Anti-fat accumulation and anti-cellulite combination.

Abstract

The invention relates to a combination containing Coleus forskohlii extract, proanthocyanidin-rich apple extract, grape marc extract and phloretin powder.

Description

Title of the invention: Anti-fat accumulation and anti-cellulite combination. Technical field.

[0001] The present invention relates to a combination containing a Coleus forskohlii extract, an apple extract rich in proanthocyanidins, a grape marc extract and a phloretin powder.

[0002] The invention also relates to a nutraceutical or topical composition containing said combination as active agent.

[0003] The invention also relates to the use of said combination or said nutraceutical or topical composition, to promote the metabolism of lipids and / or carbohydrates, and / or to reduce the accumulation of body or skin fat and / or to prevent and / or treat cellulite.

[0004] The technical field of the present invention is that of the use of plant extracts having beneficial properties for the body, in particular for the regulation of fat metabolism and / or prevention and / or treatment of cellulite. State of the art.

[0005] Body fat accumulation is a major public health problem. It is associated with an increased risk of disorders, such as overweight and obesity. It is also associated with skin conditions, such as cellulite.

[0006] Fat (lipid) metabolism is the process by which lipids (fats) are absorbed, used, and stored by the body. It is regulated by a number of factors, including hormones, diet, and physical activity.

[0007] To regulate fat accumulation, it is important to maintain a healthy lipid metabolism. This can be done by adopting a healthy lifestyle, which includes a balanced diet, regular physical activity and a healthy weight, that is, a body weight corresponding to a body mass index (BMI) between 18.5 and 25. According to the World Health Organization (WHO), a BMI below 18.5 is considered underweight, and a BMI above 25 is considered overweight or obese.

[0008] However, there is a need for products or compositions that better regulate fat metabolism. These products or compositions could be beneficial for those seeking to lose weight or maintain a healthy weight. They could also be used to improve and / or treat cellulite.

[0009] Cellulite is a condition characterized by uneven skin dotted with bumps and dimples, leaving the skin with a texture resembling orange peel. It is a common condition that primarily affects post-pubescent females and a majority of them (about 90%). It is usually seen in parts of the body with greater fat storage such as the thighs, buttocks, abdomen, and sometimes the upper arms. Cellulite is not specific to overweight people, although increased adiposity (fat accumulation) exacerbates it.

[0010] There are three types of cellulite, namely: fatty cellulite, aqueous cellulite and fibrous cellulite.

[0011] Adipose cellulite is mainly linked to an excessive accumulation of subcutaneous fat. It gives the skin a bumpy and dimpled appearance. It is only visible when the skin is pinched between the finger and index finger (hereinafter called the pinch test). It is not painful.

[0012] Aqueous cellulite is linked to excessive water retention in the skin tissues. It gives the skin a flaccid and swollen appearance. It is barely visible and has a slightly bumpy appearance on the pinch test. It is generally due to problems with venous or lymphatic circulation.

[0013] Fibrous cellulite is the most advanced form of cellulite, resulting from a prolonged presence of this condition in the affected area. It is characterized by the development of fibrous connective tissue under the skin, leading to the formation of nodules or bands, which gives a granular texture to the affected area. To the touch, it appears firm and can cause a painful sensation on the pinch test.

[0014] It is important to emphasize that cellulite is not a disease. It is described as a normal physiological condition in post-pubertal women that maximizes fat retention to ensure adequate caloric availability for pregnancy and lactation. However, it poses an aesthetic problem and can even cause considerable distress to those affected. There is therefore a need to find effective methods to combat cellulite.

[0015] The specific structure of the skin influences the appearance of cellulite. Beneath the skin's surface, adipose tissue rich in fat cells (adipocytes) is anchored by trabeculae of fibrous connective tissue, above the dermis and below the anterior fascia of the muscle. These trabeculae, also called septa, are composed mainly of collagen fibers.

[0016] The trabeculae (or septa) help maintain (i) the structural integrity of the skin; (ii) the passage of nerve fibers and blood vessels to ensure the supply of oxygen and nutrients to the skin cells; and (iii) the passage of lymphatic vessels whose function is to drain lymph (liquid containing water, various white blood cells, antibodies, nutrients, bacteria, toxins as well as cellular and metabolic waste), towards the lymph nodes and then towards the blood circulation. Furthermore, the fibrous connective tissue bands compartmentalize the fat cells into fat pockets. As the number and size of fat cells increase through the processes of adipogenesis and lipogenesis, the fat pockets swell and push against the skin, which is pulled inward by the fibrous connective tissue bands. The skin then deforms, creating bumps and dimples that give the characteristic appearance of cellulite. In addition, as the fat pockets increase in size, they exert pressure on the surrounding blood and lymphatic vessels, thus impeding the supply of oxygen and nutrients to the skin cells on one side, and disrupting the normal flow of lymph on the other.This inability to effectively drain lymph elements, such as interstitial fluids, toxins, and waste, leads to their accumulation and the formation of edema (water retention). This situation leads to asphyxiation of the cells of the surrounding tissues, disrupting the synthesis of collagen by dermal fibroblasts. In addition, it causes chronic low-grade inflammation, which in turn triggers the release of pro-inflammatory cytokines. This leads to the activation of pathways involved in the increase in the production of reactive oxygen species (ROS), thus creating a state of oxidative stress. Similarly, a high level of oxidative stress can stimulate the production of inflammatory cytokines.

[0017] Adipogenesis is a process of cellular differentiation during which preadipocytes become adipocytes. The first early differentiation gives rise to young adipocytes which, after late differentiation, give rise to mature adipocytes.

[0018] Lipogenesis refers to the set of biochemical processes involved in the biosynthesis of triglycerides and fatty acids and their storage in adipocytes. Lipogenesis is a metabolic pathway that occurs from dietary triglycerides, fatty acids and other non-lipid precursors such as dietary carbohydrates or other substrates in excess in the diet.

[0019] Furthermore, it is important to note that the configuration of the hypodermal tissue is not the same in women as in men. In women, the trabeculae framing the fat pockets are oriented in a parallel manner, perpendicular to the surface of the skin, whereas in men, these trabeculae have an oblique orientation. This difference explains why cellulite is generally a feminine manifestation. Indeed, in women, when the fat pockets increase in volume, they easily protrude into the dermis, causing the formation of bumps and dimples on the surface of the skin. In men, the oblique orientation of the trabeculae of fibrous connective tissue prevents or limits the deformation of the skin in the event of excessive development of fat cells.

[0020] The exact causes of cellulite are not fully understood, but most scientists agree that it involves localized hypertrophy (increase in volume) of fat cells, impaired microcirculation, dysregulation of collagen synthesis, persistent low-grade inflammation, and oxidative stress.

[0021] The prevalence of cellulite has led to numerous attempts at treatment.

[0022] Cellulite treatment strategies are currently based primarily on medical-aesthetic methods, for example, manual or mechanical massage techniques, invasive treatments such as mesotherapy, liposuction or subcision, and non-invasive treatments such as acoustic wave therapy or radiofrequency and / or laser therapy. Although these medical-aesthetic methods achieve satisfactory results, they are expensive, cumbersome and restrictive.

[0023] The prior art also provides anti-cellulite preparations based on plant extracts that are used topically on the skin. While current topical preparations can help improve skin texture and reduce the appearance of cellulite, they nevertheless have drawbacks related to local application. By definition, topical preparations act locally on the areas of skin to be treated, where they may be unevenly distributed, and require repeated and lengthy applications. Furthermore, they can in certain cases cause skin side effects, or even discomfort.

[0024] Alternatively, there are also oral compositions for the treatment of cellulite. Unlike topical preparations, oral compositions offer the advantage of acting from within the body to exert a cellulite-improving effect. Indeed, the metabolites and other active ingredients are particularly distributed in the skin via the bloodstream. However, no oral composition proposed so far has been sufficiently effective for treating cellulite.

[0025] In view of the above and due to the increasing demand for new products for oral administration and the attraction of consumers for products of natural origin, there is a need for new products which make it possible to stimulate the metabolism of lipids (fats) and / or to reduce the accumulation of body fat and / or to improve or treat cellulite. Presentation of the invention.

[0026] The present invention is based on the discovery that a combination containing a Coleus forskohlii extract, an apple extract rich in proanthocyanidins, a grape marc extract and a phloretin powder. The combination has several beneficial effects and in particular makes it possible to reduce the accumulation of body fat and / or to prevent and / or treat cellulite, whether adipose, aqueous and / or fibrous.

[0027] Furthermore, this combination as well as the nutraceutical composition or the topical composition containing it according to the present invention are remarkably well tolerated by the body and do not present any undesirable effects on human health. They are also inexpensive, being based on inexpensive products available on the market or simple to produce according to methods known to those skilled in the art.

[0028] The apple is a fruit produced by an apple tree which is a tree belonging to the Rosaceae family, genus Malus, more specifically the species Malus Pumula and Malus Domestica. The apple represents a rich source of polyphenols including flavonoids such as anthocyanidins and anthocyanins, proanthocyanidins, flavan-3-ols (e.g. catechin, epicatechin), and phenolic acids such as hydroxy-cinnamic acids, including chlorogenic acid. Apple skin or peel is said to contain more than twice as many polyphenols as apple flesh. The polyphenols found in apples are relatively effective antioxidants. They have also demonstrated numerous biological effects in the prevention of various chronic diseases, including cardiovascular disease, diabetes, cancers and neurodegenerative diseases.In particular, apple polyphenols may help alleviate type II diabetes through various mechanisms such as inhibition of digestive enzymes that metabolize carbohydrates, such as α-glucosidase and α-amylase (see for example the article: Barbosa, ACL, et al., 2010).

[0029] It should be recalled here that antioxidants generally derive their name from their ability to slow down or prevent the oxidation of other chemicals caused by free radicals. These chemicals can be biomolecules such as proteins, nucleic acids, carbohydrates and lipids. Free radicals include, in particular, reactive oxygen species (ROS), such as hydroxyl radicals (HO*), peroxyls (HOO*, ROO) and superoxide ions (02*-), which tend to form in cells and can cause cellular and tissue damage if not neutralized by antioxidants. In addition, an excess of free radicals can lead to a state of oxidative stress in the body. This phenomenon can have a negative impact on many aspects of health by affecting various tissues, including adipose tissue and skin tissue.In adipose tissue, excess free radicals can disrupt metabolic balance, impacting fat regulation in the body. This disruption may play a role in the development of cellulite. As for skin tissue, free radical-induced oxidative stress can damage collagen and elastin fibers, which are essential for maintaining skin firmness and elasticity. However, when . These fibers are affected by oxidative stress, they can lose their strength and natural elasticity. However, sagging skin can be a contributing factor to the appearance of cellulite.

[0030] Coleus forskohlii also called Indian Coleus, is a perennial plant belonging to The Lamiaceae family and historically used in Ayurvedic medicine to treat a variety of conditions, including inflammatory diseases, hypertension, respiratory disorders, obesity, and asthma. Its roots contain an active substance called forskolin (7[3-acetoxy-8,13-epoxy-la,6[3,9a-trihydroxylabd-14-en-ll-one; CAS No. 66575-29-9) which is a potent activator of adenylate cyclase, an enzyme that increases cyclic adenosine monophosphate (cAMP) in cells. cAMP is a key intracellular messenger in the lipolysis (fat breakdown) mechanism. An increase in cAMP promotes the breakdown of fats in fat cells and their use as fuel in the body, which can lead to increased thermogenesis and loss of body fat.Research (in vivo) has shown that forskolin can optimize body composition by increasing lean mass and metabolic activity.

[0031] Grape marc (called in English "grape marc" or "grape pomace") is a residual product obtained from the fruits (grapes) of the vine, a plant of the genus Vitis (Botanical family: Vitaceae). Typically, grape marc consists of the solid part resulting from the pressing or crushing operation of grape bunches, that is to say the skins, seeds and possibly stalks. It represents approximately 20 to 30% of the weight of grapes used to make wine or grape juice. Grape marc is a particularly rich source of polyphenolic compounds (5 to 10% by weight). These polyphenolic compounds include anthocyanins, flavonoids including flavanols (e.g., catechins) and flavonols (e.g., quercetin), phenolic acids such as hydroxycinnamic acid and gallic acid, stilbenes including resveratrol, and tannins.These polyphenol compounds are known for their broad spectrum of biological activities, including antioxidant, anti-inflammatory, anticancer, antimicrobial, antithrombotic, cardioprotective, and vasodilatory properties. Grape pomace phenolic compounds have also shown significant inhibitory activities, particularly in limiting dietary fat absorption and fat accumulation in adipose tissue (see, for example, the following articles: Hogan S., et al., 2010; Yunoki K., et al. 2008). Grape pomace is also an important source of dietary fiber, which accounts for approximately 30% to 60% of the dry weight of grape pomace. These dietary fibers include soluble fibers and insoluble fibers, such as cellulose and hemicellulose. In addition, grape pomace . Contains mono- and polyunsaturated fatty acids, essential minerals (e.g., calcium, iron, potassium, manganese, and zinc), vitamins (e.g., vitamins C and E), and a range of other health-promoting compounds. The phytochemical composition of grape pomace is influenced by various factors, including grape variety, weather conditions, terroir, and the ripeness of the grapes. For example, it is common to find that red grapes generally have a higher content of anthocyanins (a subgroup of flavonoids) than white grapes.

[0032] Phloretin [2',4',6'- trihydroxy-3-(4-hydroxyphenyl)-propiophenone; CAS No. 60-82-2] is a naturally occurring polyphenolic compound that belongs to a special family of flavonoids called dihydrochalcones and is present in many fruits and vegetables, including apple leaves (Malus sp., Rosaceae). Phloretin has many biological activities such as antioxidant, anti-inflammatory, antidiabetic, anti-obesity, antitumor, antimicrobial, hepatoprotective, and cardioprotective.

[0033] To the knowledge of the Applicant, there is currently no combination comprising an extract of Coleus forskohlii, an extract of grape marc, an apple extract rich in proanthocyanidins and a phloretin powder, and even less its use with a view to promoting the metabolism of lipids and carbohydrates and / or reducing the accumulation of fat and / or preventing and / or slowing down the formation of cellulite or reducing cellulite already present.

[0034] Thus, according to one aspect of the invention, there is provided a combination of a Coleus forskohlii extract, an apple extract rich in proanthocyanidins, a grape marc extract and a phloretin powder.

[0035] Other advantageous characteristics of the invention are listed below. Each of these characteristics can be considered alone or in combination with the remarkable characteristics defined above, and be the subject, where appropriate, of one or more divisional patent applications:

[0036] In particular, the extract of Coleus forskohlii is present in a content of at least 40% w / w combination, preferably 45 to 70% w / w combination.

[0037] The apple extract may contain at least 50% w / w apple extract, preferably at least 90% w / w apple extract of proanthocyanidins.

[0038] Preferably, the apple extract rich in proanthocyanidins is present in the combination according to the invention in a content of at least 1% w / wCombination, preferably 2 to 9% w / wCombination.

[0039] The grape marc extract may be present in the combination according to the invention in a content of at least 4% w / wCombination, preferably from 4 to 20% w / wCombination-

[0040] Preferably, the content of phloretin powder is at least 20% w / w. combination, preferably 20 to 40% p / pcombination-

[0041] In particular, the combination according to the invention contains (a) at least 40% w / w combination of Coleus forskohlii extract, (b) at least 1% w / w combination of proanthocyanidin-rich apple extract, (c) at least 4% w / w combination of grape marc extract, (d) at least 20% w / w combination of phloretin powder, the sum of (a), (b), (c) and (d) being equal to 100%.

[0042] More particularly, the combination according to the invention contains: (a') from 45 to 70% w / pcombination of the root extract of Coleus forskohlii, (b') from 2 to 9% w / p combination of apple extract rich in proanthocyanidins, (c') from 4 to 20% w / p combination of grape marc extract, (d') from 20 to 40% w / w combination of phloretin powder, the sum of (a'), (b'), (c') and (d') being equal to 100%.

[0043] More particularly still, the combination according to the invention contains: 58.39% w / pcombination of Coleus forskohlii root, 3.65% w / w combination of apple extract rich in proanthocyanidins, 8.76% w / w combination of grape marc extract, 29.20% w / w combination of phloretin powder.

[0044] According to another aspect, the invention provides a nutraceutical or topical composition comprising, as active agent, a combination according to the present invention.

[0045] The nutraceutical or topical composition according to the present invention may further comprise one or more excipients.

[0046] The nutraceutical or topical composition according to the present invention may further comprise one or more additional components such as vitamins, carotenoids, mineral salts and / or trace elements, antioxidants, one or more sources of prebiotics, one or more plant extracts capable of stimulating the metabolism of lipids and / or carbohydrates and / or of promoting blood circulation, amino acids, probiotic bacteria and / or yeasts.

[0047] Preferably, the nutraceutical composition according to the present invention is presented in a form suitable for oral administration, in particular, in the form of powders, tablets, capsules, granules, candies, chewing gums, or aqueous or oily suspensions.

[0048] In particular, the nutraceutical composition according to the present invention is a food supplement.

[0049] Preferably, the topical composition according to the present invention is presented in a form suitable for topical administration, in particular, in the form of creams, milks, ointments, lotions, serums, gels, pastes, mousses, sticks, aerosols.

[0050] Advantageously, the combination according to the present invention, or the composition nu- traceutical or topical according to the invention is intended for use in reducing the accumulation of body or skin fat and / or in preventing and / or treating cellulite. Detailed description.

[0051] The present description is given without limitation, each characteristic disclosed only in one embodiment being able to be generalized to the other embodiments presented here or in the summary of the invention presented previously. Similarly, one or more characteristics disclosed only in one embodiment can be combined with one or more other characteristics disclosed only in another embodiment.

[0052] The present invention is based on the discovery that a combination containing a Coleus forskohlii extract, an apple extract rich in proanthocyanidins, a grape marc extract and a phloretin powder, has several beneficial effects and in particular makes it possible to reduce the accumulation of body fat and / or to prevent and / or treat cellulite, the cellulite being able to be adipose, aqueous and / or fibrous.

[0053] With regard to cellulite and without wishing to be bound by a theoretical explanation and without excluding other modes of action, the inventor believes that the combination according to the present invention or the nutraceutical composition or the topical composition containing it in accordance with the invention acts to prevent or slow down the formation of cellulite and to reduce cellulite already present via one or more modes of action, in particular protection against oxidative stress, reduction of inflammation, regulation of lipid and carbohydrate metabolism, inhibition of the accumulation of fats in fat cells (adipocytes), inhibition of the differentiation, proliferation or hypertrophy of fat cells, stimulation of blood circulation, including microcirculation which is often impaired in areas affected by cellulite, and stimulation of collagen synthesis.

[0054] Indeed, the extracts of Coleus forskohlii, grape and apple marc and phloretin powder provide the combination according to the present invention with beneficial properties for preventing or slowing down the formation of cellulite and reducing cellulite already present, in particular: - antioxidant properties that can help protect cells and tissues (e.g., subcutaneous tissue) from the harmful effects of oxygen-derived free radicals, stimulate the immune system, and promote the production of collagen and elastin, which help keep skin firm and elastic, which can prevent or reduce the appearance of cellulite; - anti-inflammatory properties that help reduce inflammation, including in areas affected by cellulite; - vasodilatory properties that help stimulate and / or improve blood flow, particularly in the microcirculation. This action promotes the oxygenation of cells and tissues, including subcutaneous tissue, as well as the evacuation of toxins, cellular waste and fats through the lymphatic system. This dual benefit helps improve the quality and appearance of the skin, particularly in areas affected by cellulite; - lipid and carbohydrate metabolism regulating properties (e.g. lowering triglycerides and cholesterol) which can support metabolism while helping to limit the accumulation of fat in adipose tissue; - anti-adipogenic properties which contribute to regulating and / or inhibiting adipogenesis and / or lipogenesis, thus slowing down the differentiation, proliferation and / or increase in the size of adipocytes; - lymphangiogenic properties that stimulate the growth of new lymphatic vessels, thus contributing to the improvement of the lymphatic network. This promotes efficient drainage of lymph elements, such as interstitial fluids, and thus contributes to the maintenance of tissue homeostasis; - anti-fibrotic properties that prevent or limit excessive scar tissue formation, thereby helping to alleviate fibrosis, particularly fibrosis of adipose tissue, and potentially preventing fibrous cellulite.

[0055] On the other hand, the combination according to the present invention can be used in a composition of the food supplement type. It is chemically stable and easy to formulate for oral administration.

[0056] Definitions:

[0057] The term "cellulite" refers to one or more of the conditions selected from the group consisting of fibrous cellulite, adipose cellulite and aqueous cellulite.

[0058] The term "prevent" and its derivatives, for example "prevention", encompass all preventive measures aimed at reducing the risk of the appearance of signs, delaying their appearance and reducing the incidence of the condition in question.

[0059] The term "improve" and its derivatives, such as "enhancement," encompass the reduction of signs, the slowing of the progression of signs, and the possibility of a complete cure.

[0060] The term "treat" and its derivatives, for example "treatment," encompasses reversing, alleviating, and / or inhibiting the progression of the condition to which the term treat or one of its derivatives is applicable.

[0061] The term "polyphenols" refers to a very large family of molecules, widespread in the plant world, generally characterized by the presence of one or more aromatic hydrocarbon rings, each carrying one or more hydroxyl groups. Polyphenols are divided into subfamilies such as: (i) flavonoids, which include: take for example flavanols, flavones, flavonols or even anthocyanidins (ii), phenolic acids such as hydroxy-cinnamic acids; stilbenoids (including resveratrol) and lignans.

[0062] The term "proanthocyanidins" refers to polyphenolic compounds consisting of flavan-3-ol units (catechin or epicatechin) linked together by carbon-carbon bonds of type 4—>8 or 4—>6. Proanthocyanidins, also called condensed tannins, oligomeric proanthocyanidins, oligo-proanthocyanidins (OPC) or pro-cyanidins, are present in many plants such as apple, plum and grape seeds. Proanthocyanidins are natural glycosidase inhibitors and are distinguished by their antioxidant activity, which can be 20 to 50 times higher than that of vitamins C and E. They are of great interest in the fields of nutrition and medicine due to their numerous beneficial properties, including antioxidant, anti-inflammatory, vasodilatory, antiallergic, antibacterial, antiviral, antitumor and chemopreventive properties.They help protect the body's cells and tissues from free radicals. They are particularly beneficial for the cardiovascular system because they help protect and strengthen the components of the vessel walls, including microvessels, thus contributing to better venous circulation.

[0063] The term "nutraceutical" refers to a substance or material, whether a whole food or a component of a food, designed to be consumed by a subject and capable of conferring health benefits. Dietary supplements and natural health products are examples of nutraceuticals.

[0064] The term "nutraceutical composition" refers to a composition whose purpose is to supplement the normal diet and to constitute a concentrated source of nutrients or other substances having a nutritional or physiological effect, alone or in combination.

[0065] The term “nutritional product” designates all products having a nutritional and / or physiological effect, this includes in particular, food supplements, foods, dietetic products, etc.

[0066] The term "food supplement" refers to a product or composition which, by its addition, enhances a food. A food supplement may be added to a food before, during or after the preparation of said food. A food supplement may, however, also be ingested alone, either before, during or after a meal, or between meals.

[0067] The term "topical composition" refers to a composition formulated for topical administration. The term "topical administration" refers to application to the skin, particularly to the area of ​​skin affected by cellulite.

[0068] The term “dry extract” designates an extract free of extraction solvent or in containing only non-significant traces.

[0069] The term "standardized" is often used in the field of naturally derived nutritional products. It refers to the process of delivering a product with a specific minimum concentration of one or more plant constituents. Standardization represents the concentration level of particularly desired elements from a plant source. Methods for standardizing plant substances, including phloretin powder and extracts of apples, Coleus forskohlii, and grape pomace, are well known in the art. In addition, the measurement of particular plant constituents upon which standardization is based is also well known in the art.Typically, standardization of plant extracts such as Coleus forskohlii extract, apple extract, grape pomace extract, and phloretin powder is performed by UV spectrophotometry, high-performance liquid chromatography (HPLC), or liquid chromatography-mass spectrometry (HPLC-MS), using appropriate standards.

[0070] The term "about" when used as a modifier of a numerical range or quantity, means an approximation of the range or quantity, such that minor deviations from the range or quantity are within the scope of the invention. Such deviations are known in the art of manufacturing formulations having a number of different ingredients of varying weights and consistencies.

[0071] The expression “p / pcombinaiSon” means that the percentage of the component or ingredient considered is by weight relative to the total weight of the combination according to the invention.

[0072] The expression “p / pcompositiOn” means that the percentage of the component or ingredient considered is by weight relative to the total weight of the composition according to the invention.

[0073] The expression "p / papple extract" means that the percentage of the component considered is by weight relative to the total weight of the apple extract.

[0074] Combination of active ingredients according to the invention

[0075] The combination of active ingredients according to the invention contains an extract of Coleus forskohlii, an apple extract rich in proanthocyanidins, a grape marc extract and a phloretin powder.

[0076] In embodiments according to the invention, the combination contains, as sole active ingredients, Coleus forskohlii extract, proanthocyanidin-rich apple extract, grape marc extract and phloretin powder.

[0077] Extract from Coleus forskohlii:

[0078] Coleus forskohlii extract works in concert with phloretin, apple extract, and grape pomace extract to reduce body fat accumulation and to prevent and / or treat cellulite. It helps stimulate fat metabolism and promote the breakdown of fats in fat cells and their use as fuel in the body. In addition, it works simultaneously to reduce stress oxidative and inflammation linked to cellulite.

[0079] For the purposes of the present invention, the term "Coleus forskohlii extract" means, in particular, a dry extract of Coleus forskohlii containing forskolin (CAS number: 66575-29-9) produced by an extraction process from the root of Coleus forskohlii.

[0080] Preferably, the extract of Coleus forskohlii contains at least 10% by weight per relative to the total weight of the dry extract.

[0081] The extract of Coleus forskohlii can be prepared by different extraction processes known to those skilled in the art, implementing steps of grinding the roots of Coleus forskohlii, solid / liquid extraction, for example in a Soxhlet extractor using one or more suitable solvents such as toluene, hexane, ether, ethanol, methanol, ethyl acetate, acetone, dichloromethane, chloroform and mixtures thereof, separation of the soluble and insoluble phases by filtration, concentration and drying, and where appropriate, mixing with a matrix (e.g. dextrin and / or maltodextrin), sieving and packaging.

[0082] Examples of solid / liquid extraction include maceration, infusion, decoction, extraction using Soxhlet extractors, microwave-assisted extraction, ultrasound-assisted extraction, supercritical fluid extraction (e.g. CO2 + dipropylene glycol). Solid / liquid extraction leads to the recovery of metabolites, in particular forskolin, contained in the roots of Coleus forskohlii.

[0083] As solvent, any solvent usually used for the extraction of plants may be suitable. It is particularly preferred to use as solvent one or more solvents chosen from toluene, hexane, cyclohexane, ether, a C1-C4 alcohol such as methanol, ethanol, and isopropanol, a C1-C4 ketone such as acetone and methyl ethyl ketone, a C1-C6 ester such as ethyl acetate, isopropyl acetate and butyl acetate, and chloroform or dichloromethane.

[0084] The extraction can be carried out at room temperature or at a higher temperature, for example at reflux of the extraction solvent used.

[0085] The extraction process advantageously comprises a filtration step aimed at separating the liquid phase from the exhausted plant material (roots of Coleus forskohlii).

[0086] The extraction then filtration cycle can be repeated several times in order to exhaust the plant material (roots of Coleus forskohlii) of substances having an affinity for the extraction solvent.

[0087] Typically, the extraction process may be completed by a step of partial or total removal of the extraction solvent or solvent mixture. It is thus possible to obtain a concentrated extract free of a significant quantity of solvents or a dry residue by removing all of the extraction solvent or solvent mixture. Alternatively, the product of the extraction step may be lyophilized or atomized to present in the form of a powder.

[0088] Examples of extraction processes from Coleus forskohlii can be found in the following articles: Pritee Singh and MA Suryanarayana, 2019; and Haritha Kanne, et al., 2015).

[0089] In practice, the extract of Coleus forskohlii is presented in powder form, in particular with a particle size of less than 1000 μm, preferably less than 600 μm, preferentially less than 250 μm, and with a water content of less than 10%, preferably less than 5% by weight relative to the weight of the powder.

[0090] Advantageously, the Coleus forskohlii extract is present in the combination in a content of at least 40% w / wCombination, preferably from 45% to 70% w / wCombination-

[0091] Apple extract:

[0092] The combination according to the present invention contains an apple extract concentrated in proanthocyanidins, which enriches and strengthens the antioxidant and anti-inflammatory properties of Coleus forskohlii extract, grape marc extract and phloretin powder. This apple extract also has the ability to inhibit digestive enzymes such as lipases, α-glucosidase and α-amylase, responsible for the breakdown of fats and carbohydrates. This inhibition slows down the breakdown of nutrients, thus reducing the absorption of calories by the body, which can help limit fat deposits in the body. In addition, it has vasodilatory properties that can promote the improvement of blood flow, including at the level of microcirculation, which is often compromised in areas affected by cellulite.

[0093] For the purposes of the invention, the term "apple" means the apple fruit of the botanical genus Malus (Botanical Family: Rosaceae), for example of a species of Malus selected from the group consisting of: Malus domestica, Malus pumila, Malus sieversii, Malus sylvestris, and combinations thereof. The apple fruit may be ripe or immature (still green). More particularly, the apple fruit is immature.

[0094] For the purposes of the invention, the term "apple extract" means an extract obtained from apple fruit. It may be, for example, an extract of apple fruit or of apple fruit skin. In particular, the apple extract is (or comprises) an extract of apple fruit skin.

[0095] The apple extract is rich in proanthocyanidins such as procyanidins B1 (CAS number: 20315-25-7) and B2 (CAS number 29106-49-8). Advantageously, the apple extract used contains at least 50% w / p apple extract of proanthocyanidins. Most preferably, the apple extract contains at least 90% w / p apple extract of proanthocyanidins.

[0096] The apple extract may also comprise other polyphenolic compounds, including polyphenolic acids such as chlorogenic acid and / or dihy- drochalcones including phloridzin and / or other proanthocyanidins having a chain length of 2 to 12 monomeric units, for example 2 to 8 monomers.

[0097] Apple extracts rich in proanthocyanidins can be prepared from apple fruit or its peels by any of the extraction methods known to those skilled in the art (see for example patent documents US20050010040A1, EP0657169A1 and EP2579886A1, or the following article: Asma, U. et al., 2023).

[0098] The apple extract rich in proanthocyanidins used is preferably in powder form, in particular with a particle size of less than 1000 μm, preferably less than 600 μm, preferably less than 250 μm, and a water content of less than 10%, preferably less than 5% by weight relative to the weight of the powder.

[0099] Preferably, the combination of the invention comprises at least 1% w / w combination, preferably from 2% to 9% w / w combination of apple extract rich in proanthocyanidins.

[0100] Grape marc extract:

[0101] The combination according to the present invention contains a grape marc extract.

[0102] For the purposes of the invention, the term “grape” means the fruit of the vine of the genus Vitis (Botanical family: Vitaceae), for example of a species of Vitis selected from the group consisting of: Vitis vinifera, Vitis labrusca, Vitis riparia, Vitis rotundifolia, Vitis rupestris, Vitis aestivalis, Vitis mustangensis and combinations thereof.

[0103] For the purposes of the invention, the term "grape marc" means the residual product, in particular the skin (or film), seeds, and possibly the stalks, resulting from the operation of pressing or crushing the fruits of the vine of the genus Vitis, for example of at least one species of Vitis selected from the group consisting of: Vitis vinifera, Vitis labrusca, Vitis riparia, Vitis rotundifolia, Vitis rupestris, Vitis aestivalis, Vitis mustangensis. Typically, grape marc represents approximately 20 to 30% of the weight of the grapes used to make wine or grape juice.

[0104] The grape marc may be one or more red grape marcs, one or more white grape marcs or mixtures thereof. The grape marc may be derived from any type of grape variety known to those skilled in the art. As non-limiting examples of grape varieties, mention may be made of Merlot, Syrah, Tannat, Mourvèdre, Cabernet-Sauvignon, Cabernet-Franc, Grenache, Chardonnay, Pinot Noir and Sauvignon. Preferably, the grape marc is one or more red grape marcs.

[0105] By “grape marc extract” within the meaning of the invention a product obtained from grape marc (skins, seeds, and possibly stalks) recovered after separation of the liquid part (juice or must) of grape.

[0106] According to a first embodiment, the grape marc extract is dehydrated (raw) grape marc presented in powder form. It can be obtained according to any known process for example by recovering at least one grape marc after separation of the juice or grape must, freezing and drying, for example by freeze-drying, then grinding-sieving of the dried grape marc(s) (See for example the article by Solari-Godino, A. et al., 2017).

[0107] Grape pomace extract is an excellent source of polyphenols and dietary fiber, as previously discussed. It helps improve cellulite through one or more mechanisms. Its polyphenolic compounds, such as flavonoids, phenolic acids, and resveratrol, have the ability to reduce oxidative stress and inflammation, particularly related to cellulite. Indeed, these compounds can act by neutralizing free radicals responsible for oxidative stress and modulating inflammatory pathways, thus helping to prevent cellular damage and mitigate unwanted inflammatory responses in the body, particularly in areas affected by cellulite.Furthermore, due to their inhibitory activities, such polyphenolic compounds have the potential to limit the absorption of dietary fat and prevent excessive fat accumulation in adipose tissue, which helps prevent the formation of cellulite and / or reduce existing cellulite. Furthermore, thanks to its high dietary fiber content (at least 30% by weight), grape pomace extract may help regulate blood sugar and carbohydrate metabolism. These dietary fibers have the ability to slow the digestion and absorption of carbohydrates, thus preventing blood sugar spikes after meals. By limiting the amount of carbohydrates available to be stored as fat in fat cells (adipocytes), grape pomace extract may help prevent fat accumulation and avoid unwanted body weight gain and / or induce body weight loss.Additionally, this ability to moderate the glycemic response may also be beneficial in preventing cellulite formation and / or reducing existing cellulite. By minimizing fluctuations in blood sugar, grape pomace extract helps limit excessive fat storage in adipocytes.

[0108] According to a second embodiment, the grape marc extract is an extract concentrated in polyphenols potentially present in the grape marc (hereinafter, polyphenolic extract of grape marc).

[0109] Advantageously, the polyphenolic extract of grape marc is in powder form. It can be obtained from one or more grape marcs by extraction with one or more solvents, according to the methods for extracting grape skins or seeds known per se (see for example patent documents FR3002453A1, US6544581B1, WO2014013122, WO2017013299A1). As solvent, any solvent usually used for extracting grape seeds or skins may be suitable. It is particularly preferred to use water and / or alcohols, such as methanol or ethanol, as solvent.

[0110] Preferably, the polyphenolic extract of grape marc used contains, relative to the total weight of the dry extract, at least 50% by weight of polyphenols including at least 20% of proanthocyanidins. It may, if appropriate, be associated or mixed with one or more excipients such as dextrin and / or maltodextrin.

[0111] The dehydrated (raw) grape marc powder or the polyphenolic extract of grape marc generally has a particle size of less than 1000 μm, preferably less than 600 μm, preferentially less than 250 μm, and a water content of less than 10% by weight relative to the weight of the powder.

[0112] Preferably, the combination of the invention comprises at least 4% w / w combination, preferably from 4 to 20% w / w combination of grape marc extract.

[0113] Phloretin:

[0114] Phloretin has anti-adipogenic, antioxidant and anti-inflammatory properties. It helps limit the accumulation of fat and / or the growth of fat cells that cause cellulite. In addition, it also helps reduce oxidative stress and inflammation associated with cellulite.

[0115] For the purposes of the present invention, the term "phloretin powder" means in particular an extract containing phloretin and produced by an extraction process from the apple tree (e.g., the fruit, the leaves, the peels of the fruit). The phloretin powder can be extracted from the fruit (apple), whole or part of the fruit. Typically, the phloretin powder is obtained from apple leaves and / or peels.

[0116] Apple tree and apple fruit (Malus species) are as defined in the case of proanthocyanidin-rich apple extract.

[0117] Examples of phloretin extraction methods can be found in the article by Nakhate KT, et al., 2022.

[0118] In practice, the phloretin powder has a particle size of less than 1000 μm, preferably less than 600 μm, preferentially less than 250 μm. The moisture content of the phloretin powder may be less than 10%, preferably less than 5% by weight relative to the weight of the powder.

[0119] The phloretin powder suitable for the invention is an apple extract standardized to contain at least 90% phloretin, preferably at least 98% phloretin. It is commercially available.

[0120] Advantageously, the phloretin powder is present in the combination in a content of at least 20% w / w combination, preferably from 20% to 40% w / w combination-

[0121] A preferred combination according to the invention comprises: (a) at least 40% w / w Combination of Coleus forskohlii extract, (b) at least 1% w / w combination of proanthocyanidin-rich apple extract, (c) at least 4% w / w combination of grape marc extract, (d) at least 20% w / wCombination of phloretin powder, the sum of (a), (b), (c) and (d) being equal to 100%.

[0122] Another example of a preferred combination according to the invention comprises: (a') from 45 to 70% w / p combination of Coleus forskohlii root extract, (b') from 2 to 9% w / p combination of proanthocyanidin-rich apple extract, (c') from 4 to 20% w / p combination of grape marc extract, (d') from 20 to 40% w / p combination of phloretin powder, the sum of (a'), (b'), (c') and (d') being equal to 100%.

[0123] Yet another example of a preferred combination according to the invention comprises: 58.39% w / w combination of Coleus forskohlii root, 3.65% w / p combination of apple extract rich in proanthocyanidins, 8.76% w / p combination of grape marc extract, 29.20% w / w combination of phloretin powder.

[0124] Nutraceutical composition - Topical composition:

[0125] The combination according to the invention can be formulated in the form of a nutraceutical composition (for oral administration) or a topical composition (for topical administration). Thus, according to one embodiment, the present invention also relates to a nutraceutical composition comprising, as active agent, a combination according to the present invention. According to another embodiment, the present invention also relates to a topical composition comprising, as active agent, a combination according to the present invention.

[0126] The amount of the combination according to the invention may vary within wide limits, such as for example from 0.01% to 99.99% by weight relative to the total weight of the nutraceutical composition or the topical composition. In embodiments, the amount of the combination is at least 10% by weight relative to the total weight of the nutraceutical composition or the topical composition.

[0127] The nutraceutical composition or the topical composition in accordance with the invention may further comprise one or more suitable excipients or vehicles.

[0128] By "excipient" or "vehicles" is traditionally meant any compound or composition intended to act as a simple support or vehicle, i.e. which is not intended to have a physiological or biological activity. Examples of excipients or vehicles include, but are not limited to, diluents, binders, lubricants, dispersants, antioxidants, disintegrants, anti-caking agents, colorants, preservatives, coating agents, humectants, flavorings, etc. The excipient(s) or vehicle(s) may also consist of or comprise aqueous solvents, non-aqueous solvents (e.g. alcohols, oils, waxes, etc.), hydroalcoholic solvents (e.g. water-glycerin). Of course, a person skilled in the art will take care to choose the possible compounds or compositions of excipients as well as their proportions in such a way that the advantageous properties of the combination according to the invention or of the com nutraceutical position containing it in accordance with the invention are not or substantially not altered by the envisaged addition. The excipients are well known in the nutraceutical field and are not the subject of a detailed description here. Suitable excipients or vehicles are also described in the work entitled "Handbook of Pharmaceutical Excipients," Second Edition, American Pharmaceutical Association, 1994.

[0129] For the purposes of the present invention, the term "suitable" or "suitable" means that the excipient(s) (or vehicle) are compatible with nutraceutical or topical use of the composition in accordance with the invention, i.e. they are not toxic to humans when used in a nutraceutical or topical composition.

[0130] In embodiments, the nutraceutical composition according to the invention may further comprise one or more additional components that can promote a complementary and / or synergistic action. The additional components may be selected from vitamins (e.g., one or more vitamins A, B1, B2, B3, B5, B6, B8, B9, B12, C, D, E and / or K), mineral salts and / or trace elements (e.g., selenium, zinc, magnesium, etc.), antioxidant agents (e.g., carotenoids, resveratrol, vitamin E), one or more sources of prebiotics (e.g., acacia fiber, chicory inulin, fructooligosaccharides extracted from banana, wheat, asparagus or artichoke), one or more amino acids (e.g., glutamine, aspartic acid, arginine, lysine, etc.), one or more plant extracts capable of stimulating the metabolism of lipids and / or carbohydrates and / or promoting blood circulation (e.g. green tea extract, guarana extract, ginseng extract, turmeric extract, pepper extract, grape seed extract, vine leaf extract, blackcurrant seed extract, blackcurrant leaf extract, Griffonia simplicifolia extract, fenugreek extract, carob extract, konjac extract, yerba mate extract, etc.), probiotic bacteria (e.g. Lactobacillus acidophilus, Bifido-bacterium lactis, Lactobacillus casei, etc.), yeasts (e.g. Saccharomyces boulardii) and mixtures thereof.

[0131] The concentration, in the nutraceutical composition or in the topical composition according to the invention, of the possible additional component(s) may vary within wide limits. Of course, a person skilled in the art will take care to choose the possible additional compounds and / or their quantities, in such a way that the advantageous properties of the nutraceutical composition or the topical composition are not, or not substantially, altered by the envisaged addition. By way of example, the additional component(s) may be used in a content of between 0.001 and 30% by weight relative to the total weight of the nutraceutical composition or the topical composition.

[0132] Process - galenic - applications:

[0133] The combination according to the invention and the nutraceutical composition, or the topical composition, containing it in accordance with the invention can be obtained by any usual formulation method known in the nutraceutical or cosmetic fields. Their respective ingredients can be mixed, before shaping, in the order and under conditions easily determined by those skilled in the art.

[0134] In practice, before galenic formulation, the combination according to the invention is generally in the form of a powder with a particle size of less than 2000 μm, preferably less than 1000 μm, preferentially less than 600 μm, more preferentially less than 250 μm, and with a moisture content of less than 10%, preferentially less than 2.5% by weight relative to the weight of the powder.

[0135] When the combination is formulated as a topical composition, it may be in any of the galenic forms normally used for topical application to the skin, including in the form of an aqueous, hydroalcoholic or oily solution; an oil-in-water emulsion, water-in-oil or multiple emulsions. It may be more or less fluid and have the appearance of a cream, milk, ointment, lotion, serum, gel, paste or mousse. It may also be in solid form, such as a stick, or applied to the skin in the form of an aerosol.

[0136] When the combination is formulated as a nutraceutical composition, it may be presented in all the galenic forms normally used for oral administration, in particular in the form of powders (e.g. compressed or loose powders, powders packaged in sachets, in particular in sachets), tablets (e.g. chewable, effervescent, gastro-resistant tablets), dragees; soft or hard capsules, gelatin capsules (e.g. film-coated and / or gastro-resistant gelatin capsules), granules, hard, soft or gelled candies, chewing gums, or aqueous or oily suspensions. The preferred forms for oral administration are powders, tablets and gelatin capsules.

[0137] Such galenic forms can be produced by any usual process known to those skilled in the art.

[0138] Preferably, the nutraceutical composition according to the present invention is presented in a unit dosage form, such as a tablet, a capsule or a sachet.

[0139] When the nutraceutical composition according to the present invention is in a unit dosage form, each unit dosage form may contain the combination according to the invention in an amount containing: from 75 mg to 150 mg of Coleus forskohlii extract, from 5 mg to 30 mg of grape marc extract, from 2.5 mg to 15 mg of proanthocyanidin-rich apple extract, from 25 mg to 75 mg of phloretin powder.

[0140] The nutraceutical or topical composition according to the invention may be contained in a packaging, a wrapper or a dispensing device at the same time as instructions for use. In particular, the nutraceutical composition in the form of tablets or capsules may be presented in a sealed packaging, which may consist of a pillbox (e.g.: a pillbox made of polypropylene or high-density polyethylene), a plastic or aluminum bag, or a blister pack, in particular, a blister pack (cell) made of PVC / PVDC, covered with an aluminum film for optimal protection.

[0141] The nutraceutical composition according to the present invention can be dedicated to use as a nutritional product.

[0142] A preferred nutraceutical composition according to the invention is a food supplement.

[0143] Advantageously, the food supplement is intended to be consumed or administered orally, to provide, in a human subject, beneficial effects against cellulite and / or the accumulation of body or skin fat.

[0144] The combination according to the invention or the nutraceutical composition containing it in accordance with the invention can also be used to supplement a diet (e.g.: enrich a food product or a drink) intended to help an individual obtain the expected beneficial effects, namely: reduce the accumulation of body fat, and / or prevent and / or treat cellulite.

[0145] Also, the present invention also relates to a non-therapeutic use, by oral route and / or by topical route, of the combination according to the invention (in the form of a nutraceutical composition and / or a topical composition) for regulating the metabolism of lipids and / or carbohydrates; and / or for reducing and / or inhibiting the accumulation of interstitial fluids; and / or for limiting and / or inhibiting the expansion of adipocytes; and / or for reducing and / or inhibiting adipocyte differentiation; and / or for stimulating lymphangiogenesis; and / or for reducing the accumulation of body or skin fat; and / or for preventing the formation of cellulite and / or for reducing existing cellulite. The present invention relates in particular to the uses as defined above, in which the combination according to the invention is implemented within a nutraceutical composition or a topical composition.

[0146] The invention also relates to a non-therapeutic method for regulating lipid and / or carbohydrate metabolism; and / or for decreasing and / or inhibiting the accumulation of interstitial fluids; and / or for limiting and / or inhibiting the expansion of adipocytes; and / or for decreasing and / or inhibiting adipocyte differentiation; and / or to stimulate lymphangiogenesis; and / or to reduce the accumulation of body or skin fat; and / or to prevent the formation of cellulite and / or to reduce existing cellulite, in a human subject. In one embodiment, the non-therapeutic method according to the present invention comprises orally administering to the human subject a nutraceutical composition according to the invention. In another embodiment, the non-therapeutic method according to the present invention involves topically administering to the human subject a topical composition according to the invention. In yet another embodiment, the non-therapeutic method according to the present invention comprises both orally administering to the human subject a nutraceutical composition according to the invention and topically administering to said human subject a topical composition according to the invention.Oral and topical administration allows for a synergistic approach, exploiting the respective advantages of these modes of administration. The joint adoption of oral and topical administration promotes a synergistic approach, taking advantage of the respective advantages of these two modes of administration. The combination of these modes of administration provides a synergy that can optimize the overall effectiveness of combating the accumulation of body or skin fat and / or cellulite, acting in a complementary manner to obtain more significant results. Advantageously, the application of at least one method of the invention confers the advantages indicated above as being associated with the implementation of a combination according to the invention or a nutraceutical or topical composition containing it in accordance with the invention.

[0147] A method according to the invention can be implemented by daily administration (once, twice or more than twice) of a nutraceutical combination or composition, formulated for example in solid form (e.g. powders, capsules, soft capsules, tablets, capsules, dragees, chewing gums, etc.) or liquid form (e.g. gels, emulsions, syrups, etc.) in adequate quantity and number, depending on their form, and / or a topical composition formulated for example in the form of a cream, milk, ointment, lotion, serum, gel, paste or mousse, or in the form of a stick or aerosol.

[0148] The method according to the invention can be implemented over a period varying from one week to several weeks, or even several months. This period can also be repeated after periods of non-treatment, for several months or even several years. For example, when the nutraceutical composition according to the present invention is in a unit dosage form, as mentioned above, the administration of this nutraceutical composition can be carried out at a rate of, for example, 4 times a day, generally over a prolonged period of at least 4 weeks, or even 4 to 16 weeks, including or not including one or more periods interruption or being repeated after a period of interruption.

[0149] The present invention benefits both women and men.

[0150] The combination according to the invention and the compositions in accordance with the invention may comprise all or only part of the daily dose.

[0151] The invention also relates to a kit or a combination product for simultaneous, separate or spread out use over time, preferably a food supplement, comprising a first composition containing a portion of the components forming the combination according to the invention and a second composition comprising at least the other portion of the components forming the combination according to the invention. This food supplement may be formulated in such a way that the two compositions are in the same forms or in different forms, for example chosen from those mentioned above. Such a kit may in particular be presented in a single package.

[0152] The ingredients are mixed, before being shaped and in order, under conditions easily determined by those skilled in the art.

[0153] The content and nature of the ingredients used in the nutraceutical combination or composition, or topical composition, containing it in accordance with the invention are adjusted by those skilled in the art so as not to substantially affect the properties required for said combination and for said nutraceutical or topical compositions.

[0154] The Coleus forskohlii root extract, the apple extract concentrated in proanthocyanidins, the grape marc extract and the phloretin powder used in the context of the present invention and the methods for obtaining them are well known to those skilled in the art. Such extracts can each be combined or mixed with one or more excipients such as dextrin and / or maltodextrin.

[0155] The chemical composition of the various plant and fruit extracts used in the present invention can be determined by any analytical method known in the art, for example by high performance liquid chromatography (HPLC) optionally coupled to a mass spectrometer (MS).

[0156] The particle size of the combination and of each of its ingredients can be measured by any technique known per se. For example, using a laser particle sizer or a series of sieves standardized according to the requirements of the European Pharmacopoeia 2-1-4.

[0157] In the description and examples that follow, the ranges of values ​​labeled in the form "between ... and..." include the specified lower and upper limits.

[0158] The effect of preventing and / or reducing cellulite can be demonstrated in different ways, for example by: - In vitro and ex vivo tests, notably using ap cell models such as human adipose tissue. Ex vivo models are unique tissue models because they are derived directly from donors and retain in culture the 3D structure and all the functions of the native tissue. They are therefore representative of the physiology of human adipose tissue. These tests can determine the fibrotic activity of adipose tissue in contact with the combination of the present invention. Fibrotic activity can be performed through cytotoxicity, metabolic activity, transcriptome and secretome analysis. Cytotoxicity and metabolic activity analysis can determine the stability of the combination formula on adipose tissue. Secretome analysis can evaluate the effect of the combination on metabolism and the fibrotic response of adipose tissue. Transcriptome analysis can evaluate adipogenesis at the lymphatic network level and assess the reduction of the fibrotic phenotype. - The tests can also be supplemented by qualitative / quantitative imaging analyses allowing observation of the lymphatic network, the size and number of adipocytes as well as the extracellular matrix, with an evaluation of fibrosis on the structure of the adipose tissue before and after incubation. - In vitro and ex vivo tests on human cell models can be carried out using laboratory analytical methods (e.g.: Immunofluorescence, ELIS A (enzyme-linked immunosorbent assay), PCR (Polymerase Chain reaction).

[0159] Other characteristics and advantages of the invention will emerge more clearly from the following examples, given for illustrative and non-limiting purposes. The scope of the invention should not be limited in any way by these examples. Examples:

[0160] The phloretin powder and the extracts of Coleus forskohlii root, proanthocyanidin-rich apple and grape marc used in the examples below each have a particle size of less than 600 microns and are all commercially available.

[0161] Example 1: Evaluation of the effects of a combination according to the present invention on a 3D human adipose tissue model.

[0162] The Applicant conducted a study aimed at analyzing the metabolic function and transcriptomic effects of a grape marc extract alone, and of a combination containing grape marc extract according to the present invention at an equivalent dose (hereinafter combination A). This study was conducted on a conventional three-dimensional (3D) human adipose tissue model (see for example the following documents: PCT / EP2020 / 050720 or Dani, V, et al.; 2022).

[0163] The tested combination A contains the ingredients shown in [Table 1] below.

[0164] [Table 1], Ingredients Quantity (% w / w combination) Coleus forskohlii root extract 58.4% Apple extract rich in proanthocyanidins 3.6% Grape pomace extract 8.8% Phloretin powder 29.2%

[0165] The ex-vivo three-dimensional (3D) human adipose tissue model used comes from subcutaneous adipose tissue (abdominal region) specifically taken from a healthy donor (meeting the following criteria: female, 22 <IMC<25, âge = 65 ans), pour garantir une pertinence biologique optimale. De manière connue, ce modèle a été élaboré pour reproduire fidèlement les caractéristiques et l'organisation du tissu adipeux humain. Son architecture 3D offre une représentation précise des conditions physiologiques, permettant une exploration approfondie de la réponse métabolique et des aspects particuliers à la biologie du tissu adipeux humain. Ainsi, il est représentatif de la physiologie du tissu adipeux humain.

[0166] The combination A was suspended in 5 ml of culture medium.

[0167] The doses tested: 30 pg / ml of grape marc extract alone and 341 pg / ml of the com combination A.

[0168] Incubation period: 7 days.

[0169] Model production: Adipose tissue was mechanically dissociated to preserve the 3D structure of the adipose tissue and the integrity of the cell populations. The resulting tissue was maintained in EGM-Plus medium (EBM basal medium + EGM supplements) under 3D culture conditions at 37°C, 5% CO2> and then encapsulated in LMT agarose.

[0170] Study plan:

[0171] The culture conditions for each sample (healthy control without fibrosis, fibrotic control, anti-fibrotic molecule control, grape marc alone as well as the combination) are presented briefly in [Table 2] below.

[0172] [Table 2], Group No. Type Culture conditions 1 Healthy control Medium 2 Fibrotic control (untreated) Medium + TGF-[31 dose 5 ng / ml (fibrotic stimuli) 3 Anti-fibrotic molecule control Medium + TGF-[31 dose 5 ng / ml (fibrotic stimuli) + SB431542 dose 5 pM (anti-fibrotic) 4 Grape marc Medium + TGF-[31 dose 5 ng / ml (fibrotic stimulus) + Grape marc 30 pg / ml 5 Combination Medium + TGF-[31 dose 5 ng / ml (fibrotic stimulus) + Combination 341 pg / ml

[0173] The number of replicates (N) per sample performed for each analysis is presented in [Table 3] below.

[0174] [Table 3]. Group No. Secretome analysis (ELISA IL-6) Metabolic activity analysis (XXT D7) Transcriptome (RNA extraction at D7, RT-qPCR Fixation in 4% PFA for subsequent imaging 1 N = 4 N = 4 N = 4 N = 1 2 N = 4 N = 4 N = 4 N = 1 3 N = 4 N = 4 N = 4 N = 1 4 N = 4 N = 4 N = 4 N = 1 5 N = 4 N = 4 N = 4 N = 1

[0175] Chronology of the study:

[0176] After the production of the model, a pre-treatment was carried out with the addition of compounds with TGF-[31 in order to create a fibrotic model. The renewal by addition of compounds with TGF-[31 was carried out at D0, D3 and D6. At D7, the supernatant was recovered for XTT analysis, RNA lysis for the trans-cryptomic study, secretome analysis and fixation for imaging analyses.

[0177] The synopsis of the study is presented in [Table 4] below.

[0178] [Table 4], Type of analysis Procedures Objective METABOLIC ACTIVITY XTT test on n=4 replicates / condition at D9 Evaluation of general metabolic activity in adipose tissues. ANTI-CELLULITE ACTION TRANSCRIPTOM E Transcriptome at D7 on n=4 replicates / condition (RNA by RT-qPCR) through a set of 5 known genes / markers of the lymphatic network and the reduction of the fibrotic phenotype, simulating cellulite. terature: ADIPOQ (adiponectin), LEP (leptin), INHBA (activin A), MSCA1, LYVE-1 IMAGING VISUALIZATION OF THE ANTI-CELLULITE ACTION -Adipocyte size (Oil Red O staining): visualization and measurement of the effect of active ingredients on the reduction of fat mass in a fibrotic context -Cellular matrix labeling (immunofluorescence labeling for Laminin): visualization of the effect of active ingredients on the matrix of fibrotic adipose tissue -SHG microscopy: visualization of the reduction of collagen fiber compaction following exposure to active ingredients Visualization of the effect of active ingredients on ex vivo ExAdEx models by imaging analysis Analysis of anti-cellulite action: Secretome ELISA on n=4 replicates / condition for IL-6 secretion at the end of the incubation period Evaluation of the effect of active ingredients on the fibrotic response

[0179] Results:

[0180] An XTT test (metabolic activity measurement) (after 9 days of treatment) revealed that the combination A tested, increases metabolic activity by approximately 13% compared to grape marc alone.

[0181] The expression levels of the markers adiponectin, INHBA (activin A), MSCA1, LYVE-1 and IL-6 were examined at the same time period, by real-time quantitative polymerase chain reaction (RT-qPCR).

[0182] Adiponectin, a hormone primarily secreted by adipocytes, plays a critical role in the metabolic regulation of carbohydrates and lipids. As a key mediator between adipose tissue, liver, and muscle, it exerts notable positive effects on metabolism. Its ability to promote insulin sensitivity, regulate inflammation, and modulate fatty acid oxidation makes it a central player in metabolic balance. By promoting balanced adipocyte growth, adiponectin promotes healthy adipose tissue expansion. Plasma adiponectin levels, generally higher in healthy individuals, tend to decrease in those affected by obesity or metabolic disorders. Beyond its metabolic role, adiponectin is also distinguished by its ability to stimulate the production of antioxidant enzymes, thus helping to strengthen the body's natural defenses against oxidative stress. In addition, it exhibits an anti-inflammatory action, protecting tissues, including adipose tissue, from the harmful effects of fibrosis and inflammation.

[0183] An increase in Adiponectin expression is correlated with a favorable metabolic health status, stimulation of Lymphangiogenesis (expansion of the lymphatic network) and a reduction of adipose and aqueous cellulite.

[0184] MSCA1 (Mesenchymal Stem Cell Antigen 1), is a protein expressed by adipogenic progenitors of adipose tissue, decreased in inflammatory and fibrotic conditions. Activation of the MSCA1 marker suggests a reduction in fibrous cellulite.

[0185] INHBA (Inhibin Subunit Beta A) is a gene that codes for the beta A subunit of activin, a protein better known as activin A. This protein is expressed by fibrogenic progenitors in response to stimulation by pro-inflammatory cytokines, themselves produced by macrophages that invade adipose tissue. It is a characteristic marker of pro-inflammatory fibroblasts, whose proliferation increases in inflammatory and fibrotic conditions, without differentiation into adipocytes. Induction of the INHBA marker is at the origin of a deregulation of adipose tissue homeostasis, thus promoting the transition to a fibrotic state.Furthermore, an increase in the expression of the MSCA1 marker and at the same time a decrease in the expression of the INHBA (Activin A) marker suggest an anti-fibrotic effect, and consequently, a reduction in fibrous cellulite and an improvement in adipose tissue homeostasis with a balance between fibroblast-like cellular progenitors and adipocytes towards a healthy adipose tissue condition.

[0186] LYVE-1 (Lymphatic Vessel Endothelial Hyaluronan Receptor 1), a protein encoded by a gene of the same name, plays a key role in the regulation of lymphatic vessels and interstitial fluid composition. This protein acts as a specific receptor for hyaluronic acid, a macromolecule involved in tissue water retention. When the LYVE-1 gene is functioning optimally, it actively participates in the elimination of hyaluronic acid, thus contributing to the maintenance of tissue water balance. An improvement in LYVE-1 gene expression indicates an optimization of lymphangiogenesis (growth of the lymphatic network) and / or the transport of interstitial fluid through the lymphatic network, thus suggesting a potential effect in reducing aqueous cellulite.

[0187] IL-6: pro-inflammatory cytokine produced in response to various inflammatory and fibrotic stimuli. Role in the emergence of cellulite. It also contributes to a disruption of metabolic homeostasis (contributes to insulin resistance and metabolic disorders linked to weight gain)

[0188] Laminin: protein of the extracellular matrix of many tissues, including adipose tissue, playing a role in structural support and in the phenomena of cell migration and adhesion. In response to various inflammatory and fibrotic stimuli (in a context of cellulite, overweight, etc.), an alteration of the laminin mesh (compaction, heterogeneous distribution) is often observed.

[0189] Transcriptomic analysis revealed that the tested combination A promotes the activation of the examined markers Adiponectin, LYVE-1 and MSCA1 and inhibits the expression of INHBA (Activin A).

[0190] Transcriptomic analysis also revealed that the tested combination A induced a 47% increase in adiponectin expression compared to grape marc tested alone.

[0191] Secretome analysis revealed that the tested combination A promotes the decrease in the secretion of the pro-inflammatory cytokine IL-6 on human adipose tissue models by -88% compared to grape marc alone.

[0192] By stimulating the expression of Adiponectin (a molecule beneficial to metabolism and the promotion of lymphangiogenesis), by increasing the general metabolic activity of adipose tissue and decreasing the secretion of the pro-inflammatory cytokine IL-6, combination A may have greater efficacy in reducing the cellulite phenotype compared to grape marc alone.

[0193] Combination A allows to reestablish a homogeneous distribution of Laminin, with a signal which surrounds the adipocytes.

[0194] Combination A also makes it possible to reduce the presence of fibrillar collagen by approximately 60% compared to grape marc alone (at an equivalent dose)

[0195] - Under the conditions of the study, combination A shows its superiority over marc grape as an anti-fibrotic active, which suggests a better capacity as a product for cellulite reduction.

[0196] Imaging analysis revealed the decrease in adipocyte size for combination A.

[0197] Combination A induces a reduction in adipocyte size of approximately 30% compared to grape marc alone at an equivalent dose.

[0198] Conclusion:

[0199] Overall, the combination described in the present invention demonstrates the ability to improve lipid and carbohydrate metabolism, to stimulate lymphangiogenesis with an improvement in the lymphatic network, to induce an improvement in adipose tissue homeostasis by balancing fibroblast and adipocyte type cellular progenitors towards a healthy adipose tissue condition, to cause an effect anti-fibrotic, to reduce the presence and quantity of fibrotic fibrillar collagen, to improve the structure of Laminin and finally, to reduce adipocyte size. These different effects could be beneficial for the regulation of lipid and carbohydrate metabolism, the management of interstitial fluid accumulation, and the reduction of cellulite (adipose, aqueous or fibrous cellulite).

[0200] Example 2: Nutraceutical composition in powder form.

[0201] A nutraceutical composition was prepared in powder form, then packaged in a sachet (e.g., in a stick), by mixing the ingredients presented in [Table 5] below.

[0202] [Table 5]. Ingredients Quantity (per sachet) Coleus forskohlii root extract 75 mg to 150 mg Proanthocyanidin-rich apple extract 2.5 mg to 15 mg Grape pomace extract 5 mg to 30 mg Phloretin powder 25 mg to 75 mg

[0203] Recommended dosages for the reduction of fat accumulation and for the prevention and / or treatment of cellulite: 2 doses (sachets) per day (midday and / or evening) to be mixed with a glass of water (approximately 200 mL) and taken before meal(s).

[0204] Example 3: Nutraceutical composition in the form of tablets.

[0205] A nutraceutical composition in tablet form was prepared, comprising the combination shown in [Table 6] below.

[0206] [Table 6], Ingredients Amount (per tablet) Coleus forskohlii root extract 100 mg Proanthocyanidin-rich apple extract 6.25 mg Grape pomace extract 15 mg Phloretin powder 50 mg

[0207] This nutraceutical composition, in the form of tablets, also includes conventional excipients including flavorings.

[0208] Recommended dosages for the reduction of fat accumulation and for the prevention or treatment of cellulite: 2 tablets per day (midday or evening) to be taken with a glass of water, 30 minutes before meals.

[0209] Bibliographic references: Asma, U.; Morozova, K.; Ferrentino, G.; Scampicchio, Mr. Apples and Apple By- Products: Antioxidant Properties and Food Applications. Antioxidants 2023,12, 1456. Barbosa, A. C. L., Pinto, M. D. S., Sarkar, D., Ankolekar, C., Greene, D., and Shetty, K. (2010). Varietal influences on antihyperglycemia properties of freshly harvested apples using in vitro assay models. J. Med. Food 13, 1313-1323. doi: 10.1089 / jmf.2009.0273. Csilla Zsuzsanna David, Judit Hohmann, Andrea Vasas. Chemistry and Pharmacology of Cyperaceae Stilbenoids: A Review. Molécules 2021, 26 (9), 794. Dani, V., Bruni-Favier, S., Chignon-Sicard, B., Loubat, A., Doglio, A., Dani, C. et al.. Régulation of Adipose Progenitor Cell Expansion in a Novel Micro-Physiological Model of Human Adipose Tissue Mimicking Fibrotic and Pro-Inflammatory Microen-vironments. Cells 2022, 11(18), 2798; doi.org / 10.3390 / cellsl 1182798. Haritha Kanne, Narayan Pandurang Burte, V. Prasanna, and Ravi Gujjula., Extraction and elemental analysis of Coleus forskohlii extract, Pharmacognosy Research, 2015; 7; 237-241. Hogan S, Canning C, Sun S, Sun X, Zhou K. Effects of grape pomace antioxidant extract on oxidative stress and inflammation in diet induced obese mice. Journal of Agricultural and Food Chemistry. 2010;58(21):11250-11256. Nakhate KT, Badwaik H, Choudhary R, Sakure K, Agrawal YO, Sharma C, Ojha S, Goyal SN. Therapeutic Potential and Pharmaceutical Development of a Multitargeted Flavonoid Phloretin. Nutrients. 2022 Sep 2;14(17):3638. doi: 10.3390 / nul4173638. PM1D: 36079895; PMC1D: PMC9460114. Pritee Singh and M. A. Suryanarayana, Effect of Solvents and Extraction Methods on Forskolin Content from Coleus forskholii Roots, Indian Journal of Pharmaceutical Sciences; 2019, 1136-1140. Solari-Godino A., Lindo-Rojas I. & Pandia-Estrada S.l (2017) Détermination of phenolic compounds and évaluation of antioxidant capacity of two grapes residues (Vitis vinifera) of varieties dried: Quebranta (red) and Torontel (white), Cogent Food & Agriculture, 3:1, 1361599, DOI: 10.1080 / 23311932.2017.1361599. Valeria Marinelli, Lucia Padalino, Amalia Conte , Matteo Alessandro Del Nobile and Karlis Briviba. Red Grape Marc Flour as Food Ingrédient in Durum Wheat Spaghetti: Nutritional Evaluation and Bioaccessibility of Bioactive Compounds; Food Science and Technology Research, 2018, 24 (6), 1093-1100. Yu, J., & Ahmedna, M. (2013). Functional components of grape pomace: Their composition, biological properties and potential applications. International Journal of Food Science and Technology 2013, 48, 221-237. Yunoki K, Sasaki G, Tokuji Y, et al. Effect of dietary wine pomace extract and oleanolic acid on plasma lipids in rats fed high-fat diet and its DNA microarray analysis. Journal of Agricultural and Food Chemistry. 2008;56(24):12052-12058.

Claims

Claims

1. Combination containing Coleus forskohlii extract, proanthocyanidin-rich apple extract, grape marc extract and phloretin powder.

2. Combination according to claim 1, characterized in that the extract of Coleus forskohlii is present in a content of at least 40% w / w combination, preferably 45 to 70% w / w combination-

3. Combination according to claim 1 or 2, characterized in that the apple extract contains at least 50% w / p apple extract, preferably at least 90% w / p apple extract of proanthocyanidins.

4. Combination according to any one of claims 1 to 3, characterized in that the apple extract rich in proanthocyanidins is present in a content of at least 1% w / w combination, preferably 2 to 9% w / w combination*

5. Combination according to any one of claims 1 to 4, characterized in that the grape marc extract is present in a content of at least 4% w / w combination, preferably 4 to 20% w / w Combination-

6. Combination according to any one of claims 1 to 5, characterized in that the phloretin powder is present in a content of at least 20% w / wcombmaiSOn, preferably 20 to 40% w / wCOmbinaiSOn-

7. Combination according to any one of claims 1 to 6, characterized in that it contains: (a) at least 40% w / w combination of Coleus forskohlii extract, (b) at least 1% w / w combination of apple extract rich in proanthocyanidins, (c) at least 4% w / w combination of grape marc extract, (d) at least 20% w / w combination of phloretin powder, the sum of (a), (b), (c) and (d) being equal to 100%.

8. Combination according to any one of claims 1 to 7, characterized in that it contains: (a') from 45 to 70% w / p combination of Coleus forskohlii root extract, (b') from 2 to 9% w / p combination of apple extract rich in proanthocyanidins, (c') from 4 to 20% w / p combination of grape marc extract, (d') from 20 to 40% w / p combination of phloretin powder, the sum of (a'), (b'), (c') and (d') being equal to 100%.

9. A combination according to any one of claims 1 to 8, ca- characterized in that it contains: 58.39% w / p combination of Coleus forskohlii root, 8.76% w / p combination of grape marc extract, 3.65% w / p combination of apple extract rich in proanthocyanidins, 29.20% w / p combination of phloretin powder.

10. Nutraceutical or topical composition, characterized in that it comprises, as active agent, a combination according to any one of claims 1 to 9.

11. Nutraceutical or topical composition according to claim 10, characterized in that it further comprises one or more excipients.

12. Nutraceutical or topical composition according to claim 10 or 11, characterized in that it further comprises one or more additional components such as vitamins, carotenoids, mineral salts and / or trace elements, antioxidants, one or more sources of prebiotics, one or more plant extracts capable of stimulating the metabolism of lipids and / or carbohydrates and / or promoting blood circulation, amino acids, probiotic bacteria and / or yeasts.

13. Nutraceutical composition according to any one of claims 10 to 12, characterized in that it is presented in a form suitable for oral administration, in particular, in the form of powders, tablets, capsules, granules, candies, chewing gums, or aqueous or oily suspensions.

14. A nutraceutical composition according to any one of claims 10 to 13, which is a dietary supplement.

15. Topical composition according to any one of claims 10 to 12, characterized in that it is presented in a form suitable for topical administration, in particular, in the form of creams, milks, ointments, lotions, serums, gels, pastes, mousses, sticks, aerosols.

16. A combination according to any one of claims 1 to 9, or the nutraceutical composition according to any one of claims 10 to 14, or the topical composition of any one of claims 10 to 12 or 15, for use in promoting the metabolism of lipids and / or carbohydrates, and / or in reducing the accumulation of body or skin fat, and / or in preventing and / or treating cellulite.