WO2012072670A2

WO2012072670A2 - Use of a slimming combination

Info

Publication number: WO2012072670A2
Application number: PCT/EP2011/071360
Authority: WO
Inventors: Albert Duranton
Original assignee: L'oreal
Priority date: 2010-12-03
Filing date: 2011-11-30
Publication date: 2012-06-07
Also published as: FR2968213B1; FR2968213A1; WO2012072670A3

Abstract

The invention relates to the use of a combination of at least two active principles chosen from (i) a lotus extract, (ii) a pomegranate (Punica granatum) extract and (iii) a xanthine base chosen from caffeine, theophylline, theobromine and paraxanthine, or a plant extract rich in xanthine base, as slimming agent. It also relates to a cosmetic treatment method for preventing or reducing the increase in the volume of adipose tissue and/or the formation of fatty lumps.

Description

USE OF A SLIMMING COMBINATION

The present invention relates to an improved combination of active principles and to the use thereof as slimming agent and/or for the cosmetic treatment and/or the prevention of signs resulting from an excessive synthesis of lipids or from a decrease in vitality of the adipocytes.

Plumpness and/or excess weight are related to the response of certain cells, known as adipocytes, to excessive food consumption, which cells store this excess of ingested calories in the form of triglycerides. These triglycerides are synthesized in vivo by the adipocytes themselves, according to reactions of enzymatic type (lipogenesis), from the free fatty acids present in the blood in the form of lipoproteins and from the glucose provided in particular by certain foods. The release of the fatty acids from the lipoproteins takes place with the help of an enzyme, lipoprotein lipase, present in the adipocytes or via cell receptors for HDL, LDL or VLDL lipoproteins. The conversion of the glucose results either in the formation of glycerol or in the formation of free fatty acids via a specific enzyme, acetyl-CoA carboxylase, which converts the glucose to acetyl-CoA, which participates in the synthesis of the fatty acids. In point of fact, at the same time, the triglycerides thus formed and then stored in the adipocyte cells can also decompose (lipolysis), still under the action of specific enzymes, triglyceride lipases, which are present in these same cells and which are capable of being activated by cyclic AMP. Cyclic AMP is regulated by adenylate cyclase and is capable of being hydrolysed to 5ΆΜΡ by phosphodiesterase. This lipolysis mechanism results in the release of fatty acids, on the one hand, and of glycerol and/or of glycerol mono- and/or diesters, on the other hand.

The fatty acids thus released can then either diffuse into the body, to be consumed or converted therein in various ways, or can be recaptured by the adipocytes so as to regenerate triglycerides by lipogenesis.

In the case of an excessively rich diet or for people not pursuing physical exercise, a substantial imbalance is established in the body between lipogenesis (formation of triglycerides by enzymatic reaction between fatty acids and glycerol) and lipolysis (enzymatic decomposition of triglycerides to give fatty acids and glycerol), that is to say, more specifically, if the amounts of fat formed by lipogenesis become significantly and continually greater than those which are removed by lipolysis, then accumulation of triglycerides occurs in the adipocytes, which, if it becomes excessive, may be gradually reflected by deformation of the skin brought about by the thickening of the hypodermis in which the adipocytes are found.

Cellulite is composed of adipocytes in a collagen network. Located under the skin, they gorge themselves on fat and become hypertrophied. The swollen adipocytes compress the blood and lymphatic vessels, which results in poor removal of toxins and liquids, a state of hypoxia. This causes the adipocytes to suffer and, along with this, their subcutaneous supporting tissue also suffers. At a macroscopic level, this is reflected by the appearance of cellulite with that well-known and uneven appearance: "orange peel", which remains even in individuals, in particular women, who have lost weight. The surface of the skin becomes uneven and to a greater or lesser extent has a flabby or jelly-like consistency, finally giving the figure an unsightly general appearance.

Abdominal subcutaneous adipocytes are less sensitive than those of other parts of the body, i.e. visceral adipocytes, to the physiological regulation of lipolysis, in particular to the lipolytic effects of physical exercise which constitutes the main "natural" regulation of lipolysis (Dicker A et al., Horm. Metab. Res., 2009, Van Harmelen V. et al., Int. J. Obes. Relat. Metab. Disord., 1997).

This phenomenon is accentuated with age, the adipocytes becoming less reactive, and the lipolytic response to physical efforts decreases when the adipocytes lose their vitality. This is observed in particular for subcutaneous adipocytes of the abdominal region (Imbeault P. et al., J. Clin. Endocrinol. Metab., 2001).

Furthermore, it has been discovered that, when the cell vitality of adipocytes is reduced, in particular because of oxidative stress (cigarette smoke, sunlight, i.e. long wavelength UVA, visible, infrared), lipid accumulation or more generally through chronological ageing, the homeostasis of cutaneous structures dependent thereon is detrimentally affected.

From a biochemical viewpoint, it is known that adipocytes secrete numerous mediators which may have trophic effects or, on the contrary, detrimental pro-inflammatory effects in the surrounding tissues (Andersson C.X. et al., Diabetes Metab. Res. Rev., 2008). Thus, in cellulitis, it is suspected that the hypoxia of the adipocytes which results from the engorging of the adipose tissue induces the secretion of cytokines, such as IL-6 (Wang B. et al., Pflugers Arch., 2007). It is recognized that a low degree of chronic inflammation accompanies the accumulation of the adipocytes and this in return detrimentally affects their operation with in particular a decreased lipolysis. IL-6 would be one of the factors involved in this self-maintained phenomenon. The secretion of IL-6 characterizes cell ageing of the adipocyte (Horrillo R. et al., J. Immunol., 2010; Zoico E. et al., Biogerontology, 2010). In the connective tissue, IL-6 is an important factor in the induction of the secretion of metalloproteases by the fibroblasts, in particular MMP1 resulting in the destruction of collagen (Martin R. et al., Eur. J. Dermatol., 2008; Wisithphrom K. et al., J. Endod., 2006). The differential change in the cell vitality of subcutaneous adipocytes thus has numerous aesthetic repercussions.

These aesthetic disorders mean that there exists a need in cosmetics for compounds which act on subcutaneous adipocytes in order to improve their cell vitality when it is detrimentally effected.

In view in particular of the great unpleasantness, both physical and aesthetic, and sometimes psychological, which is caused in individuals who are affected by it, adiposity is nowadays a disorder which is increasingly poorly endured or accepted.

Solutions have been provided in the prior art for intervening in the metabolism of the fatty acids, which is one of the favoured targets in the control of this adipocytic lipid accumulation. This metabolism can be adjusted:

- either by blocking the transportation of the glucose inside the adipocyte, which results in a reduction in the capture of the fatty acids by the adipocyte,

- or by inhibition of lipoprotein lipase,

- or by activation of triglyceride lipase (or hormone-sensitive lipase), generally by stimulating cyclic AMP, generally by activation of adenylate cyclase, or by causing it to accumulate by inhibition of phosphodiesterase.

Other biological routes have been explored for acting on the mechanism of lipogenesis and/or of lipolysis. Thus, the proposal has been made to use neuropeptide Y (NPY) receptor antagonists, neuropeptide Y being a neuromediator involved in a number of physiological processes, the involvement of which in the regulation of lipolysis it has been possible to demonstrate (P. Valet, J. Clin. Invest., 1990, 85, 291-295). Use may also be made of oc₂ receptor antagonists or

receptor agonists. The cosmetic compositions provided to date for the purpose of treating adiposity thus comprise compounds, referred to as slimming compounds, which act on one or more of the mechanisms mentioned above. Among these, mention may more particularly be made of xanthine bases (i.e. xanthine derivatives), such as theophylline, caffeine, theobromine and 1-hydroxyalkylxanthines and their compatible salts (see the document FR-A-2 617 401), which are phosphodiesterase inhibitors, nicotinic acid derivatives, such as more particularly oc-tocopherol nicotinate and hexyl nicotinate (see the document EP-A-371 844), substances referred to as oc₂ blockers, capable of blocking oc₂ receptors at the surface of the adipocytes, such as, for example ginkgo biloba (see the document FR-A-2 669 537 or US 5, 194,259), and

receptor agonists, such as alverine and its salts. AMPK is present in all the cells of the body and it plays therein the role of energy gauge. AMPK (or 5' adenosine monophosphate-activated protein kinase) is a heterotrimeric enzyme composed of an a catalytic subunit having kinase activity and of 2 β and γ regulating subunits. The activity of the AMPK depends on the variation in the AMP/ATP ratio which characterizes the energy level of the cell (the ATP being hydrolysed to give AMP in order to "deliver" the necessary energy to all the biochemical processes of the cell). It is present in 2 forms, phosphorylated or nonphosphorylated, the phosphorylated form being the active form.

When it is activated in response to an energy demand or to a stress of the cell, the AMPK enhances the energy-generating processes, such as glycolysis, and inhibits the nonessential consuming processes, thus allowing the cell to survive. The preservation of the cell energy status is involved in the maintenance of the longevity of the species and the control of signs of ageing. Thus, the compounds capable of enhancing the activity of AMPK currently form the subject of great interest in the treatment of age-related clinical signs.

The AMPK activity corresponds to the cell concentration of phosphorylated AMPK. Thus, it is advisable to have the highest possible levels of phosphorylated protein in order to have this high activity. As regards more particularly the aesthetic disorders resulting from the detrimental change in the cell vitality of adipocytes, it is known that AMPK is involved in lipolysis and the oxidation of lipids (Koh H.J. et al., Biochem. J., 2007, Gaidhu M.P. et al., J. Lipid Res., 2009) and that it increases the reactivity of adipocytes to physical exercise. Lipolysis, when it is strongly stimulated, is a phenomenon which can detrimentally affect the cell vitality of adipocytes (and thus their long-term reactivity). Thus, the effects of the activation of AMPK on lipolysis are paradoxical: a strong and temporary activation results in a reduction in lipolysis, in order to preserve the vitality of the adipocytes (Ruderman N., J. Biol. Chem., 2008), and a prolonged activation results in an increase in lipolysis (Gaidhu M.P., J. Lipid Res., 2009). It is also known, from US 20100081643, KR 10681439 and WO 2006001278, that the activation of AMPK makes it possible to reduce the expression of the enzymes involved in the synthesis of lipids, such as acetyl-CoA carboxylase (ACC). This makes it possible to use AMPK activators to reduce obesity.

The importance of the detrimental change in mitochondria, organelles which supply the cell with energy, in the ageing and cell death of adipocytes is also known (Berneburg M. et al., J. Dtsch. Dermatol. Ges., 2010) and it is also known that activation of AMPK makes possible the biogenesis of mitochondria.

Thus, the advantage is conceived of using AMPK activators to reduce the production of lipids in adipocytes and to maintain the vitality of subcutaneous adipocytes, to, according to the skin region and/or the effect expected, prevent suffering of the supporting tissue of the adipocytes and thus reduce cellulite, increase the reactivity of the adipocytes to physical exercise in order to holistically reduce excessive adiposity. Unexpectedly, it has been found, in the context of the present invention, that combinations of active principles otherwise known are capable of synergistically stimulating the activity of AMPK, in particular the production of phosphorylated AMPK by human adipocytes. These combinations will thus be of particular use in combating excess weight, in particular localized excess weight and lipodystrophy, and preventing or reducing the phenomenon of cellulite.

Such combinations act both on the mechanisms of synthesis and of decomposition of lipids in cutaneous and subcutaneous tissues and by reducing the associated inflammatory phenomena and the detrimental change in the supporting connecting tissue. This is why a subject-matter of the present invention is the use of a combination of at least two active principles chosen from (i) a lotus extract, (ii) a pomegranate (Punica granatum) extract and (iii) a xanthine base chosen from caffeine, theophylline, theobromine and paraxanthine, or a plant extract rich in xanthine base, as slimming agent.

These combinations make it possible to combat unattractive excess weight, in particular local lipodystrophia, for the purpose of obtaining a general effect or, on the contrary, a local effect of slimming and/or refining the body or the face.

Combinations of particular use according to the invention comprise at least one lotus extract and one pomegranate (Punica granatum) extract.

According to a specific embodiment, the combination comprises the 3 types of active principles.

Combinations of particular use according to the invention comprise at least two active principles chosen from:

(i) a lotus extract,

(ii) a pomegranate (Punica granatum) extract, and

(iii) caffeine or its derivatives, or a caffeine-rich plant extract.

The use according to the invention is advantageously a cosmetic use; the term "cosmetic" is understood to mean intended to improve the aesthetic appearance of the skin or its appendages.

The extracts can be obtained by any method for the preparation of a plant extract known to a person skilled in the art. In particular, the extract can be obtained by maceration of the part of the plant in water, or in a solvent composed of a mixture of water and of an organic solvent, for example water/alcohol, or water/acetone, or water/propylene glycol, or water/butylene glycol. The plant/solvent ratio can vary, for example and without limitation, from 1/4 to 1/20. Advantageously, the preparation of the extract begins with the grinding of the parts of the plant, followed by maceration in the extraction solvent for several hours. The extraction can be carried out with stirring in order to improve the performance thereof. The extraction can be carried out at ambient temperature or by increasing the temperature, for example to 50°C or to 60°C. Once the extraction has been carried out, the solution is filtered.

The solution thus obtained can be concentrated by any process known to a person skilled in the art. Likewise, the solution obtained can be lyophilized by any conventional lyophilization method; a powder is thus obtained.

The term "lotus extract" is understood to mean any extract of a plant from the Nelumbonaceae family, in particular from the Nelumbo genus. Use may in particular be made of extracts of Nelumbo nucifera, or of other species, such as Nelumbo lutea (American yellow lotus) or Nelumbo pentapetala. Advantageously, the extract is an extract of Nelumbo nucifera or sacred lotus. Extracts can originate from different parts of the plant and will be chosen in particular from extracts of flowers, fruits, seeds, roots, leaves or stems. Extracts which are particularly suitable in the invention are extracts of leaves or the green part of the plant. The lotus extract will be chosen in particular from extracts of leaves or stems of Nelumbo nucifera, Nelumbo lutea and/or Nelumbo pentapetala.

The extraction of a starting material from Nelumbo nucifera can be obtained with a water/ butylene glycol mixture in a proportion variant from 90/10 to 10/90 v/v, preferably 50/50 v/v. A dry matter content obtained is of the order of 5 to 12 g/l.

Advantageously, the lotus extract is an aqueous, alcoholic or aqueous/alcoholic extract of leaves.

Preferably, the lotus extract of use for the invention has a concentration of total polyphenols of 0.4 to 0.8 g/l.

Such extracts of leaves of Nelumbo nucifera are sold by Silab.

Caffeine (or 1 ,3,7-trimethylxanthine) is present in the seeds, leaves and fruits of various plants. The most widely used natural products containing caffeine are coffee, tea and, to a lesser extent, cocoa. Mate and guarana are other sources of caffeine. Other methylxanthines with a xanthine base on which one or more hydrogens in the 1 , 3 and 7 positions is/are replaced by a methyl group or extracts comprising them can also be used according to the invention. These include: caffeine (1 ,3,7-trimethylxanthine), theophylline (1 ,3-dimethylxanthine), theobromine (3,7-dimethylxanthine), paraxanthine (1 ,7- dimethylxanthine) and xanthine.

The caffeine-rich plant extract can be chosen in particular from coffee, tea and/or cocoa extracts.

Advantageously, the caffeine-rich plant extract is a coffee extract comprising at least 1 %, in particular at least 3%, of caffeine (by weight). The pomegranate tree (Punica granatum) is a small deciduous fruit tree originating in Asia, in particular in the Middle East, belonging to the family Punicaceae. It can reach approximately 5 m, but often it reaches no more than 1.50 m. The flowers are orange-red or scarlet, single or double, depending on the variety. The fruits are edible and have been consumed since antiquity. Synonyms are Punica spinosa, Punica florida or Granatum puniceum.

Its fruit is the pomegranate of orange-red colour, with a hard and tough pericarp (or skin), containing, in from 6 to 12 membranous loculi, numerous triangular seeds with a translucent and juicy aril. The pomegranate is rich in vitamins (B, C and D) and in polyphenols in the form of tannins. The skin extracts have been proposed for their astringent, antibacterial and antihaemorrhagic properties.

According to an advantageous embodiment of the invention, the pomegranate extract is an extract of the fruit of Punica granatum, or of its fractions.

The extraction starting from pomegranate fruit, including the seed, can result, according to a specific embodiment, in the preparation of an essential oil. The extract in the form of a concentrated solution, as well as the extract in the form of a powder and as well as the extract in the form of an essential oil, can be used in a medium appropriate for application to the skin or its appendages.

The oil from the seeds, rich in conjugated linolenic acid (pucinic acid) and in conjugated linoleic acid, is thus particularly suitable for the invention. Use is made in particular of an oil extracted from pomegranate seeds comprising at least 0.0008% of conjugated linoleic acid.

A preferred pomegranate extract is a pomegranate seed oil extract, comprising conjugated linoleic acid (at least 5% by weight of the extract). .

This is because it has now been found that such active principles, which, taken in isolation, have no or little effect on the production of AMPK by adipocytes, are capable, in combination, of stimulating the activation of AMPK by adipocytes, and thus of improving the vitality of these cells.

AMPK is involved at two levels in:

- the lipolytic effect of exercise (Koh H.J. et al., Biochem. J., 2007),

- the maintenance of the vitality of adipocytes when they are highly stimulated, by reducing the depletion in ATP which accompanies intense lipolysis (Gauthier M.S., Ruderman N.B., J. Biol. Chem., 2008), with as a consequence a reduced detrimental change in the adipocytes themselves and in the subcutaneous supporting tissue for these adipocytes.

In particular, the combination has a synergistic effect on the activation of AMPK in abdominal adipocytes.

This synergy is advantageous, the impact of the activation of AMPK on lipolysis being known, which can be demonstrated by the increase in the release of nonesterified fatty acids (NEFAs). The combination furthermore reduces the release of mediators, such as IL-6 and MCP-1 , and thus improves the signs associated with the deformation of the supporting tissue.

The effect of the combination is also synergistic with regard to the oxidation of the lipids, corresponding to the production of energy by consumption of the fats in the adipose tissue. This effect mimics the effect of moderate physical exercise on lipolysis, in particular in abdominal adipose tissue.

The combination has a synergistic effect on the activity of the adipocytes. Within the meaning of the invention, the effect displayed by the combination is described as synergistic insofar as it proves to be greater than that expected from the simple superposition of the respective effects of each of its constituents.

The invention relates in particular to the cosmetic use of a synergistic combination of at least two agents chosen from (i) a lotus extract, (ii) a pomegranate (Punica granatum) extract and (iii) caffeine or a caffeine-rich plant extract.

Advantageously, use is made of a combination which comprises at least (i) one lotus extract, (ii) one pomegranate (Punica granatum) extract and (iii) caffeine or a caffeine-rich plant extract.

More advantageously, use is made of a combination which comprises at least one lotus extract, one pomegranate (Punica granatum) extract and one caffeine-rich plant extract.

The amounts of the various active principles will be adjusted by a person skilled in the art according to the effect desired, the type of starting material and the formulation.

Combinations exhibiting an improved stimulating effect on AMPK, of synergistic type, will comprise, for example, a ratio of the lotus extracts to the xanthine base or the extract comprising it, in particular caffeine or coffee extracts, of between 200 (lotus): 1 (xanthine base) and 10: 1 , in particular approximately 5:1.

The ratio of the pomegranate extracts to the lotus extracts is preferably between 1 (pomegranate):200 (lotus) and 1 :10 in the combinations according to the invention, in particular approximately 1 :5. Likewise, for the implementation of the invention, the ratio of the pomegranate extracts to the coffee extracts and/or caffeine is preferably between 1 :5 and 5:1 , in particular 1 :1. Preferably, the ratio of the pomegranate extracts to the coffee extracts and/or caffeine is preferably between 1 :5 and 1 : 1.

According to one of the embodiments of the invention, pomegranate extract/lotus extract/ caffeine or caffeine-rich extract ratios are between 0.5: 1 :10 and 0.5:1 : 1 , in particular approximately 1 :5: 1.

According to other embodiments of the invention, pomegranate extract/lotus extract/caffeine or caffeine-rich extract ratios are between 1 :5: 1 and 1 :200:5; preferably between 1 :5: 1 and 1 : 10:5.

Preferably, the ratios defined hereinabove are weight ratios between the raw extracts present in the claimed combinations. Advantageously, the combinations according to the invention will be present in a composition suitable for topical application to the skin or superficial body growths, comprising a physiologically acceptable medium. It is in particular a cosmetic or dermatological composition. According to one of the embodiments, the combinations according to the invention will be used orally and will be present in a composition suitable for oral administration.

The concentrations of each of the constituents of the combination can vary in particular from 0.0001 % to 40% by weight, with respect to the total weight of the composition, in particular from 0.0001 % to 0.01 % or from 0.01 to 10%. Concentrations giving good results are concentrations from 0.008% to 7%, in particular from 0.04% to 5%, more particularly from 0.4 to 4%.

Preferably, the combination of active principles is present in the composition comprising a physiologically acceptable medium at a concentration of between 0.001 % and 40%, with respect to the total weight of the composition.

For each of the natural constituents, the total extract will be used at one of these concentrations chosen within this range for its compatibility in terms of the cosmetic formulation and will be adjusted by a person skilled in the art according to the type of formulation chosen, the method of administration and the effect desired, so as to supply the effective concentration of the combination to the skin cells; the activity with regard to AMPK will in particular be confirmed in vitro as described in the examples with a dilution to 1/100 of this concentration and a maximum non-cytotoxic concentration of the mixture.

By way of indication, use may be made, according to the invention, of a lotus extract, such as a Nelumbo nucifera extract, which is a water/butylene glycol 50/50 extract with a concentration from at least 1 up to 12 g/l by weight of dry matter. From 0.4 to 0.8 g/l of total polyphenols (gallic acid) can be assayed. Such an extract can be used in the composition according to the invention at a concentration from at least 0.05% up to 10% by weight, in particular from 0.1 to 5% by weight and preferably from 1 to 4% by weight, with respect to the total weight of the composition.

As set out above, use may be made, as pomegranate extract, of all or part of the plant, for example a 100% seed oily extract dissolved in glycerol. It can be used at a concentration of 0.005 to 15% by weight, in particular of 0.05 to 5% by weight, more particularly of 0.1 to 3% by weight, with respect to the total weight of the composition according to the invention. Notably, said pomegranate extract is used at a concentration of less than 0.03%, more preferably of less than 0.02% by weight, still preferably of less than 0.01 % by weight, with respect to the total weight of the composition according to the invention. In particular, said pomegranate extract may be used at a concentration of 0.005 to 0.03% by weight, in particular of 0.006 to 0.02% by weight, in particular of 0.007 to 0.01 % by weight, more particularly of about 0.008% by weight, with respect to the total weight of the composition according to the invention.

As regards the xanthine or methylxanthine bases, such as caffeine, or any extract capable of contributing xanthine bases, the concentration of caffeine in the composition according to the invention will be from at least 0.05 up to 10% by weight, in particular from 0.5 to 5% by weight, with respect to the total weight of the composition, indeed even from 0.5 to 3% by weight. It is possible, for example, to take a concentrated aqueous solution comprising 15 g/l of caffeine, which also includes xanthine bases, such as theobromine, 7- methylxanthine or paraxanthine, and to dilute this solution of 15 g/l of caffeine to 0.5 to 5% in the composition which will be administered to the subject according to the invention. Preferably, the concentration of caffeine in the final composition applied to the subject will be greater than or equal to 0.0008% and in particular greater than or equal to 0.008%, with respect to the total weight of the composition.

Preferably, the compositions according to the invention additionally comprise a physiologically acceptable medium. The term "physiologically acceptable" is understood to mean in particular compatible with the skin, mucous membranes and superficial body growths and not causing undesirable effects when brought into contact with the various parts of the body. For oral administration, the composition of the invention can be provided in all the appropriate forms, particularly in the form of a solution to be taken orally, a syrup, a tablet, including a sugar-coated tablet, a capsule, including a hard gelatin capsule, a nutritional food or a nutritional supplement.

The said composition can additionally comprise at least one appropriate excipient suitable for oral administration.

A composition suitable for topical administration can be provided in particular in the form of an aqueous, aqueous/alcoholic or oily solution, of a dispersion of the solution type or dispersion of the lotion or serum type, of an emulsion with a liquid or semiliquid consistency of the milk type, obtained by dispersion of a fatty phase in an aqueous phase (O/W) or vice-versa (W/O), of a suspension or emulsion with a soft, semisolid or solid consistency of the cream, aqueous gel or anhydrous type, or of a microemulsion, of microcapsules, of microparticles or of vesicular dispersions of ionic and/or nonionic type. The pH of a composition according to the invention, when it comprises at least one aqueous phase (e.g.: aqueous solutions, emulsions, and the like), is preferably between 4 and 9, preferably between 4 and 7, advantageously from 5 to 6 and in particular 5.5.

The composition according to the invention can be more or less fluid and can have the appearance of a white or coloured cream, of an ointment, of a milk, of a lotion, of a serum, of a paste or of a foam. It can also be applied to the skin in the aerosol form. It can also be provided in the solid form, for example in the stick form. It can be used as a care product but also as a toiletry product.

This composition can constitute a mask, a cleansing, protecting, treating or care cream for the face or for the body (for example day creams, night creams, make-up removing creams, foundation creams or antisun creams), a make-up removing milk or a lotion, gel or foam for caring for the skin, such as a cleansing lotion.

Generally, any composition of the invention can be applied to the skin (over any skin region of the body).

In a known way, a cosmetic composition can also comprise adjuvants normal in the cosmetics field, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic additives, preservatives, antioxidants, solvents, fragrances, fillers, UV screening agents (sunscreens), odour absorbers and colouring materials. The contents and the natures of the ingredients employed in the compositions of the invention are adjusted by a person skilled in the art so as not to substantially affect the effect of the combination under consideration according to the invention. The compositions according to the invention can comprise additional cosmetic active principles and in particular at least one other active principle chosen from desquamating agents, firming agents, agents which promote the synthesis of the constituents of the extracellular matrix or which inhibit their decomposition, moisturizing agents, aquaporin modulators, agents active with regard to microcirculation, or agents active with regard to the energy metabolism of the cells.

Use is made in particular of:

- active principles which act with regard to microcirculation (vasoprotective or vasodilatory agents), such as flavonoids, Ginkgo biloba extracts, ruscogenins, esculosides, the aescin extracted from the horse chestnut, nicotinates, hesperidin methyl chalcone, butcher's broom, or essential oils of lavender or rosemary;

- firming active principles, such as Centella asiatica and Siegesbeckia extracts, which stimulate the synthesis of collagen, silicon, amadorine, vitamin C and its derivatives and retinol and its derivatives, or yeast extracts, such as Saccharomyces cerevisiae extracts; - desquamating agents which are particularly suitable for the invention will be chosen from hydroxylated acids, in particular a- or β-hydroxylated acids, in particular salicylic acid and its derivatives, 5-(n-octanoyl)salicylic acid, jasmonic acid and its derivatives, in particular 3-hydroxy-2-pentylcyclopentaneacetic acid, and their cosmetically acceptable salts. Compositions suitable for the implementation of the invention can thus comprise, in addition to the synergistic combination defined above, at least one agent chosen from butcher's broom extracts, Ginkgo biloba extracts, Saccharomyces cerevisiae extracts, 5-(n-octanoyl)salicylic acid and/or salicylic acid and their salts, and green coffee extracts. The amounts of these various additional active principles will be adjusted by a person skilled in the art but are generally from approximately 0.01 to 15% by weight, preferably from 0.1 to 10% by weight, with respect to the total weight of the composition.

Another subject-matter of the invention is a cosmetic treatment method intended to prevent or reduce the increase in the volume of adipose tissue and/or the formation of fatty lumps and/or a slimming method comprising the application, over all or part of the body, of a composition comprising at least one combination as defined above. Application can be carried out in particular over regions subject to lipodystrophia, such as the abdomen, the top of the thighs or arms, or certain regions of the face, such as the bottom of the face. This application can advantageously be carried out after a physical effort or in complementing a slimming course of treatment. It can be repeated over time, several times daily and for several weeks or several months.

The treatment can also be carried out in complementing a treatment for obesity.

According to a specific embodiment, a combination comprising at least one active principle from each of the 3 categories: (i) a lotus extract, (ii) a pomegranate (Punica granatum) extract and (iii) a xanthine base chosen from caffeine, theophylline, theobromine and paraxanthine, or a plant extract rich in xanthine base, is applied to the skin or its appendages.

According to a more specific embodiment, a combination comprising at least one active principle from each of the 3 categories: (i) a lotus extract, (ii) a pomegranate (Punica granatum) extract and (iii) caffeine or a caffeine-rich plant extract, is applied to the skin or its appendages.

The term "skin" is understood to mean, within the meaning of the invention, the whole of the covering of the body and in particular the skin, mucous membranes and scalp.

The term "superficial body growths" is understood to mean all of the skin appendages and in particular the nails, hair and non-scalp hairs.

The combination according to the invention can be used to prevent or combat cellulite and/or to refine the figure or the outlines of the face.

According to an alternative form, a subject-matter of the invention is a cosmetic method for preventing or reducing the increase in the volume of adipose tissue and/or the formation of fatty lumps and/or a slimming method for a subject, in which the said subject ingests a combination of at least two of the active principles as defined above, in particular a combination of at least one lotus extract and one pomegranate extract, more particularly at least the 3 active principles, or a composition comprising them.

In the methods according to the invention, the synergistic combination or the composition comprising it will preferably be applied topically to the skin, mucous membranes and/or keratinous fibres. Application can be daily or twice daily. Results may be observed from the start of the application of the cosmetic treatment. However, the cosmetic treatment according to the invention can be prolonged for several weeks, indeed even several months. The combination or the composition comprising it will preferably be applied to the regions of skin affected by the disorders which it is desired to combat.

In the case of the active principles or compositions according to the invention being taken orally, oral administration can be daily or several times daily, for example in the morning and evening. It can be continued for several weeks and/or several months, according to the effects desired.

The methods and the uses according to the invention make it possible to improve the vitality of subcutaneous adipocytes and/or to improve their metabolism. The implementation of the invention is thus particularly suitable for reducing or preventing fatty lumps, persistent plumpness and cellulite, in particular on the stomach and waist of the individuals treated. In addition, the invention makes it possible to improve the firmness of the regions of skin affected and to combat slackening and the orange-peel appearance. The method and the use of a combination according to the invention has in addition an action in reinforcing the beneficial lipolytic effect of a moderate long-term physical activity.

The use of a combination and/or the method according to the invention have an improved effect with regard to the inhibition of lipogenesis, the increase in lipolysis and the reduction in the production of MMP (matrix metal loprotease), thus preventing detrimental changes to the collagen.

The invention will be illustrated in more detail in the following examples.

In these examples, reference will be made to the appended figure, which represents the amount of phosphorylated AMPK measured in the cytosol of adipocytes cultured in the presence of different active principles.

Example 1 :

In the present study, an attempt has been made to confirm, in differentiated human preadipocytes, whether an AMPK activator is capable of interacting with a lipolysis activator at the nonesterified fatty acids and of reducing possible releases of IL-6 and MCP-1 , and also its effects on the formation of reactive oxygen species (ROSs). The test was thus carried out according to the following scheme:

1) A dose/effect test on isoproterenol was carried out, on abdominal subcutaneous adipocytes, with regard to:

- the secretion of interleukin-6 (it is known that adrenergic stimulation induces the secretion of IL-6 (Fasshauer M. et al., Horm. Metab. Res., 2003)) and monocyte chemoattractant protein-1 ,

- the production of activated oxygen species, which results from the oxidation of fatty acids at the mitochondria in adipocytes (Carriere A. et al., J. Biol. Chem., 2004), under our experimental conditions, where a very low adrenergic activation is mimicked.

2) At the lowest active dose of isoproterenol, the adipocytes were pretreated with an AICAR positive reference AMPK activator (validation of the experimental model) in order to test whether the adipocytes have become more sensitive with regard to some of these parameters.

3) The AMPK-activating products were tested under these conditions where the effect of the isoproterenol is "enhanced" by an activation of the AMPK with AICAR.

The compounds tested are as follows:

- Nelumbo nucifera lotus extract, water/butylene glycol 50/50, taken at 0.4% and 1 % in the culture medium;

- Caffeine, anhydrous powder, used at 0.008% and 0.04% in the culture medium;

- Pomegranate seed oily extract, at 0.008% and 0.04% in the culture medium.

The level of expression of the P-AMPK was analysed by Western blotting:

At the end of the incubation, the proteins were extracted, quantified, then separated by electrophoresis on 10% polyacrylamide gel and then transferred on to a nitrocellulose membrane.

The phospho-AMPK (P-AMPK) and AMPK proteins were visualized using specific antibodies, themselves visualized using an anti-immunoglobulin peroxidase conjugate. After washing in PBS-Tween, the peroxidase activity was visualized by the ECL (electrochemiluminescence) method. The densitometry measurements on a chemiluminescence scanner (Fuji LAS 3000) are obtained with Multigauge software (Fujifilm).

In this test, an increase in the phosphorylated form of AMPK (active form of the enzyme) is evaluated with respect to the effect of the AICAR reference (5-amino-4- imidazolecarboxamide-riboside):

Effects on the phosphorylation of AMPK

Effects of the various treatments on the phosphorylation of the APMK in differentiated human preadipocytes.

In bold, marked difference with respect to the differentiated control. Under the experimental conditions, the differentiated control exhibits a content of phosphorylated AMPK of 599 AU/mm².

The AICAR positive reference made it possible to increase this content to 751 AU/mm². This result validates the test. The pomegranate extract tested alone did not modify the content of phosphorylated AMPK in the preadipocytes, whatever the concentration tested.

The combination lotus extract + caffeine did not show a very marked effect on the phosphorylation of the AMPK under these conditions.

The combination pomegranate + lotus + caffeine at the lower concentration (at 0.008% + at 0.04% + at 0.008%) brings about a large increase in the content of phosphorylated AMPK (706 AU/mm²).

This synergistic effect is thus equivalent to the AICAR reference with the combination of compounds, each taken at a very low concentration. It may be concluded that, with these combinations, an AMPK-activating effect is obtained, whereas this effect remains weak when the compounds are taken in isolation, even at higher concentrations.

Example 2: Search for a synergistic effect between AMPK activators and lipolysis activator with regard to the release of IL-6, and MCP-1 and with regard to the production of ROSs. The ROSs are measured with the DCF probe added in the final 30 minutes of culturing. The intensity of fluorescence (ex 485 nm, em 538 nm) is proportional to the concentration of ROSs and is quantified with a fluorometer (Spectramax, Molecular Devices).

The amounts of NEFAs, IL-6, and MCP-1 in the supernatants are measured with assay kits (according to the instructions of the suppliers) respectively Oxoid 46551 , R&D Systems ref DY206, and R&D Systems ref DY279. a) Basal release/effect of the isoproterenol

The effects of the isoproterenol on the basal release of NEFA, IL-6, MCP-1 and ROSs by differentiated human preadipocytes.

* : Statistically significant difference with respect to the control.

The isoproterenol brings about an increase in the release of NEFAs by the cultured preadipocytes (dose-dependent increase) but has not significantly modified the release of IL-6 and MCP-1.

The isoproterenol increases the production of ROSs with respect to the control, which corresponds to inducing oxidation of the fatty acids in the mitochondria (Carriere A. et al., J. Biol. Chem., 2004). b) Effect of the combination of active principles

Effect of the isoproterenol on the release of NEFAs, IL-6, MCP-1 and ROSs by differentiated human preadipocytes pre-treated for 8 h with the combination Pomegranate + lotus extract + caffeine or the AICAR reference.

* and ** : Statistically significant difference with respect to the control. The AICAR has brought about an increase in the release of IL-6 after stimulation with isoproterenol (+70% of the control), but has been shown to be without significant effect on MCP-1 and the ROSs.

The combination has made it possible to inhibit in a statistically significant way the release of IL-6 and MCP-1 with respect to the preadipocytes treated solely with the isoproterenol. An advantageous soothing effect is thus recorded.

The effect of the isoproterenol on the production of ROSs (and thus the oxidation of the lipids) is enhanced with the combination. Conclusion

Under the experimental conditions of this study, it is shown that the combination Pomegranate + lotus + caffeine exerts a synergistic effect on the activation of AMPK. Their combination at these inactive concentrations is synergistic, with levels of activity equivalent to the AICAR pharmacological reference.

A synergistic effect with isoproterenol is also found: this activation improves the response of the abdominal subcutaneous adipocytes to a minimal adrenergic stimulation, mimicking the effect of moderate physical exercise. Example 3: Compositions for topical application The 3 following compositions A, B and C are prepared according to the invention.

The compositions will be applied every day, twice daily, to the stomach, legs, hips and/or waist.

Claims

I- Use of a combination of at least (i) one lotus extract and (ii) one pomegranate (Punica granatum) extract, as slimming agent

2- Use according to claim 1 , characterized in that the combination further comprises at least one xanthine base chosen from caffeine, theophylline, theobromine and paraxanthine, or a plant extract rich in xanthine base.

3- Use according to Claim 1 or 2, characterized in that at least one lotus extract is chosen from extracts of leaves or stems of Nelumbo nucifera, Nelumbo lutea and/or Nelumbo pentapetala.

4- Use according to either of the preceding claims, characterized in that the lotus extract is an aqueous, alcoholic or aqueous/alcoholic extract of leaves.

5- Use according to one of the preceding claims, characterized in that at least one xanthine base is caffeine.

6- Use according to at least one of the preceding claims, characterized in that at least one caffeine-rich plant extract is chosen from coffee, tea, cocoa, mate and/or guarana extracts.

7- Use according to one of the preceding claims, characterized in that the caffeine- rich plant extract is a coffee extract comprising at least 1 % of caffeine.

8- Use according to at least one of the preceding claims, characterized in that at least one pomegranate extract is an extract of the fruit of Punica granatum, or of its fractions.

9- Use according to Claim 7, characterized in that at least one pomegranate extract is a pomegranate seed oil extract comprising conjugated linoleic acid.

10- Use according to any one of the preceding claims, characterized in that the combination has a synergistic effect on the activation of AMPK in adipocytes and/or preadipocytes.

I I- Use according to at least one of the preceding claims, characterized in that the ratio of the lotus extracts to the xanthine base is between 200:1 and 10: 1.

12- Use according to at least one of the preceding claims, characterized in that the ratio of the lotus extracts to the pomegranate extracts is between 200:1 and 10:1.

13- Use according to at least one of the preceding claims, characterized in that the ratio of the pomegranate extracts to the xanthine base is between 1 :5 and 5:1.

14- Use according to at least one of the preceding claims, characterized in that the combination comprises at least one lotus extract, one pomegranate (Punica granatum) extract and caffeine or a caffeine-rich plant extract. 15- Use according to any one of the preceding claims, characterized in that the combination of active principles is present in a composition comprising a physiologically acceptable medium at a concentration of between 0.001 % and 40%, with respect to the total weight of the composition.

16- Use according to any one of the preceding claims, characterized in that the lotus extract is from 0.0001 % to 40% and the pomegranate extract and the xanthine base or the plant extract rich in xanthine base are each at a concentration of between 0.0001 % and 40%, with respect to the total weight of the composition.

17- Cosmetic treatment method for preventing or reducing the increase in the volume of adipose tissue and/or the formation of fatty lumps, characterized in that a combination of at least one lotus extract and one pomegranate (Punica granatum) extract, or a composition comprising it, as are defined in one of Claims 1 to 16, is applied to the skin or its appendages.

18- Cosmetic treatment method according to claim 17, characterized in that the combination further comprises at least one caffeine-rich plant extract.