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WO2025081253A1 - Nutraceutical composition for protecting against liver conditions and method for producing same - Google Patents

Nutraceutical composition for protecting against liver conditions and method for producing same Download PDF

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Publication number
WO2025081253A1
WO2025081253A1 PCT/BR2024/050475 BR2024050475W WO2025081253A1 WO 2025081253 A1 WO2025081253 A1 WO 2025081253A1 BR 2024050475 W BR2024050475 W BR 2024050475W WO 2025081253 A1 WO2025081253 A1 WO 2025081253A1
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Prior art keywords
composition according
solution
composition
group
liver
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French (fr)
Portuguese (pt)
Inventor
Carlos ANDRES RODRIGUEZ PANTANALI
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Psm Pantanali Servicos Medicos Ltda
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Psm Pantanali Servicos Medicos Ltda
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A61K31/05—Phenols
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/12—Ketones
    • A61K31/122—Ketones having the oxygen directly attached to a ring, e.g. quinones, vitamin K1, anthralin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
    • A61K31/202—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids having three or more double bonds, e.g. linolenic
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/455—Nicotinic acids, e.g. niacin; Derivatives thereof, e.g. esters, amides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/04—Sulfur, selenium or tellurium; Compounds thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/02—Nutrients, e.g. vitamins, minerals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines

Definitions

  • the present invention belongs to the field of nutraceutical compositions and suspensions aimed at protecting health conditions. More specifically, the present invention relates to a nutraceutical composition for protecting liver conditions, which are ischemia and reperfusion of this organ and apoptosis, as well as the method for producing such composition.
  • Nutraceuticals are natural bioactive or chemical compounds that, in addition to playing a nutritional role, improve health, cure diseases or have preventive properties. They are dietary supplements and, from a nutritional point of view, nutraceuticals are a source of nutrients (lipids, carbohydrates, vitamins, proteins, minerals) and non-nutrients (prebiotics, probiotics, phytochemicals, enzyme regulators).
  • Nutraceuticals can be extracted from plant and animal foods, concentrated and administered in a suitable pharmaceutical form, with the aim of improving health, in dosages that exceed those obtained from normal foods.
  • nutraceuticals have antioxidant, anti-inflammatory, antibacterial, antiviral and antifungal activities, as well as evidence that they act as modulators of the immune response, angiogenesis and cell death. These effects are possible due to the multiple mechanisms of action of nutraceuticals
  • nutraceuticals are a growing therapy due to their nutritional and therapeutic benefits, some of which have already been studied and shown promising results, such as when using components such as resveratrol, quercetin, omega-3 fatty acids, selenium, ginger and avocado, separately or in different combinations.
  • components such as resveratrol, quercetin, omega-3 fatty acids, selenium, ginger and avocado, separately or in different combinations.
  • synergistic effects of nutraceutical combinations The concept of synergism was introduced by Liu et al., and arises from the overlapping of positive physical and chemical effects of one or another addition, causing a greater final sum effect of the components combined with each other.
  • composition comprising a synergistic effect of components combined with each other is still required, since such a combination is not foreseen in the state of the art.
  • the objective of the present invention is to provide a nutraceutical composition with a synergistic effect for the protection of liver conditions, which are ischemia and reperfusion of this organ.
  • Another objective of the present invention is to provide a pharmaceutical composition that favors the reduction in gene expression and caspase-3 protein, as well as reduction of TNF-a protein in liver tissue.
  • the present invention relates to a nutraceutical composition for protection against liver conditions, more specifically conditions related to liver ischemia and reperfusion, comprising: between 0.5 and 1.0 g/L of at least one polyphenol; between 0.5 and 1.0 g/L of each of at least four flavonoids; between 0.5 and 1.0 g/L of each of at least two carotenoids; between 0.2 and 0.8 g/L of at least one antioxidant mineral; between 0.4 and 1.0 g/L of at least one polyunsaturated fatty acid; between 0.5 and 1.0 g/L of at least one plant extract; between 1.0 and 1.5 g/L of avocado; between 0.8 and 1.5 g/L of at least one branched-chain amino acid; between 4.0 and 6.0 g/L of at least one amide; in a pharmaceutically acceptable liquid vehicle.
  • At least one polyphenol is resveratrol.
  • At least one flavonoid is quercetin, one is isoflavonoid, one is anthocyanidin, and one is cannaflavin.
  • At least one carotenoid is astaxanthin and another is lycopene.
  • At least one antioxidant mineral is selenium, more specifically chelated selenium.
  • At least one polyunsaturated fatty acid is omega-3.
  • At least one plant extract is ginger extract.
  • At least one amide is nicotinamide.
  • the pharmaceutically acceptable carrier is a carboxymethyl cellulose syrup.
  • Liver conditions are due to liver ischemia and reperfusion and apoptosis.
  • the pharmaceutical forms are chosen from suspension, solution, emulsion, capsule, tablet, dragee, granule or powder.
  • the present invention relates to a method for producing a nutraceutical composition for protecting against liver conditions, which comprises obtaining a first composition by the steps of: (a) obtaining a pharmaceutically acceptable liquid vehicle; (b) adding at least one polyunsaturated fatty acid to an oil under stirring until complete solubilization; (c) combining the contents of steps (a) and (b) under stirring until complete incorporation to obtain the first solution.
  • step (d) adding separately: at least one polyphenol, at least four flavonoids, at least two carotenoids, at least one antioxidant mineral, at least one polyunsaturated fatty acid, at least one plant extract, avocado powder, at least one branched-chain amino acid, at least one minus one amide to a container; (e) homogenize the components of step (d) in a liquid vehicle to obtain the second solution.
  • the first solution is added to the second solution, and both are stirred until completely homogenized.
  • Step (a) comprises mixing EDTA, sodium benzoate, deionized water and carboxymethyl cellulose until completely dissolved.
  • At least one oil of step (b) is a vegetable oil, preferably sunflower oil.
  • Step (c) comprises adding honey.
  • the liquid vehicle (d) is deionized water in a quantity sufficient for complete solubilization of the components, and the pharmaceutically acceptable liquid vehicle from step (a).
  • Figure 1 comprises graphs relating to the serum transaminase levels of each group: (a) AST - aspartate aminotransferase; (b) ALT - alanine aminotransferase.
  • Figure 2 comprises graphs relating to inflammatory mediators: (a) IL-1 p, (b) IL-6, (c) IL-10 and (d) TNF-a.
  • Figure 3 comprises a graph relating to MDA in the liver tissue of each group.
  • Figure 4 comprises graphs relating to gene expression of genes related to apoptosis in all groups: (a) BAX, (b) BCL-2, (c) CASPASE-8 and (d) CASPASE-3 genes.
  • Figure 5 shows TUNEL assay of liver tissues from different groups.
  • Figure 6 shows immunohistochemistry of cleaved caspase-3 protein in liver tissue from different groups.
  • Figure 7 shows immunohistochemistry of TNF-a protein in liver tissue from different groups.
  • Figure 8 shows histological and immunohistochemical analyses for different groups.
  • Figure 9 corresponds to a process flowchart of the method according to the present invention.
  • the present invention relates to a nutraceutical composition for protecting liver conditions, more specifically aimed at protecting liver ischemia and reperfusion.
  • the nutraceutical composition is based on a synergistic effect achieved by combining different components capable of bringing about advantageous effects for the protection of said clinical condition.
  • the synergistic composition has the combination of the following elements in their respective concentration ranges, in g/L, in relation to the volume of the composition obtained:
  • the polyphenol present in the composition must be resveratrol.
  • the flavonoid present in the composition may be at least one of the isoflavones, preferably quercetin.
  • At least one carotenoid is astaxanthin and another is lycopene.
  • the antioxidant mineral present in the composition is selenium, more specifically chelated selenium.
  • the polyunsaturated fatty acid present in the composition is omega-3, and may be in powder form.
  • the plant extract present in the composition may be one of saffron, ginger, or a mixture thereof, preferably being ginger extract.
  • the avocado is used in powder form, with avocado leaves in dry powder form being preferably used.
  • the branched chain amino acid present in the composition can be replaced by one of the saponins, with leucine being the preferred component.
  • the amide present in the composition is nicotinamide (niacin).
  • the pharmaceutically acceptable liquid carrier is carboxymethyl cellulose syrup, for example CMC syrup 0.5% (sold by Bio Identica compounding pharmacy, S ⁇ o Jose, Santa Catarina, Brazil).
  • the pharmaceutical form of the nutraceutical composition according to the present invention is preferably the solution, but it can also be any other liquid (suspension or emulsion) or solid (capsule, tablet, dragee, granule and powder).
  • concentrations of each of the components in the composition are the same, regardless of their pharmaceutical forms.
  • the present invention relates to a method for producing a nutraceutical composition for protection against liver conditions as previously described.
  • the method according to the present invention comprises obtaining a first solution by the steps of:
  • (d) add separately: at least one polyphenol, at least four flavonoids, at least two carotenoids, at least one antioxidant mineral, at least one polyunsaturated fatty acid, at least one plant extract, avocado powder, at least one amino acid, at least one amide to a container,
  • step (e) homogenize the components of step (d) in a liquid vehicle to obtain the second solution.
  • the first solution is added to the second solution, and both are stirred until completely homogenized.
  • step (a) of obtaining a pharmaceutically acceptable liquid carrier comprises mixing EDTA, sodium benzoate, deionized water and carboxymethyl cellulose until complete dissolution.
  • the preparation of the 0.5% CMC syrup is carried out in the usual manner: EDTA and sodium benzoate are placed in deionized water at 70 ° C; then, CMC is stirred and pulverized until its complete solubilization, obtaining the 0.5% CMC syrup.
  • each component of the composition is calculated considering the lowest effective oral dose, in 1.0 ml of solution. For this purpose, use if the EC50 of each one, which is the antioxidant concentration required to obtain 50% inhibition of free radicals.
  • the correction factor of each raw material (given by the supplier) is applied to this value and multiplied by the total volume of the solution, giving the concentration in mg/ml, as follows:
  • At least one polyunsaturated fatty acid is added to an oil under agitation until complete solubilization, and at least the oil used is a vegetable oil, preferably sunflower oil.
  • the omega-3 will be incorporated into the sunflower oil.
  • step (a) Combine the contents of steps (a) and (b) under stirring until completely incorporated to obtain the first solution, and during such step (c) honey can be added in order to make the solution more palatable, increase the concentration of the compounds used, their oral bioavailability, and, consequently, increase the bioactivity of each nutraceutical compound.
  • the method of the present invention also involves obtaining a second solution.
  • the following are added separately: at least one polyphenol, at least four flavonoids, at least two carotenoids, at least one antioxidant mineral, at least one polyunsaturated fatty acid, at least one plant extract, powdered avocado leaves, at least one amino acid and at least one amide to a container.
  • an inorganic salt such as NaCl, can be added, followed by the complete homogenization of all components.
  • the liquid vehicle CMC is added, thus obtaining the second solution.
  • the first solution is added to the second solution, and both are stirred until completely homogenized, thus obtaining the nutraceutical composition according to the present invention.
  • the rats received nutraceutical solution for seven consecutive days before being subjected to hepatic IR.
  • the animals received nutraceutical solution for seven consecutive days.
  • the rats underwent median laparotomy and the liver was only manipulated.
  • Example 1 represented by the following table, of nutraceutical composition according to the present invention was used, in the pharmaceutical form of solution.
  • Table 1 Concentration of nutraceutical compounds, in g/L, used in the solution
  • Rats in the IR group exhibited a significant increase in serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels compared to the CONTROL and NUT groups.
  • the NUT + IR group showed a significant increase in serum AST and ALT levels compared to the CONTROL group, as shown in Figure 1.
  • Data presented are mean ⁇ SE; *p ⁇ 0.05 vs. CONTROL group; A p ⁇ 0.05 vs. NUT group.
  • the CONTROL group exhibited a significantly higher level of MDA in liver tissue when compared to the IR, NUT+IR and SHAM groups. The same was also observed in the NUT group compared to the NUT+IR group as shown in Figure 3 which represents MDA in liver tissue of each group.
  • the data presented are mean ⁇ SE; * p ⁇ 0.05 vs. IR group; • p ⁇ 0.05 vs. NUT + IR group; A p ⁇ 0.05 vs. SHAM group.
  • Apoptosis Gene Expression BAX, BCL-2, CASPASE 8 and CASPASE 3: [0079] The gene expression of BAX and BCL-2 was significantly higher in the NUT + IR group compared to the CONTROL and SHAM groups and similar to the NUT and IR groups. The latter group exhibited a significant increase in BAX gene expression compared to the CONTROL, NUT and SHAM groups. Among the gene expression of CASPASES, there was only a difference with CASPASE 3. The gene expression of CASPASE 3 was significantly lower in the NUT + IR group than in the IR group, which in turn had a significantly higher value compared to the CONTROL and SHAM groups, as shown in Figure 4.
  • Figure 4 represents the gene expression of apoptosis-related genes in all groups: (a) BAX, (b) BCL-2, (c) CASPASE 8 and (d) CASPASE 3. Data presented are mean ⁇ SE; *p ⁇ 0.05 vs. CONTROL group; p ⁇ 0.05 vs. NUT + IR group; • p ⁇ 0.05 vs. NUT group; A p ⁇ 0.05 vs. SHAM group.
  • the TUNEL method was used to determine liver cell apoptosis.
  • the NUT+IR group exhibited a significant decrease in the percentage of apoptosis compared to the IR group.
  • the IR group had a significantly higher percentage of apoptosis than the CONTROL, NUT, and SHAM groups ( Figure 5).
  • Figure 5 represents semiquantification of TUNEL in liver cells from different groups: (a) data shown are mean ⁇ SE; *p ⁇ 0.05 vs. CONTROL group; ⁇ p ⁇ 0.05 vs. NUT + IR group; ⁇ p ⁇ 0.05 vs. NUT group; *p ⁇ 0.05 vs. SHAM group. Arrows and highlighted boxes indicate TUNEL-positive cells in each group: (b) CONTROL; (c) IR; (d) NUT + IR; (e) NUT; and (f) SHAM. All images were acquired at 50x magnification and highlighted boxes at 400x magnification, with scale bars at 500 ⁇ m and 50 ⁇ m, respectively. [0085] Cleaved Caspase-3 Protein.
  • Figure 6 shows immunohistochemistry of cleaved caspase-3 protein in liver tissue from different groups: (a) data shown are mean ⁇ SE; *p ⁇ 0.05 vs. CONTROL group; ⁇ p ⁇ 0.05 vs. NUT + group; A p ⁇ 0.05 vs. SHAM group. Arrows and highlighted boxes indicate cleaved caspase-3 immunopositive cells in each group: (b) CONTROL; (c) IR; (d) NUT + IR; (e) NUT; and (f) SHAM. All images were acquired at 50x magnification and highlighted boxes at 400x magnification, with scale bars at 500 ⁇ m and 50 ⁇ m, respectively.
  • the NUT group had a significantly higher percentage compared to all other groups.
  • the IR group had a significantly higher percentage of TNF-a compared to the NUT + IR and SHAM groups ( Figure 7).
  • the IR group (score 37) presented a significantly higher level of liver injury when compared to the NUT + IR group (score 25). It was also observed that the immunohistochemical analysis of caspase-3 showed a marked presence in the field related to IR injury by hematoxylin-eosin (HE), demonstrating a correlation between the histological and immunohistochemical findings, as shown in Figure 8.
  • HE hematoxylin-eosin
  • Figure 8 shows histological and immunohistochemical analysis: (a) total histological score of the liver of each group; *p ⁇ 0.05 vs. IR group. (b) HE (Hematoxylin-eosin — 600x): photomicrograph of liver tissue showing ischemic changes (ballooning, apoptosis, destrabeculation, and sinusoid congestion), (c) Caspase-3 (Immunohistochemistry - 600x): photomicrograph of the liver parenchyma showing positivity in the cytoplasm (intracytoplasmic brown granular pattern).
  • nutraceutical composition [0093] Based on all the tests performed, the effect of the nutraceutical composition on apoptosis was observed, and its possible mechanism of action was then investigated and it was discovered that it causes a decrease in gene expression and caspase-3 protein in liver tissue.
  • caspase inhibitor involves a new target to protect the liver not only from IR injury, but also from other diseases that have apoptosis in their pathophysiology, such as Alcoholic Liver Disease, Non-Alcoholic Fatty Liver Disease, Hepatocellular Carcinoma, Viral Hepatitis B and C, Cholestatic Liver Disease and Sarcopenia.
  • pan-caspase inhibitor IDN-6556 inhibits caspase-3 activation and reduces sinusoidal endothelial cell apoptosis when used as an additive.
  • composition according to the present invention is capable of acting in the protection of hepatic conditions and favoring a reduction in gene expression and caspase-3 protein, as well as a reduction in TNF-a protein in hepatic tissue.

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Abstract

The present invention relates to a nutraceutical composition for protecting against liver conditions, i.e., ischaemia and reperfusion, as well as to a method for producing said composition. The nutraceutical composition benefits from a synergistic effect of various components for protecting the liver against ischaemia and reperfusion, reducing apoptosis and histological lesions.

Description

COMPOSIÇÃO NUTRACÊUTICA PARA PROTEÇÃO DE CONDIÇÕES HEPÁTICAS E MÉTODO PARA PRODUÇÃO DA MESMA NUTRACEUTICAL COMPOSITION FOR PROTECTION OF LIVER CONDITIONS AND METHOD FOR PRODUCTION OF SAME

[001] A presente invenção pertence ao campo das composições e suspensões nutracêuticas voltadas a proteções de condições de saúde. Mais especificamente, a presente invenção se refere a uma composição nutracêutica para proteção de condições hepáticas, que são a isquemia e reperfusão deste órgão e a apoptose, bem como o método para produção de tal composição. [001] The present invention belongs to the field of nutraceutical compositions and suspensions aimed at protecting health conditions. More specifically, the present invention relates to a nutraceutical composition for protecting liver conditions, which are ischemia and reperfusion of this organ and apoptosis, as well as the method for producing such composition.

ESTADO DA TÉCNICA STATE OF THE TECHNIQUE

[002] Os nutracêuticos são compostos bioativos ou químicos naturais que, além de desempenharem um papel nutricional, melhoram a saúde, curam doenças ou têm propriedades preventivas. São suplementos dietéticos e, do ponto de vista nutricional, os nutracêuticos são uma fonte de nutrientes (lipídios, carboidratos, vitaminas, proteínas, minerais) e não nutrientes (prebióticos, probióticos, fitoquímicos, reguladores enzimáticos). [002] Nutraceuticals are natural bioactive or chemical compounds that, in addition to playing a nutritional role, improve health, cure diseases or have preventive properties. They are dietary supplements and, from a nutritional point of view, nutraceuticals are a source of nutrients (lipids, carbohydrates, vitamins, proteins, minerals) and non-nutrients (prebiotics, probiotics, phytochemicals, enzyme regulators).

[003] Os nutracêuticos podem ser extraídos de alimentos vegetais e animais, concentrado e administrado de forma farmacêutica adequada, com o objetivo de melhorar a saúde, em dosagens que excedem aquelas obtidas a partir de alimentos normais. [003] Nutraceuticals can be extracted from plant and animal foods, concentrated and administered in a suitable pharmaceutical form, with the aim of improving health, in dosages that exceed those obtained from normal foods.

[004] Estudos in vitro e in vivo forneceram evidências de que os nutracêuticos têm atividades antioxidantes, anti-inflamatórias, antibacterianas, antivirais e antifúngicas, bem como evidências de que atuam como moduladores da resposta imune, angiogênese e morte celular. Esses efeitos são possíveis devido aos múltiplos mecanismos de ação dos nutracêuticos [004] In vitro and in vivo studies have provided evidence that nutraceuticals have antioxidant, anti-inflammatory, antibacterial, antiviral and antifungal activities, as well as evidence that they act as modulators of the immune response, angiogenesis and cell death. These effects are possible due to the multiple mechanisms of action of nutraceuticals

[005] Neste contexto, a isquemia e reperfusão hepática são, ainda, um grande problema não resolvido na prática clínica e, são, portanto buscados métodos, composições e tratamentos envolvendo tal condição clínica. Neste sentido, os nutracêuticos são uma terapia em ascensão por seus benefícios nutricionais e terapêuticos, sendo alguns deles já estudados e mostraram resultados promissores, como ao utilizar componentes como resveratrol, quercetina, ácidos graxos ômega-3, selênio, gengibre e abacate, separadamente ou em diferentes combinações. [006] Atualmente, a atenção tem sido focada nos efeitos sinérgicos das combinações nutracêuticas. O conceito de sinergismo foi introduzido por Liu et al., e decorre da sobreposição de efeitos positivos físicos e químicos de uma ou outra adição, provocando um efeito somatório final maior dos componentes combinados entre si. [005] In this context, hepatic ischemia and reperfusion are still a major unresolved problem in clinical practice and, therefore, methods, compositions and treatments involving such clinical condition are sought. In this sense, nutraceuticals are a growing therapy due to their nutritional and therapeutic benefits, some of which have already been studied and shown promising results, such as when using components such as resveratrol, quercetin, omega-3 fatty acids, selenium, ginger and avocado, separately or in different combinations. [006] Currently, attention has been focused on the synergistic effects of nutraceutical combinations. The concept of synergism was introduced by Liu et al., and arises from the overlapping of positive physical and chemical effects of one or another addition, causing a greater final sum effect of the components combined with each other.

[007] Ainda é necessária uma composição compreendendo um efeito sinérgico de componentes combinados entre si, já que tal combinação não é prevista no estado da técnica. [007] A composition comprising a synergistic effect of components combined with each other is still required, since such a combination is not foreseen in the state of the art.

OBJETIVOS DA INVENÇÃO OBJECTIVES OF THE INVENTION

[008] É objetivo da presente invenção prover uma composição nutracêutica de efeito sinérgico para proteção de condições hepáticas, que são a isquemia e reperfusão deste órgão. [008] The objective of the present invention is to provide a nutraceutical composition with a synergistic effect for the protection of liver conditions, which are ischemia and reperfusion of this organ.

[009] É mais um dos objetivos da presente invenção prover uma composição nutracêutica capaz de diminuir a apoptose e a lesão histológica causada pela isquemia e reperfusão hepática. [009] It is another objective of the present invention to provide a nutraceutical composition capable of reducing apoptosis and histological damage caused by hepatic ischemia and reperfusion.

[0010] É outro objetivo da presente invenção prover uma composição farmacêutica que favoreça a redução na expressão gênica e da proteína caspase-3, bem como redução da proteína TNF-a em tecido hepático. [0010] Another objective of the present invention is to provide a pharmaceutical composition that favors the reduction in gene expression and caspase-3 protein, as well as reduction of TNF-a protein in liver tissue.

[0011] É, também, objetivo da presente invenção prover um método para produção da respectiva composição nutracêutica. [0011] It is also an objective of the present invention to provide a method for producing the respective nutraceutical composition.

SUMÁRIO DA INVENÇÃO SUMMARY OF THE INVENTION

[0012] Este sumário é fornecido para apresentar uma seleção de conceitos que são descritos mais detalhadamente na seção descrição detalhada. [0012] This summary is provided to present a selection of concepts that are described in more detail in the detailed description section.

[0013] Neste contexto, a presente invenção se refere a uma composição nutracêutica para proteção de condições hepáticas, mais especificamente condições relacionadas à isquemia e reperfusão do fígado, compreendendo: entre 0,5 e 1 ,0 g/L de pelo menos um polifenol; entre 0,5 e 1 ,0 g/L de cada um de pelo menos quatro flavonoides; entre 0,5 e 1 ,0 g/L de cada um de pelo menos dois carotenoides; entre 0,2 e 0,8 g/L de pelo menos um mineral antioxidante; entre 0,4 e 1 ,0 g/L de pelo menos um ácido graxo poli-insaturado; entre 0,5 e 1 ,0 g/L de pelo menos um extrato vegetal; entre 1 ,0 e 1 ,5 g/L de abacate; entre 0,8 e 1 ,5 g/L de pelo menos um aminoácido de cadeia ramificada; entre 4,0 e 6,0 g/L de pelo menos uma amida; em um veículo líquido farmaceuticamente aceitável. [0013] In this context, the present invention relates to a nutraceutical composition for protection against liver conditions, more specifically conditions related to liver ischemia and reperfusion, comprising: between 0.5 and 1.0 g/L of at least one polyphenol; between 0.5 and 1.0 g/L of each of at least four flavonoids; between 0.5 and 1.0 g/L of each of at least two carotenoids; between 0.2 and 0.8 g/L of at least one antioxidant mineral; between 0.4 and 1.0 g/L of at least one polyunsaturated fatty acid; between 0.5 and 1.0 g/L of at least one plant extract; between 1.0 and 1.5 g/L of avocado; between 0.8 and 1.5 g/L of at least one branched-chain amino acid; between 4.0 and 6.0 g/L of at least one amide; in a pharmaceutically acceptable liquid vehicle.

[0014] Pelo menos um polifenol é resveratrol. [0014] At least one polyphenol is resveratrol.

[0015] Pelo menos um flavonóide é quercetina, um é isoflavonoide, outro é antocianidina e outro é canaflavina. [0015] At least one flavonoid is quercetin, one is isoflavonoid, one is anthocyanidin, and one is cannaflavin.

[0016] Pelo menos um carotenoide é astaxantina e outro é licopeno. [0016] At least one carotenoid is astaxanthin and another is lycopene.

[0017] Pelo menos um mineral antioxidante é selênio, mais especificamente selênio quelado. [0017] At least one antioxidant mineral is selenium, more specifically chelated selenium.

[0018] Pelo menos um ácido graxo poli-insaturado é ômega-3. [0018] At least one polyunsaturated fatty acid is omega-3.

[0019] Pelo menos um extrato vegetal é extrato de gengibre. [0019] At least one plant extract is ginger extract.

[0020] O abacate é proveniente de folhas do abacate em pó. Pelo menos um aminoácido de cadeia ramificada é leucina. [0020] Avocado comes from powdered avocado leaves. At least one branched-chain amino acid is leucine.

[0021] Pelo menos uma amida é nicotinamida. [0021] At least one amide is nicotinamide.

[0022] O veículo farmaceuticamente aceitável é um xarope de carboximetilcelulose. [0022] The pharmaceutically acceptable carrier is a carboxymethyl cellulose syrup.

[0023] As condições hepáticas são por isquemia e reperfusão do fígado e apoptose. [0023] Liver conditions are due to liver ischemia and reperfusion and apoptosis.

[0024] As formas farmacêuticas é uma escolhida dentre suspensão, solução, emulsão, cápsula, comprimido, drágea, grânulo ou pó. [0024] The pharmaceutical forms are chosen from suspension, solution, emulsion, capsule, tablet, dragee, granule or powder.

[0025] Adicionalmente, a presente invenção se refere a método para produção de uma composição nutracêutica para proteção de condições hepáticas que compreende obter uma primeira composição pelas etapas de: (a) obter um veículo líquido farmaceuticamente aceitável; (b) adicionar pelo menos um ácido graxo poli-insaturado a um óleo sob agitação até completa solubilização; (c) juntar os conteúdos das etapas (a) e (b) sob agitação até completa incorporação para obter a primeira solução. E obter uma segunda solução pelas etapas de: (d) adicionar separadamente: pelo menos um polifenol, pelo menos quatro flavonóides, pelo menos dois carotenoides, pelo menos um mineral antioxidante, pelo menos um ácido graxo poli-insaturado, pelo menos um extrato vegetal, abacate em pó, pelo menos um aminoácido de cadeia ramificada, pelo menos uma amida a um recipiente; (e) homogeneizar os componentes da etapa (d) em um veículo líquido para obter a segunda solução. [0025] Additionally, the present invention relates to a method for producing a nutraceutical composition for protecting against liver conditions, which comprises obtaining a first composition by the steps of: (a) obtaining a pharmaceutically acceptable liquid vehicle; (b) adding at least one polyunsaturated fatty acid to an oil under stirring until complete solubilization; (c) combining the contents of steps (a) and (b) under stirring until complete incorporation to obtain the first solution. And obtaining a second solution by the steps of: (d) adding separately: at least one polyphenol, at least four flavonoids, at least two carotenoids, at least one antioxidant mineral, at least one polyunsaturated fatty acid, at least one plant extract, avocado powder, at least one branched-chain amino acid, at least one minus one amide to a container; (e) homogenize the components of step (d) in a liquid vehicle to obtain the second solution.

[0026] A primeira solução é adicionada à segunda solução, sendo ambas agitadas até completa homogeneização. [0026] The first solution is added to the second solution, and both are stirred until completely homogenized.

[0027] A etapa (a) compreende misturar EDTA, benzoato de sódio, água deionizada e carboximetilcelulose até completa dissolução. [0027] Step (a) comprises mixing EDTA, sodium benzoate, deionized water and carboxymethyl cellulose until completely dissolved.

[0028] Pelo menos um óleo da etapa (b) é um óleo vegetal, preferencialmente óleo de girassol. [0028] At least one oil of step (b) is a vegetable oil, preferably sunflower oil.

[0029] A etapa (c) compreende adicionar mel. [0029] Step (c) comprises adding honey.

[0030] O veículo líquido (d) é água deionizada em quantidade suficiente para completa solubilização dos componentes, e o veículo líquido farmaceuticamente aceitável da etapa (a). [0030] The liquid vehicle (d) is deionized water in a quantity sufficient for complete solubilization of the components, and the pharmaceutically acceptable liquid vehicle from step (a).

BREVE DESCRIÇÃO DAS FIGURAS BRIEF DESCRIPTION OF THE FIGURES

[0031] A presente invenção será detalhadamente descrita a seguir com base nas Figuras 1 a 9, descritas nesta seção. [0031] The present invention will be described in detail below based on Figures 1 to 9, described in this section.

[0032] A Figura 1 compreende gráficos referentes aos níveis séricos de transaminases de cada grupo: (a) AST - aspartato aminotransferase; (b) ALT - alanina aminotransferase. [0032] Figure 1 comprises graphs relating to the serum transaminase levels of each group: (a) AST - aspartate aminotransferase; (b) ALT - alanine aminotransferase.

[0033] A Figura 2 compreende gráficos referentes a Mediadores inflamatórios: (a) IL-1 p, (b) IL-6, (c) IL-10 e (d) TNF-a. [0033] Figure 2 comprises graphs relating to inflammatory mediators: (a) IL-1 p, (b) IL-6, (c) IL-10 and (d) TNF-a.

[0034] A Figura 3 compreende gráfico referente ao MDA no tecido hepático de cada grupo. [0034] Figure 3 comprises a graph relating to MDA in the liver tissue of each group.

[0035] A Figura 4 compreende gráficos referentes a Expressão gênica de genes relacionados à apoptose em todos os grupos: (a) BAX, (b) BCL-2, (c) CASPASE-8 e (d) genes CASPASE-3. [0035] Figure 4 comprises graphs relating to gene expression of genes related to apoptosis in all groups: (a) BAX, (b) BCL-2, (c) CASPASE-8 and (d) CASPASE-3 genes.

[0036] A Figura 5 mostra ensaio TUNEL de tecidos hepáticos dos diferentes grupos. [0036] Figure 5 shows TUNEL assay of liver tissues from different groups.

[0037] A Figura 6 mostra imuno-histoquímica da proteína caspase-3 clivada no tecido hepático de diferentes grupos. [0038] A Figura 7 mostra imuno-histoquímica da proteína TNF-a no tecido hepático de diferentes grupos. [0037] Figure 6 shows immunohistochemistry of cleaved caspase-3 protein in liver tissue from different groups. [0038] Figure 7 shows immunohistochemistry of TNF-a protein in liver tissue from different groups.

[0039] A Figura 8 mostra análises histológica e imuno-histoquímica para diferentes grupos. [0039] Figure 8 shows histological and immunohistochemical analyses for different groups.

[0040] A Figura 9 corresponde a um fluxograma de processo do método de acordo com a presente invenção. [0040] Figure 9 corresponds to a process flowchart of the method according to the present invention.

DESCRIÇÃO DETALHADA DA INVENÇÃO DETAILED DESCRIPTION OF THE INVENTION

[0041] A presente invenção será detalhadamente descrita a seguir, incluindo seus dados experimentais e completa base para interpretação e reprodução da invenção proposta. [0041] The present invention will be described in detail below, including its experimental data and complete basis for interpretation and reproduction of the proposed invention.

[0042] Neste contexto, a presente invenção se refere a uma composição nutracêutica para proteção de condições hepáticas, mais especificamente voltada para proteção de isquemia e reperfusão hepática. A composição nutracêutica é baseada em um efeito sinérgico alcançado pela combinação de diferentes componentes capazes de trazer efeitos vantajosos para a proteção de dita condição clínica. [0042] In this context, the present invention relates to a nutraceutical composition for protecting liver conditions, more specifically aimed at protecting liver ischemia and reperfusion. The nutraceutical composition is based on a synergistic effect achieved by combining different components capable of bringing about advantageous effects for the protection of said clinical condition.

[0043] A composição sinérgica conta com a combinação dos seguintes elementos em suas respectivas faixas de concentração, em g/L, em relação ao volume da composição obtida: [0043] The synergistic composition has the combination of the following elements in their respective concentration ranges, in g/L, in relation to the volume of the composition obtained:

- entre 0,5 e 1 ,0 g/L de pelo menos um polifenol, - between 0.5 and 1.0 g/L of at least one polyphenol,

- entre 0,5 e 1 ,0 g/L de cada um de pelo menos quatro flavonoides,- between 0.5 and 1.0 g/L of each of at least four flavonoids,

- entre 0,5 e 1 ,0 g/L de cada um de pelo menos dois carotenoides,- between 0.5 and 1.0 g/L of each of at least two carotenoids,

- entre 0,2 e 0,8 g/L de pelo menos um mineral antioxidante, - between 0.2 and 0.8 g/L of at least one antioxidant mineral,

- entre 0,4 e 1 ,0 g/L de pelo menos um ácido graxo poli-insaturado,- between 0.4 and 1.0 g/L of at least one polyunsaturated fatty acid,

- entre 0,5 e 1 ,0 g/L de pelo menos um extrato vegetal, - between 0.5 and 1.0 g/L of at least one plant extract,

- entre 1 ,0 e 1 ,5 g/L de abacate, - between 1.0 and 1.5 g/L of avocado,

- entre 0,8 e 1 ,5 g/L de pelo menos um aminoácido de cadeia ramificada,- between 0.8 and 1.5 g/L of at least one branched-chain amino acid,

- entre 4,0 e 6,0 g/L de pelo menos uma amida, em um veículo líquido farmaceuticamente aceitável. [0044] De acordo com uma concretização da presente invenção, o polifenol presente na composição tem que ser o resveratrol. - between 4.0 and 6.0 g/L of at least one amide, in a pharmaceutically acceptable liquid vehicle. [0044] According to an embodiment of the present invention, the polyphenol present in the composition must be resveratrol.

[0045] De acordo com uma concretização da presente invenção, o flavonóide presente na composição pode ser pelo menos um dentre as isoflavonas, sendo preferencialmente a quercetina. [0045] According to an embodiment of the present invention, the flavonoid present in the composition may be at least one of the isoflavones, preferably quercetin.

[0046] De acordo com uma concretização da presente inveção, pelo menos um carotenoide é astaxantina e outro é licopeno. [0046] According to an embodiment of the present invention, at least one carotenoid is astaxanthin and another is lycopene.

[0047] De acordo com uma concretização da presente invenção, o mineral antioxidante presente na composição é o selênio, mais especificamente o selênio quelado. [0047] According to an embodiment of the present invention, the antioxidant mineral present in the composition is selenium, more specifically chelated selenium.

[0048] De acordo com uma concretização da presente invenção, o ácido graxo poli-insaturado presente na composição é ômega-3, podendo ser na forma de pó. [0048] According to an embodiment of the present invention, the polyunsaturated fatty acid present in the composition is omega-3, and may be in powder form.

[0049] De acordo com uma concretização da presente invenção, o extrato vegetal presente na composição pode ser um dentre açafrão, gengibre, ou mistura destes, sendo preferencialmente o extrato de gengibre. [0049] According to an embodiment of the present invention, the plant extract present in the composition may be one of saffron, ginger, or a mixture thereof, preferably being ginger extract.

[0050] De acordo com uma concretização da presente invenção, o abacate é utilizado na forma de pó, sendo preferencialmente utilizadas as folhas do abacate em forma de pó seco. [0050] According to an embodiment of the present invention, the avocado is used in powder form, with avocado leaves in dry powder form being preferably used.

[0051] De acordo com uma concretização da presente invenção, o aminoácido de cadeia ramificada presente na composição pode ser substituído por uma entre as saponinas, sendo leucina o componente preferencial. [0051] According to an embodiment of the present invention, the branched chain amino acid present in the composition can be replaced by one of the saponins, with leucine being the preferred component.

[0052] De acordo com uma concretização da presente invenção, a amida presente na composição é nicotinamida (niacina). [0052] According to an embodiment of the present invention, the amide present in the composition is nicotinamide (niacin).

[0053] De acordo com uma concretização da presente invenção, o veículo líquido farmaceuticamente aceitável é o xarope de carboximetil celulose, por exemplo xarope CMC 0,5% (comercializado por Bio Idêntica farmácia de manipulação, São José, Santa Catarina, Brasil). [0053] According to one embodiment of the present invention, the pharmaceutically acceptable liquid carrier is carboxymethyl cellulose syrup, for example CMC syrup 0.5% (sold by Bio Identica compounding pharmacy, São José, Santa Catarina, Brazil).

[0054] A forma farmacêutica da composição nutracêutica de acordo com a presente invenção é, preferencialmente, a solução, mas também pode ser qualquer outra forma farmacêutica líquida (suspensão ou emulsão) ou sólida (cápsula, comprimido, drágea, grânulo e pó). As concentrações de cada um dos componentes na composição são as mesmas, independentemente de suas formas farmacêuticas. [0054] The pharmaceutical form of the nutraceutical composition according to the present invention is preferably the solution, but it can also be any other liquid (suspension or emulsion) or solid (capsule, tablet, dragee, granule and powder). The concentrations of each of the components in the composition are the same, regardless of their pharmaceutical forms.

[0055] Adicionalmente, a presente invenção se refere a um método para produção de uma composição nutracêutica para proteção de condições hepáticas conforme anteriormente descrita. [0055] Additionally, the present invention relates to a method for producing a nutraceutical composition for protection against liver conditions as previously described.

[0056] Neste contexto, o método de acordo com a presente invenção compreende obter uma primeira solução pelas etapas de: [0056] In this context, the method according to the present invention comprises obtaining a first solution by the steps of:

(a) obter um veículo líquido farmaceuticamente aceitável, (a) obtain a pharmaceutically acceptable liquid carrier,

(b) adicionar pelo menos um ácido graxo poli-insaturado a um óleo sob agitação até completa solubilização, (b) adding at least one polyunsaturated fatty acid to an oil under stirring until complete solubilization,

(c) juntar os conteúdos das etapas (a) e (b) sob agitação até completa incorporação para obter a primeira solução. (c) combine the contents of steps (a) and (b) under stirring until completely incorporated to obtain the first solution.

[0057] E obter uma segunda solução pelas etapas de: [0057] And obtain a second solution by the steps of:

(d) adicionar separadamente: pelo menos um polifenol, pelo menos quatro flavonóides, pelo menos dois carotenoides, pelo menos um mineral antioxidante, pelo menos um ácido graxo poli-insaturado, pelo menos um extrato vegetal, abacate em pó, pelo menos um aminoácido, pelo menos uma amida a um recipiente, (d) add separately: at least one polyphenol, at least four flavonoids, at least two carotenoids, at least one antioxidant mineral, at least one polyunsaturated fatty acid, at least one plant extract, avocado powder, at least one amino acid, at least one amide to a container,

(e) homogeneizar os componentes da etapa (d) em um veículo líquido para obter a segunda solução. (e) homogenize the components of step (d) in a liquid vehicle to obtain the second solution.

[0058] A primeira solução é adicionada à segunda solução, sendo ambas agitadas até completa homogeneização. [0058] The first solution is added to the second solution, and both are stirred until completely homogenized.

[0059] De acordo com o método da presente invenção, a etapa (a) de obter um veículo líquido farmaceuticamente aceitável compreende misturar EDTA, benzoato de sódio, água deionizada e carboximetil celulose até completa dissolução. Neste contexto, realiza-se o preparo do xarope CMC a 0,5% de maneira habitual: EDTA e benzoato de sódio são colocados em água deionizada a 70eC; em seguida, agita-se e pulveriza-se CMC até a sua completa solubilização, obtendo-se o xarope CMC 0,5%. [0059] According to the method of the present invention, step (a) of obtaining a pharmaceutically acceptable liquid carrier comprises mixing EDTA, sodium benzoate, deionized water and carboxymethyl cellulose until complete dissolution. In this context, the preparation of the 0.5% CMC syrup is carried out in the usual manner: EDTA and sodium benzoate are placed in deionized water at 70 ° C; then, CMC is stirred and pulverized until its complete solubilization, obtaining the 0.5% CMC syrup.

[0060] A concentração de cada componente da composição é calculada pensando-se na menor dose efetiva via oral, em 1 ,0ml de solução. Para tanto, utiliza- se a EC50 de cada um, que é a concentração antioxidante necessária para se obter inibição de 50% dos radicais livres. A este valor é aplicado o fator de correção de cada matéria-prima (dado pelo fornecedor) e multiplicado pelo volume total da solução, dando a concentração em mg/ml, da seguinte forma: [0060] The concentration of each component of the composition is calculated considering the lowest effective oral dose, in 1.0 ml of solution. For this purpose, use if the EC50 of each one, which is the antioxidant concentration required to obtain 50% inhibition of free radicals. The correction factor of each raw material (given by the supplier) is applied to this value and multiplied by the total volume of the solution, giving the concentration in mg/ml, as follows:

EC50 x Fator de Correção x Volume Total = X mg/ml EC50 x Correction Factor x Total Volume = X mg/ml

[0061] Ainda de acordo com a etapa (b) do método da presente invenção, adiciona-se pelo menos um ácido graxo poli-insaturado a um óleo sob agitação até completa solubilização, e pelo menos 0 óleo utilizado é um óleo vegetal, preferencialmente óleo de girassol. Preferencialmente, 0 ômega-3 será incorporado ao óleo de girassol. [0061] Still according to step (b) of the method of the present invention, at least one polyunsaturated fatty acid is added to an oil under agitation until complete solubilization, and at least the oil used is a vegetable oil, preferably sunflower oil. Preferably, the omega-3 will be incorporated into the sunflower oil.

[0062] Juntar os conteúdos das etapas (a) e (b) sob agitação até completa incorporação para obter a primeira solução, sendo que durante tal etapa (c) mel pode ser adicionado visando a tornar a solução mais palatável, aumentar a concentração dos compostos utilizados, a biodisponibilidade oral deles, e, por conseguinte, aumentar a bioatividade de cada composto nutracêutico. [0062] Combine the contents of steps (a) and (b) under stirring until completely incorporated to obtain the first solution, and during such step (c) honey can be added in order to make the solution more palatable, increase the concentration of the compounds used, their oral bioavailability, and, consequently, increase the bioactivity of each nutraceutical compound.

[0063] O método da presente invenção conta, ainda, com a obtenção de uma segunda solução. Para tanto, são adicionados separadamente: pelo menos um polifenol, pelo menos quatro flavonóides, pelo menos dois carotenóides, pelo menos um mineral antioxidante, pelo menos um ácido graxo poli-insaturado, pelo menos um extrato vegetal, folhas do abacate em pó, pelo menos um aminoácido e pelo menos uma amida a um recipiente. Pode-se, por fim, adicionar um sal inorgânico, como NaCI, seguindo-se com a completa homogeneização de todos os componentes. A esta mistura, adiciona-se 0 veículo líquido CMC, obtendo-se, então, a segunda solução. [0063] The method of the present invention also involves obtaining a second solution. To this end, the following are added separately: at least one polyphenol, at least four flavonoids, at least two carotenoids, at least one antioxidant mineral, at least one polyunsaturated fatty acid, at least one plant extract, powdered avocado leaves, at least one amino acid and at least one amide to a container. Finally, an inorganic salt, such as NaCl, can be added, followed by the complete homogenization of all components. To this mixture, the liquid vehicle CMC is added, thus obtaining the second solution.

[0064] Por fim, a primeira solução é adicionada à segunda solução, sendo ambas agitadas até completa homogeneização, obtendo-se, então, a composição nutracêutica de acordo com a presente invenção. [0064] Finally, the first solution is added to the second solution, and both are stirred until completely homogenized, thus obtaining the nutraceutical composition according to the present invention.

Dados e experimentos Data and experiments

[0065] Com 0 intuito de testar os resultados vantajosos obtidos pela composição nutracêutica de acordo com a presente invenção, diversos ensaios laboratoriais foram realizados, tendo como objetivo avaliar vias de sinalização da lesão de IR (isquemia e reperfusão) do fígado. [0065] In order to test the advantageous results obtained by the nutraceutical composition according to the present invention, several laboratory tests were carried out, aiming to evaluate signaling pathways of IR (ischemia and reperfusion) injury of the liver.

[0066] Neste sentido, foram estudados lesão hepatocelular, mediadores inflamatórios, apoptose por TUNEL, expressão gênica de genes relacionados à apoptose, proteínas TNF-a e caspase-3 no tecido hepático e histologia. [0066] In this sense, hepatocellular injury, inflammatory mediators, apoptosis by TUNEL, gene expression of genes related to apoptosis, TNF-a and caspase-3 proteins in liver tissue and histology were studied.

[0067] Os resultados foram comparados entre cinco grupos: CONTROLE - sem intervenção; IR - ratos submetidos à IR hepática; NUTRACÊUTICOS + IR (NUT + IR) - ratos que receberam a solução nutracêutica por gavagem por 7 dias e foram submetidos a IR hepática; NUTRACÊUTICOS (NUT) - ratos que receberam a solução nutracêutica por 7 dias; e SHAM - ratos submetidos apenas à manipulação hepática. [0068] Os ratos foram alocados em cinco grupos. No grupo CONTROLE (n = 8), os ratos não foram submetidos a nenhum procedimento cirúrgico. No Grupo IR (n = 8), os animais foram submetidos à IR hepática. No grupo NUT + IR (n = 8), os ratos receberam solução nutracêutica por sete dias consecutivos antes de serem submetidos à IR hepática. No grupo NUT (n = 8), os animais receberam solução nutracêutica durante sete dias consecutivos. E no grupo SHAM (n = 5), os ratos foram submetidos à laparotomia mediana e o fígado foi apenas manipulado. [0067] The results were compared between five groups: CONTROL - no intervention; IR - rats subjected to hepatic IR; NUTRACEUTICALS + IR (NUT + IR) - rats that received the nutraceutical solution by gavage for 7 days and underwent hepatic IR; NUTRACEUTICALS (NUT) - rats that received the nutraceutical solution for 7 days; and SHAM - rats subjected only to hepatic manipulation. [0068] The rats were allocated into five groups. In the CONTROL group (n = 8), the rats were not subjected to any surgical procedure. In the IR group (n = 8), the animals underwent hepatic IR. In the NUT + IR group (n = 8), the rats received nutraceutical solution for seven consecutive days before being subjected to hepatic IR. In the NUT group (n = 8), the animals received nutraceutical solution for seven consecutive days. And in the SHAM group (n = 5), the rats underwent median laparotomy and the liver was only manipulated.

[0069] Os dados foram analisados estatisticamente utilizando o software GraphPad Prism (versão 9.5.2). A análise de variância unidirecional (ANOVA) foi usada para avaliar as diferenças entre os grupos testados, seguido pelos testes de comparações múltiplas de Tukey. Os resultados não paramétricos de transaminases e expressão gênica foram analisados pelo teste de Kruskal-Wallis, seguido pelo teste de Dunn. [0069] Data were statistically analyzed using GraphPad Prism software (version 9.5.2). One-way analysis of variance (ANOVA) was used to evaluate differences between tested groups, followed by Tukey's multiple comparisons tests. Nonparametric results of transaminases and gene expression were analyzed by the Kruskal-Wallis test, followed by Dunn's test.

[0070] Os dados categóricos da lesão histológica hepática foram analisados utilizando a estatística do qui-quadrado. Os resultados são apresentados como médias ± erro padrão (EP). O valor de p < 0,05 foi considerado estatisticamente significativo. [0070] Categorical data of liver histological injury were analyzed using chi-square statistics. Results are presented as means ± standard error (SE). A p-value < 0.05 was considered statistically significant.

[0071] Para os testes, o Exemplo 1 , representado pela tabela a seguir, de composição nutracêutica de acordo com a presente invenção foi utilizado, na forma farmacêutica de solução. Tabela 1 - Concentração dos compostos nutracêuticos, em g/L, utilizados na solução

Figure imgf000012_0001
[0071] For the tests, Example 1, represented by the following table, of nutraceutical composition according to the present invention was used, in the pharmaceutical form of solution. Table 1 - Concentration of nutraceutical compounds, in g/L, used in the solution
Figure imgf000012_0001

[0072] Lesão hepatocelular: [0072] Hepatocellular injury:

[0073] Os ratos do grupo IR exibiram um aumento significativo nos níveis séricos de aspartato aminotransferase (AST) e alanina aminotransferase (ALT) em comparação com os grupos CONTROLE e NUT. O grupo NUT + IR apresentou aumento significativo nos níveis séricos de AST e ALT comparado ao grupo CONTROLE, como mostra a Figura 1 . Os dados apresentados são média ± EP; *p < 0,05 vs. CONTROLE grupo; A p < 0,05 vs. grupo NUT. [0073] Rats in the IR group exhibited a significant increase in serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels compared to the CONTROL and NUT groups. The NUT + IR group showed a significant increase in serum AST and ALT levels compared to the CONTROL group, as shown in Figure 1. Data presented are mean ± SE; *p < 0.05 vs. CONTROL group; A p < 0.05 vs. NUT group.

[0074] Mediadores inflamatórios: [0074] Inflammatory mediators:

[0075] Como mostra a Figura 2, não houve diferença em termos de IL-113, IL-6 e IL-10 entre os grupos. O nível sérico de TNF-a foi significativamente aumentado no grupo IR em comparação com o grupo SHAM. Os dados apresentados são média ± EP; *p < 0,05 vs. Grupo SHAM. [0075] As shown in Figure 2, there was no difference in terms of IL-113, IL-6, and IL-10 between the groups. The serum level of TNF-a was significantly increased in the IR group compared with the SHAM group. Data presented are mean ± SE; *p < 0.05 vs. SHAM group.

[0076] Peroxidação lipídica: [0076] Lipid peroxidation:

[0077] O grupo CONTROLE exibiu um nível significativamente mais alto de MDA no tecido hepático quando comparado aos grupos IR, NUT + IR e SHAM. O mesmo também foi observado no grupo NUT em comparação ao grupo NUT + IR, como mostra a Figura 3, que representa MDA no tecido hepático de cada grupo. Os dados apresentados são média ± EP; * p < 0,05 vs. IR grupo; • p < 0,05 vs. grupo NUT + IR; A p < 0,05 vs. grupo SHAM. [0077] The CONTROL group exhibited a significantly higher level of MDA in liver tissue when compared to the IR, NUT+IR and SHAM groups. The same was also observed in the NUT group compared to the NUT+IR group as shown in Figure 3 which represents MDA in liver tissue of each group. The data presented are mean ± SE; * p < 0.05 vs. IR group; • p < 0.05 vs. NUT + IR group; A p < 0.05 vs. SHAM group.

[0078] Expressão Gênica de Apoptose: BAX, BCL-2, CASPASE 8 e CASPASE 3: [0079] A expressão gênica de BAX e BCL-2 foi significativamente maior no grupo NUT + IR comparado aos grupos CONTROLE e SHAM e semelhante aos grupos NUT e IR. O último grupo exibiu um aumento significativo na expressão gênica de BAX em comparação com os grupos CONTROLE, NUT e SHAM. Dentre a expressão gênica de CASPASES, houve apenas uma diferença com CASPASE 3. A expressão gênica de CASPASE 3 foi significativamente menor no grupo NUT + IR do que no grupo IR, que por sua vez teve um valor significativamente maior em comparação aos grupos CONTROLE e SHAM, como mostra a Figura 4. [0078] Apoptosis Gene Expression: BAX, BCL-2, CASPASE 8 and CASPASE 3: [0079] The gene expression of BAX and BCL-2 was significantly higher in the NUT + IR group compared to the CONTROL and SHAM groups and similar to the NUT and IR groups. The latter group exhibited a significant increase in BAX gene expression compared to the CONTROL, NUT and SHAM groups. Among the gene expression of CASPASES, there was only a difference with CASPASE 3. The gene expression of CASPASE 3 was significantly lower in the NUT + IR group than in the IR group, which in turn had a significantly higher value compared to the CONTROL and SHAM groups, as shown in Figure 4.

[0080] A Figura 4 representa a expressão gênica de genes relacionados à apoptose em todos os grupos: (a) BAX, (b) BCL-2, (c) CASPASE 8 e (d) CASPASE 3. Os dados apresentados são média ± EP; *p < 0,05 vs. grupo CONTROLE; p < 0,05 vs. grupo NUT + IR; • p < 0,05 vs. grupo NUT; A p < 0,05 vs. grupo SHAM. [0080] Figure 4 represents the gene expression of apoptosis-related genes in all groups: (a) BAX, (b) BCL-2, (c) CASPASE 8 and (d) CASPASE 3. Data presented are mean ± SE; *p < 0.05 vs. CONTROL group; p < 0.05 vs. NUT + IR group; • p < 0.05 vs. NUT group; A p < 0.05 vs. SHAM group.

[0081] Imunohistoquímica: Apoptose, Caspase-3 Clivada e Proteínas TNF-a no Fígado: [0081] Immunohistochemistry: Apoptosis, Cleaved Caspase-3 and TNF-a Proteins in the Liver:

[0082] Apoptose -Método TUNEL: [0082] Apoptosis -TUNEL Method:

[0083] O método TUNEL foi utilizado para determinar a apoptose das células do fígado. O grupo NUT + IR exibiu uma diminuição significativa na porcentagem de apoptose em comparação com o grupo IR. Além disso, o grupo IR teve uma percentagem significativamente maior de apoptose do que os grupos CONTROLE, NUT e SHAM (Figura 5). [0083] The TUNEL method was used to determine liver cell apoptosis. The NUT+IR group exhibited a significant decrease in the percentage of apoptosis compared to the IR group. Furthermore, the IR group had a significantly higher percentage of apoptosis than the CONTROL, NUT, and SHAM groups (Figure 5).

[0084] A Figura 5 representa semiquantificação do TUNEL nas células hepáticas dos diferentes grupos: (a) os dados mostrados são média ± EP; *p < 0,05 vs. grupo CONTROLE; A p < 0,05 vs. grupo NUT + IR; □ p < 0,05 vs. grupo NUT; *p < 0,05 vs. grupo SHAM. Setas e caixas destacadas indicam células positivas para TUNEL em cada grupo: (b) CONTROLE; (c) IR; (d) NUT + IR; (e) NUT; e (f) SHAM. Todas as imagens foram obtidas com ampliação de 50x e caixas destacadas com ampliação de 400x, com barras de escala de 500 pm e 50 pm, respectivamente. [0085] Proteína Caspase-3 Clivada-. [0084] Figure 5 represents semiquantification of TUNEL in liver cells from different groups: (a) data shown are mean ± SE; *p < 0.05 vs. CONTROL group; □ p < 0.05 vs. NUT + IR group; □ p < 0.05 vs. NUT group; *p < 0.05 vs. SHAM group. Arrows and highlighted boxes indicate TUNEL-positive cells in each group: (b) CONTROL; (c) IR; (d) NUT + IR; (e) NUT; and (f) SHAM. All images were acquired at 50x magnification and highlighted boxes at 400x magnification, with scale bars at 500 μm and 50 μm, respectively. [0085] Cleaved Caspase-3 Protein.

[0086] A análise imuno-histoquímica mostrou que a proteína caspase-3 clivada em tecido hepático foi significativamente menor no grupo NUT + IR do que no grupo IR. Isso significa que a solução nutracêutica foi capaz de diminuir tanto a expressão gênica quanto a proteína caspase-3 na lesão por IR. O grupo IR teve um nível de proteína caspase-3 clivada significativamente maior em comparação aos grupos CONTROLE e SHAM, como evidencia a Figura 6. [0086] Immunohistochemical analysis showed that cleaved caspase-3 protein in liver tissue was significantly lower in the NUT+IR group than in the IR group. This means that the nutraceutical solution was able to decrease both gene expression and caspase-3 protein in IR injury. The IR group had a significantly higher cleaved caspase-3 protein level compared to the CONTROL and SHAM groups, as evidenced in Figure 6.

[0087] A Figura 6 mostra imuno-histoquímica da proteína caspase-3 clivada no tecido hepático dos diferentes grupos: (a) os dados mostrados são média ± EP; *p < 0,05 vs. grupo CONTROLE; □ p < 0,05 vs. grupo NUT +; A p < 0,05 vs. grupo SHAM. Setas e caixas destacadas indicam células imunopositivas à caspase-3 clivadas em cada grupo: (b) CONTROLE; (c) IR; (d) NUT + IR; (e) NUT; e (f) SHAM. Todas as imagens foram obtidas com ampliação de 50x e caixas destacadas com ampliação de 400x, com barras de escala de 500 pm e 50 pm, respectivamente. [0087] Figure 6 shows immunohistochemistry of cleaved caspase-3 protein in liver tissue from different groups: (a) data shown are mean ± SE; *p < 0.05 vs. CONTROL group; □ p < 0.05 vs. NUT + group; A p < 0.05 vs. SHAM group. Arrows and highlighted boxes indicate cleaved caspase-3 immunopositive cells in each group: (b) CONTROL; (c) IR; (d) NUT + IR; (e) NUT; and (f) SHAM. All images were acquired at 50x magnification and highlighted boxes at 400x magnification, with scale bars at 500 μm and 50 μm, respectively.

[0088] Proteína TNF-a [0088] TNF-a protein

[0089] Em relação à proteína TNF-a no tecido hepático, o grupo NUT apresentou porcentagem significativamente maior em comparação com todos os outros grupos. O grupo IR teve maior percentual significativo de TNF-a em comparação aos grupos NUT + IR e SHAM (Figura 7). [0089] Regarding TNF-a protein in liver tissue, the NUT group had a significantly higher percentage compared to all other groups. The IR group had a significantly higher percentage of TNF-a compared to the NUT + IR and SHAM groups (Figure 7).

[0090] Histologia-. [0090] Histology-.

[0091] De acordo com o escore histológico utilizado, o grupo IR (escore 37) apresentou nível de lesão hepática significativamente maior quando comparado ao grupo NUT + IR (escore 25). Também foi observado que a análise imuno-histoquímica da caspase-3 mostrou presença acentuada no campo relacionado à lesão de IR pela hematoxilina-eosina (HE), demonstrando uma correlação entre os achados histológicos e imuno-histoquímicos, como mostra a Figura 8. [0091] According to the histological score used, the IR group (score 37) presented a significantly higher level of liver injury when compared to the NUT + IR group (score 25). It was also observed that the immunohistochemical analysis of caspase-3 showed a marked presence in the field related to IR injury by hematoxylin-eosin (HE), demonstrating a correlation between the histological and immunohistochemical findings, as shown in Figure 8.

[0092] A Figura 8 mostra Análise histológica e imuno-histoquímica: (a) pontuação histológica total do fígado de cada grupo; *p < 0,05 vs. grupo IR. (b) HE (Hematoxilina- eosina — 600x): fotomicrografia de tecido hepático mostrando alterações isquêmicas (balonização, apoptose, destrabeculação e congestão de sinusoides), (c) Caspase-3 (Imunohistoquímica - 600x): fotomicrografia do parênquima hepático mostrando positividade no citoplasma (padrão granulado marrom intracitoplasmático). [0092] Figure 8 shows histological and immunohistochemical analysis: (a) total histological score of the liver of each group; *p < 0.05 vs. IR group. (b) HE (Hematoxylin-eosin — 600x): photomicrograph of liver tissue showing ischemic changes (ballooning, apoptosis, destrabeculation, and sinusoid congestion), (c) Caspase-3 (Immunohistochemistry - 600x): photomicrograph of the liver parenchyma showing positivity in the cytoplasm (intracytoplasmic brown granular pattern).

[0093] Com base em todos os ensaios realizados, observou-se o efeito da composição nutracêutica na apoptose, tendo sido, então, investigado seu possível mecanismo de ação e descobriu-se que causa uma diminuição na expressão gênica e na proteína caspase-3 no tecido hepático. Esses fatos podem caracterizar a composição nutracêutica de acordo com a presente invenção como um inibidor de caspases, que envolve um novo alvo para proteger o fígado não só da lesão de IR, mas também de outras doenças que possuem a apoptose na sua fisiopatologia, como Doença Hepática Alcoólica, Doença Hepática Gordurosa Não Alcoólica, Carcinoma Hepatocelular, Hepatites Virais B e C, Doenças Hepáticas Colestáticas e Sarcopenia. [0094] Nesse sentido, existem alguns compostos em estudo como o IDN-6556 e o F573. O inibidor pan-caspase IDN-6556 inibe a ativação da caspase-3 e reduz a sinusoidal apoptose de células do endotélio quando usado como aditivo. [0093] Based on all the tests performed, the effect of the nutraceutical composition on apoptosis was observed, and its possible mechanism of action was then investigated and it was discovered that it causes a decrease in gene expression and caspase-3 protein in liver tissue. These facts can characterize the nutraceutical composition according to the present invention as a caspase inhibitor, which involves a new target to protect the liver not only from IR injury, but also from other diseases that have apoptosis in their pathophysiology, such as Alcoholic Liver Disease, Non-Alcoholic Fatty Liver Disease, Hepatocellular Carcinoma, Viral Hepatitis B and C, Cholestatic Liver Disease and Sarcopenia. [0094] In this sense, there are some compounds under study such as IDN-6556 and F573. The pan-caspase inhibitor IDN-6556 inhibits caspase-3 activation and reduces sinusoidal endothelial cell apoptosis when used as an additive.

[0095] Observa-se, portanto, que a composição de acordo com a presente invenção é capaz de agir na proteção de condições hepáticas e favorecer uma redução na expressão gênica e da proteína caspase-3, bem como redução da proteína TNF-a em tecido hepático. [0095] It is therefore observed that the composition according to the present invention is capable of acting in the protection of hepatic conditions and favoring a reduction in gene expression and caspase-3 protein, as well as a reduction in TNF-a protein in hepatic tissue.

[0096] Tendo explicado e descrito detalhadamente os aspectos funcionais e construtivos da presente invenção, esclarece-se que os aspectos e características descritos não são limitativos, podendo ser variados em formas e realizações, ainda seguindo as características essenciais desta invenção. [0096] Having explained and described in detail the functional and constructive aspects of the present invention, it is clarified that the aspects and characteristics described are not limitative, and may be varied in forms and embodiments, while still following the essential characteristics of this invention.

Claims

REIVINDICAÇÃO CLAIM 1. Composição nutracêutica para proteção de condições hepáticas caracterizada por compreender: 1. Nutraceutical composition for protection against liver conditions characterized by comprising: - entre 0,5 e 1 ,0 g/L de pelo menos um polifenol, - between 0.5 and 1.0 g/L of at least one polyphenol, - entre 0,5 e 1 ,0 g/L de cada um de pelo menos quatro flavonoides, - between 0.5 and 1.0 g/L of each of at least four flavonoids, - entre 0,5 e 1 ,0 g/L de cada um de pelo menos dois carotenoides, - between 0.5 and 1.0 g/L of each of at least two carotenoids, - entre 0,2 e 0,8 g/L de pelo menos um mineral antioxidante, - between 0.2 and 0.8 g/L of at least one antioxidant mineral, - entre 0,4 e 1 ,0 g/L de pelo menos um ácido graxo poli-insaturado, - between 0.4 and 1.0 g/L of at least one polyunsaturated fatty acid, - entre 0,5 e 1 ,0 g/L de pelo menos um extrato vegetal, - between 0.5 and 1.0 g/L of at least one plant extract, - entre 1 ,0 e 1 ,5 g/L de abacate, - between 1.0 and 1.5 g/L of avocado, - entre 0,8 e 1 ,5 g/L de pelo menos um aminoácido de cadeia ramificada,- between 0.8 and 1.5 g/L of at least one branched-chain amino acid, - entre 4,0 e 6,0 g/L de pelo menos uma amida, em um veículo líquido farmaceuticamente aceitável. - between 4.0 and 6.0 g/L of at least one amide, in a pharmaceutically acceptable liquid vehicle. 2. Composição, de acordo com a reivindicação 1 , caracterizada pelo fato de que pelo menos um polifenol é resveratrol. 2. Composition according to claim 1, characterized in that at least one polyphenol is resveratrol. 3. Composição, de acordo com a reivindicação 1 , caracterizada pelo fato de que pelo menos um flavonóide é quercetina, um é isoflavonoide, outro é antocianidina e outro é canaflavina. 3. Composition according to claim 1, characterized in that at least one flavonoid is quercetin, one is isoflavonoid, another is anthocyanidin and another is cannaflavin. 4. Composição, de acordo com a reivindicação 1 , caracterizada pelo fato de que pelo menos um carotenoide é astaxantina e outro é licopeno. 4. Composition according to claim 1, characterized in that at least one carotenoid is astaxanthin and another is lycopene. 5. Composição, de acordo com a reivindicação 1 , caracterizada pelo fato de que pelo menos um mineral antioxidante é selênio, mais especificamente selênio quelado. 5. Composition according to claim 1, characterized in that at least one antioxidant mineral is selenium, more specifically chelated selenium. 6. Composição, de acordo com a reivindicação 1 , caracterizada pelo fato de que pelo menos um ácido graxo poli-insaturado é ômega-3. 6. Composition according to claim 1, characterized in that at least one polyunsaturated fatty acid is omega-3. 7. Composição, de acordo com a reivindicação 1 , caracterizada pelo fato de que pelo menos um extrato vegetal é extrato de gengibre. 7. Composition according to claim 1, characterized in that at least one plant extract is ginger extract. 8. Composição, de acordo com a reivindicação 1 , caracterizada pelo fato de que o abacate é proveniente de folhas do abacate em pó. 8. Composition according to claim 1, characterized in that the avocado comes from powdered avocado leaves. 9. Composição, de acordo com a reivindicação 1 , caracterizada pelo fato de que pelo menos um aminoácido de cadeia ramificada é leucina. 9. Composition according to claim 1, characterized in that at least one branched chain amino acid is leucine. 10. Composição, de acordo com a reivindicação 1 , caracterizada pelo fato de que pelo menos uma amida é nicotinamida. 10. Composition according to claim 1, characterized in that at least one amide is nicotinamide. 11 . Composição, de acordo com a reivindicação 1 , caracterizada pelo fato de que pelo menos o veículo farmaceuticamente aceitável é um xarope de carboximetilcelulose. 11. Composition according to claim 1, characterized in that at least the pharmaceutically acceptable vehicle is a carboxymethyl cellulose syrup. 12. Composição, de acordo com a reivindicação 1 , caracterizada pelo fato de que as condições são por isquemia e reperfusão do fígado e apoptose. 12. Composition according to claim 1, characterized in that the conditions are liver ischemia and reperfusion and apoptosis. 13. Composição, de acordo com a reivindicação 1 , caracterizada pelo fato de que é fornecida em formas farmacêuticas de suspensão, solução, emulsão, cápsula, comprimido, drágea, grânulo ou pó. 13. Composition according to claim 1, characterized in that it is supplied in pharmaceutical forms of suspension, solution, emulsion, capsule, tablet, dragee, granule or powder. 14. Método para produção de uma composição nutracêutica para proteção de condições hepáticas conforme definida por qualquer uma das reivindicações 1 a 11 , caracterizado pelo fato de que compreende obter uma primeira composição pelas etapas de: 14. A method for producing a nutraceutical composition for protecting against liver conditions as defined by any one of claims 1 to 11, comprising obtaining a first composition by the steps of: (f) obter um veículo líquido farmaceuticamente aceitável, (f) obtain a pharmaceutically acceptable liquid carrier, (g) adicionar pelo menos um ácido graxo poli-insaturado a um óleo sob agitação até completa solubilização, (g) adding at least one polyunsaturated fatty acid to an oil under stirring until complete solubilization, (h) juntar os conteúdos das etapas (a) e (b) sob agitação até completa incorporação para obter a primeira solução; e obter uma segunda solução pelas etapas de: (h) combine the contents of steps (a) and (b) under stirring until completely incorporated to obtain the first solution; and obtain a second solution by the steps of: (i) adicionar separadamente: pelo menos um polifenol, pelo menos quatro flavonóides, pelo menos dois carotenoides, pelo menos um mineral antioxidante, pelo menos um ácido graxo poli-insaturado, pelo menos um extrato vegetal, abacate em pó, pelo menos um aminoácido de cadeia ramificada, pelo menos uma amida a um recipiente, (i) separately add: at least one polyphenol, at least four flavonoids, at least two carotenoids, at least one antioxidant mineral, at least one polyunsaturated fatty acid, at least one plant extract, avocado powder, at least one branched chain amino acid, at least one amide to a container, (j) homogeneizar os componentes da etapa (d) em um veículo líquido para obter a segunda solução, em que a primeira solução é adicionada à segunda solução, sendo ambas agitadas até completa homogeneização. (j) homogenize the components of step (d) in a liquid vehicle to obtain the second solution, in which the first solution is added to the second solution, and both are stirred until completely homogenized. 15. Método, de acordo com a reivindicação 14, caracterizado pelo fato de que a etapa (a) compreende misturar EDTA, benzoato de sódio, água deionizada e carboximetilcelulose até completa dissolução. 15. Method according to claim 14, characterized in that step (a) comprises mixing EDTA, sodium benzoate, deionized water and carboxymethyl cellulose until complete dissolution. 16. Método, de acordo com a reivindicação 14, caracterizado pelo fato de que pelo menos um óleo da etapa (b) é um óleo vegetal, preferencialmente óleo de girassol. 16. Method according to claim 14, characterized in that at least one oil of step (b) is a vegetable oil, preferably sunflower oil. 17. Método, de acordo com a reivindicação 14, caracterizado pelo fato de que a etapa (c) compreende adicionar mel. 17. The method of claim 14, wherein step (c) comprises adding honey. 18. Método, de acordo com a reivindicação 14, caracterizado pelo fato de que o veículo líquido (d) é água deionizada em quantidade suficiente para completa solubilização dos componentes, e o veículo líquido farmaceuticamente aceitável da etapa (a). 18. Method according to claim 14, characterized in that the liquid vehicle (d) is deionized water in a quantity sufficient for complete solubilization of the components, and the pharmaceutically acceptable liquid vehicle from step (a).
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