Anti-fat accumulation and anti-cellulite combination.

Abstract

The invention relates to a combination containing an extract of Coleus forskohlii, an apple extract rich in proanthocyanidins, an extract of grape pomace and a 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 an extract of Coleus forskohlii, an apple extract rich in proanthocyanidins, an extract of grape marc and a phloretin powder.

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

[0003] The invention also relates to the use of said combination or 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 properties beneficial to the body, in particular for the regulation of fat metabolism and / or the prevention and / or treatment of cellulite. State of the art.

[0005] The accumulation of body fat 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 formulations that better regulate fat metabolism. These products or formulations 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, giving the skin an orange-peel texture. It is a common condition that primarily affects post-pubescent women, and a majority of them (approximately 90%). It is generally observed in areas of the body with greater fat storage, such as the thighs, buttocks, abdomen, and sometimes the upper arms. Cellulite is not specific to overweight individuals, although increased adiposity (fat accumulation) exacerbates it.

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

[0011] Adipose cellulite is primarily linked to an excessive accumulation of subcutaneous fat. It gives the skin a bumpy, dimpled appearance. It is only visible when the skin is pinched between the middle finger and forefinger (hereinafter referred to as 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 flabby and swollen appearance. It is barely visible and presents a slightly bumpy appearance on the pinch test. It is generally due to venous or lymphatic circulation problems.

[0013] Fibrous cellulite is the most advanced form of cellulite, resulting from the 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 the affected area a granular texture. It feels firm to the touch and may cause pain upon pinching.

[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 calorie availability for pregnancy and breastfeeding. However, it poses an aesthetic problem and can even cause considerable distress to those affected. Therefore, there is 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 primarily of collagen fibers.

[0016] The trabeculae (or septa) help to 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 skin cells; and (iii) the passage of lymphatic vessels whose function is to drain lymph (a fluid containing water, Various white blood cells, antibodies, nutrients, bacteria, toxins, and cellular and metabolic waste products are transported to the lymph nodes and then into the bloodstream. Meanwhile, fibrous connective tissue structures compartmentalize fat cells into fat pockets. As the number and size of fat cells increase through adipogenesis and lipogenesis, these fat pockets swell and press against the skin, which is pulled inward by the fibrous connective tissue. The skin then becomes deformed, creating the bumps and dimples that give cellulite its characteristic appearance. Furthermore, as the fat pockets increase in size, they exert pressure on the surrounding blood and lymphatic vessels, thus hindering the supply of oxygen and nutrients to the skin cells on one hand, and disrupting normal lymphatic circulation on the other.This inability to efficiently drain lymphatic components, such as interstitial fluids, toxins, and waste products, leads to their accumulation and the formation of edema (fluid retention). This situation causes asphyxiation of surrounding tissue cells, disrupting collagen synthesis by dermal fibroblasts. Furthermore, it triggers chronic low-grade inflammation, which in turn activates the release of pro-inflammatory cytokines. This leads to the activation of pathways involved in increased 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 cell differentiation during which preadipocytes become adipocytes. The first early differentiation gives rise to young adipocytes which, after a later differentiation, give rise to mature adipocytes.

[0018] Lipogenesis refers to all the biochemical processes involved in the biosynthesis of triglycerides and fatty acids and their storage in adipocytes. Lipogenesis is a metabolic pathway that uses dietary triglycerides, fatty acids, and other non-lipid precursors such as dietary carbohydrates or other substrates in excess of 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 surrounding the fat pockets are oriented parallel to, and perpendicular to, the skin surface, whereas in men, these trabeculae have an oblique orientation. This difference explains why cellulite is generally a female condition. 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 skin surface. In men, the orientation The oblique trabeculae of fibrous connective tissue prevent or limit skin deformation in cases of excessive development of fat cells.

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

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

[0022] Current cellulite treatment strategies are based primarily on Medical-aesthetic methods include, for example, manual or mechanical massage techniques, invasive treatments such as mesotherapy, liposuction, or subcision, and non-invasive treatments such as acoustic wave therapy, radiofrequency, and / or laser therapy. While these medical-aesthetic methods can achieve satisfactory results, they are expensive, time-consuming, and restrictive.

[0023] The prior art also provides anti-cellulite preparations based on plant extracts that are applied topically to 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 prolonged applications. Furthermore, they can, in some cases, cause skin side effects or even discomfort.

[0024] Alternatively, there are also oral compositions intended for the treatment of cellulite. Unlike topical preparations, oral compositions offer the advantage of acting from within the body to improve the appearance of cellulite. This is because the metabolites and other active ingredients are readily distributed throughout the skin via the bloodstream. However, no oral composition proposed to date has been sufficiently effective in treating cellulite.

[0025] In view of the above and because of the increasing demand for new orally administered products and the appeal of natural products to consumers, there is a need for new products that stimulate lipid (fat) metabolism and / or reduce body fat accumulation and / or improve or treat cellulite. Presentation of the invention.

[0026] The present invention is based on the discovery that a combination containing an extract of Coleus forskohlii, an apple extract rich in proanthocyanidins, An extract of grape marc and phloretin powder. The combination has several beneficial effects and notably helps 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 topical composition containing it according to the present invention, is remarkably well tolerated by the body and presents no adverse effects on human health. They are also inexpensive, being based on inexpensive products available on the market or simple to produce using methods known to those skilled in the art.

[0028] The apple is a fruit produced by an apple tree, which belongs to the Rosaceae family, genus Malus, more specifically the species Malus pumula and Malus domestica. The apple is a rich source of polyphenols, including flavonoids such as anthocyanins and anthocyanins, proanthocyanidins, flavan-3-ols (e.g., catechin, epicatechin), and phenolic acids such as hydroxycinnamic acids, notably chlorogenic acid. The apple peel contains more than twice as many polyphenols as the 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 diseases, diabetes, cancers, and neurodegenerative diseases.In particular, apple polyphenols may help alleviate type II diabetes through various mechanisms such as the 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 is worth recalling 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 reactive oxygen species (ROS), such as hydroxyl radicals (HO*), peroxyl radicals (HOO*, ROO), and superoxide ions (O2*-), which tend to form in cells and can cause cellular and tissue damage if not neutralized by antioxidants. Furthermore, an excess of free radicals can lead to a state of oxidative stress in the body. This phenomenon can negatively impact 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 the regulation of fats in the body. This disruption can play a role. a role in the development of cellulite. As for skin tissue, oxidative stress induced by free radicals 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. And sagging skin can be a contributing factor to the appearance of cellulite.

[0030] Coleus forskohlii, also known as Indian Coleus, is a perennial plant belonging to the large Lamiaceae family and has historically been used in Ayurvedic medicine to treat various 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 an essential intracellular messenger in the mechanism of lipolysis (fat breakdown). 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.In vivo research has shown that forskolin can optimize body composition by increasing lean mass and metabolic activity.

[0031] Grape pomace (also called grape marc or grape pomace) is a residual product obtained from the fruit (grapes) of the grapevine, a plant of the genus Vitis (Botanical family: Vitaceae). Typically, grape pomace consists of the solid part resulting from the pressing or crushing of the grape bunches, i.e., the skins, seeds, and possibly stems. It represents approximately 20 to 30% of the weight of the grapes used to make wine or grape juice. Grape pomace is a particularly rich source of polyphenolic compounds (5 to 10% by weight). Among these polyphenolic compounds are 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 renowned for their broad spectrum of biological activities, among which their antioxidant, anti-inflammatory, anticancer, antimicrobial, antithrombotic, cardioprotective, and vasodilatory properties stand out. The phenolic compounds in grape pomace have also shown significant inhibitory activities, particularly with regard to limiting the absorption of dietary fats and the accumulation of fat in adipose tissue (see, for example, the following articles: Hogan S., et al., 2010; Yunoki K., et al., 2008). Grape pomace is also a... Grape pomace is an important source of dietary fiber, comprising approximately 30% to 60% of its dry weight. This dietary fiber includes both soluble and insoluble fibers, such as cellulose and hemicellulose. Furthermore, grape pomace contains monounsaturated 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 components. The phytochemical composition of grape pomace is influenced by various factors, including the grape variety, weather conditions, terroir, and the degree of grape ripeness. For example, red grapes typically have a higher anthocyanin content (a subgroup of flavonoids) compared to white grapes.

[0032] Phloretin [2',4',6'-trihydroxy-3-(4-hydroxyphenyl)-propiophenone; CAS No. 60-82-2] is a natural polyphenolic compound belonging to a special family of flavonoids called dihydrochalcones and is found in many fruits and vegetables, particularly apple leaves (Malus sp. Rosaceae). Phloretin possesses numerous biological activities, including antioxidant, anti-inflammatory, antidiabetic, anti-obesity, antitumor, antimicrobial, hepatoprotective, and cardioprotective effects.

[0033] To the Applicant's knowledge, there is currently no combination comprising an extract of Coleus forskohliL, an extract of grape marc, an extract of apple rich in proanthocyanidins and a powder of phloretin, 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, a combination of an extract of Coleus forskohlii, an apple extract rich in proanthocyanidins, an extract of grape marc and a phloretin powder is proposed.

[0035] Other advantageous features of the invention are listed below. Each of these features may be considered alone or in combination with the notable features defined above, and may, where appropriate, be the subject 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 from 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 at a level of at least 1% w / wcombinaiSOn, preferably from 2 to 9% w / wcombinaiSOn-

[0039] Grape marc extract may be present in the combination according to the invention at a content of at least 4% w / wcombinaiSON, preferably from 4 to 20% w / wcombinaiSON.

[0040] Preferably, the phloretin powder content is at least 20% w / pcombinaiSOn-, preferably from 20 to 40% w / pcombinaiSOn-

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

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

[0043] More specifically, the combination according to the invention contains: 58.39% w / pcombination of Coleus forskohlii root, 3.65% w / w 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 proposes a nutraceutical or topical composition comprising, as an 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 suitable for stimulating lipid and / or carbohydrate metabolism and / or 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, foams, sticks, aerosols

[0050] Advantageously, the combination according to the present invention, or the nutraceutical or topical composition according to the invention, is intended for use to reduce the accumulation of body or skin fat and / or to prevent and / or treat cellulite. Detailed description.

[0051] The present description is given by way of non-limiting example, each feature described in only one embodiment being generalizable to the other embodiments presented herein or in the summary of the invention presented previously. Similarly, one or more features described in only one embodiment may be combined with one or more other features described in only another embodiment.

[0052] The present invention is based on the discovery that a combination containing an extract of Coleus forskohlii, 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, 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 according to 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, including protection against oxidative stress, reduction of inflammation, regulation of lipid and carbohydrate metabolism, inhibition of fat accumulation in fat cells (adipocytes), inhibition of the differentiation, proliferation or hypertrophy of fat cells, stimulation of blood circulation, including microcirculation which is often altered in areas affected by cellulite, and stimulation of collagen synthesis.

[0054] Indeed, the extracts of Coleus forskohlii, grape pomace and apples and phloretin powder contribute to the combination according to the present invention beneficial properties for preventing or slowing the formation of cellulite and reducing existing cellulite, including: - 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 maintain firm and elastic skin, thus preventing or reducing the appearance of cellulite; - anti-inflammatory properties that help reduce inflammation, including in areas affected by cellulite; - Vasodilatory properties that 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 elimination of toxins, cellular waste, and fats through the lymphatic system. This dual benefit helps improve the quality and appearance of the skin, especially in areas affected by cellulite; - properties that regulate lipid and carbohydrate metabolism (e.g., lowering triglycerides and cholesterol) that can support metabolism while helping to limit fat accumulation in adipose tissue; - anti-adipogenic properties which help to regulate and / or inhibit adipogenesis and / or lipogenesis, thus helping to curb 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 components, such as interstitial fluids, and thus participates in maintaining tissue homeostasis; - anti-fibrotic properties that prevent or limit excessive scar tissue formation, thus helping to reduce fibrosis, particularly fibrosis of adipose tissue, and potentially prevent fibrous cellulite.

[0055] Furthermore, the combination according to the present invention can be used in a dietary supplement composition. It is chemically stable and easy to formulate for oral administration.

[0056] Definitions

[0057] The term "cellulite" refers to one or more of the selected conditions in 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 decreasing the incidence of the condition in question.

[0059] The term "improve" and its derivatives, such as "improvement", 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”, encompass the reversal, relief, and / or inhibition of 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 hydrocarbon aromatic rings, each bearing one or more hydroxyl groups. Polyphenols are divided into subfamilies such as: (i) flavonoids, which include, for example, flavanols, flavones, flavonols, or anthocyanidins; (ii) phenolic acids such as hydroxycinnamic 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 the 4->8 or 4->6 type. Proanthocyanidins, also called condensed tannins, oligomeric proanthocyanidins, oligo-proanthocyanidins (OPC) or procyanidins, are present in many plants such as apples, plums 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 because of their many beneficial properties, including their antioxidant, anti-inflammatory, vasodilatory, anti-allergic, antibacterial, antiviral, antitumor and chemo-preventive properties.They help protect the body's cells and tissues against 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” refers to all products having a nutritional and / or physiological effect, including in particular food supplements, foods, dietetic products, etc.

[0066] The term "food supplement" refers to a product or composition which, by its addition, enhances the value of a food. A food supplement may be added to a food before, during, or after its preparation. However, a food supplement may 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” refers to an extract free from extraction solvent or containing it only in insignificant trace amounts.

[0069] The term “standardized” is often used in the field of natural 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. Processes for standardizing plant substances, including phloretin powder and extracts of apples, Coleus forskohlii, and grape pomace, are well known in the relevant literature. Furthermore, the measurement of specific plant constituents on which standardization is based is also well known in the relevant literature.Typically, the standardization of plant extracts such as Coleus forskohlii extract, apple extract, grape pomace extract and phloretin powder is carried out by UV spectrophotometry, high-performance liquid chromatography (HPLC) or liquid chromatography coupled to mass spectrometry (HPLC-MS), using appropriate standards.

[0070] The term "approximately," when used as a modifier of a numerical range or quantity, denotes 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 / p apple extract" means that the percentage of the component in question is by weight relative to the total weight of the apple extract.

[0074] Combination of assets 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 the only 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 conjunction with phloretin, apple extract, and grape pomace extract to reduce the accumulation of body fat and to prevent and / or treat cellulite. It helps stimulate fat metabolism and promote the breakdown of fat in fat cells and its use as fuel in the body. In addition, it works simultaneously to reduce oxidative stress and inflammation associated with cellulite.

[0079] For the purposes of this invention, "Coleus forskohlii extract" includes, in particular, a dry extract of Coleus forskohlii containing forskolin (CAS No. 66575-29-9) produced by an extraction process from Coleus forskohlii root.

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

[0081] The extract of Coleus forskohlii can be prepared by various extraction processes known to those skilled in the art, involving 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, ultrasonic-assisted extraction, and supercritical fluid extraction (e.g., CO2 + dipropylene glycol). Solid / liquid extraction leads to the recovery of metabolites, particularly forskolin, contained in the roots of Coleus forskohlii.

[0083] As a solvent, any solvent commonly used for plant extraction may be suitable. In particular, 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 Ci-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 by refluxing the extraction solvent used.

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

[0086] The extraction and 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 can be completed by a step of partial or total removal of the extraction solvent or solvent mixture. This allows obtaining a concentrated extract free of a significant amount of solvent or a dry residue by removing all of the extraction solvent or solvent mixture. Alternatively, the product of the extraction step can be lyophilized or spray-dried to obtain a powder form.

[0088] Examples of Coleus forskohlii extraction processes 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 pm, preferably less than 600 pm, preferably less than 250 pm, and a water content of less than 10%, preferably less than 5% by weight relative to the weight of the powder.

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

[0091] Apple extract

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

[0093] For the purposes of this invention, "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 their combinations. The apple fruit can be ripe or immature (still green). More specifically, the apple fruit is immature.

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

[0095] 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 / w apple extract of proanthocyanidins. Most preferably, the apple extract contains at least 90% w / w apple extract of proanthocyanidins.

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

[0097] Proanthocyanidin-rich apple extracts can be prepared from the apple fruit or its peels by any of the extraction processes 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 proanthocyanidin-rich apple extract used is preferably in powder form, in particular with a particle size of less than 1000 pm, preferably less than 600 pm, preferably less than 250 pm, 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 / wcombinaiSOn, preferably from 2% to 9% w / wCOmbinaiSOn of proanthocyanidin-rich apple extract.

[0100] Extract of grape marc n:

[0101] The combination according to the present invention contains an extract of grape pomace.

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

[0103] For the purposes of this invention, "grape pomace" means the residual product, in particular the skin (or pellicle), seeds, and possibly the stems, resulting from the pressing or crushing of grapes of the genus Vitis, for example from 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 pomace represents about 20 to 30% of the weight of the grapes used to make wine or grape juice.

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

[0105] For the purposes of this invention, "grape marc extract" means a product obtained from grape marc (skins, seeds, and possibly stems) recovered after the separation of the liquid part (juice or must) of the grape.

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

[0107] Grape marc extract is an excellent source of polyphenols and dietary fiber, as previously mentioned. 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 associated with cellulite. Indeed, these compounds can act by neutralizing free radicals responsible for oxidative stress and by modulating inflammatory pathways, thus helping to prevent cellular damage and mitigate undesirable inflammatory responses in the body, particularly in areas affected by cellulite.Furthermore, thanks to their inhibitory activities, such polyphenolic compounds have the potential to limit the absorption of dietary fats and prevent excessive fat accumulation in adipose tissue, thus helping to prevent the formation of cellulite and / or reduce existing cellulite. In addition, due to its high dietary fiber content (at least 30% by weight), grape pomace extract can help regulate blood sugar and carbohydrate metabolism. This dietary fiber slows down the digestion and absorption of carbohydrates, thereby preventing post-meal blood sugar spikes. By limiting the amount of carbohydrates available to be stored as fat in fat cells (adipocytes), grape pomace extract can help prevent fat accumulation and avoid unwanted weight gain. induce body weight loss. Furthermore, this ability to moderate the glycemic response may also be beneficial in preventing the formation of cellulite and / or reducing existing cellulite. By minimizing blood sugar fluctuations, grape pomace extract helps limit excessive fat storage in adipocytes.

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

[0109] Advantageously, the polyphenolic grape pomace extract is in powder form. It can be obtained from one or more grape pomaces by extraction with one or more solvents, according to methods for extracting grape skins or seeds known per se (see, for example, patent documents FR3002453A1, US6544581B1, WO2014013122, WO2017013299A1). Any solvent commonly used for extracting grape seeds or skins is suitable. Water and / or alcohols, such as methanol or ethanol, are particularly preferred solvents.

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

[0111] Dehydrated (raw) grape pomace powder or polyphenolic grape pomace extract generally has a particle size of less than 1000 pm, preferably less than 600 pm, preferably less than 250 pm, 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 / wcombinaiSOn, preferably from 4 to 20% w / wCOmbinaiSOn 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 this invention, "phloretin powder" means, in particular, an extract containing phloretin produced by an extraction process from the apple tree (e.g., the fruit, leaves, or peels). The phloretin powder can be extracted from the fruit (apple), either whole or in part. Typically, phloretin powder is obtained from apple leaves and / or peels.

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

[0117] Examples of phloretin extraction processes 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, preferably 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 adapted to 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 at a concentration of at least 20% w / w combinaiSOn, preferably from 20% to 40% w / w combinaiSOn-

[0121] A preferred combination according to the invention comprises: (a) at least 40% w / wCOmbinaiSOn of the Coleus forskohlii extract, (b) at least 1% w / wcombinaiSOn of the proanthocyanidin-rich apple extract, (c) at least 4% w / wcombinaiSOn of the grape marc extract, (d) at least 20% w / wcombinaiSOn of the 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 includes: (a') 45 to 70% w / wcombinaiSOn of Coleus forskohlii root extract, (b') 2 to 9% w / wCOmbinaiSOn of proanthocyanidin-rich apple extract, (c') 4 to 20% w / wcombinaiSOn of grape marc extract, (d') 20 to 40% w / wCOmbinaiSOn 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 includes: 58.39% w / w combination of Coleus forskohlii root, 3.65% w / w combination of proanthocyanidin-rich apple extract, 8.76% w / w combination of grape pomace extract, 29.20% w / w combination of phloretin powder.

[0124] Nutraceutical composition - Topical composition:

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

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

[0127] The nutraceutical composition or topical composition according to the invention may further comprise one or more suitable excipients or vehicles.

[0128] The term "excipient" or "vehicle" traditionally refers to any compound or composition intended to act as a simple support or vehicle, that is, not intended to have any 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.), or hydroalcoholic solvents (e.g., water and glycerin).Naturally, a person skilled in the art will take care to select any excipient compounds or compositions, as well as their proportions, in such a way that the advantageous properties of the combination according to the invention or of the nutraceutical composition containing it according to the invention are not, or are not substantially, altered by the intended addition. Excipients are well known in the nutraceutical field and are not described in detail here. Suitable excipients or vehicles are also described in the book entitled "Handbook of Pharmaceutical Excipients," Second Edition, American Pharmaceutical Association, 1994.

[0129] For the purposes of this invention, "appropriate" or "suitable" means that the excipient(s) (or vehicle(s) are compatible with nutraceutical or topical use of the composition according to 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 may 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 several amino acids (e.g., glutamine, aspartic acid, arginine, lysine, etc.), one or more plant extracts suitable for stimulating lipid and / or carbohydrate metabolism 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, Bifidobacterium lactis, Lactobacillus casei, etc.), yeasts (e.g., Saccharomyces boulardii) and mixtures thereof

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

[0132] Process - galenic - applications:

[0133] The combination according to the invention and the nutraceutical composition, or the topical composition containing it according to the invention, can be obtained by any conventional formulation method known in the nutraceutical or cosmetic fields. Their respective ingredients can be mixed, prior to formulation, in the order and under conditions readily 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 pm, preferably less than 1000 pm, preferably less than 600 pm, more preferably less than 250 pm, and with a moisture content of less than 10%, preferably 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 dosage forms normally used for topical application to the skin, including aqueous, hydroalcoholic, or oily solutions; oil-in-water, 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 foam. It may also be in solid form, such as a stick, or be applied to the skin as an aerosol.

[0136] When the combination is formulated as a nutraceutical composition, it may be in any of the dosage forms normally used for oral administration, including powders (e.g., compressed or loose powder, powders packaged in sachets, particularly single-dose sachets), tablets (e.g., chewable, effervescent, or gastro-resistant tablets), coated tablets, soft or hard capsules, softgels, hard capsules (e.g., film-coated and / or gastro-resistant capsules), granules, hard, soft, or gelled candies, chewing gum, or aqueous or oily suspensions. Powders, tablets, and softgels are preferred forms for oral administration.

[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 in a unit dosage form, such as a tablet, a capsule or a sachet-dose.

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

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

[0141] The nutraceutical composition according to the present invention can be dedicated to use as a nutrition 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 according to the invention can also be used to supplement a diet (e.g., enriching a food product or beverage) intended to help an individual achieve the expected beneficial effects, namely: reducing the accumulation of body fat, and / or preventing and / or treating cellulite.

[0145] The present invention also relates to a non-therapeutic use, by oral and / or topical route, of the combination according to the invention (in the form of a nutraceutical composition and / or a topical composition) to regulate lipid and / or carbohydrate metabolism; and / or to decrease and / or inhibit the accumulation of interstitial fluids; and / or to limit and / or inhibit the expansion of adipocytes; and / or to decrease and / or inhibit 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. 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 fluid; and / or for limiting and / or inhibiting adipocyte expansion; and / or for decreasing 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, in a human subject. In one embodiment, the non-therapeutic method according to the present invention comprises the oral administration to the human subject of a nutraceutical composition according to the invention. In another embodiment, the non-therapeutic method according to the present invention involves the topical administration to the human subject of a topical composition according to the invention.In yet another embodiment, the non-therapeutic method according to the present invention comprises both the oral administration to a human subject of a nutraceutical composition according to the invention and the topical administration to said human subject of 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 combined adoption of oral and topical administration promotes a synergistic approach, taking advantage of the respective benefits of these two modes of administration. The combination of these modes of administration offers a synergy that can optimize the overall effectiveness in 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... a method of the invention confers the advantages indicated above as being associated with the implementation of a combination according to the invention or of 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, softgels, tablets, caplets, dragees, chewing gums, etc.) or liquid form (e.g. gels, emulsions, syrups, etc.) in adequate quantity and number, depending on their form, and / or of a topical composition formulated for example in the form of a cream, milk, ointment, lotion, serum, gel, paste or foam, or in the form of a stick or aerosol.

[0148] The method according to the invention can be implemented over a period ranging 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. By way of example, when the nutraceutical composition according to the present invention is in a unit-dose form, as mentioned above, the administration of this nutraceutical composition can be carried out, for example, 1 to 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 of 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 according to the invention may include all or only part of the daily dose.

[0151] The invention also relates to a kit or combination product for simultaneous, separate, or staggered use, preferably a food supplement, comprising a first composition containing some of the components forming the combination according to the invention and a second composition comprising at least the other part of the components forming the combination according to the invention. This food supplement may be formulated such that the two compositions are in the same form 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 shaping and in the order, under conditions easily determined by a person skilled in the art.

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

[0154] The Coleus forskohlii root extract, the proanthocyanidin-concentrated apple extract, the grape pomace extract, and the phloretin powder used in the context of the present invention, and the processes for obtaining them, are well known to those skilled in the art. Each of these extracts can 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) possibly coupled to a mass spectrometer (MS).

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

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

[0158] The cellulite prevention and / or reduction effect can be demonstrated in various ways, for example by: - In vitro and ex vivo tests, particularly using appropriate 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 assessed through cytotoxicity analysis, metabolic activity analysis, or transcriptome and secretome analysis. Cytotoxicity and metabolic activity analysis allows for the determination of the stability of the combination's formula on adipose tissue. Secretome analysis allows for the evaluation of the combination's effect on metabolism and the fibrotic response of adipose tissue.Transcriptome analysis allows for the evaluation of adipogenesis at the lymphatic network level and the assessment of the reduction of the fibrotic phenotype. - The tests can also be complemented 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 assessment 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 immunoabsorption assay), PCR (Polymerase Chain reaction).

[0159] Other features and advantages of the invention will become clearer from the following examples, which are given by way of illustration and not limitation. The scope of the invention shall not be limited in any way by these examples. Examples:

[0160] Phloretin powder and 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 to analyze 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 classic 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 / pccombination) Coleus forskohlii root extract 58.4% Proanthocyanidin-rich apple extract 3.6% Grape pomace extract 8.8% Phloretin powder 29.2% 1. #

[0165] The three-dimensional (3D) ex vivo model of human adipose tissue used is derived from subcutaneous adipose tissue (abdominal region) specifically harvested 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] 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 combination A.

[0168] Incubation period: 7 days.

[0169] Production of model j. The adipose tissue was mechanically dissociated to preserve the 3D structure of the adipose tissue and the integrity of the cell populations. The tissue thus obtained was maintained in EGM-Plus medium (EBM basal medium + EGM supplements) under 3D culture conditions at 37°C, 5% CO2, 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 and 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 stimulant) 3 Anti-fibrotic Molecule Control Medium + TGF-

[31] dose 5 ng / ml (fibrotic stimulant) + SB431542 dose 5 pM (anti-fibrotic) 4 Grape Pomace Medium + TGF-

[31] dose 5 ng / ml (fibrotic stimulant) + Grape Pomace 30 pg / ml 5 Combination Medium + TGF-

[31] dose 5 ng / ml (fibrotic stimulant) + 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. Secretory Analysis (IL-6 ELISA) Metabolic Activity Analysis (XXT J7) Transcriptome (RNA extraction on J7, RT-qPCR) Fixation in PF A 4% 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 4N=4N=4N=4N=1 5N=4N=4N=4N=1

[0175] Study Timeline

[0176] After the model was produced, a pre-processing was carried out with the addition of Compounds containing TGF-

[31] were used to create a fibrotic model. Replenishment by adding compounds containing TGF-

[31] was performed on days 0, 3, and 6. On day 7, the supernatant was collected for XTT analysis, RNA lysis for transcriptomic studies, 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 METABLIC ACTIVITY XTT test on n=4 replicas / condition at day 9 Evaluation of general metabolic activity in adipose tissue. ANTICELLULITIS ACTION TRANSCRIPTOME Transcriptome at day 7 on n=4 replicas / condition (RNA by RT-qPCR) across a set of 5 genes / markers known from the literature: A DIPOQ (adiponectin), LEP (leptin), INHBA (activin A), MSCA1, LYVE-1 Evaluation of the lymphatic network and the reduction of the fibrotic phenotype, simulating cellulitis.VISUALIZATION BY IMAGING OF THE ANTI-CELLULITE ACTION -Adipocyte size (Oil Red 0 staining): visualization and measurement of the effect of the active ingredients on the reduction of fat mass in a fibrotic context -Cell matrix labeling (immunofluorescence labeling for Laminin): visualization of the effect of the active ingredients on the matrix of fibrotic adipose tissue -SHG microscopy: visualization of the reduction of the compaction of collagen fibers following exposure to the active ingredients Visualization of the effect of the active ingredients on ex vivo Ex AdEx 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 the active ingredients on the fibrotic response.

[0179] Results _

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

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

[0182] Adiponectin, a hormone primarily secreted by adipocytes, plays a crucial role in the metabolic regulation of carbohydrates and lipids. As a key mediator between adipose tissue, the liver, and muscles, it exerts significant positive effects on metabolism. Its ability to enhance insulin sensitivity, regulate inflammation, and modulate fatty acid oxidation makes it a central player in metabolic balance. By promoting balanced adipocyte growth, adiponectin supports 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 notable for its ability to stimulate the production of antioxidant enzymes, thereby helping to strengthen the body's natural defenses against oxidative stress.Furthermore, it exhibits 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 state, 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 in adipose tissue, which are decreased under 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 more commonly 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 under inflammatory conditions and fibrotic cells, without differentiation into adipocytes. Induction of the INHBA marker leads to dysregulation of adipose tissue homeostasis, thus promoting the transition to a fibrotic state. Furthermore, an increase in MSCA1 marker expression and a simultaneous decrease in INHBA (Activin A) marker expression suggest an anti-fibrotic effect, and consequently, a reduction in fibrotic cellulite and an improvement in adipose tissue homeostasis with a balance between fibroblast-like cell progenitors and adipocytes, leading to 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 regulating 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 functions optimally, it actively participates in the elimination of hyaluronic acid, thus contributing to the maintenance of tissue fluid balance. Enhanced LYVE-1 gene expression indicates optimized lymphangiogenesis (growth of the lymphatic network) and / or interstitial fluid transport across the lymphatic network, suggesting a potential effect in reducing aqueous cellulite.

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

[0188] Laminin: an extracellular matrix protein found in many tissues, including adipose tissue, playing a structural support role and contributing to cell migration and adhesion. In response to various inflammatory and fibrotic stimuli (in the context of cellulite, overweight, etc.), alterations in the laminin network (compaction, heterogeneous distribution) are 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 combination A tested induces a 47% increase in adiponectin expression compared to grape marc tested alone.

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

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

[0193] Combination A allows for the restoration of a homogeneous distribution of Laminin, with a signal that surrounds the adipocytes.

[0194] Combination A also reduces the presence of fibrillar collagen by approximately 60% compared to grape pomace alone (at an equivalent dose)

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

[0196] Imaging analysis revealed a 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, stimulate lymphangiogenesis with an improvement in the lymphatic network, induce improved adipose tissue homeostasis by balancing fibroblast and adipocyte progenitor cells toward a healthy adipose tissue state, produce an anti-fibrotic effect, decrease the presence and amount of fibrotic fibrillar collagen, improve laminin structure, and finally, decrease adipocyte size. These various effects could be beneficial for regulating lipid and carbohydrate metabolism, managing interstitial fluid accumulation, and reducing cellulite (adipose, aqueous, or fibrous cellulite).

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

[0201] A nutraceutical composition in powder form was prepared and then packaged in sachet-dose (e.g., in stick form) by mixing the ingredients shown 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 reducing 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 tablet form.

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

[0206] [Table 6], Ingredients Quantity (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 tablet form, also includes conventional excipients including flavorings.

[0208] Recommended dosages for reducing fat accumulation and for preventing or treating 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, M. Apples and Apple By-Products: Antioxidant Properties and Food Applications. Antioxidants 2023, 72, 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 / jmf2009.0273. 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 Microenvironments. Célis 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. PMID: 36079895; PMCID: 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

Demands

1. Non-therapeutic use of a combination of: (a) at least 40% w / wcombinaiSOn of an extract of Coleus forskohlii, (b) at least 1% w / wcombinaiSOn of an extract of apple rich in proanthocyanidins, (c) at least 4% w / wcombinaiSOn of an extract of grape marc, (d) at least 20% w / wcombinaiSon of a phloretin powder, the sum of (a), (b), (c) and (d) being equal to 100%, for the prevention or reduction of cellulite in a human subject.

2. Non-therapeutic use according to claim 1, wherein the extract of Coleus forskohlii is present in the combination at a content of 45 to 70% w / wcombinaiSOn

3. Non-therapeutic use according to claim 1 or 2, wherein the apple extract present in the combination contains at least 50% w / w apple extract, preferably at least 90% w / w apple extract of proanthocyanidins.

4. Non-therapeutic use according to any one of claims 1 to 3, wherein the proanthocyanidin-rich apple extract is present in the combination at a concentration of 2 to 9% w / w combination*

5. Non-therapeutic use according to any one of claims 1 to 4, wherein the grape pomace extract is present in the combination at a content of 4 to 20% w / w combination.

6. Non-therapeutic use according to any one of claims 1 to 5, wherein the phloretin powder is present in the combination at a content of 20 to 40% w / wCOmbinaiSOn-

7. Non-therapeutic use according to any one of claims 1 to 6, wherein the combination contains: (a') 45 to 70% w / w combination of Coleus forskohlii root extract, (b') 2 to 9% w / w combination of proanthocyanidin-rich apple extract, (c') 4 to 20% w / w combination of grape pomace extract, (d') 20 to 40% w / w combination of phloretin powder, the sum of (a'), (b'), (c') and (d') being equal to 100%.

8. Non-therapeutic use according to any one of claims 1 to 7, wherein the combination contains: 58.39% w / w Coleus forskohlii root combination, 8.76% w / w grape marc extract combination, 3.65% w / w proanthocyanidin-rich apple extract combination, 29.20% w / w phloretin powder combination.

9. Non-therapeutic use according to any one of claims 1 to 8, which further aims to stimulate lipid and / or carbohydrate metabolism, and / or reduce the accumulation of body or skin fat.

10. Combination as defined in any one of claims 1 to 8.

11. Nutraceutical composition, characterized in that it comprises, as an active agent, a combination according to claim 10.

12. Nutraceutical composition according to claim 11, 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.

13. Nutraceutical composition according to claim 11 or 12, which is a food supplement.

14. Topical composition, characterized in that it comprises, as an active agent, a combination according to claim 10.

15. Topical composition according to claim 14, 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, foams, sticks, aerosols.