslipidemia is a major risk factor for coronary artery disease. In obese patients, the estimated prevalence of hypertriglyceridemia is twice that of non-obese individuals. In addition, the atherogenic combination of hypertriglyceridemia and high LDL and low HDL is more prevalent in obese and overweight patients.

[0253] Serum analysis and blood analysis

[0254] Serum fasting blood glucose, serum total cholesterol, serum triglycerides were analyzed using enzymatic colorimetric assays (QCA, Barcelona, ​​Spain). Serum fasting blood insulin was analyzed using a mouse insulin ELISA kit (Mercodia, Uppsala, Sweden), and serum HDL levels and LDL levels were analyzed using EnzyChrom™ AF HDL and LDL / VLDL assay kits (BioAssay System, Hayward, CA, USA).

[0255] 16 control mice were maintained on a standard diet (D12328, Research Diets, New Brunswick, NJ, USA), and 32 animals (atherogenic group) were fed with a HFHFr diet (HFHC: D12331, Research Diets) supplemented with 23.1 g / L fructose and 18.9 g / L sucrose in the drinking water. The mice were maintained on these diets under free conditions for 20 weeks. In the last 4 weeks of the experiment (weeks 16 to 20), the atherogenic mice were randomly divided into two groups: 16 mice were maintained under the same feeding conditions as before, and the other 16 mice were treated with the histidine amino acid combination explained in Example 1 above.

[0256] In an experiment with n=48 mice, circulating levels of high-density lipoprotein (HDL), low-density lipoprotein (LDL), and the LDL / HDL ratio were also determined. Figure 5 As shown, it can be seen that the group receiving the supplement (histidine AA) showed higher HDL levels compared to mice receiving the HFHFr diet ( Figure 5 (A) ), and serum LDL levels were significantly lower in mice fed the HFHFr diet compared with those in mice fed the HFHFr diet ( Figure 5 (B) ), while the LDL / HDL ratio was within the mean of the control ( Figure 5 (C)). The data confirmed that the supplement was able to reduce levels of so-called "bad cholesterol" (LDL), increase so-called "good cholesterol" (HDL), and reduce the risk of atherosclerosis.

[0257] Example 3. AA supplementation reduces visceral fat and obesity parameters

[0258] Adipose tissue has unique heterogeneity. Subcutaneous adipose tissue (scWAT) accounts for the vast majority of total body fat storage, and visceral adipose tissue (vWAT) accounts for a smaller part of total body fat storage, but the excess of vWAT has a more serious risk than the excess of scWAT, wherein scWAT is reported to play a protective role. In this regard, obesity is defined as excessive accumulation or abnormal distribution of body fat, which can also be complicated by related diseases, such as type 2 diabetes, hepatic steatosis, cardiovascular disease, stroke, dyslipidemia, hypertension, and certain types of cancer. Although obesity is usually caused by excessive energy supply relative to energy consumption, its etiology is complex, including genetic factors, physiological factors, environmental factors, social factors, psychological factors and economic factors, which interact to promote the development of obesity. This excessive energy supply promotes the expansion of adipose tissue depots, which is mediated by the growth of adipocyte size (hypertrophy) and the increase of adipocyte number (hyperplasia).

[0259] In an experiment with the same number of mice as in Example 1, body weight, weight gain and other parameters were also determined to elucidate the ability of the supplement (AA) to reduce obesity risk and visceral fat.

[0260] Materials and methods

[0261] Histological analysis of EWAT and BAT

[0262] The EWAT and BAT parts were fixed in buffered formalin (4% formaldehyde, 4g / L NaH2PO4, 6.5g / L Na2HPO4; pH 6.8) and cut at a thickness of 5 μm and stained with hematoxylin-eosin (H&E). BAT IWAT and EWAT images were taken (magnification 4 times) using a microscope (ECLIPSE Ti; Nikon, Tokyo, Japan) connected to a digital aiming 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). In order to avoid any bias in the analysis, the study had a double-blind design to prevent the reviewers from understanding any data from mice during the histopathological analysis. The adipocyte area was quantified by the Adiposoft plug-in to assess the EWAT status between groups. The lipid droplet quantification in BAT was analyzed by the Droplet Finder plug-in.

[0263] RNA extraction and quantitative polymerase chain reaction

[0264] According to the manufacturer's instructions, TriPure reagent (Roche Diagnostic, Sant Cugat-Del Valles, Barcelona, ​​Spain) is used to extract the homogenate from EWAT, IWAT and BAT for total RNA. Nanophotometer (Implen GmbH, Munich, Germany) is used to measure RNA concentration and purity. High-capacity RNA-to-cDNA test kit (Applied Biosystems, Wilmington, DE, USA) is used to convert RNA into cDNA. Before incubation on Lightcycler® 480 II (Roche Diagnostics GmbH, Mannheim, Germany) using commercial LightCycler480 Sybr green I master, cDNA is diluted with 1:10. Table 2 shows the primer list used, which has been previously described in other studies and verified using Primer-Blast software (National Center for Biotechnology Information, Bethesda, MD, USA). 36b4 is used as housekeeping gene.

[0265] BAT temperature measurement

[0266] The temperature around BAT was visualized using a high-resolution infrared camera (FLIR Systems) and analyzed using a specific software package (FLIR-Tools-Software, FLIR; Kent, UK) as described previously79. For each image, the area around BAT was demarcated, and the mean temperature of the skin area was calculated as the average of 3 images / animal.

[0267] Table 2. Sequences of oligonucleotides used in RT-PCR.

[0268]

[0269] result

[0270] from Figure 6 As can be seen from the data presented, mice fed a HFHFr diet supplemented with the quadruple combination of the present invention (histidine, cysteine, serine and carnosine) did not change their initial and final body weights after 4 weeks of treatment, in contrast to mice fed a HFHFr diet alone, whose final body weights were significantly higher than their initial body weights ( Figure 6 (A)). In addition, obese animals treated with the quadruple combination of the present invention stopped gaining weight in the second week of treatment ( Figure 6 (C)). In addition, when epididymal white adipose tissue (EWAT), retroperitoneal adipose tissue (RWAT), and mean adipose tissue weights were determined, those mice fed the HFHFr diet supplemented with the quadruple combination had reduced fat mass compared to unsupplemented mice ( Figure 6 (B) ). Reduction of hypertrophic visceral adipose tissue by quadruple combination treatment was confirmed at the microscopic level by reduction in adipocyte size, increased number of smaller adipocytes, and decreased number of larger adipocytes ( Figure 6 (D)). In addition, the quadruple combination treatment modulated brown adipose tissue (BAT), reducing hypertrophy in this fat depot and decreasing the size of brown adipocytes ( Figure 6 (E) These effects on BAT were accompanied by increases in mRNA expression of key thermogenic genes (Ucp1, Fgf21, Pgc1a, Prdm16, Dio2, and Dio3), insulin-sensitive adipokine (Adipoq), and other genes involved in fatty acid oxidation (Cpt1b) and glucose uptake (Glut1 and Glut4) ( Figure 6 (G)) Increased Ucp1 mRNA levels after quadruple combination treatment were confirmed at the protein level, indicating a higher ability to burn fatty acids ( Figure 6 (H)). Indeed, this ability was assessed by thermography, which observed higher temperatures in the BAT region of animals treated with the quadruple combination ( Figure 6 (I))

[0271] Example 4. In vitro experiments

[0272] In vitro experiments were performed using a hepatocyte cell line (HepG2) to test the effects of different supplemental amino acid combinations 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 lipase, HSL; and lipid droplet envelope protein, LPIN), and lipogenesis (fatty acid synthase, FASN). Gene expression was determined by qPCR as previously indicated (see Materials and Methods in Example 1). Table 3 shows a 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 results in a decreased rate of fatty acid β-oxidation.

[0273] Table 3. Sequences of oligonucleotides used in RT-PCR.

[0274]

[0275] Data such as Figure 7As shown, the expression level (mRNA) for any supplemented medium is indicated. The supplements 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 tetrahistidine + cysteine ​​+ serine + carnosine (4AA). The concentrations tested were as follows: histidine 10 mmol / L, carnosine 5 mmol / L, cysteine ​​5 mmol / L, and serine 5 mmol / L. These concentrations were constant across the different combinations. Figure 7 Figure 5 shows mean ± SEM; n = 3 to 4 per condition.

[0276] like Figure 7 As shown, the best performance was obtained by supplementation with 4AA. Compared with the control group, the triple combination was also effective (all combinations contained histidine), which showed an increase in the expression of the mentioned enzymes compared to the expression caused by the single amino acids. It is also worth noting that the triple combination containing histidine, serine and cysteine ​​also showed a significant increase. In general, these data can confirm that the combination of at least three of histidine, cysteine, serine and carnosine, especially the triple combination and the quadruple combination containing histidine, unexpectedly increased the levels of the analyzed enzymes, which serves as evidence of their positive role in those diseases or conditions involving fat accumulation. It is also worth noting that the quadruple combination showed better synergy even in the presence of carnosine, while when carnosine was added together with some other amino acids, the compound showed a trend of reduced effect.

[0277] Example 5: AA supplementation improves metabolic changes associated with menopause

[0278] Materials and methods

[0279] Forty 24-week-old rats were ovariectomized according to the procedure described in Koebele, SV and Bimonte-Nelson, H. A, 2016. Another 10 rats of the same age underwent sham surgery. Two to three weeks after surgery, the rats were randomly divided into four experimental groups for an 8-week period of the following experiments:

[0280] -SHAM, simulated surgery

[0281] -OVX: OVX rats receiving a placebo daily

[0282] -OVX-E2: OVX rats treated with subcutaneous injections of estrogen (17β-estradiol) every two weeks. OVX-E2 rats received subcutaneous injections of 25 µg / kg body weight of 17β-estradiol (Sigma-Aldrich, St. Louis, MO) in corn oil vehicle solution every two weeks.

[0283] -OVX+4AA: OVX rats supplemented with 4AA daily at a dose of 105 mg / kg body weight of histidine, carnosine, serine, and 245 mg / kg body weight of cysteine.

[0284] Body composition analysis: At the beginning and end of the study, lean body mass and fat body mass were measured (in grams) without anesthesia using an EchoMRI-700™ device (Echo Medical Systems, LLC, Houston, USA). The measurements were repeated three times at 8 am under free conditions and the results were expressed as a percentage of body weight.

[0285] Obesity Index: The adiposity index was calculated as the sum of the weights of the IWAT depot, MWAT depot, and RWAT depot in grams and expressed as a percentage of body weight.

[0286] Serum leptin level: Serum leptin levels were determined using a rat ELISA kit (EZRL-83K / Millipore, Barcelona, ​​Spain).

[0287] Glucose levels and insulin levels: Glucose levels 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, Uppsala, Sweden).

[0288] 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).

[0289] result

[0290] Previous studies using ovariectomized (OVX) rats have shown that these animals exhibit weight gain, obesity, and insulin resistance impairment compared to sham-operated rats (controls).

[0291] The results disclosed herein indicate that treatment with the tetrahistidine+cysteine+serine+carnosine (4AA) combination is effective in ameliorating metabolic changes associated with menopause.

[0292] Example 6. Relationship between plasma histidine and steatosis and the human intestinal microbiota and Hut gene

[0293] Materials and methods

[0294] Metagenomic analysis

[0295] For metagenomic analysis, those families, genera, or species with less than 10 counts in 10% of the samples were excluded. Differential abundance analysis of taxa associated with circulating histidine levels, hepatic steatosis, or HAA treatment was performed using the Analysis of the Microbiome with Bias Correction (ANCOM-BC) approach. This approach accounts for bias caused by different sampling fractions between samples by adding sample-specific biases estimated from observed data to linear regression models. Log-scaled linear regression models were similar to log-ratio transformations to account for the compositional nature of metagenomic datasets, while the bias term served as bias correction. Models for all humans were adjusted for age, sex, BMI, and country. P values ​​were adjusted for multiple comparisons using sequential goodness-of-fit as implemented in the ‘SGoF’ R package. Unlike the FDR approach, which reduces its statistical power with the number of hutG, hutH, hutI, and hutU, respectively.

[0296] Fecal genomic DNA extraction and whole-genome shotgun sequencing (metagenomics)

[0297] Fecal shotgun sequencing data from n = 73 patients in the discovery cohort were generated. Briefly, total DNA was extracted from frozen human feces using the QIAampDNA stool mini kit (Qiagen) with a slight modification of 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 per sample was prepared using the Bioscientific PCR free library kit according to the manufacturer's instructions and sequenced on a HiSeq 2500 (Illumina) using 2 × 150 paired-end sequencing chemistry. The input fastq files obtained were decompressed, filtered, and 3'-end trimmed by quality using the prinseq-lite-0.20.4 program, and overlapping pairs were joined using FLASH-1.2.11. Then the fastq files were converted to fast files, and human host reads were removed by aligning reads to the reference human genome (GRCh38.p11, Dec 2013) using Bowtie 2 with the end-to-end option and the very sensitive option. Then, non-host reads were assembled into contigs by MEGAHIT v1.1.2, and these reads were aligned to the contigs using Bowtie 2 for functional analysis. Unassembled reads were attached to the contigs. Then, Prodigal v2.6.342 was used to predict coding regions. HMMER was used to perform functional annotations against the 2016 version of the Kyoto Encyclopedia of Genes and Genomes (KEGG) database to obtain the functional subcategories, pathways, and annotations of genes. R 3.1.0 was used to filter the best annotations and assign orf annotations to each read, and R 3.1.0 was used to count the aligned reads, add the categories and their coverage, and finally construct the abundance matrix. Taxon annotation of non-human sequence fragments was performed using the greedy mode on Kaiju v1.6.2. Lineage information addition, taxon counts, and abundance matrix generation for all samples were performed using R packages.

[0298] Bacterial genomic DNA isolation, 16S rRNA sequencing and analysis (metagenomics)

[0299] Bacterial genomic DNA was isolated from mouse fecal samples using the MBP DNA Soil Extraction Kit. Genomic DNA was normalized to 5 ng / µL with EB (10 mmol / L Tris-HCl) and libraries were created. Briefly, after the first PCR and cleanup using KAPA purification magnetic beads (Roche catalog number 07983298001), a second PCR master mix was composed using index primers P7 and P5 of the Nextera XT Index Kit v2 (Illumina catalog number: FC-131-2001 to FC-131-2004). After the PCR reaction, the library was quantified using the Quant-iT dsDNA Assay Kit, High Sensitivity Kit (catalog number 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 molarity. The pool was run on an Illumina MiSeq instrument (600 cycles) using the MiSeq® Reagent Kit v3 (Illumina catalog number FC-102-3003), following Illumina recommended denaturation and loading recommendations, including a 20% PhiX spike-in control (PhiX Control v3 Illumina catalog number FC-110-3001). Raw data were analyzed locally on the MiSeq using MiSeq reporter. For 16S sequencing analysis, LotuS1.36 used the analysis pipeline with short amplicon mode and default quality filtering. The raw 16S rRNA gene sequencing fragments were quality filtered to ensure a minimum length of 170 bp, no more than 8 identical nucleotides, no ambiguous bases, an average quality >= 27, a cumulative error less than 0.5, and the deduplication filter was set to "6:1,3:2". The cleaned sequencing fragments were clustered into amplicon sequencing variants (ASVs) using DADA2, and chimeric ASVs were removed using DADA2 de novo chimera check. The remaining phiX sequencing fragments were filtered by aligning the ASVs with the phiX reference genome. ASV classification was performed using the LotuS2 LCA algorithm against the Silva138.1 reference database. On average, 5500±3108 sequencing fragments of cyanobacterial origin were assigned to each sample. Further data analysis used the R statistical language version 3.00 (The R Foundation, https: / / www.r-project.org / ), and all data were normalized using rtk software. The 16S raw data can be found at https: / / 1drv.ms / u / s!ApVezdktX3aIlZVJXYWok9s2qhgfvg?e=VDyr2c Found in.

[0300] Primer design for HutH and HutG microbiota functions

[0301] Multiple sequences of prokaryotic species were uploaded from the database into the ARDEP program to design degenerate primers that can amplify the conserved regions of the HutH and HutG genes and cover all species used for primer design. Based on the k-mer algorithm, the primer length was set to k and all sequences were divided into k-mers. The analysis platform allows sequence length statistics, length filtering, and redundancy removal to be performed on the sequence database using tools located in the "Sequence Processing" section. After performing sequence database quality control, ARDEP for primer design was run in the "Primer Processing" section to obtain a list of possibilities for different primer sets. The number of sequences of taxa and functional groups covered by the primer set was counted using the "Covered Taxonomy Calculator" option. The best solution was selected considering the basic properties of the primers (Tm, GC%, primer length).

[0302] result

[0303] Microbial-host interactions may contribute to the development of metabolic diseases, and molecular networks linking the gut microbiome to hepatic steatosis have recently been elucidated.

[0304] Gut microbiome in nonalcoholic fatty liver disease (NAFLD) Fig.11 b) and plays a key role in the pathophysiology of other diseases and conditions such as obesity, IBD, visceral adipose tissue, and unhealthy metabolic profiles.

[0305] The families Xanthomonadaceae, Rickettsiaceae, and Alteromonadales of the Proteobacteria phylum were increased in hepatic steatosis. At the genus level, Bilophila and Campylobacter were also described as increased in NAFLD.

[0306] Histidine can be utilized by many bacteria and is an important source of carbon, nitrogen, and / or energy for Proteobacteria. The histidine utilization (Hut) pathway involves the elimination of ammonia from histidine to produce urocanic acid, imidazolide propionate, and iminomethylglutamate. Fig.11 c)

[0307] To assess the potential role of the gut microbiota in regulating plasma histidine levels, a shotgun metagenomic analysis of stool samples from 73 patients from the discovery cohort was performed. Analysis of the microbiome with bias correction (ANCOM-BC) was used to account for the compositional nature of the metagenomic dataset and to identify differentially enriched taxa associated with circulating histidine levels, controlling for age, BMI, sex, and country.

[0308] There was a strong negative correlation between histidine levels and membership in the Proteobacteria. Fig.11 a) is also closely related to the degree of hepatic steatosis. Circulating histidine levels are also strongly positively correlated with Cyanobacteria, Corynebacteriales (Actinobacteria), and Marinelabiliales (Bacteroidetes) ( Fig.11 a).

[0309] Circulating histidine levels also correlated positively with indicators of healthy gut bacteria known to produce anti-inflammatory metabolites, such as Faecalibacterium, Bifidobacterium, and Odoribacter ( Fig.11 a).

[0310] Consistent with these results, microbial Hut gene levels were found to be elevated in patients with higher degrees of steatosis, whereas microbial Hut gene levels decreased after HAA supplementation (“HAA” as used herein corresponds to the composition comprising histidine, cysteine, serine, and carnosine used in the previous examples), suggesting an effect of HAA administration on the associated dysbiosis.

[0311] Patients with steatosis <33% and those with hepatic steatosis >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, hydration of urocanic acid to imidazole propionate, and cleavage of the imidazole ring to obtain iminomethylglutamate ( Fig.11 d).

[0312] These results suggest that supplementation with histidine-related amino acids (HAA) improves NAFLD and reduces liver inflammation in mice.

[0313] Here we show that HAA supplementation in NAFLD-induced mice reduces the expression of bacterial histidine utilization genes (hutH and hutG), which are increased in NAFLD patients ( Fig.11 e).

[0314] Together, these results suggest that plasma histidine levels are negatively correlated with dysregulated bacterial families in NAFLD. This microbiome imbalance, known as dysbiosis, can be counteracted by HAA administration.

[0315] Further aspects / embodiments of the invention can be found in the following clauses:

[0316] 1. A pharmaceutical composition or dietary supplement composition comprising one or more than one pharmaceutically or nutraceutically acceptable excipient or carrier, and a therapeutically or nutraceutically effective amount of at least three of the following substances: histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two; cysteine ​​or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, and carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two; and / or a therapeutically or nutraceutically effective amount of a peptide comprising an amino acid sequence containing at least three of histidine, cysteine, serine and carnosine.

[0317] 2. The composition according to clause 1, comprising one or more than one pharmaceutically or nutraceutically acceptable excipients or carriers, and therapeutically or nutraceutically effective amounts of at least three of the following substances: histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either; cysteine ​​or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, and carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either.

[0318] 3. A composition according to any one of clauses 1 to 2, wherein the therapeutically or nutraceutically effective amount of histidine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either of the two, is a dosage of 10 mg per Kg of subject to be administered to 250 mg per Kg of subject; the therapeutically effective amount of cysteine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either of the two, is a dosage of 30 mg per Kg of subject to be administered to 550 mg per Kg of subject; the therapeutically effective amount of serine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either of the two, is a dosage of 10 mg per Kg of subject to be administered to 250 mg per Kg of subject; and the therapeutically effective amount of carnosine, or a pharmaceutically or nutraceutically acceptable salt thereof, or a solvate of either of the two, is a dosage of 10 mg per Kg of subject to be administered to 200 mg per Kg of subject.

[0319] 4. A composition according to any one of clauses 1 to 3, comprising therapeutically or nutraceutically effective amounts of at least three of the following substances: L-histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either; L-cysteine ​​or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, L-serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, and L-carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either.

[0320] 5. The composition according to any one of clauses 1 to 4, wherein the pharmaceutically or nutraceutically acceptable salts of histidine and cysteine ​​are hydrochloride of L-histidine and hydrochloride of L-cysteine.

[0321] 6. The composition according to any one of clauses 1 to 5, wherein the pharmaceutically acceptable salt of histidine is L-histidine monohydrochloride monohydrate.

[0322] 7. A composition according to any one of clauses 1 to 6, comprising a therapeutically or nutraceutically effective amount of histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two; a therapeutically or nutraceutically effective amount of cysteine ​​or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two; a therapeutically or nutraceutically effective amount of serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two; and a therapeutically or nutraceutically effective amount of carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, and one or more pharmaceutically or nutraceutically acceptable excipients or carriers.

[0323] 8. A composition according to clause 7, comprising:

[0324] - a therapeutically or health food effective amount of L-histidine monohydrochloride monohydrate;

[0325] - a therapeutically or health food effective amount of L-cysteine ​​hydrochloride;

[0326] - a therapeutically or health food effective amount of L-serine; and

[0327] - L-carnosine in an effective amount for treatment or health food,

[0328] and one or more than one excipient or carrier acceptable in medicine or health food.

[0329] 9. A food in solid form or liquid form comprising a pharmaceutical composition or dietary supplement composition as defined in any one of clauses 1 to 8.

[0330] 10. The food according to clause 9, which is a beverage.

[0331] 11. A pharmaceutical composition or dietary supplement composition as defined in any one of clauses 1 to 8, or a food as defined in any one of clauses 9 to 10, for use in therapy.

[0332] 12. A pharmaceutical composition or dietary supplement composition or food for use according to clause 11 for preventing and / or treating a disease selected from fatty liver disease, obesity, atherosclerosis and a combination thereof.

[0333] 13. A pharmaceutical composition or dietary supplement composition or food for use according to any one of clauses 11 to 12 for preventing and / or treating fatty liver disease selected from non-alcoholic fatty liver disease and alcoholic fatty liver disease.

[0334] 14. A pharmaceutical composition 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 with simple steatosis (NAFLD) and non-alcoholic steatohepatitis (NASH).

[0335] Reference List

[0336] -Koebele, SV & Bimonte-Nelson, HA “Modeling menopause: The utility of rodents in translational behavioral endocrinology research”.Maturitas 2016, 87, 5-17.

[0337] -Mardinoglu et al., "Personal model-assisted identification of NAD+ andglutathione metabolism as intervention target in NAFLD", Mol Syst Biol-2017, vol. no. 13:916

[0338] - Mong et al., “Histidine and carnosine alleviated hepatic steatosis in mice consumed high saturated fat diet”, European Journal of Pharmacology 653 (2011) 82-88

[0339] - 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.

Claims

1. A pharmaceutical composition or dietary supplement composition comprising one or more than one pharmaceutically or health food acceptable excipient or carrier, and the following substances: (i) a therapeutically or health food effective amount of cysteine ​​or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of at least two of the following substances: histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two; and / or a therapeutically or health food effective amount of a peptide comprising an amino acid sequence containing cysteine ​​and at least two of histidine, serine and carnosine; or, alternatively, (ii) a therapeutically or health food effective amount of N-acetylcysteine ​​(NAC) or a pharmaceutically or health food acceptable salt or a solvate of either of the two, and a therapeutically or health food effective amount of at least two of the following substances: histidine or a pharmaceutically or health food acceptable salt or a solvate of either of the two, serine or a pharmaceutically or health food acceptable salt or a solvate of either of the two, and carnosine or a pharmaceutically or health food acceptable salt or a solvate of either of the two; and / or a therapeutically or health food effective amount of a peptide comprising an amino acid sequence containing NAC and at least two of histidine, serine and carnosine.

2. The composition according to claim 1, comprising one or more than one pharmaceutically or health food acceptable excipient or carrier, and the following substances: (i) a therapeutically or health food effective amount of cysteine ​​or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two; and / or a therapeutically or health food effective amount of a peptide comprising an amino acid sequence containing cysteine, histidine, serine and carnosine; or, alternatively, (ii) a therapeutically or nutraceutically effective amount of NAC or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, a therapeutically or nutraceutically effective amount of histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, a therapeutically or nutraceutically effective amount of serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, and a therapeutically or nutraceutically effective amount of carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two; and / or a therapeutically or nutraceutically effective amount of a peptide comprising an amino acid sequence containing NAC, histidine, serine and carnosine.

3. The composition according to claim 1, comprising one or more than one pharmaceutically or health food acceptable excipient or carrier, and the following substances: (i) a therapeutically or health food effective amount of cysteine ​​or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of at least two of the following substances: histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two; or, alternatively, (ii) a therapeutically or healthily effective amount of N-acetylcysteine ​​(NAC) or a pharmaceutically or healthily acceptable salt thereof or a solvate of either of the two, and a therapeutically or healthily effective amount of at least two of the following substances: histidine or a pharmaceutically or healthily acceptable salt thereof or a solvate of either of the two, serine or a pharmaceutically or healthily acceptable salt thereof or a solvate of either of the two, and carnosine or a pharmaceutically or healthily acceptable salt thereof or a solvate of either of the two.

4. The composition according to any one of claims 1 to 3, comprising one or more than one pharmaceutically or health food acceptable excipient or carrier, and the following substances: (i) a therapeutically or health food effective amount of cysteine ​​or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two; or, alternatively, (ii) a therapeutically or health food effective amount of NAC or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of histidine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, a therapeutically or health food effective amount of serine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two, and a therapeutically or health food effective amount of carnosine or a pharmaceutically or health food acceptable salt thereof or a solvate of either of the two.

5. The composition according to any one of claims 1 or 3, comprising one or more than one pharmaceutically or nutraceutically acceptable excipient or carrier, and the following substances: a therapeutically or nutraceutically effective amount of N-acetylcysteine ​​(NAC) or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, and a therapeutically or nutraceutically effective amount of at least two of the following substances: histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two, and carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either of the two.

6. The composition according to any one of claims 1 to 5, comprising one or more than one pharmaceutically or nutraceutically acceptable excipient or carrier, and the following substances: a therapeutically or nutraceutically effective amount of NAC or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, a therapeutically or nutraceutically effective amount of histidine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, a therapeutically or nutraceutically effective amount of serine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either, and a therapeutically or nutraceutically effective amount of carnosine or a pharmaceutically or nutraceutically acceptable salt thereof or a solvate of either.

7. The composition according to any one of claims 1 to 4, wherein the therapeutically effective amount of cysteine ​​or its pharmaceutically or nutraceutically acceptable salt or solvate of either is a dose of 2 mg per kg to 550 mg per kg of subject; the therapeutically or nutraceutically acceptable amount of histidine or its pharmaceutically or nutraceutically acceptable salt or solvate of either is a dose of 7.5 mg per kg to 250 mg per kg of subject; the therapeutically effective amount of serine or its pharmaceutically or nutraceutically acceptable salt or solvate of either is a dose of 1.3 mg per kg to 250 mg per kg of subject; and the therapeutically effective amount of carnosine or its pharmaceutically or nutraceutically acceptable salt or solvate of either is a dose of 4.5 mg per kg to 200 mg per kg of subject.

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

9. The composition according to any one of claims 1 to 6, wherein the therapeutically effective amount of NAC or its pharmaceutically or nutraceutically acceptable salt or solvate of either is a dosage of 2 mg per kg of subject to be administered to 550 mg per kg of subject; the therapeutically or nutraceutically acceptable amount of histidine or its pharmaceutically or nutraceutically acceptable salt or solvate of either is a dosage of 7.5 mg per kg of subject to be administered to 250 mg per kg of subject; the therapeutically effective amount of serine or its pharmaceutically or nutraceutically acceptable salt or solvate of either is a dosage of 1.3 mg per kg of subject to be administered to 250 mg per kg of subject; and the therapeutically effective amount of carnosine or its pharmaceutically or nutraceutically acceptable salt or solvate of either is a dosage of 4.5 mg per kg of subject to be administered to 200 mg per kg of subject.

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

11. The composition according to any one of claims 1 to 10, wherein the pharmaceutically or nutraceutically acceptable salt of histidine is L-histidine hydrochloride.

12. The composition according to any one of claims 1 to 11, wherein the pharmaceutically acceptable salt of histidine is L-histidine monohydrochloride monohydrate.

13. The composition according to any one of claims 1 to 4, 7, 8, 11 or 12, wherein the pharmaceutically or nutraceutically acceptable salt of cysteine ​​is L-cysteine ​​hydrochloride.

14. A composition according to any one of claims 1 to 6 or 9 to 12, comprising: - a therapeutically or health food effective amount of L-histidine monohydrochloride monohydrate; - A therapeutically or nutraceutically effective amount of NAC; - a therapeutically or health food effective amount of L-serine; and - L-carnosine in an effective amount for treatment or health food, and one or more than one excipient or carrier acceptable in medicine or health food.

15. A composition according to any one of claims 1 to 4 and 7, 8, 11 to 13, comprising: - a therapeutically or health food effective amount of L-histidine monohydrochloride monohydrate; - a therapeutically or health food effective amount of L-cysteine ​​hydrochloride; - a therapeutically or health food effective amount of L-serine; and - L-carnosine in an effective amount for treatment or health food, and one or more than one excipient or carrier acceptable in medicine or health food.

16. A food in solid form or liquid form comprising a pharmaceutical composition or a dietary supplement composition as defined in any one of claims 1 to 15. The food according to claim 16 , which is a beverage.

18. A pharmaceutical or dietary supplement composition as defined in any one of claims 1 to 15, or a food product as defined in any one of claims 16 to 17, for use in therapy.

19. The pharmaceutical composition or dietary supplement composition or food for use according to claim 18, for preventing and / or treating a disease selected from the group consisting of fatty liver disease, obesity, overweight, atherosclerosis, dysbiosis and combinations thereof.

20. The pharmaceutical composition or dietary supplement composition or food for use according to any one of claims 18 to 19 for preventing and / or treating fatty liver disease selected from non-alcoholic fatty liver disease and alcoholic fatty liver disease.

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

22. A pharmaceutical composition or dietary supplement composition or food for use according to claim 18 for preventing and / or treating symptoms associated with menopause in female mammals, particularly female humans.

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

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