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    • 专利摘要:
    Composition for the reduction of metabolic decline associated with aging and/or the treatment of disorders related to lipid metabolism. The present invention is directed to compositions comprising Rosmarinus officinalis extract and other plants, as well as nutritional supplements comprising said compositions, as well as to the use of said compositions for the reduction of metabolic decline and of pathologies resulting from said decline. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2739133A1
    申请号:ES201830740
    申请日:2018-07-20
    公开日:2020-01-29
    发明作者:Rada Guillermo Reglero；Olivares Carlos Fernando Torres；De Molina Ana Ramirez；Urbieta María Tabernero；Miguel Del Campo Enrique De；Gomez De Segura Ignacio Alvarez；Aramburu Carlota Largo；Ramiro Mónica Santamaria；Paredes Gonzálo Polo；Perez Daniel Ruiz
    申请人:Fund Imdea Alimentacion；Fundacion Para La Investigacion Biomedica Del Hospital Univ de la Paz；Igenfarma S L；Universidad Autonoma de Madrid；Fundacion para la Investigacion Biomedica del Hospital Universitario La Paz；
    IPC主号:
    专利说明:

    [0001]
    [0002] COMPOSITION FOR REDUCTION OF METABOLIC DECLIVE ASSOCIATED WITH AGING AND / OR TREATMENT OF DISORDERS
    [0003]
    [0004] FIELD OF THE INVENTION
    [0005] The present invention is directed to compositions comprising Rosmarinus officinalis extract and other plants, as well as nutritional supplements comprising said compositions, and to the use of said compositions for the reduction of metabolic decline and of pathologies resulting from said decline.
    [0006]
    [0007] BACKGROUND OF THE INVENTION
    [0008] In the Mediterranean diet, fruits play an essential role as sources of bioactive compounds and nutrients. For example, grape seeds ( Vitis vinifera) are an important natural source of oligomeric polyphenols and proanthocyanidins. Most investigations of oligomeric polyphenols and proanthocyanidins focus on the study of the ability to counteract the negative effect of oxidative stress. Similarly, the fruits of pomegranate ( Púnica granatum) have been widely studied for their high content in phytochemicals, minerals and vitamins. It stands out for its content in elagitannins and punicalagina, antioxidant compounds with potential anticancer effect (Vini R et al, Biofactors, 2015, 41 (2): 78-89),
    [0009] The consumption of herbal infusions shows an unequal pattern in Mediterranean areas. While the European areas its consumption is limited and reduced compared to other types of beverages, in the Mediterranean areas corresponding to North Africa the consumption of infusions is widespread, forming part of the Islamic tradition. Among the most consumed infusions are tea ( Camellia sinensis) and hibiscus flower or Jamaican water ( Hibiscus sabdariffa): Both infusions contain high amounts of antioxidant compounds, the presence of polyphenols such as catechins being characteristic of tea, while tea Hibiscus infusions are characterized by their intense red color produced by their anthocyanin content. Both plants have been studied for their beneficial effects on health, such as anticancer (Conde VR et al, Anticancer agents med chem, 2015, 15 (1), 26-36) or antihypertensives (Serba C, J Hypertens, 2015, 33 (6 ) 119-27). Curcumin is the yellow pigment obtained from the seeds of turmeric ( Curcuma longa), a popular spice of Hindu cuisine present in the preparation of curry. Turmeric is a perennial herb plant with very branched rhizomes of intense yellow color and deep aroma. The geographical distribution of these plants ranges from Polynesia to Southeast Asia and is used in the production of spices. Turmeric consumption is widely established in Asia, reaching an average intake in India about 200 mg / day. It has been described that extracts of turmeric with high concentrations of high availability curcumin have the ability to reduce joint pain and improve motor function in patients with osteoarthritis (Belcaro G et al, Alt Med Rev, 2010, 15 (4), 337 -44). The potential of turmeric extract in improving visual capacity has also been described (Allegri P et al, Clinical Ophthalmology, 2010, 4, 1201-1206).
    [0010] Among the species traditionally used in the Mediterranean diet also highlights the use of rosemary ( Rosmannus officinalis). Among the properties of this plant stands out the powerful antioxidant effect, which has been attributed to some of its components, such as carnosol, carnosic and rosmarinic acids, and flavonoids. Therefore, different diets provide compounds with high potential in health promotion. However, various circumstances, such as lifestyle, eating habits or culinary preferences, mean that part of the population does not benefit from the potential healthy effects of plants and spices. On the other hand, the content of these compounds is normally reduced, which means that the doses obtained within a balanced diet do not reach the minimum effective. It is therefore advisable in certain states and / or pathologies the administration of supplements or drugs. In this sense, the challenge in preventive and therapeutic treatments is to achieve high efficiency in the intervention on the molecular mechanisms to which they are directed, with the greatest safety and acceptance for the patient. In recent years, consumers have increased their acceptance and demand for natural products, extracts or fractions of raw materials of plant or animal origin. The safety and acceptability is even greater when these extracts or fractions are obtained through clean physical technologies.
    [0011] On the other hand, it is always desirable that a composition be able to prevent or treat different aspects of a pathological profile, especially in the case of patients suffering from different diseases or pathologies that require treatment as a whole, as occurs for example in cases of patients suffering from metabolic syndrome or during metabolic decline associated with aging (Schaab et al, PloS One, 2012, 7 (4): e34787).
    [0012] Therefore, the present invention aims to provide an effective and safe natural composition in the treatment of diseases or metabolic disorders, which is especially capable of alleviating different types of said diseases or metabolic disorders simultaneously through different mechanisms.
    [0013]
    [0014] SUMMARY OF THE INVENTION
    [0015] The present inventors have surprisingly discovered that compositions comprising Rosmarinus officinalis extract , and additional extracts of certain plants achieve that objective. In particular, it has been unexpectedly observed that said compositions act globally, in different tissues and systemically simultaneously.
    [0016] Therefore, in a first aspect, the present invention relates to a composition comprising Rosmarinus officinalis extract , and at least one additional extract selected from: Vitis vinífera seed extract, Curcuma longa rhizome extract, leaf extract Camellina sinensis, Hibiscus sabdariffa calyx extract , and Punica granatum fruit extract .
    [0017] In a second aspect, the invention relates to the composition of the first aspect for use in the prevention or treatment of one or more diseases or metabolic disorders.
    [0018]
    [0019] DESCRIPTION OF THE FIGURES
    [0020] Figure 1. Effect of the composition of the invention on adipocyte differentiation and expression of PPARy and C / EBPa.
    [0021] Figure 2. Gene expression of A) Adam 10; Vegfb and Ucp1 in white adipose tissue and B) Srrt, Vegfb and Lepr in brown adipose tissue. (CC): control; (CT) treated.
    [0022] Figure 3. Average weight of the livers after the intervention. Asterisks indicate significant difference. The strip indicates the range of values obtained in the group of healthy animals.
    [0023] Figure 4. Increase in gene expression of genes involved in liver cell proliferation processes of animals treated with the composition of the invention.
    [0024] Figure 5. Modulation of gene expression of genes involved in metabolic homeostasis of lipids and carbohydrates in liver of animals treated with the compound of the invention.
    [0025] Figure 6: Evolution of the average weight of the animals according to the experimental group. Asterisks indicate significant differences (p <0.05) with respect to the fat diet control group.
    [0026] Figure 7: Anthropometric determinations. Total gain of body weight and weight of fatty deposits in retroperitoneal and epididymal area in control animals and treated with the compound of the invention.
    [0027]
    [0028] DETAILED DESCRIPTION OF THE INVENTION
    [0029] The authors of the present invention have observed that, surprisingly, a new composition comprising extracts obtained from the Rosmarinus officinalis plant and certain other plants can be used for the reduction of processes related to metabolic decline, especially that associated to aging Specifically, the compositions have a powerful global effect on weight gain, both preventive and palliative. Moreover, at the systemic level it is effective in reducing circulating lipids and glucose, as well as an improvement in the antioxidant capacity of plasma and a surprising reduction in circulating leptin concentrations. Extracts from other plants are preferably selected from: Vitis vinífera seed extract, Curcuma longa rhizome extract, Camellina sinensis leaf extract, Hibiscus sabdariffa calyx extract , and Punica granatum fruit extract .
    [0030] In a preferred embodiment, the extracts of said certain other plants are preferably selected from: Vitis vinífera seed extract and rhizome extract of Turmeric longa. More preferably, the composition comprises Rosmarinus officinalis extract, Vitis vinífera seed extract and Rhizome extract of Turmeric longa.
    [0031] In another preferred embodiment, the extracts of said other certain plants are preferably selected from: Camellina sinensis leaf extract, Hibiscus sabdariffa calyx extract , and Punica granatum fruit extract . More preferably, the composition comprises Rosmarinus officinalis extract, Camellina sinensis leaf extract, Hibiscus sabdariffa calyx extract , and Punica granatum fruit extract.
    [0032] Additionally, an unexpected effect is observed in target organs of metabolic regulation that are adipose tissue (white and brown) and liver. Surprisingly, the compositions of the invention demonstrate a positive effect by promoting the expression of vascularization markers of both white adipose tissue and brown, a parameter of great relevance in the prevention of metabolic diseases. The biological importance of this parameter is critical, especially when the energy demand is lower than the energy obtained by the diet, such as during aging (due to a decrease in metabolic rate) or obesogenic states (due to an increase in energy intake) . As a consequence of the excess energy, the body promotes an increase in the volume of adipose tissue. When this increase is maintained over time, the existing vascularization is insufficient for the correct oxygenation of the adipocytes. Localized hypoxia generates inflammatory processes and cell death that ends up triggering insulin resistance. Therefore, the correct vascularization of adipose tissue is proposed as a new therapeutic route in the maintenance of metabolic homeostasis.
    [0033] In addition to this, the compositions of the invention provide two important effects on the liver. On the one hand, a beneficial effect on liver tissue regeneration, keeping the size of the organ within the levels considered normal. The biological importance of this parameter is key in aging processes, when cell renewal is diminished. On the other hand, at the hepatic level an effect of beneficial metabolic modulation, both at the level of lipid metabolism and glycogen homeostasis. The biological importance of this parameter is high and is related to a better tolerance to metabolic stress and energy homeostasis.
    [0034] As used in the present invention, the terms Rosmarinus officinalis and rosemary are interchangeable. The composition of the invention comprises a rosemary extract obtained from the inflorescences and upper parts, such as the leaves, of the bush. Rosemary extract can be obtained by conventional conventional methods known in the art. Among the different methods of rosemary extraction, solvent extraction, pressing extraction or supercritical fluid extraction are used.
    [0035] Some of the major compounds that are present in the extracts obtained from Rosmarinus officinalis are for example, but not limited to, rosmanol, rosmadial, epirosmanol, isorosmanol, carnosol, rosmaridiphenol, rosmariquinone, carnosic acid, rosmarinic acid, methyl carnosate, flavonoids such as genkwanina, escutellareina and other compounds such as verbenone, camphor, borneol and 1,8-cineole. Carnosol, which together with carnosic acid and rosmarinic acid is one of the The most potent antioxidants of these compounds, is the component usually the majority component reaching 90% of the extracts.
    [0036] Preferably, the extract comprises between 15 and 35%, between 20 and 30%, or between 24 and 26%, by weight of phenolic diterpenes (preferably including more than 10% carnosic acid) with respect to the total weight of the extract, preferably with respect to the weight Total dry extract. In the context of the present invention, the dry weight of an extract is the weight of said extract when it comprises less than 5%, preferably less than 1%, by weight, or between 5 and 0.5%, preferably between 1 and 0.1%, by weight of the sum of solvents that have been used in the extraction of the extract and water.
    [0037] In the case of solvent extraction, the material (the bush or part thereof) is preferably ground, macerated or chopped before being subjected to extraction with the solvent, thus allowing greater contact area between the solid and the solvent. It is preferably performed at ambient temperature and pressure. The process can be executed in batch (batch) or continuously (such as percolation, leaching and soxhlet extraction). The most commonly used solvents are ethanol, methanol, isopropanol, hexane, cyclohexane, toluene, xylene, ligroin, ethyl ether, isopropyl ether, ethyl acetate, acetone, chloroform, etc. The next step is to separate the solvent from the extract by evaporation, vacuum distillation, molecular distillation or by extraction by supercritical fluids.
    [0038] A second method is pressing extraction, where the plant material is subjected to pressure, either in batch presses or continuously, using for example the following equipment: high or low pressure worm, expeller extractor, centrifugal extractor, decanter extractor or press rollers.
    [0039] As for the extraction under supercritical conditions, the critical point corresponds to the conditions of temperature and pressure, for a gas or a vapor, above which the substance can no longer be "liquefied" by pressure increase. of the liquid and / or vapor phase are the same, ie there is no visible or measurable differentiation between gas and liquid.In a particularly preferred embodiment of the invention, the rosemary extract of the compositions of the invention is obtained by fluid extraction. supercritical, preferably using supercritical carbon dioxide (CO2) Carbon dioxide (CO2) under these conditions has low viscosity, low surface tension, high diffusion coefficient, which leads to high contact with the surface of the material and can penetrate small pores and slits of the same which ensures a Good extraction efficiency in a short time. In the final part of the process there is a total elimination of the solvent and it is carried out at a low temperature, the loss of volatile substances is reduced and the formation of strange flavors and odors is avoided. In a more preferred embodiment of the invention, the rosemary extract of said compositions is obtained with supercritical carbon dioxide, and using co-solvents such as an alcohol, for example ethanol, preferably in amounts of 10% alcohol or less with respect to the total weight of the sum of CO2 and alcohol. In an even more preferred embodiment, the extraction is carried out with carbon dioxide at a temperature of carbon dioxide greater than 30 ° C, or between 30 ° C and 70 ° C, or more preferably 31.10 ° C or higher , or between 31.10 ° C and 70 ° C; and at a carbon dioxide pressure of 73.86 bar or higher, preferably between 200 and 300 bar. In other words, Rosmarinus officinalis extract is a supercritical CO2 or supercritical-alcoholic CO2 extract. In one embodiment, the extraction is carried out with a fixed bed leaching equipment using isopropyl alcohol as a solvent, to subsequently separate the solvent from the extract by vacuum distillation or by extraction by supercritical fluids. For this, it is necessary to have the rosemary leaves ground and dried before extraction. The resulting extract, green and highly viscous, is usually subjected to concentration by molecular distillation.
    [0040] Alternatively, the extracts can be obtained from commercial sources, such as the Rosemary Antoxidant extract 25 (type No. 027.020) product from FLAVEX Naturextrakte GmbH.
    [0041]
    [0042] As used herein, the terms "grape" and "Vitis vinifera" as well as "seed" and "nugget" are used interchangeably. Grape seed extracts are known in the state of the art. They are generally made up of a mixture of polyphenols, more specifically flavonoids (monomers) and their respective proanthocyanidins (dimer or higher). In one embodiment, the flavonoids are the (+) catechin, (-) epicatechin and its gallates, while the proanthocyanidins are the polymeric derivatives of these (dimer or higher).
    [0043] Examples of polymer derivatives of (-) epicatechin are dimers, trimers, tetramers, pentamers, hexamers, heptamers, and preferably dimers, trimers, tetramers.
    [0044] In one embodiment the grape seed extract comprises between 20 and 35%, or between 25 and 30%, by weight of flavonoids plus proanthocyanidins with respect to the total weight of the extract, preferably with respect to the total dry weight of the extract. In another embodiment, the grape seed extract comprises between 20 and 35%, or between 25 and 30%, by weight of proanthocyanidins with respect to the total weight of the extract, preferably with respect to the total dry weight of the extract.
    [0045] In one embodiment the grape seed extract comprises between 0.1 and 10% by weight of the sum of the pentamers, hexamers and heptamers of the (-) epicatechin, and their respective gallic salts, with respect to the weight of the sum of all flavonoids and proanthocyanidins present in the extract; even more preferably this amount is between 3 and 7%, and particularly preferably between 4 and 6%.
    [0046] In a particular preferred embodiment, the amounts by weight of the different components are as follows, always with respect to the total weight of flavonoids and proanthocyanidins present in the extract:
    [0047] 5-25%, preferably 15%: (+) - catechin plus (-) - epicatechin;
    [0048] 60-90%, preferably 80%: sum of gallate, dimers, trimers, tetramers of (-) - epicatechin and gallates thereof;
    [0049] 1-20%, preferably: 5% sum of pentamers, hexamers and heptamers of (-) - epicatechin and gallates thereof;
    [0050] with the proviso that the sum of the percentages of the components adds up to 100%. Vitis vinifera seed extracts can be prepared by extracting and purifying fresh or dried seeds. The extraction can be carried out with different polar solvents such as water, acetone, ethyl acetate, alcohols (such as methanol, ethanol, butanol) or mixtures thereof. In the context of the present invention, the term "polar" solvent refers to a solvent with a dipole moment of more than 1.70 D, while if the value is lower than indicated it is a "non-polar" solvent. GB 1541469 A describes the extraction of grape seeds using a mixture of acetone and water. EP 0348781 A2 similarly refers to the extraction in water and organic solvent such as alcohols or acetone, and further describes additional stages of enrichment of the extract by filtration. In other words, Vitis vinífera seed extract is an extract of polar solvent.
    [0051] In a preferred embodiment, the grape seed extract forms a complex with at least one phospholipid. Such complexes can be prepared according to the procedures described in ES 2056943 T3, the content of which is incorporated herein in its entirety.
    [0052] In one embodiment, the complexes are formed by reacting the extract of Vitis vinifera with phospholipids (of plant, animal or synthetic origin), which respond with the general formula:
    [0053]
    [0054]
    [0055]
    [0056]
    [0057] in which R and R1, identical or different, represent a group -C (= O) alkyl, which is obtained by reaction of the alcohol where R and R1 are H, with natural fatty acids such as palmitic, stearic, oleic acids , linoleic and linolenic; while OR2 represents an aliphatic amino alcohol group, and preferably is - (CH2) 2-N + - (CH 3 ) 3 ; - (CH 2 ) 2 -NH 2 , or - (CH 2 ) - (C (COOH) H) -NH 2 , as described in ES 2056943 T3.
    [0058] Preferred phospholipids are soy or egg lecithin, brain and ox skin bacon or phospholipids and phosphatidylcholines, phosphatidylserines and phosphatidylethanolamines, whose esters derive from the aforementioned fatty acids.
    [0059] In these complexes, the molar ratio of phospholipid to combined flavonoid and proanthocyanidin molecules is generally between 0.5 and 3, preferably 1.
    [0060] The complexes are prepared by adding the phospholipids on the extract comprising the flavonoids and proanthocyanidins in an inert solvent, preferably apolar such as dioxane. The complexes are removed after the solvent for example by evaporation. The phospholipid polar head adheres to the flavonoids and proanthocyanidins, while the fat-soluble part (body and tail) envelops the material adhered to the choline. The result is that a small microsphere or cell is produced, with which the valuable components of the herbal extract are protected from destruction by digestive secretions and intestinal bacteria.
    [0061] In one embodiment the complex comprising the grape seed extract comprises between 20 and 35%, or between 25 and 30%, by weight of flavonoids plus proanthocyanidins with respect to the total weight of the complex, preferably with respect to the total dry weight of the complex. In another embodiment the complex comprising the grape seed extract comprises between 20 and 35%, or between 25 and 30%, by weight of proanthocyanidins with respect to the total weight of the complex, preferably with respect to the total dry weight of the complex.
    [0062] Grape seed extracts can also be obtained from commercial sources, such as the product Leucoselect Phytosome de Indena S.p.A.
    [0063]
    [0064] As used herein, the terms "turmeric", "curcumin" and "Turmeric longa" as well as "root" and "rhizome" are used interchangeably.
    [0065] Turmeric longa root extract can be prepared by procedures known in the art. Preferably, the root includes the primary rhizomes and the several multiple cylindrical secondary rhizomes that grow downward from the primary rhizomes. The root includes curcuminoid compounds. In one embodiment, the curcuminoid compounds are curcumin, demethoxycurcumin and bisdesmethoxycurcumin.
    [0066] The root is preferably treated in a manner that provides curcuminoids in a uniform and easily accessible condition. Such treatments include boiling, curing and subsequent drying of the root, after which the curcominoids are separated. The separation includes any separation process such as precipitation, extraction, filtration or distillation.
    [0067] In a preferred embodiment, the separation step is an extraction performed by adding one or more organic solvents to the curcuminoid-containing material. The addition of any suitable solvent to the material containing curcuminoid serves to carry out an extraction. Examples of suitable solvents are aromatic hydrocarbons, aliphatic hydrocarbons such as petroleum ether, heptane, pentane, hexane; chlorinated hydrocarbons such as ethylene dichloride, dichloromethane, trichlorethylene; ketones such as acetone; esters such as ethyl acetate; alcohols such as methyl alcohol, ethyl alcohol, isopropyl alcohol and n-butanol, or mixtures thereof with water; or any mixture thereof. In one embodiment, the solvent is polar. In another embodiment the solvent is apolar.
    [0068] In another embodiment the extraction is a supercritical extraction using carbon dioxide as a solvent, as described above for the Rosmarinus officinalis extract .
    [0069] In one embodiment, the extract obtained is a liquid form, a semi-liquid form or a solid form.
    [0070] In one embodiment, the extract comprises between 15 and 25%, or between 18 and 22%, by weight of curcuminoids with respect to the total weight of the extract, preferably with respect to the total dry weight of the extract.
    [0071] In one embodiment, the proportion of the three curcuminoids mentioned above in the extract is: 50-70% curcumin, 10-30% demethoxycurcumin and 10-30% bis-demethoxycurcumin; in weight with respect to the combined weight of the three curcuminoids, with the condition that the percentage totals 100%.
    [0072] In a preferred embodiment, the Curcuma longa root extract forms a complex with phospholipids. Such complexes can be prepared according to the procedures described in US 2009/0131373 A1, the content of which is incorporated herein in its entirety.
    [0073] In one embodiment, the complexes are formed by reaction of the Curcuma longa root extract with phospholipids (of plant or synthetic origin). Preferably, the phospholipid is soy lecithin, such as phosphatidyl choline, phosphatidyl serine and phosphatidyl ethanolamine.
    [0074] Preferably, the complexes are prepared by adding the phospholipid to the extract, preferably hydroalcoholic, more preferably ethanolic, comprising the curcuminoids, in a protic solvent, preferably in an alcoholic solvent. Preferably, the addition is carried out under reflux and stirring. The suspension resulting from the addition is concentrated under reduced pressure, and the resulting residue is dried.
    [0075] The molar ratio of phospholipids to curcuminoids is in the range of 10: 1 to 1:10, preferably 5: 1 to 1: 2, and more preferably 2: 1.
    [0076] Preferably, the complex comprises between 15 and 25%, or between 18 and 22%, by weight of curcuminoids with respect to the total weight of the complex, preferably with respect to the total dry weight of the complex.
    [0077] As used herein, the terms "tea", "green tea" and "Camellina sinensis" as well as "infusion" and "leaf extract or" leaves "are used interchangeably. Green tea leaf extracts are known in the state of the art. They are generally made up of a mixture of polyphenols, especially flavonoids and polyphenolic proanthocyanidins, more specifically of catechins and their respective derivatives such as epigallocatechin gallate. In one embodiment, the flavonoids are the (+) catechin, (-) epicatechin and its gallates, while the proanthocyanidins are the polymeric derivatives of these (dimer or higher). Examples of polymer derivatives of (-) epicatechin are dimers, trimers, tetramers, pentamers, hexamers, heptamers, and preferably dimers, trimers, tetramers. In one embodiment, the tea leaf extract comprises between 40 and 65%, or between 35 and 60%, by weight of polyphenols with respect to the total weight of the extract, preferably with respect to the total dry weight of the extract. In one embodiment the green tea leaf extract comprises between 0.1 and 15% by weight of the sum of the pentamers, hexamers and heptamers of the (-) epicatechin, and their respective gallic salts, with respect to the weight of the sum of all flavonoids and proanthocyanidins present in the extract; even more preferably this amount is between 3 and 7%, and particularly preferably between 4 and 6%. Camellia sinensis leaf extracts can be prepared by extracting the leaves by different polar solvents such as water, acetone, ethyl acetate, alcohols (such as methanol, ethanol, butanol) or mixtures thereof, and more preferably with water. , that is to say that Camellina sinensis leaf extract is preferably an aqueous extract. More specifically, the leaves, preferably dried, are ground and subjected to extraction to obtain an extract. The solvent is removed from the extract and the resulting residue is dried, preferably by lyophilization, preferably until a powder is obtained. Therefore, in one embodiment, the extract is preferably in powder form.
    [0078] As used herein, the terms "hibiscus", "hibiscum", "jamaica flower" and "Hibiscus sabdariffa" as well as "infusion", "water" and "extract" as well as "calyx" and "flower "Are used interchangeably. Hibiscus calyx extracts are known in the state of the art. They are generally constituted by a mixture of phenolic compounds and organic acids, more specifically phenolic acids such as protocatechic acid and anthocyanins such as delphinidin- 3-O-sambubioside, delfinidin-3-O-glucoside (mirtillin) and cyanidin-3-O-sambubioside As for organic acids, the presence of hydroxycitric acid and hibiscus acid is characteristic. Jamaica comprises between 4 and 6.5%, or between 4.5 and 6%, by weight of anthocyanins with respect to the total weight of the extract, preferably with respect to the total dry weight of the extract. amaica comprises between 0.01 and 5% by weight of the sum of the pentamers, hexamers and heptamers of the (-) epicatechin, and their respective gallic salts, with respect to the weight of the sum of all the flavonoids and proanthocyanidins present in the extract; even more preferably this amount is between 0.1 and 1%, and particularly preferably between 0.3 and 0.6%. Hibiscus sabdariffa calyx extracts can be prepared extracting and purifying the calyces, preferably dried, by different polar solvents such as water, acetone, ethyl acetate, alcohol (such as methanol, ethanol, butanol, preferably ethanol) or mixtures thereof, and preferably water, alcohol or a combination thereof, and even more preferably a combination of water and alcohol, preferably the alcohol is ethanol. In a preferred embodiment, the Hibiscus flower calyx extract is obtained by a liquid solid extraction using a mixture of ethanol and water, preferably from 1:10 to 10: 1 and more preferably from 1: 1. Subsequently the solvent of the extract can be reduced or eliminated. Therefore, in a preferred embodiment, the hibiscus sabdariffa calyx extract is an aqueous-alcoholic extract. As used herein, the terms "pomegranate" and "Punica granatum" as well as "fruit" and "fruit" are used interchangeably. Pomegranate fruit extracts are known in the state of the art. They are generally made up of a mixture of phenolic compounds, among which anthocyanins (mono- and di-glycosides of cyanidine, pelargonidine and delfinidine) and hydrolysable tannins (galotannins, elagitannins, and galaxyl esters) stand out. Specifically this fruit is rich in punicalagina and punicalin. These molecules contain monomers of ellagic acid esterified among themselves and with other phenolic compounds. In one embodiment the pomegranate extract comprises 30 and 55%, or between 35 and 50%, by weight of ellagic acid with respect to the total weight of the extract, preferably with respect to the total dry weight of the extract. In one embodiment, the pomegranate extract comprises between 1.5 and 5% by weight of the sum of the phenolic compounds and their respective gallic salts, with respect to the weight of the sum of all the flavonoids and proanthocyanidins present in the extract; even more preferably this amount is between 2.0 and 4.0%, and particularly preferably between 2.5 and 3.5%. Pomegranate fruit extracts can be prepared by extracting the fruits by different polar solvents such as water, acetone, ethyl acetate, alcohols (such as methanol, ethanol, butanol, preferably ethanol) or mixtures thereof, and preferably water, alcohol. or a combination thereof. Subsequently, the solvent of the extract can be reduced or eliminated, and preferably the extract is dried, for example, by lyophilization, preferably until a powder is obtained. Therefore, in a preferred embodiment, the Punica granatum fruit extract is an aqueous, alcoholic, or aqueous-alcoholic extract, preferably in powder form.
    [0079] The components of the compositions of the invention are present in a therapeutically effective amount so that the desired effect is achieved, that is, the prevention or treatment of a metabolic disease or disorder.
    [0080] As used herein, the term "treatment" or referrals thereof includes eradication, elimination, reversal, relief, modification or control of the disease or disorder.
    [0081] As used herein, the term "prevention" or referrals thereof refers to preventing or minimizing the occurrence of the disease or disorder.
    [0082] The compositions of the present invention can be formulated for oral, topical, mucosal (e.g., nasal, pulmonary, sublingual, vaginal, buccal or rectal), parenteral (e.g., subcutaneous, intravenous, bolus, intramuscular or intra-arterial injection administration). ), or transdermal to a patient.
    [0083] The compositions of the present invention preferably include one or more pharmaceutically acceptable excipients. Pharmaceutically acceptable means that the excipient is physiologically tolerated, usually meaning that it is not toxic when used for the treatment of a disease or an in humans or animals.
    [0084] The term "excipient" refers to components of a drug compound other than the active substance. They preferably include a "carrier, adjuvant and / or vehicle". Carriers are ways in which substances are incorporated to improve the administration and efficacy of drugs. Drug carriers are used in drug delivery systems such as controlled release technology to prolong drug actions in vivo, decrease drug metabolism and reduce drug toxicity. Carriers are also used in designs to increase the effectiveness of drug delivery to the target sites of pharmacological actions. Adjuvant is a substance added to a finished product formulation that affects the action of the active substance in a predictable manner. Vehicle is an excipient or a substance, preferably without therapeutic action, used as a means to provide volume for the administration of medications. Such carriers, adjuvants or pharmaceutical vehicles may be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like, excipients, disintegrants, wetting agents or diluents.
    [0085] The composition of the invention will preferably be administered orally. Oral dosage forms can be solid or liquid. Solid forms include for example capsules, for example sustained-release capsules, tablets, for example sustained-release tablets, chewable tablets, sublingual tablets, effervescent tablets, pills, suspensions, powders, granules and gels. Liquid forms include, for example, drops, syrups or solutions.
    [0086] In a particularly preferred embodiment of the present invention, the compositions of the invention are in capsule form.
    [0087] In such oral dosage forms, the active compounds may be mixed with conventional excipients known in the art such as binders, for example syrup, gum arabic, gelatin, sorbitol, gum tragacanth or polyvinylpyrrolidone; fillers, for example lactose, sucrose, starch, calcium phosphate, sorbitol or glycine; tablet preparation lubricants, for example magnesium stearate; disintegrants, for example starch, polyvinylpyrrolidone, sodium starch glycolate or microcrystalline cellulose; or pharmaceutically acceptable wetting agents such as sodium lauryl sulfate. In the case of capsules, tablets, effervescent tablets and pills, the dosage forms may also comprise buffering agents. Soft gelatin capsules can be prepared to contain a mixture of the composition of the invention and vegetable oil. The hard gelatin capsules may contain granules of the composition of the invention in combination with a solid, powdery carrier such as lactose, sucrose, sorbitol, mannitol, potato starch, corn starch, amylopectin, cellulose derivatives and gelatin. Tablets and pills can be prepared with enteric coatings. A review of the different forms of administration of active ingredients, of the excipients to be used and their manufacturing procedures can be found in the Treaty of Farmacia Galenica, C. Faulí i Trillo, Luzán 5, S.A. de Ediciones, 1993. In a preferred embodiment, the administration form are soft gelatin capsules, which comprise, in addition to the composition of the invention, water, gelatin, glycerin and vegetable oil. In a preferred embodiment, the vegetable oil is virgin olive oil.
    [0088] In a particular embodiment, the composition of the invention is administered in the form of a nutritional supplement. By "nutritional supplement", as used herein, all additional dietary intake that is consumed orally and that contains one or more nutrients intended to supplement feeding. Generally nutritional supplements are distributed in the form of different presentations, such as tablets, capsules, soft capsules, liquids (syrups or drinks) and powders. Said nutritional supplement may contain conventional excipients, such as those described above, and may be prepared by conventional methods.
    [0089] The compositions of the present invention comprise different components, which, when present, are preferably in the following amounts by weight with respect to the total weight of the composition: 1 to 40%, preferably 2.5 to 30% Rosmarinus officinalis extract ; 5 to 15%, preferably 6 to 10% Vitis vinifera seed extract ; 5 to 15%, preferably 6 to 10% Curcuma longa rhizome extract ; 5 to 18%, preferably 7 to 12% green tea leaf extract; 5 to 15%, preferably 7 to 11% hibiscus calyx extract; 3 to 12%, preferably 5 to 7% pomegranate fruit extract. Preferably, these percentages refer to the dry weight of each extract, or to an extract comprising water or solvent in an amount of maximum 10%, or maximum 5% or maximum 1% with respect to the weight of the extract. In another embodiment, these percentages refer to the dry weight of each extract, or to an extract comprising water or solvent in an amount of between 1 and 10%, or between 1 and 5%, or between 5 and 10% with respect to the Extract weight
    [0090] In a particular embodiment, the total amount of Rosmarinus officinalis extract used in the composition of the invention is between 20-500 mg, the total amount of grape seed extract is between 25-200 mg, the total amount of rhizome extract of turmeric is between 25-200 mg, the amount of green tea leaf extract is between 85-110 mg, the amount of hibiscus calyx extract is between 75-90 mg, and / or the amount of fruit extract from Pomegranate is between 55-75 mg.
    [0091] In a particular embodiment, pharmaceutical formulations, preferably capsules, comprising the composition of the invention contain 25-1000 mg of a composition according to the present invention. Preferably, other ingredients used in said capsules are water, gelatin, glycerin and / or vegetable oil.
    [0092] The exact dose of the composition to be used will depend on the severity of the disease or disorder, and should be decided at the discretion of the patient's circumstances. In a preferred embodiment, said recommended dose is 1 to 3 capsules daily. In a particular embodiment, the composition of the invention comprises, in addition to Rosmarinus officinalis extract, grape seed extract and turmeric rhizome extract, at least one of the ingredients selected from the group: vitamin A, vitamin E, vitamin D3, Vitamin B9, Vitamin B6, Vitamin B12, Vitamin C, Vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B7, vitamin K, calcium, iron, zinc, magnesium, phosphorus, copper, iodine, selenium, lecithin (preferably soy or sunflower, more preferably soy), fish oil , Garcinia cambogia extract, Irvingia gabonensis extract , Softisan, beeswax. In another embodiment, the composition further comprises docosahexanoic acid and / or isoflavones.
    [0093] In a more particular embodiment, the composition of the invention comprises, in addition to Rosmarinus officinalis extract, grape seed extract and turmeric rhizome extract, at least one or all of the ingredients selected from the group: fish oil (preferably from tuna), vitamin E, vitamin B12, vitamin B6, vitamin D3, Softisan, beeswax, lecithin (preferably soy or sunflower, more preferably soy).
    [0094] As used herein, "fish oil" refers to oil obtained from any species of fish, and preferably from salmon, tuna, mackerel, herring, sea bass, sea bass, halibut, catfish, sardine, shark, shrimp, clam, or any combination of them, and even more preferably tuna.
    [0095] As used herein, "Softisan" refers to esters of glycerin and: of natural fatty acids, such as caprylic or capric acid; of isostearic acid; and / or adipic acid.
    [0096] As used herein, "beeswax" refers to a mixture of saturated and unsaturated monoesters, hydrocarbons, free fatty acids, free fatty alcohols and other minor substances produced by the worker bee.
    [0097] Irvingia gabonensis extract is preferably an extract obtained with organic solvent, preferably apolar, such as hexane. More preferably, it is an extract obtained from the leaves, preferably ground, through a Soxhlet extraction using a solvent of said type.
    [0098] Garcinia cambogia extract is preferably an aqueous-alcoholic, aqueous or alcoholic extract. In particular, water, alcohol (preferably methanol) or a mixture thereof is added to the fruit peels, and subsequently sodium or potassium hydroxide is added. The mixture thus obtained is filtered to remove solids, and dried to obtain the extract.
    [0099] The following table indicates the preferred amounts of each of said ingredients, which are to be considered individually and in combination:
    [0100]
    [0101]
    [0102]
    [0103]
    [0104] In a particular embodiment, the composition of the invention further comprises Rosmarinus officinalis extract, Camellina sinensis leaf extract, Hibiscus sabdariffa calyx extract , and Púnica granatum fruit extract , at least one of the ingredients selected from the group: vitamin A, vitamin E, vitamin D3, vitamin B9, vitamin B6, vitamin B12, vitamin C, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B7, vitamin K, calcium, iron, zinc, magnesium, phosphorus, copper , iodine, selenium, lecithin (preferably soy or sunflower, more preferably sunflower), fish oil, Garcinia cambogia extract, Irvingia extract Gabonensis , Softisan, beeswax. In another embodiment, the composition further comprises docosahexanoic acid and / or isoflavones.
    [0105] In a more particular embodiment, the composition of the invention comprises, in addition to Rosmarinus officinalis extract, Camellina sinensis leaf extract, Hibiscus sabdariffa calyx extract , and Púnica granatum fruit extract , at least one or all of the ingredients selected from the group: Garcinia cambogia extract, Irvingia gabonensis extract , vitamin E, vitamin B6, vitamin B12, Softisan, lecithin beeswax (preferably soy or sunflower, more preferably sunflower). The following table indicates the preferred amounts of each of said ingredients, which are to be considered individually and in combination:
    [0106]
    [0107]
    [0108]
    [0109]
    [0110] Therapeutic uses
    [0111] The present invention is directed to the compositions of the invention for use in the prevention or treatment of a metabolic disease or disorder. In the context of the present invention, a disease or metabolic disorder refers to an alteration in normal energy functioning resulting in a pathological state. Preferably, the disease or metabolic disorder is a disease or disorder related to the lipid levels of a subject, in particular characterized by high levels thereof.
    [0112]
    [0113] In a particular embodiment, the metabolic disease or disorder is a disease or disorder related to the triglyceride levels of a subject, particularly characterized by high levels thereof.
    [0114]
    [0115] In a particular embodiment, the metabolic disease or disorder is a disease or disorder related to the cholesterol levels of a subject, in particular characterized by high levels thereof.
    [0116]
    [0117] In a particular embodiment, the metabolic disease or disorder is a disease or disorder related to the levels of free fatty acids of a subject, in particular characterized by high levels thereof.
    [0118]
    [0119] Examples of such diseases are: dyslipidemias or dyslipoproteinemias, such as hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, hyperglyceridemia, hypertriglyceridemia. These can be primary (not associated with other diseases), usually of genetic origin and family transmission, or secondary, that is, linked to other pathological entities, such as metabolic syndrome or obesity.
    [0120]
    [0121] Preferably, the disease or metabolic disorder is obesity. Obesity is understood as the meaning given in the International Classification of Diseases (ICD) -10 of the World Health Organization. Therefore, the present invention is directed in a preferred embodiment to the compositions of the invention for use in the prevention or treatment of obesity.
    [0122] Obesity and diseases and disorders of lipid levels can in turn play an important role in the appearance and progression of the so-called metabolic syndrome. The metabolic syndrome is a set of physiological, biochemical, clinical and metabolic factors that include insulin resistance, excess abdominal fat, atherogenic dyslipidemia, endothelial dysfunction, arterial hypertension, hypercoagulability status and chronic stress. The metabolic syndrome according to the context of the present invention will in any case have the meaning given in ICD-10 of the World Health Organization. Therefore, the present invention is directed in a preferred embodiment to the compositions of the invention for use in the prevention or treatment of metabolic syndrome.
    [0123]
    [0124] Obesity and therefore the metabolic syndrome are characterized by an increase in the oxidative stress of accumulated fats. This oxidative stress increases the proliferation of preadipocytes and their subsequent differentiation, modulated by transcription factors sensitive to free radicals. Therefore, there is a feedback cycle in the increase of adipose tissue that increases oxidative stress, which in turn promotes the synthesis of lipids such as triglycerides and adipocyte differentiation, thus promoting and amplifying the base of the obesity, and consequently of the metabolic syndrome.
    [0125]
    [0126] In the adipose tissue hypoxia and resulting inflammation and apoptosis are also observed when the proliferation of the tissue is greater than normal and the vascularization of the tissue is not sufficient to keep it adequately oxygenated. As mentioned above, the present inventors have observed that the compositions of the present invention are capable of inducing the expression of vascularization markers in adipose tissue. Therefore, the present invention is also directed to the compositions of the invention for use in the prevention or treatment of hypoxia, inflammation and / or apoptosis of adipose tissue, more particularly associated with the aforementioned diseases or disorders (disease or disorder related to lipid levels, obesity, metabolic syndrome). In a particular embodiment, the adipose tissue is white adipose tissue. In another particular embodiment, the adipose tissue is brown adipose tissue.
    [0127]
    [0128] Obesity and diseases and disorders of lipid levels can in turn generate cardiovascular diseases or disorders. Therefore, the present invention is also directed to the treatment of such diseases or cardiovascular disorders. Examples of diseases or cardiovascular disorders are: diseases ischemic heart disease, such as angina pectoris, myocardial infarction, thrombosis or thrombosis-derived embolism, Dressler's syndrome; pulmonary heart and pulmonary circulation diseases, such as pulmonary thromboembolism; cardiomyopathy; cerebrovascular diseases, such as cerebral infarction; diseases of arteries, arterioles and capillaries, such as atherosclerosis; vein diseases, lymphatic vessels and lymph nodes, such as thrombosis and thrombophlebitis, or varicose veins. In particular, the cardiovascular disease or disorder is atherosclerosis. The meaning of these pathological denominations will be that included in the ICD-10 of the World Health Organization.
    [0129] In a particular embodiment, the compositions of the present invention are used for the prevention of one of the above mentioned diseases or disorders in a subject which has a genetic predisposition to said disease or disorder, such as in cases of family transmission.
    [0130]
    [0131] In another particular embodiment, the compositions of the present invention are used for the prevention of the disease or disorder in a subject which has a non-genetic predisposition to said disease or disorder. Examples of non-genetic predispositions are cardiac revascularization surgery, percutaneous transluminal coronary angioplasty, cancers where genes associated with increased lipid activity are over-expressed (for example, prostate, breast, colon or lung cancers in which over-expresses the FASN gene), hypothyroidism, kidney diseases, beta blocker treatments, contraceptive treatments, diuretic treatments, steroid treatments (especially glucocorticoids), or modulator treatments estrogenic receptors (especially tamoxifen), or the abuse of substances such as alcohol or diets high in fat and free sugars.
    [0132]
    [0133] As mentioned above, the inventors have also observed that the compositions of the invention have a direct effect on glucose levels, reducing them, as well as an indirect effect through the prevention of hypoxia, inflammation and / or apoptosis of adipose tissue. That would lead to insulin resistance. Therefore, in another embodiment, the present invention is also directed to the compositions of the present invention for use in the prevention or treatment of a disease or metabolic disorder related to the glucose levels of a subject, in particular characterized by high levels thereof. The present The invention is also directed to the compositions of the present invention for use in the prevention or treatment of insulin resistance.
    [0134]
    [0135] Preferably, the present invention is directed to the compositions of the present invention for use in the prevention or treatment of diabetes, and more preferably of type II diabetes. Diabetes refers to diabetes mellitus as defined in the ICD-10 of the World Health Organization and includes the following diseases derived from diabetes: diabetic coma, diabetic ketoacidosis, diabetic nephropathy, diabetic retinopathy, diabetic neuropathy, diabetic angiopathy , diabetic arthropathy.
    [0136]
    [0137] It has been observed that the compositions of the present invention are particularly active in the liver. On the one hand, in accordance with what is described above, the compositions of the invention are capable of regulating the lipid and glycogen metabolism in the liver, so they are particularly suitable for use in the prevention or treatment of a metabolic disease or disorder. liver characterized by an excessive level of lipids (especially free fatty acids or triglycerides) or glucose. On the other hand, in addition to being active in the regulation of lipid metabolism and glycogen in the liver, the compositions of the present invention induce the expression of genes involved in hepatic proliferation and homeostasis processes.
    [0138]
    [0139] Therefore, in a preferred embodiment, the hepatic metabolic disease or disorder is fatty liver (also known as hepatic steatosis). In the context of the present invention, fatty liver means the meaning given in ICD-10 of the World Health Organization. In a particular embodiment, the fatty liver is alcoholic. In a different embodiment, it is non-alcoholic.
    [0140]
    [0141] In some cases, hepatic steatosis results in steatohepatitis, and in the worst cases in liver fibrosis and / or cirrhosis. Therefore, the present invention is also directed to the use of the compositions of the invention to treat these diseases.
    [0142] On the other hand, the present inventors have also discovered that the compositions of the present invention are capable of alleviating the loss of regenerative capacity of the liver and the resulting loss of volume of the latter. This occurs in cases such as aging. Loss of liver volume leads to a reduction in liver perfusion, crucial in first-pass metabolism, and can lead, for example, to complications such as inappropriate biliary function including reduced bile flow and bile acid secretion. Therefore, this The invention is also directed to the compositions of the invention for use in the prevention or treatment of loss of regenerative capacity or liver volume.
    [0143]
    [0144] In a particularly preferred embodiment of the invention, prevention or treatment of any of the above-mentioned diseases or disorders is achieved through vascularization of adipose tissue, in particular in white and / or brown adipose tissue. More specifically, it is achieved through the induction of Vegfb gene expression.
    [0145]
    [0146] In a particularly preferred embodiment, the invention relates to the compositions of the invention for use in the prevention or treatment of more than one of the metabolic diseases or disorders of those described above. In the context of the present invention, the occurrence of more than one of said diseases or metabolic disorders is considered a metabolic decline.
    [0147]
    [0148] The case of aging deserves particular attention. Scientific advances have increased human life expectancy in the last century. In the European Union, the average life expectancy in 2060 is estimated to be 84.8 years for men and 89.1 for women. This fact, together with the decrease in birth rates, increases the demographic pressure, so that the mechanisms to increase the number of "healthy years" are of vital importance both at the individual and the population level (European Commission The 2015 Ageing Report: Economic and budgetary projections for the 28 EU Member States (2013-2060). "Healthy years" means the number of years that a person of a certain age will live without moderate or severe health problems or without developing a disability (EUROSTAT. Europe in figures, Luxembourg, Office for Official Publications of the European Communities; 2008). From a biological point of view, aging or senescence is the set of morphological and physiological modifications that appear as a consequence of the action of time on living beings, which implies a decrease in the ability to adapt in each of the organs, devices and systems, as well as the ability to respond to harmful agents that affect the individual. Aging is largely characterized by a metabolic decline that leads to a loss of liver, kidney, coronary and / or cardiac functions. These alterations are characterized by the appearance of diseases or metabolic disorders such as those described above. Normally the decline Metabolic by aging is multifactorial, that is to say that it involves the appearance of more than one of said diseases or metabolic disorders.
    [0149] Therefore, in a particularly preferred embodiment, the invention relates to the compositions of the invention for use in the treatment of one or more diseases or metabolic disorders of those described above, wherein said diseases or metabolic disorders are related to the aging. Alternatively, the present invention relates to the compositions of the invention for use in the treatment of one or more diseases or metabolic disorders of those described above, in human subjects 50 years of age or older, 60 or over years of age, 70 years of age or older, or 80 years of age or older.
    [0150] In one embodiment, the aging metabolic decline is caused by a physiological reduction in the production of sex hormones associated with age. In order to determine if there has been a reduction in the metabolic decline associated with aging, the expression of some of the markers mentioned below in the experimental part would be measured. In a preferred embodiment, plasma markers of metabolic risk would be measured. Therefore, one skilled in the art would know that if a decrease in circulating lipids, glucose and leptin is detected accompanied by an increase in the antioxidant capacity of the plasma, it would mean a metabolic systemic protective effect and therefore a positive effect of the compositions of the invention. On a global level, the ability to maintain body weight indices in limits established by normality is considered beneficial, in the same way as the ability to maintain body fat accumulations in a controlled manner without exceeding the limits related to metabolic risks. In a complementary manner, if a protective effect is observed in key tissues in energy homeostasis, such as white and brown adipose tissue and liver, it would mean a metabolic protective effect and therefore a positive effect of the compositions of the invention.
    [0151] In another related aspect, the invention is directed to the use of the compositions of the invention for the preparation of a medicament for the prevention or treatment of a metabolic disease or disorder. In more specific embodiments, the disease or metabolic disorder will be any of those mentioned above. In another related aspect, the invention is directed to a method of treatment comprising administering, to a subject in need, the compositions of the invention, to prevent or treat a metabolic disease or disorder. In more embodiments In particular, the disease or metabolic disorder will be any of those mentioned above.
    [0152] The following examples serve to illustrate the invention and should not be considered as limiting the scope thereof.
    [0153]
    [0154]
    [0155] A series of tests were carried out to know the biological effect of the following composition according to the invention in capsule form:
    [0156]
    [0157]
    [0158] Effect of the administration of the composition of extracts of rosemary, grape seed, and turmeric rhizome in the plasma of animals subjected to metabolic stress
    [0159]
    [0160] For the analysis of the protective effect of the composition an animal model of chronic metabolic stress was designed. For this, the production of endogenous testosterone was suppressed by castration and a high fat diet was administered. During the five weeks after the surgical process, all animals were subjected to a high-fat diet (18% dry weight). The animals were then divided into a control group (n = 8) and a treated group (n = 8), which was given the 5% composition over the next five weeks. After the intervention period, the average weight of the animals treated with the composition was 7% lower than that of the control group (426 ± 24 g versus 456 ± 34 g) although this difference did not reach statistical significance (p = 0.059) .
    [0161] In the plasmas collected at the beginning (t0) and end of the study (sacrifice) the levels of circulating lipids and glucose were evaluated. All these results can be seen in table 2.
    [0162]
    [0163]
    [0164]
    [0165] Table 2: Lipid profile and circulating glucose at the end of the study and difference between onset. P value less than 0.05 indicates statistically significant difference between the control and treated group. The non-difference is represented by ns (not significant). A ^ f indicates the difference observed between the initial and final value of the intervention.
    [0166] The results in plasma indicate a clear regulatory action of the metabolism on the part of the composition. Both the final concentrations of free fatty acids and triglycerides are significantly lower in the group of treated animals than in the control group (68% in the case of free fatty acids and 46% in triglycerides). In addition, comparing the variations of the measures taken at the beginning and at the end of the study, a significant decrease in total cholesterol concentrations and a significant reduction in the increase in circulating glucose were observed. A tendency was observed in the reduction of LDL cholesterol levels (p = 0.09). HDL levels instead were increased, but this increase was not significant.
    [0167] The concentrations of insulin, leptin and proinflammatory cytokines (IL1 p, Il6, TNFa and PAI1) and the antioxidant capacity of plasma were also analyzed. All these results are summarized in table 3.
    [0168]
    [0169]
    [0170]
    [0171] Table 3: Metabolic markers, inflammation and oxidative stress at the end of the study and their variation from the beginning and end of the intervention. P value less than 0.05 indicates statistically significant difference between the control and treated group. No difference It is represented by ns (not significant). A¡ f ^ indicates the difference observed between the initial and final value of the intervention.
    [0172] The results show a clear effect in increasing the antioxidant capacity of plasma. The animals treated with the combination of rosemary, grape seed and turmeric rhizome presented greater antioxidant capacity both at the end of the study and at the level of start-to-end variation. Plasma concentrations of hormones involved in energy metabolism, such as insulin and leptin, were determined. After the intervention, while insulin levels were similar in both groups, animals treated with the composition had significantly lower levels of leptin. Leptin is a hormone of 167 amino acids that participates in the regulation of appetite and energy and whose circulating concentrations are increased in obese individuals.
    [0173] No significant differences were observed in the concentrations of circulating cytokines (IL1beta, IL6 and TNFalfa).
    [0174] In summary these results show that the combination of extracts of rosemary, grape seed and turmeric rhizome is able to improve circulating metabolic markers and increase the antioxidant capacity of plasma without affecting circulating cytokine levels.
    [0175]
    [0176]
    [0177] Effect of the administration of the composition of extracts of rosemary, grape seed and turmeric rhizome in white and brown adipose tissue. In vitro and in vivo studies .
    [0178]
    [0179] In order to evaluate the effect of the compound on the differentiation of preadipocytes, a study was carried out with the mouse fibroblast cell line (3T3 L1) derived from an embryonic state. These cells initiate differentiation of pre-adipocytes to mature adipocytes when they reach confluence and inhibition of contact growth. Once the confluence was reached, the growth medium was replaced by the so-called "induction medium": 10% fetal bovine serum, 0.25 pM Dexamethasone, 0.5 mM IBMX (1-methyl-3- (2-methylpropyl ) -7H-purine-2,6-dione) and 10 pg / ml of insulin After 48 hours in the presence of the induction medium, it was replaced by "differentiation medium" (DMEM, 10% FBS, 5 pDg / ml insulin) for the next six days. The compound was added in a concentration range between 50 pg / ml and 150 pg / ml. Quantitative PCR was analyzed. PPARy and C / EBPa expression levels (both markers of differentiated adipocytes).
    [0180] As shown in Figure 1, the effect of the composition studied on the progression of adipocyte differentiation was evaluated.
    [0181] As can be seen in the images of figure 1, the size of the lipid drops and the presence of mature adipocytes was smaller in those cells treated with the composition. The effectiveness of this composition was observed from a concentration of 150 mg / ml or greater. Analysis of PPARy and C / EBPa expression levels (both markers of differentiated adipocytes) by quantitative PCR indicates a dose-related reducing effect on the expression of C / EBPa.
    [0182] In order to evaluate the effects of the composition in white and brown adipose tissue, the gene expression of samples obtained in an animal model of metabolic stress was evaluated, consisting of the administration of a high-fat diet in castrated animals. During the five weeks after the surgical process, all animals were subjected to a high-fat diet (18% dry weight). The animals were then divided into a control group (n = 8) and a treated group (n = 8), which was given the 5% composition over the next five weeks. At the end of the study, the fatty deposits of the retroperitoneal area were weighed and no differences were observed between the control and treated animals. Brown adipose tissue was collected from the interscapular area at the end of the intervention. No significant differences were seen in the average weight of brown adipose tissue.
    [0183] As shown in Figure 2, in these samples the levels of gene expression of different genes involved in metabolic regulation were determined.
    [0184] In white adipose tissue it was observed that the treatment significantly increased the gene expression of ADAM10 that corresponds to domain 10 of a family of metallopeptidase which are membrane proteins with adhesion capacity plus a protease domain. The gene encoding ADAM has the ability to cleave large numbers of proteins including TNFa and E-caderin. In mouse adipocytes it has been described that ADAM10 is the main protease for the cleavage of the Leptin membrane receptor, to give rise to its soluble version. This process is highly increased in cases of lipotoxicity and apoptosis, which can trigger a blockage of circulating circulating leptin (Schaab et al, 2012 PloS One. 4: e34787).
    [0185] In both white and brown adipose tissue an increase in the gene expression of Vegfb vascular growth factor was observed in the treated animal group, indicating a greater potential vascularization of adipose tissue. Although in white adipose tissue the difference does not become statistically significant (p = 0.07), the increase in Vegfb gene expression in adipose tissue may have an important biological function. This gene encodes vascular endothelial growth factor, which regulates the formation of new vessels and is involved in endothelial cell physiology. Animal studies have shown that Vegfb overexpression protects against hypercaloric diets and the development of obesity and insulin resistance (Elias et al, 2012, Diabetes 61 (7), 1801-1813). In hypercaloric diets, the rapid increase in adipose tissue volume causes vascularization to be insufficient for proper oxygenation. Localized hypoxia generates inflammatory processes and cell death that ends up triggering insulin resistance. The correct vascularization of adipose tissue is proposed as a new therapeutic pathway in metabolic diseases associated with obesity. It has also been proposed that overexpression of Vegfb in adipose tissue could be related to white fat browning, promoting thermogenesis, in addition to an anti-inflammatory effect by chemotactic activity for M2 anti-inflammatory macrophages (Elias, Franckhauser and Bosch 2013 Adipocyte. 2,109 ).
    [0186] In the case of brown adipose tissue, the increase in Vegfb gene expression was statistically significant. It has been described that Vegfb plays an important role in the uptake of circulating lipids, which in the case of brown adipose tissue would be partly destined to thermogenesis (Hashimoto et al, 2016, Biochem. Biophys. Res. Commun. 478 (4), 1746-1750). In addition, vascularization is key in the maintenance of brown adipose tissue and it has been described that vascular dysfunction is a cause of the loss of brown adipose tissue and its conversion into white.
    [0187] In white adipose tissue, a non-significant decrease in the gene expression of UCP1 was observed, a member of the mitochondrial anion transporter family that plays a fundamental role in energy dissipation from ATP phosphorylation and is expressed only in brown adipose tissue . The control group has higher levels of expression of this protein, which could be due to an increase in white fat tissue browning. It has been described that factors present in the animal model used increase the expression of Ucp1, such as castration (Hashimoto et al, 2016, Biochem Biophys Res Commun. 4, 1746) or high fat diet (García-Ruiz et al, 2015 Int. J. Obes. 39 (11), 1619-1629). On the other hand, the gene expression of UCP1 does not directly imply an increase in thermogenesis and adipocyte bioenergetics (Keipert and Jastroch 2014, Biochim. Biophys. Acta BBA 1837 (7), 1075-1082).
    [0188] In brown adipose tissue a significant increase in the gene expression of the Serrate gene was observed, which codes for the homologous RNA effector molecule, also known as arsenic resistance protein 2. This gene plays a role in gene silencing mediated by microRNAs.
    [0189] In this same tissue, leptin receptor gene expression levels (Lepr) were higher in the treated group than in the control group, although the differences were not significant. In peripheral tissues, Lepr expression is related to an increase in basal metabolism, influences reproductive function, regulation of p-pancreatic cells and insulin secretion. It is pro-angiogenic and affects innate and adaptive immunity. In white adipose tissue, Lepr expression levels were also increased by treatment (without being significant; data not shown).
    [0190]
    [0191]
    [0192] Effect of administration of the composition of rosemary extract, grape seed, and turmeric rhizome in liver
    [0193]
    [0194] For the analysis of the protective effect of the composition against hepatic metabolic stress, an animal model of metabolic damage was designed by administering a high-fat diet and suppressing the production of endogenous testosterone by castration. During the five weeks after the surgical process, all animals were subjected to a high-fat diet (18% dry weight). The animals were then divided into a control group (n = 8) and a treated group (n = 8), which was given the 5% composition over the next five weeks. A group of control animals, without fat diet or castration, was maintained as a reference to normal values. After the intervention time the animals were sacrificed and the whole liver that was weighed and processed was obtained.
    [0195] As Figure 3 indicates, the compound was able to reverse the effect of the intervention (castration). While the group of control animals had a liver that was less than normal, the animals treated with the compound had weights in the normal range. This difference was statistically significant (p <0.001) and was maintained by correcting the weight of the liver with the total weight of the animals.
    [0196] Gene expression analysis of liver tissue revealed that several genes involved in hepatic proliferation and homeostasis processes were also regulated by treatment. As Figure 4 shows, the gene expression of Serrate and Adam 10 increased after treatment. Serrate is a mediator of the production of microRNAs during cell proliferation. On the other hand, Adam10 is a gene that encodes a protein involved in hepatic homeostasis and the regulation of the Notch signaling pathway (organ development and tissue integrity). The expression of bile transporters is dependent on Adam10 (Müller M et al, 2016, Oncotarget.
    [0197] 7 (14), 17431-41).
    [0198] In addition, as shown in Figure 5, the gene expression of several genes involved in lipid and glycogen metabolism was modified after treatment indicating an improvement in the hepatic response to a high fat diet. The gene expression data at the hepatic level corroborates the plasma lipid and glucose levels set forth in Example 1. Gene expression of genes involved in lipid metabolism was significantly lower in the group of treated animals compared to controls. For example, Srebf1 is a transcription factor involved in LDL uptake and sterile biosynthesis. Insulin induces its expression, which is associated with an increase in plasma and liver triglyceride levels. After treatment with the composition, Srrbf1 gene expression was reduced.
    [0199] Similarly, a reduction in the gene expression of Lpl, a gene that encodes lipoprotein lipase, was also observed. This enzyme hydrolyzes triglycerides, chylomicrons and VLDLs to produce fatty acids and monoacrylic glycerol, a process necessary to incorporate circulating lipids into different tissues. This enzyme is specifically regulated in different tissues at different levels since it is involved in important energy regulation processes.
    [0200] Fasn gene expression encoding an enzyme involved in the synthesis of long-chain saturated fatty acids was also reduced. It has been described that the high fat diet promotes the expression of this gene, while testosterone reduces it, so the animal model used would have increased expression levels of this gene. As shown in Figure 5, treatment with the compound significantly reduced Fasn expression, exerting protection against the development of steatosis.
    [0201] After treatment, the expression of CD36 was significantly increased, which is related to glycogen homeostasis (Garbacz et al, 2016, Mol Cell Biol.).
    [0202] In diets high in fat CD36 is under-expressed, which induces steatosis and insulin resistance. It has been proposed as a potential target against that for the control of metabolic syndrome.
    [0203]
    [0204]
    [0205] Effect of the administration of the composition of extracts of rosemary, pomegranate fruit, hibiscus calyx and green tea leaf in the prevention and treatment of metabolic syndrome.
    [0206]
    [0207] A series of tests were carried out to know the biological effect of the following composition according to the invention in capsule form:
    [0208]
    [0209]
    [0210] For the analysis of the protective effect of the composition against the development of obesity and metabolic syndrome, a study based on the administration of high fat diet was designed. For this, after adaptation to the experimental conditions 40 animals (healthy C57BL6 male mice of 6 weeks and 17-20 grams of weight) were weighed and subsequently distributed randomly in the experimental groups • Normal diet control: 10 weeks with A04 diet
    [0211] • Control fat diet: 10 weeks with fat diet (TD03307)
    [0212] • Preventive treatment: 10 weeks with fat diet (TD03307) 15% composition
    [0213] • Palliative treatment: 5 weeks with fat diet (TD03307) 5 weeks with fat diet 15% composition
    [0214] During the intervention, the weight gain of the animals and the food intake were monitored. After the intervention time, the animals were sacrificed and a blood sample was obtained, as well as the whole liver and the fatty tissue deposits corresponding to the epididymal and retroperitoneal areas that were weighed and processed.
    [0215] As Figure 6 shows, differences in the weight of the animals were observed throughout the study, these differences being significant with respect to the control with high fat diet (p <0.05; indicated with an asterisk). The group of animals that initiated the consumption of the compound along with the fatty diet (blue color) from the beginning of the study maintained body weight levels similar to the normal diet group and significantly lower than the group of animals with a fat diet. On the other hand, the animals that initially consumed fat diet (orange color) increased their weight in a similar way to the fat diet control group. However, after the first 5 weeks of study, by incorporating the compound into their diet, they quickly reduced their weight, reaching values similar to the normal control group and significantly lower than the fat diet animal group. At the end of the study, the total weight gain for each animal was evaluated, by means of the calculation made with the measurements of the end of the study minus the weight with which each animal began the intervention. As Figure 7 shows, differences in the total weight gain of the animals were observed from the beginning to the end of the study. It is observed that the groups to which the composition was administered increased their weight in a similar way to those observed in the group of animals that consumed a normal diet. The Weight gain was 50% lower than that observed in the fat diet control group, although the caloric content of the diets was similar (5400 kcal / kg) much higher than the normal diet (2900 kcal / kg). Intake data show fluctuations in intakes throughout the study, but at the global level no significant differences were observed between the groups.
    [0216] Figure 7 shows the weight of white adipose tissues collected in the retroperitoneal and epididymal areas. In both deposits it is observed that the compound of the invention is capable of counteracting the deleterious effect of the fatty diet, and maintains the volume of fatty deposits similar to those observed in the group of healthy control animals. The effect is slightly more marked in the group of animals that consumed the compound of the invention since the beginning of the study, which could indicate the efficacy of the product in the control of fatty deposits and their mobilization as energy sources.
    权利要求:
    Claims (24)
    [1]
    1. Composition comprising Rosmarinus officinalis extract , and at least one additional extract selected from: Vitis vinifera seed extract, Curcuma longa rhizome extract, Camellina sinensis leaf extract, Hibiscus sabdariffa calyx extract , and extract of Púnica granatum fruit .
    [2]
    2. Composition according to claim 1, comprising Rosmarinus officinalis extract, Vitis vinífera seed extract and Rhizome extract of Turmeric longa.
    [3]
    3. Composition according to claim 1, comprising Rosmarinus officinalis extract, Camellina sinensis leaf extract, Hibiscus sabdariffa calyx extract , and Punica granatum fruit extract .
    [4]
    4. Composition according to any of the preceding claims, wherein the Rosmarinus officinalis extract is present in an amount of 1 to 40% by weight with respect to the total weight of the composition.
    [5]
    5. Composition according to any one of claims 1, 2, or 4, wherein the Vitis vinífera seed extract is present in an amount of 5 to 15% by weight with respect to the total weight of the composition.
    [6]
    6. Composition according to any one of claims 1, 2, 4 or 5, wherein the Curcuma longa rhizome extract is present in an amount of 5 to 15% by weight with respect to the total weight of the composition.
    [7]
    7. Composition according to any of claims 1, 3 or 4, wherein the Camellina sinensis leaf extract is present in an amount of 5 to 18% by weight with respect to the total weight of the composition.
    [8]
    8. Composition according to any of claims 1, 3, 4 or 7, wherein the Hibiscus sabdariffa calyx extract is present in an amount of 5 to 15% by weight with respect to the total weight of the composition.
    [9]
    9. Composition according to any of claims 1, 3, 4, 7 or 8, wherein the fruit extract of Punica granatum is present in an amount of 3 to 12% by weight with respect to the total weight of the composition.
    [10]
    10. Composition according to any of the preceding claims, wherein the composition further comprises at least one of the ingredients selected from the group consisting of vitamin A, vitamin E, vitamin D3, vitamin B9, vitamin B6, vitamin B12, vitamin C, vitamin B1, vitamin B2, Vitamin B3, Vitamin B5, Vitamin B7, Vitamin K, calcium, iron, zinc, magnesium, phosphorus, copper, iodine, selenium, lecithin, fish oil, Garcinia cambogia extract, Irvingia gabonensis extract , softisan and beeswax.
    [11]
    11. Composition as defined in any of the preceding claims, for use as a medicament.
    [12]
    12. Composition for use according to claim 11, for use in the prevention or treatment of a metabolic disease or disorder.
    [13]
    13. Composition for use according to claim 12, wherein the disease or metabolic disorder is a disease or disorder related to the lipid levels of a subject.
    [14]
    14. Composition for use according to claim 12, wherein the disease or metabolic disorder is hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, hyperglyceridemia, or hypertriglyceridemia.
    [15]
    15. Composition for use according to claim 12, wherein the disease or metabolic disorder is obesity.
    [16]
    16. Composition for use according to claim 12, wherein the metabolic disease or disorder is the metabolic syndrome.
    [17]
    17. Composition for use according to claim 12, wherein the disease or metabolic disorder is fatty liver.
    [18]
    18. Composition for use according to claim 11, for use in the prevention or treatment of diabetes, diabetic coma, diabetic ketoacidosis, diabetic nephropathy, diabetic retinopathy, diabetic neuropathy, diabetic angiopathy or diabetic arthropathy.
    [19]
    19. Composition for use according to claim 11, for use in the prevention or treatment of angina pectoris, myocardial infarction, thrombosis or thrombosis-derived embolism, Dressler's syndrome, pulmonary thromboembolism, cardiomyopathy, cerebral infarction, atherosclerosis, thrombophlebitis, or varicose veins.
    [20]
    20. Composition for use in the prevention or treatment of more than one disease or disorder as defined in claims 12 to 19.
    [21]
    21. Composition for use according to any of claims 11 to 20, wherein the composition is administered orally.
    [22]
    22. Composition for use according to any of claims 11 to 21, wherein the composition is in capsule form.
    [23]
    23. Composition for use according to any of claims 11 to 22, wherein the composition is administered to a human subject 50 years of age or older.
    [24]
    24. Nutritional supplement comprising a composition as defined in any of claims 1 to 10.
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    同族专利:
    公开号 | 公开日
    ES2739133B2|2020-06-08|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    ES2182708A1|2001-07-05|2003-03-01|Velasquez Rene Gabriel Bustos|Diuretic wine dissolving accumulated fat consists of vine, thyme, rosemary and juniper with additives and citrus juice|
    US20090004334A1|2007-06-29|2009-01-01|Vijaya Nair|Dietary nutritional supplements for heal thcare|
    WO2011068812A1|2009-12-04|2011-06-09|Colgate-Palmolive Company|Oral compositions containing a combination of natural extracts and related methods|
    WO2015099616A1|2013-12-23|2015-07-02|Vitiva Proizvodnja In Storitve D.D.|Mixture for reducing body mass|
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