EP1581178A1

EP1581178A1 - Cosmetic composition based on cirsimarin

Info

Publication number: EP1581178A1
Application number: EP03815708A
Authority: EP
Inventors: Alain Geloen; Frédéric Résidence Cadenelle DEMARNE; Catherine Girotti
Original assignee: Gattefosse SA
Current assignee: Gattefosse SA
Priority date: 2003-01-09
Filing date: 2003-12-30
Publication date: 2005-10-05
Also published as: CA2512275A1; JP2006514082A; FR2849775A1; FR2849775B1; BR0317925A; AU2003303900A1; US20060115502A1; WO2004069217A1

Abstract

The invention relates to the use of the active molecule of formula (I), wherein R = H or R = glucose, for the production of a composition used to activate lipolysis.

Description

COSMETIC COMPOSITION BASED ON CIRSIMARINE

The invention relates to a cosmetic composition based on cirsimarine or cirsimarine derivatives, intended in particular for the cosmetic treatment of cellulite.

Adipose tissue is made up of cells called adipocytes, most of whose cell space is occupied by triglycerides. The amount of triglycerides in an organism depends on both the size and the number of fat cells. In other words, hypertrophy of fat cells (increase in size) and hyperplasia of these cells (increase in number) are important parameters indicating an increase in fat mass. In vivo, hypertrophy and hyperplasia of adipose cells are closely linked mechanisms. It has been shown that the increase in the number of adipocytes is first preceded by an increase in the size of the adipocytes up to a critical threshold, which triggers the recruitment of new adipose cells. For the treatment of cutaneous cellulite, the control of the only size parameter is sufficient. Indeed, the subcutaneous adipocytes are located in fatty chambers delimited by connective tissue. When the size of the adipocytes increases, the fatty chambers are deformed and tensions are exerted on the connective tissue delimiting them. These tensions make appear, on the surface of the skin, a wrinkling in appearance of orange peel characteristic of cellulite. From this observation, it appears that a solution to treat the phenomenon of cellulite is to reduce the size of the adipocytes. To do this, it is necessary to catabolize, that is to say to degrade, the triglycerides contained in the adipocytes, phenomenon called "lipolysis".

The regulation of lipolysis in adipocytes is a particularly complex phenomenon illustrated schematically in Figure 1.

The main lipolytic agents present in the organism are neurotransmitters of the catecholamine type, respectively adrenaline and noradrenaline, neuro-transmitters of the sympathetic nervous system. These catecholamines act on several types of receptors, essentially the β-adrenergic receptors and the α-adrenergic receptors present in the extracellular medium on the surface of the adipocyte membrane.

Adenylate cyclase plays an important role in the regulation of lipolysis. When activated, adenylate cyclase degrades adenosine triphosphate (ATP) to cyclic adenosine monophosphate (cAMP), which then transforms an inactive protein kinase A into active phosphorylated protein A, hydrolyzing triglycerides to di, then mono glycerides. In other words, the higher the intracellular concentration of cAMP, the more active the lipolysis. This therefore means that to stimulate lipolysis, it is necessary to increase the intracellular concentration of cAMP. However, this concentration is subject to several regulations.

First, cAMP is permanently degraded by a phosphodiesterase activated by insulin (anti-lipolytic agent) into 5'-AMP, which in turn is degraded to adenosine. At the same time, adenosine can leave the cell towards the extracellular medium where it binds to membrane type A adenosine receptors, which are themselves coupled to adenylate cyclase via the inhibitory protein G. In other words, adenosine has an inhibitory effect on the activity of adenylate cyclase, and therefore of lipolysis. So that means that lipolysis is permanently inhibited and that when you stimulate lipolysis with stimulants, you are actually stimulating a system that is permanently inhibited.

Also, the Applicant's idea is not to directly stimulate lipolysis, but to lift the permanent inhibition which is exerted on lipolysis by blocking the adenosine A1 receptors, present on the surface of adipocytes.

Caffeine and theophylline are molecules reported to be adenosine A1 receptor antagonists, caffeine being known for its lipolytic activity. However, the difficulty faced by the Applicant is that all the molecules having antagonist properties of the A1 receptors do not have a lipolytic activity such that one can envisage a cosmetic application. This is demonstrated by the documents NOGOWSKI "Genistein-Induced Changes in lipid metabolism of ovariectomized rats", Armais of Nutrition and Metabolism, 42 (1-2): 360-366 and JACOBSON "Interactions offlavones and other phytochemicals with adenosine receptors", Adv. Exp. Med. Biol., 505: 163-171, in which it is stated that genistein, a molecule belonging to the flavonoid family, identified as an Al receptor antagonist, had negligible lipolytic activity.

The problem which the invention proposes to solve is to search for a molecule antagonist of the A1 receptors which has an improved lipolytic effect compared to known molecules, such as in particular caffeine.

As part of his research, the Applicant discovered that the molecule with the following formula:

GOLD

with R = Glucose, or R = H was able, at very low doses, to lift the permanent inhibition acting on lipolysis.

When the radical R is a glucose, the molecule used in the invention corresponds to cirsimarine. Cirsimarine is a flavonoid belonging to the class of flavones. It is a glycosilated flavone whose crude formula is C ₂₃ H ₂₄ 0n corresponding to the CAS number 13020-19-04. It is also known under the names:

- 5-hydroxy-6,7-dimethoxyflavone 4'-O-glucoside, - 5-hydroxyapigenine 6,7-dimethyl ether 4 'glycoside,

- Scutellareine 6,7-dimethyl ether 4'-0-glucoside,

- Cirsitakaoside,

- Cirsimaritine 4'-0-glucoside.

Cirsimarine is found naturally in a very limited number of plants. They are found in particular in Teucrium arduini (family Lamiaceae), Clerodendrum mandarinorum (family Verbenaceae), Scoparia dulcis (family Scrophulariaceae), Cirsium maritimum (family Compositae) and Cirsium pendulum. A more widespread plant in which we find cirsimarine is a creeping herb called Microtea debilis belonging to the family of Phytolaccaceae, an annual herb native to South America. This herb is known for its use in traditional medicine in the form of dried plant powder, administered orally to treat proteinuria.

In this regard, the document "Adenosine-1 Active Ligands: Cirsimarin, a Flavone Glycoside from Microtea debilis", John A. Hasrat, Luc Pieters, Magda Claeys, Arnold Vlietinck, J. Nat. Prod. 1997, 60, 638-641 highlights the antagonistic properties of the cirsimarine Al receptors and explains that the effect on the proteinuria of this molecule is due to the interaction of cirsimarine with the Al receptors, present on the surface of kidney cells. Nothing in this document indicates the possible use of this molecule in a composition intended to activate lipolysis.

However, the Applicant has demonstrated this property, the probable mechanism of action of cirsimarine corresponding to the blocking of Al receptors, present on the surface of adipocytes. When the radical R is a hydrogen atom, the molecule used in the invention corresponds to cirsimaritine. This molecule is known by the following names:

- Scutellareine 4,7-dimethyl ether,

- 5,4'-dihydrixy 6, 7-dymethoxy flavone,

- Cirsistakaogenine,

- Scrophuleine,

- 5-hydroxyapigenin 6,7-dimethyl ether.

Cirsimaritine is reported in many cultivated plants such as Salvia tomentosa (family Lamiaceae), Salvia officinalis, Lippia citriodora (family Verbenaceae), but also in the wild in Sideretis sventenii (family Lamiaceae), Ocimum gratissimum, variety gratissimum (Lamiaceae family), this list is not exhaustive.

In other words and according to a first characteristic, the invention relates to the use of the molecule of the following formula:

with R = glucose or R = H, for the manufacture of a composition intended to activate lipolysis.

As already said, the activation of lipolysis would not be exerted by the direct stimulation of lipolysis, but by lifting the permanent inhibition exerted on lipolysis by blocking the Al receptors present on the surface of adipocytes. This use can be therapeutic or non-therapeutic.

In the context of the invention, the use is above all non-therapeutic by localized topical application of cirsimarine or cirsimaritine, for the treatment of cellulite.

The invention therefore relates to the use of the above-mentioned molecule in a cosmetic composition intended for the topical treatment of cellulite.

The Applicant has moreover found that the aforementioned molecule stimulates the synthesis of the components of the extracellular matrix. More precisely, it increases the synthesis of structural proteins (collagen, elastin), the synthesis of adhesion molecules (collagen, laminin, nidogen, integrin, cadherin, ...) as well as the synthesis of polysaccharides (glycosaminoglycans and proteogly canes) . The invention therefore also relates to the use of the aforementioned molecule for the manufacture of a composition intended to stimulate the synthesis of the components of the extracellular matrix.

Also part of the invention are cosmetic compositions comprising as active ingredient, cirsimarine or cirsimaritine corresponding to the formulas mentioned above.

To be effective, the concentration of cirsimarine or cirsimaritine in the cosmetic composition is between 0.0005 and 10% by weight, advantageously between 0.05 and 5% by weight.

In a particular embodiment, cirsimarine is in the form of a dry or liquid plant extract, preferably from Microtea debilis. When the extract is in dry form, it represents between 0.005 and 20%, advantageously between 0.1 and 10% by weight of the composition. When the extract is in liquid form, it represents between 0.1 and 20%, advantageously between 0.5 and 10% by weight of the composition. In practice, the extraction is carried out from the whole plant, dried and then ground, in a polar solvent which can be used in a topical cosmetic application, therefore in aqueous, alcoholic or glycolic media. Generally, the polar solvent is chosen from the group comprising water, ethanol, glycols such as propylene glycol, butylene glycol, alone or as a mixture, ethanol however remaining one of the preferred solvents.

The Applicant has moreover demonstrated the existence of a synergy between cirsimarine or cirsimaritine and xanthic bases, such as for example caffeine, on lipolysis.

In an advantageous embodiment, the cosmetic composition of the invention therefore also contains a xanthic base, in particular caffeine.

In practice, caffeine represents between 0.1 and 10% of the composition, by weight.

The composition will also be generally formulated in the form, for example, of gel, milk, cream, serum, microemulsion, etc.

The composition according to the invention can be in all the galenical forms normally used for topical application to the skin or the hair, in particular in the form of an aqueous solution, of an oil-in-water or water-in- emulsion. oil or multiple, of a silicone emulsion, of a microemulsion or nanoemulsion, of an aqueous gel.

This composition can be more or less fluid and have the appearance of, among other things, a white or colored cream, an ointment, a milk, a lotion, a serum, a gel. The composition of the invention may contain the adjuvants usual in the cosmetic and dermatological fields, such as fats, emulsifiers and co-emulsifiers, hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active agents, preservatives, antioxidants, solvents, fragrances, fillers, hydrophilic and lipophilic filters, dyes, neutralizers, penetrating agents, and polymers.

The amounts of these various adjuvants are those conventionally used in the fields considered, and for example from 0.01 to 30% of the total weight of the composition. These adjuvants, depending on their nature, can be introduced into the fatty phase or into the aqueous phase.

As fats which can be used in the invention, it is possible to use mineral oils, oils of animal origin (lanolin), synthetic oils (isopropyl myristate, octyldodecyl, isostearyl isostearate, decyl oleate, isopropyl palmitate), silicone oils ( cyclomethicone, dimethicone) and fluorinated oils. Fatty alcohols, fatty acids, waxes and gums and in particular gums and silicone elastomers can be used as fats.

As emulsifiers and coemulsifiers which can be used in the invention, mention may, for example, be made of polyglycerol and fatty acid esters, sucrose and fatty acid esters, sorbitan and fatty acid esters, acid esters fatty and sorbitan oxyethylenated, ethers of fatty alcohol and PEG, esters of glycerol and fatty acid, all yl sulfates, alkyl ether sulfates, alkyl phosphates, alkyl polyglucosides, dimethicone copolyols.

As hydrophilic gelling agents, there may be mentioned in particular include carboxyvinyl polymers (carbomer), acrylic copolymers such as acrylate / alkylacrylate, the polyacrylamid ^'es, polysaccharides such as xanthan gum, guar gum, natural gums such than cellulose gum and derivatives, clays and copolymers of 2-acrylamido-2-methyl lpropane acid. As lipophilic gelling agents, mention may be made of modified clays such as bentones, metal salts of fatty acids, hydrophobic silica and ethylcellulose.

The cosmetic composition can also contain active ingredients. As active agents, depigmentants, emollients, moisturizers, antiseborrheics, anti-acne, keratolytic and / or desquamating agents, anti-wrinkle and tightening agents, draining agents, anti-irritants, can be used in particular. soothing agents, slimming agents such as xanthic bases (caffeine), vitamins and their mixtures, and matting agents.

In the event of incompatibility between them or with the extract of Microtea debilis, the active agents indicated above and / or the extract of Microtea debilis can be incorporated into spherules, in particular ionic or nonionic vesicles and / or nanoparticles (nanocapsules and / or nanospheres), so as to isolate them from each other in the composition.

As preservatives which can be used according to the invention, mention may be made of benzoic acid, its salts and its esters; sorbic acid and its salts; parabens, their salts and esters; triclosan; imidazolidinyl urea; phenoxyethanol; DMDM hydantoin; diazolidinyl urea; chlorphenesin.

As antioxidants which can be used according to the invention, mention may be made of chelating agents such as EDTA and its salts.

As solvents which can be used according to the invention, mention may be made of water, ethanol, glycerin, propylene glycol, butylene glycol, sorbitol.

As fillers which can be used according to the invention, mention may be made of talc, kaolin, mica, serecite, magnesium carbonate, aluminum silicate, magnesium silicate, organic powders such as nylon. As filters which can be used according to the invention, mention may be made of the UVA and UVB filters conventionally used such as benzophenone-3, butyl methoxydibenzoyl methane, octocrylene, octyl methoxycinnamate, 4-methylbenzylidene camphor, octyl salycylate, tacephthalylidene dicamphor sulfanic acid, and drometrizole trisiloxane. Mention will also be made of physical filters TiO ₂ and ZnO in their micrometric and nanometric forms.

As coloring materials which can be used according to the invention, mention may be made of lipophilic dyes, hydrophilic dyes, pigments and nacres usually used in cosmetic or dermatological compositions, and their mixtures.

As neutralizers which can be used according to the invention, mention may be made of sodium hydroxide, triethanolamine, aminomethyl propanol and potassium hydroxide.

As penetrating agents which can be used according to the invention, there may be mentioned alcohols and glycols (ethanol, propylene glycol), ethoxydiglycol, alcohols and fatty acids (oleic acid), fatty acid esters, dimethyl isosorbide.

The composition according to the invention can be used as a care product (for example a slimming product), as a cleansing product, and / or as a skin makeup product, as a sun protection product, or as a hair product, for example as a shampoo or after shampoo.

The invention also relates to a cosmetic treatment method for cellulite, comprising locally applying an effective amount of the cosmetic composition topically.

The invention and the advantages which result therefrom will emerge clearly from the following exemplary embodiment in support of the appended figures. Figure 1 is a diagram illustrating the regulation of lipolysis in adipocytes.

FIG. 2 represents the lipolytic activity of cirsimarine compared to control molecules (caffeine, theophylline, noradrenaline). FIG. 3 represents the lipolytic activity of the cirsimarine / caffeine association with respect to caffeine alone.

Example 1: making an extract of Microtea debilis

The whole dried plant comes from South America. This plant is ground until a powder is obtained.

The extraction of the crushed plant is carried out in an ethanol mixture 96.2 ° H ₂ 0 (80/20); volume / volume at room temperature with magnetic stirring, protected from light, for 6 hours.

The extract is then filtered on a nylon filter and then on a cellulose membrane (up to 0.22 microns).

EXAMPLE 2 Lipolytic Activity of Cirsimarine

This in vitro test on isolated adipocytes shows the lipolytic activity of cirsimarine extracted from Microtea debilis.

2.1 / Materials and methods Most of the reagents used come from SIGMA-ALDRICH a. Positive references and products to be evaluated

"Noradrenaline (syn. Norepinephrine NE): this molecule, of molar mass 319.3 g, is evaluated at the final concentration of 1 μM in KREBS-RINGER buffer at 4% albumin." Caffeine: this molecule, of mass 194.2 g molar, is evaluated at the final concentration of 0.5 mM in KREBS-RINGER buffer at 4% albumin. "Theophylline: this molecule with a molar mass of 180.2 is evaluated at the final concentration of 0.5 mM in KREBS-RINGER buffer at 4% albumin.

"Cirsimarine: this molecule of molar mass 476.44 is tested at 0.5 mM and 0.1 lmM. This molecule is first of all dissolved in a mixture of solvents: 0.2 M NaOH / DMSO (95/5; v / v).

b. Reaction medium

The reactions take place in a buffer solution of KREBS-RINGER bicarbonate containing 4% of defatted albumin (mas se / volume), pH ^: = 7.4.

The defatted albumin will fix the fatty acids released during lipolysis; this precaution is important because they retio-inhibit lipolysis. The buffer solution is brought to 37 ° C.

vs. Preparation of adipocytes and implementation of the reaction

The epididymal adipose tissue is removed from rats. It is placed in KREBS-RINGER bicarbonate buffer at 4% albumin and at pH 7.4, to which 188 units of collagenase / ml will have been added to digest the collagen network which supports the adipose tissue.

The digestion lasts about one hour at 37 ° C with stirring. It is stopped by strongly diluting the collagenase by successive rinses of the adipocyte suspension with KREBS-RINGER buffer at 4% albumin.

On the MALLASSEZ slide, 8 counts of the adipocyte suspension obtained are then carried out. The average of these 8 counts is used to distribute the adipocyte solution in the reaction tubes at the concentration of approximately 150,000 cells per ml. The reaction tubes used are 2 mL EPPENDORF ^® tubes. If A is the volume in μL of adipocyte solution necessary to have the concentration of 150,000 cells / ml and B the volume in μL of mother solution of the molecule to be tested, we first put in the EPPENDORF® tube (1 000-AB) μL of KREBS-RINGER buffer at 4% albumin, then the B μL of stock solution of the molecule to be tested. The volume of the tube is then made up to 1 ml by delicately depositing on the surface the volume necessary for the solution of adipocytes A.

All tests are repeated 3 times. For the reaction to take place, the tubes are placed in a water bath at 37 ° C., with gentle lateral stirring. The reaction is stopped after one hour, by immersing the tubes in ice.

To find out the basal lipolysis of adipocytes in the KREBS-RINGER buffer as well as the basal lipolysis of adipocytes in the KREBS-RINGER buffer containing the solvent for dissolving cirsimarine, control tubes are produced. Eight 2mL EPPENDORF tubes are prepared with A μL of adipocyte solution and (1000-A) μL of KREBS-RINGER buffer; four tubes are prepared with A μL of adipocyte solution, B μL of solvent for dissolving cirsimarine and (1,000-A-B) μL of KREBS-RINGER buffer.

Four of the eight tubes containing the KREBS-RINGER buffer and the adipocyte solution are placed directly on ice (tubes 00). The remaining four tubes (0 Ref tubes) as well as the four tubes containing the KREBS-RINGER buffer, the adipocyte solution and the cirsimarine dilution solvent (0 solvent tubes) are placed in a water bath and treated as reaction tubes.

Lipolytic activity is measured by the amount of free fatty acids; a NEFA C colorimetric assay kit (Non Esterified Fatty Acid / Colorimetric) sold by WAKO Chemical GmbH is used for this. 2.2 / Results

The amount of free fatty acids present in tubes 0 minus that present in tubes 00 corresponds to the basal lipolysis of the adipocytes in the buffer. The quantity of free fatty acids present in the tubes 0 solvent minus that present in the tubes 00 corresponds to the basal lipolysis of the adipocytes in the buffer containing the solvent for dissolving cirsimarine. The quantity of fatty acids released in the reaction tubes minus that present in the 00 tubes corresponds to the lipolysis induced by the molecules studied. The results appear in Figure 2.

10

2.3 / Conclusions

As shown ^'in this figure, cirsimarin has proved lipolytic activity in vitro, under the conditions of the test.

] 5 The diluting solvent for Microtea debilis extract has no effect on basal lipolysis.

Example 3 Cosmetic Composition

0 Examples of slimming formulas Body slimming gel

Slimming Body Milk

O / W emulsion

W / O emulsion

Micro em ulsion

W / O / W multiple emulsion

Solar cream

Shampoo

Example 4 Synergistic Action of Cirsimarine and Caffeine

In this example, the lipolytic activity of the cirsimarine / caffeine association is compared with caffeine alone on human adipocytes isolated under the same conditions as in Example 2. The results are shown in FIG. 3.

As this figure shows, the release of fatty acids is much higher with a given amount (0.11 mM) of the cirsimarine / caffeine association compared to the same amount of caffeine (0.11 mM).

Claims

BFVFNDICATIONS

/ Use of the active molecule of the following formula

with R = H or. glucose, for the manufacture of a composition intended to activate lipolysis.

/ Use according to claim 1, characterized in that when the radical R is a glucose molecule, the molecule corresponds to cirsimarine.

/ Use according to claim 1, characterized in that when the radical R is a hydrogen atom, the molecule corresponds to cirsimaritine.

/ Use according to one of claims 1 to 3 in a cosmetic composition intended for the topical treatment of cellulite.

/ Use according to claim 1 for the manufacture of a composition intended to stimulate the synthesis of the components of the extracellular matrix. 6 / Cosmetic composition characterized in that it contains, as active ingredient, the molecule of the following formula:

GOLD

with R = Hou R = glucose.

Composition according to Claim 6, characterized in that when the radical R is a glucose, it is cirsimarine.

8 / Composition according to claim 6, characterized in that when the radical R is a hydrogen, it is cirsimaritine.

9 / cosmetic composition according to claim 6, characterized in that the concentration of active ingredients is between 0.0005 and 10% by weight.

10 / cosmetic composition according to claim 6, characterized in that the active agent is in the form of a plant extract.

1 1 / Composition according to claim 10, characterized in that the plant extract is an extract of Microtea debilis.

12 / cosmetic composition according to claim 10, characterized in that when the extract is in dry form, it represents between 0.005 and 20% by weight of the composition. / Cosmetic composition according to claim 10, characterized in that when the extract is in liquid form, it represents between 0.1 and 20% by weight of the composition.

/ Cosmetic composition according to claim 10, characterized in that the extract is an extract of whole plant.

/ Cosmetic composition according to claim 6, characterized in that it also contains a xanthic base.

/ Cosmetic composition according to claim 15, characterized in that the xantic base is caffeine.

/ Composition according to Claim 16, characterized in that the caffeine represents between 0.1 and 10% by weight of the composition

/ Cosmetic composition according to claim 6, characterized in that it also contains a cosmetically acceptable vehicle.

/ A method of treating cellulite consisting in locally applying an effective amount of the cosmetic composition which is the subject of claims 6 to 18, topically.