EP1280542A1 - Use of a cell extract of at least a plant of the family pontederiaceae as anti-pollution agent - Google Patents

Abstract

The invention concerns the use of a cell extract of at least a plant of the family Pontederiaceae as anti-pollution or depollution agent, compositions containing at least one of said extracts, and a cosmetic treatment method using said cell extracts or said compositions.

Description

 Use of an extract of cells from at least one plant of the family Pontederiaceae as an anti-pollution agent

The subject of the present invention is the use of extracts of cells from at least one plant of the family Pontederiaceae as anti-pollution agent and depollution agent as well as compositions containing at least one of these extracts, The invention also has relates to a cosmetic treatment process using cell extracts or compositions. Metal ions are necessary for the body in the form of traces as essential nutrients. For example, several functions involving polypeptides such as enzymatic, structural and immunological functions require metal cofactors.

However, other metal ions, in particular heavy metal ions when in non-physiological concentrations, can alter these functions. Thus, overexposure to metals from the environment can lead to toxic effects.

Ecological studies conducted in industrialized countries show that the quantities of metals present in the atmosphere are increasing. This leads to an increase in the levels of heavy metals in the tissues of organisms following the ingestion of contaminated foods and exposure to metals from the atmosphere.

The effects of the accumulation of heavy metals can be extremely dangerous and their toxicity is due in part to the alteration of the tertiary and quaternary structures of proteins, which leads to a reduction in their catalytic activity. The altered proteins can become antigenic and trigger an immune response. They are then recognized by the body as foreign polypeptide agents and can cause autoimmune responses. Another mechanism responsible for the toxic effects of metals is the competitive substitution of natural physiological co-factors by heavy metals at non-physiological concentrations. So the Control of heavy metals that pollute the atmosphere is essential to prevent diseases related to exposure to metals.

Due to the increasing contamination of the environment by heavy metals and their ubiquitous presence in the ecosystem, the skin, the hair and the accessible mucous membranes represent the largest contact surface and therefore favor the accumulation of metals and their subsequent absorption in the body.

Certain metals and metallic compounds present in industrial manufacturing, chemicals, jewelry, clothing, medicinal products, dyes and cleaning products are involved in primary irritation reactions, allergy reactions and carcinogenicity in the skin tissue.

The metals particularly incriminated in the environment are copper, cobalt, zinc, manganese, mercury and nickel, lead and cadmium.

Skin rash caused by metal dermatitis is a problem encountered in people exposed to high amounts of certain metal ions. Exposure to nickel in the environment is largely due to the frequent use of this metal in articles of jewelry, watch straps and buttons on clothing. Raising awareness of nickel with the development of dermatitis is an industrial hazard in certain professions. Furthermore, the deposit of minerals on the hair is an inevitable phenomenon.

Hair is a strong absorbent for metals. The fixation is so strong that once these fixed metals are captured by the anionic sites of the fiber, they are difficult to elute. The degree of fixation of metals on the hair generally depends on several factors such as the size of the fiber, its porosity, and the exposure time. Metals such as copper, lead and iron can interfere with chemical treatments such as coloring and perming the hair.

Some cosmetic products contain metals such as magnesium, copper or iron. The absorption of these metals by keratin fibers can interfere with chemical treatments such as coloring, bleaching or perm effects. This interference can lead to color problems or precipitation as explained in American patent US-5, 635, 167.

Certain heavy metals have been shown to penetrate the skin and accumulate (A.B .G. Landsdown. Critical Reviews in Toxicology. 1995, 25: 397-462). At high concentrations they can induce:

- oxidation mechanisms on membrane lipids,

- direct cytotoxicity, capable of leading to cell necrosis,

- an alkylation of the cellular nucleqphiles which can be at the origin of phenomena of sensitization or carcinogenesis.

(SJ Stochs and D. Bagchi. Free Radical Biology and Medicine. 1995, 18: 321-336.; ME Figueiredo Pereira et al., The Journal of Biological Chemistry. 1998, 21: 12703-12709,; NL Acan et al. , 1995, Biochemical and Molecular Medicine, 54: 33-37.)

Cosmetic and therapeutic treatments have already been envisaged by protecting the tissues by:

- compounds with sulfur groups which behave as heavy metal sequestrants such as metallothionein in patent EP 0 557 042 A1, and

- amino acid compounds with sulfur groups in patent application EP 0 914 815 Al.

Furthermore, patent application GB 2333705 A mentions the use of ethylenediamine disuccinic acid in compositions for the treatment of skin irritations by heavy metals.

The problem is therefore to protect the skin against metallic particles floating in the air and their deleterious effects encountered in urban pollution by selecting active principles which are effective in combating these effects and / or preventing the penetration of metals into the skin or keratin materials and by introducing these active ingredients into cosmetic formulations. It has now been found, quite surprisingly, that the use of extract of cells from at least one plant of the family Pontederiaceae makes it possible to protect the keratin materials, the skin and the integuments against the effects of a certain category of air pollutants such as heavy metals.

The subject of the present invention is the use of an extract of cells from at least one plant of the Pontederiaceae family as an anti-pollution or depollution cosmetic agent.

Cosmetic anti-pollution agent is understood to mean an agent which protects the skin and keratinous materials so as to prevent, attenuate and / or eliminate the deleterious effects of metals.

The present invention also relates to cosmetic compositions containing such cellular extracts as well as a method of cosmetic anti-pollution treatment or depollution using these compositions or the cellular extracts of Pontederiacae.

The Applicant has discovered that this model of plants accumulates heavy metals with a broad spectrum, thus covering at best all of the heavy metals deemed dangerous for the skin.

The subject of the invention is the use of an extract of cells from at least one plant of the Pontederiaceae family as an anti-pollution or depollution cosmetic agent.

The Pontederiaceae family has several genera including

VEichhornia crassipes also called water hyacinth, this genus contains about seven species including E. azurea and E. crassipes. They are floating grasses with a creeping rhizome, aquatic or amphibious with sheathing floating or submerged leaves.

The present invention relates more particularly to the use of cell extracts from Eichhornia crassipes. The cellular extracts are preferably extracts from root or leaf cells.

Any extraction method known to a person skilled in the art can be used according to the invention. We can in particular quote the extracts alcoholics, especially ethanolic and / or methanolic, and hydro-alcoholic extracts.

One can also use an extract prepared by the method described in French patent application No. 95-02379. Thus, in a first step the plant material is ground in a cold aqueous solution, in a second step the particles in suspension are removed from the aqueous solution obtained from the first step, and in a third step the aqueous solution obtained from the second step. This aqueous solution corresponds to the extract. On the other hand, the first step can advantageously be replaced by a simple freezing operation of the plant tissues (for example at -20 ° C.), followed by an aqueous extraction repeating the second and third steps described above.

Preferably, according to the invention, an ethanolic extract is used.

Whatever the method of preparation of the extract, it can then be lyophilized for possible conservation. An example of preparation of extract usable according to the invention is given elsewhere in the examples. The subject of the present invention is the use of at least one cell extract as an anti-pollution or depollution cosmetic agent in cosmetic or dermatological compositions.

These cell extracts can be used as antipollution or depollution agents, preferably as anti-pollution or depollution cosmetic agents, to protect keratin materials, dander and skin, from the harmful effects of heavy metals.

The integuments represent the nails, the eyelashes, the mucous membranes, the eyebrows and the hairs in general.

These cell extracts can be used as anti-pollution agents, preferably as anti-pollution or depollution cosmetic agents, to improve cellular respiration and / or reduce flaking and / or to avoid tarnishing or dirtying the keratin materials, the skin. or the integuments. Another object of the present invention is the preparation of a cosmetic composition with at least one cell extract defined beforehand.

Said composition can contain from 0.01 to 10% and preferably 0.1 to 5% by weight of cellular extract relative to the total weight of the composition.

This composition can also contain at least one other anti-pollution compound.

This can in particular be chosen from anthocyanins and / or its derivatives, compounds containing a thioether, sulfoxide or sulfone function, ergothionein and / or its derivatives, metal chelators such as, for example, derivatives of the N, N'-dibenzyl ethylene diamine acid

N, N'-diacetic, antioxidants.

Among the antioxidants, polyphenols and, among others, ellagic acid will be chosen.

The cosmetic compositions used according to the invention also comprise a cosmetically acceptable medium which more particularly consists of water and / or optionally a cosmetically acceptable organic solvent. They can be chosen from the group consisting of hydrophilic organic solvents, amphiphilic solvents, lipophilic organic solvents or their mixtures.

Among the hydrophilic organic solvents, mention may be made, for example, of lower, linear or branched mono-alcohols having from 1 to 8 carbon atoms such as ethanol, propanol, butanol, isopropanol, isobutanol; polyethylene glycols having from 6 to 80 ethylene oxides, polyols such as propylene glycol, isoprene glycol, butylene glycol, glycerol, sorbitol, mono- or dialkyls of isosorbide whose alkyl groups have from 1 to 5 carbon atoms like dimethyl isosorbide, glycol ethers like diethylene glycol mono-methyl or mono-ethyl ether and propylene glycol ethers like dipropylene glycol methyl ether.

As amphiphilic organic solvents, there may be mentioned polyols such as propylene glycol derivatives (PPG), such as esters polypropylene glycol and fatty acids, PPG and fatty alcohols such as PPG-23 oleyl ether and PPG-36 oleate.

As lipophilic organic solvents, there may be mentioned, for example, fatty esters such as diisopropyl adipate, dioctyl adipate, alkyl benzoates.

The organic solvents are preferably chosen from mono- or polyfunctional alcohols, optionally oxyethylenated polyethylene glycols, polypropylene glycol esters, sorbitol and its derivatives, isosorbide dialkyls, glycol ethers and polypropylene glycol ethers, fatty esters.

The organic solvents can represent from 5 to 98% of the total weight of the composition.

So that the compositions of the invention are more pleasant to use, softer on application, more nourishing and more emollient, it is possible to add a fatty phase to the medium of these compositions.

The fatty phase preferably represents from 0 to 50% by total weight of the composition.

This fatty phase can comprise one or more oils preferably chosen from the group consisting of:

- volatile or non-volatile, linear, branched or cyclic silicones, organo-modified or not, water-soluble or liposoluble,

- mineral oils such as paraffin and petrolatum oil, - oils of animal origin such as perhydrosqualene,

- oils of vegetable origin such as sweet almond oil, avocado oil, castor oil, olive oil, joj oba oil, sesame oil, peanut oil, macadamia oil, grape seed oil, rapeseed oil, coconut oil, - synthetic oils such as Purcellin oil, isoparaffins,

- fluorinated and perfluorinated oils,

- fatty acid esters such as Purcellin oil. It can also contain as fat (one) or more fatty alcohols, fatty acids or waxes (paraffin, polyethylene wax, Carnauba, beeswax).

In a known manner, the compositions used in the invention can also contain adjuvants customary in the cosmetic field such as gelling agents and / or thickening agents, aqueous or lipophilic, hydrophilic or lipophilic active agents, preservatives, antioxidants, perfumes, emulsifiers, hydrating agents, pigmenting agents, depigmenting agents, keratolytic agents, vitamins, emollients, sequestrants, surfactants, polymers, alkalizing or acidifying agents, fillers, anti-free radical agents , ceramides, sunscreens, especially ultraviolet, insect repellents, slimming agents, coloring matters, bactericides, anti-dandruff. The quantities of these various adjuvants are those conventionally used in the fields considered.

Of course, those skilled in the art will take care to choose the optional compound (s) to be added to the composition according to the invention, so that the advantageous properties intrinsically attached to the composition according to the invention are not, or substantially not, altered by the proposed addition.

The compositions used according to the invention can be presented in all the galenical forms normally used for a topical application, in particular in the form of an aqueous, hydroalcoholic or oily solution, of an oil-in-water or water-in-oil emulsion or multiple, of an aqueous or oily gel, of a liquid, pasty or solid anhydrous product or of an oil dispersion in an aqueous phase using spherules, these spherules possibly being polymeric nanoparticles such as nanospheres and nanocapsules, or better, lipid vesicles of ionic and / or nonionic type.

The compositions used in the present invention may be more or less fluid and have the appearance of a white cream or colored, an ointment, a milk, a lotion, a serum, a paste, a foam or a solid.

They can optionally be applied to the skin in the form of an aerosol. They can also be in solid form, and for example in the form of a stick.

They can be used as a care product and / or as a makeup product.

The compositions of the invention can have a pH of between 3 and 8, and preferably between 5 and 7.

The present invention also relates to the use of a cellular extract in or for the preparation of an antipolluting or depolluting composition.

The subject of the invention is the use of such a cosmetic composition for the protection of keratin materials, the integuments and the skin, of the harmful effects of heavy metals but also for improving cellular respiration and / or reducing flaking and / or avoiding tarnish or soil keratin materials, dander and skin. Another object of the invention consists in a cosmetic treatment process intended to obtain protection of the organism against the effects of pollution, consisting in applying to the keratinous matter, the integuments or the skin, a cosmetically effective amount of at least one cell extract as defined above. Another cosmetic treatment method according to the invention, intended to obtain protection of the organism against the effects of pollution, consists in applying to the keratin material, the integuments or the skin, a cosmetic composition according to the invention such as defined above. These cosmetic treatment methods can be followed by rinsing after application to the keratinous material, the integuments or the skin of the cell extracts or cosmetic compositions according to the invention as defined above. The examples which follow are intended to illustrate the invention, without however being limiting in nature.

Preparation of plant extracts.

Protocol 1: 185 g of fresh roots are introduced into 800 ml of distilled water or of MilliQ quality. Coarse grinding is carried out in a food processor for 5 minutes. A fine grinding completes this: Turax for 10 minutes. The separable part is separated by centrifugation 8000 G / 20 minutes or front cascade filtration. The final filtration is carried out on a Whatman GFD filter. 621 g of filtrate are thus recovered then lyophilized and give 1.26 g of lyophilisate.

Protocol 2: Leaves + Roots: 52.8 g, scissor cutting, grinding with liquid nitrogen in a mortar, taken up in 200 ml of H ₂ O MilliQ, magnetic stirring at 900 rpm for 10 minutes, 180 ml collected then filtered on Whatman GFF gave 417 mg of lyophilisate.

Protocol 3: 12 feet of hyacinth were washed with water and then coarsely wrung. Passed through a knife grinder (cutting robot), 700 g of ground material were obtained. Addition of 700 ml of H ₂ O, then 300 ml of H ₂ O MilliQ. New passage in 5-minute cutting robot, centrifugation 2,078,000 G filtration

Whatmann GFD then GFF and lyophilization: 5.43 g of lyophilisate are thus obtained.

Demonstration of the protective effect on keratinocytes cultured in vitro of a hyacinth extract vis-à-vis a representative heavy metal: cadmium.

Principle:

Heavy metals such as cadmium, nickel, lead, Mercury, etc. , can cause cytotoxicity on the cells of different organs including the skin. The cytotoxic effect of cadmium on human keratinocytes in culture was evaluated by a technique using a fluorescent probe to measure the level of intracellular glutathione (JC Fernandez Checa et al. 1990, R. Kannan et al. 1993).

Seeding cells:

The study was carried out on a monolayer culture of human keratinocytes from surgical plasties. The cells are seeded on D-3 in 96-well culture dishes at the rate of 25,000 cells / cm ² in 100 μl of culture medium (medium defined without calf serum, NR2, Biofluids). The incubations are carried out in an oven at 37 ° C in a humid atmosphere and enriched with 5% CO ₂ .

Treatment with the pollutant in the presence of the protector: The cells are treated for 24 hours with increasing concentrations (0, 10, 25, 50, 75, 100, 150 and 200 μM) of cadmium chloride (CdCl ₂ ) alone, so as to determine its cytotoxicity. At the same time, a treatment is carried out under the same conditions, but in the presence of hyacinth extract (0.125%). Incubations are carried out in an oven at 37 ° C,% CO ₂ in a humid atmosphere.

Cell viability measurement:

At the end of the contact period, mark the intacellular gluthation using a specific fluorescent probe: monochlorobimane (MCB).

Briefly, after treatment, rinse the cells with buffer (PBS) in order to remove the treatment solutions,

Add 100 μl per well of a solution of MCB to ImM for one hour at room temperature and protected from light. Remove the MCB, and add 200 μl / well of PBS buffer. Read with a spectrofluorimeter (excitation: 380 nm; emission: 485 nm). The fluorescence obtained is proportional to the amount of gluthation conjugated to MCB.

Calculate the concentration of CdCl ₂ resulting in a 50% drop in the gluthation rate (= LC50).

Results

Number of independent experiences: 3

Number of repetitions per experiment: 4

The results given in concentration causing a 50% drop in cell viability are shown in the following table:

Cytotoxicity of cadmium chloride towards human keratinocytes in culture, in the presence or absence of hyacinth extract.

Conclusion

Cadmium chloride alone has significant toxicity with an IC.50 of 43.7 μM. In the presence of hyacinth extract (0.125%), the cytotoxicity of cadmium chloride decreases significantly (which corresponds to an increase in the IC.50): 61 μM.

From an in vitro biological model using human keratinocytes, this experiment shows that an agent representative of a category of air pollutants (heavy metals) such as cadmium, under these experimental conditions leads to high toxicity and that the hyacinth extract significantly protects the cells against the toxicity of this pollutant.

EXAMPLES OF FORMULATIONS

Example 1: According to the usual preparation techniques, the constituents below are mixed to prepare an emulsion.

COMPOSITION FOR TOPICAL APPLICATION

cell extract dLEichhornia crassipes 3 g oxyethylenated polyethylene glycol 3 g per 50 moles of ethylene monodiglyceryl stearate 3 g petroleum jelly oil 24 g cetyl alcohol 5 g water qs 100 g

Example 2: In the same way, an emulsion is prepared according to a conventional technique, from the following compounds:

cell extract AEichhornia crassipes 4 g sorbitol 10 g glyceryl isostearate 4 g paraffin oil 24 g vitamin E 1 g glycerol 3 g water qs 100 g Example 3: From the constituents below, the following composition is formulated:

cell extract 1 g of Eichhornia crassipes

jojoba oil 13 g parabenzoxy benzoate 0.05g methyl and isopropyl potassium sorbate 0.3 g cyclopentadimethylsiloxane 10 g ellagic acid 0.01g stearic acid g polyethyleneglycol stearate g vitamin E 1 g glycerol 3 g water qs 100 g

Example 4: TABLETS

cellular extract of Eichhornia crassipes 0.1 g starch 0.14 g bicalcium phosphate 0.02 g lactose 0.06 g magnesium stearate 0.005 g

After compaction, a 0.2g tablet is obtained

Example 5: ORAL SUSPENSION

cellular extract of Eichhornia crassipes 0.1 g glycerol 0.5 g 70% sorbitol 0.5 g

L (+) ergothionein where R = H, R '= Ethyl, R "= H 0.01g and X = NH2

methyl parahydroxybenzoate 0,04g aroma qs purified water qs 5 g

Example 6: INJECTABLE BULB

cellular extract 0, 1 g of Eichhornia crassipes

water for injection qsp 3 g

Claims

1. Use of at least one cellular extract of at least one plant from the Pontederiaceae family as an anti-pollution or depollution cosmetic agent. 2. Use according to claim 1, characterized in that the cell extract comes from Eichhornia crassipes cells 3. Use according to claim 1 or 2, characterized in that the cells used come from the root or from the leaf. 4. Use of at least one cell extract according to claims 1 to 3, as an anti-pollution or depollution cosmetic agent in cosmetic or dermatological compositions. 5. Use of at least one cell extract according to claims 1 to 3, as an anti-pollution or depollution cosmetic agent making it possible to protect keratin materials, dander and the skin, from the harmful effects of heavy metals. 6. Use of at least one cellular extract according to claims 1 to 3, as an anti-pollution or depollution cosmetic agent making it possible to improve cellular respiration and / or to reduce flaking and / or to avoid tarnishing or dirty keratinous matter, dander and skin. 7. Cosmetic composition, characterized in that it contains, in a cosmetically acceptable medium, at least one cellular extract of at least one plant from the Pontederiaceae family. 8. Composition according to claim 7, characterized in that the cells come from Eichhornia crassipes. 9. Composition according to claim 7 or 8, characterized in that the extract is an extract of root or leaf cells. 10. Composition according to any one of claims 7 to 9, characterized in that said composition contains from 0.01 to 10% and preferably 0.1 to 5% by weight of cellular extract relative to the total weight of the composition. 11. Composition according to any one of claims 7 to 10, characterized in that said composition also contains at least one other anti-pollution compound. 12. Composition according to Claim 11, characterized in that the said anti-pollution compound is chosen from anthocyanins and / or its derivatives, the compounds containing a thio-ether, sulfoxide or sulfone function, ergothionein and / or its derivatives, metal chelators such as, for example, N, N'-dibenzyl ethylene diamine N, N'-diacetic acid derivatives, antioxidants. 13. Composition according to claim 12, characterized in that the antioxidants chosen are preferably polyphenols. 14. Composition according to claim 13, characterized in that the polyphenol chosen is preferably ellagic acid. 15. Composition according to any one of claims 7 to 14, characterized in that the cosmetically acceptable medium consists of water and / or at least one organic solvent. 16. Composition according to claim 15, characterized in that the organic solvent is chosen from the group consisting of hydrophilic organic solvents, lipophilic organic solvents, amphiphilic solvents or their mixtures. 17. Composition according to any one of claims 15 to 16, characterized in that the organic solvents are preferably chosen from mono- or polyfunctional alcohols, optionally oxyethylenated polyethylene glycols, polypropylene glycol esters, sorbitol and its derivatives, dialkyls isosorbide, glycol ethers and polypropylene glycol ethers, fatty esters. 18. Composition according to claim 15 or 17, characterized in that the organic solvent (s) represent from 5 to 98% of the total weight of the composition. 19. Composition according to any one of claims 7 to 18, characterized in that it also comprises at least one fatty phase. 20. Composition according to Claim 19, characterized in that the fatty phase represents from 0 to 50% of the total weight of the composition. 21. Composition according to any one of claims 7 to 20, characterized in that it additionally contains at least one additive chosen from, the group consisting of conventional aqueous or lipophilic gelling agents and / or thickeners, hydrophilic active agents or lipophilic, preservatives, antioxidants, perfumes, emulsifiers, hydrating agents, pigmenting agents, depigmentants, keratolytic agents, vitamins, emollients, sequestrants, surfactants, polymers, alkalizing agents or acidifiers, fillers, free anti-radical agents, ceramides, sun filters, in particular ultraviolet, insect repellents, slimming agents, coloring matters, bactericides, anti-dandruff. 22. Composition according to any one of claims 7 to 21, characterized in that it is in the form of an aqueous, hydroalcoholic or oily solution, of an oil-in-water or water-in-oil emulsion or multiple, an aqueous or oily gel, a liquid, pasty or solid anhydrous product or an oil dispersion in an aqueous phase using spherules. 23. Composition according to any one of claims 7 to 22, characterized in that the composition has the appearance of a white or colored cream, an ointment, a milk, a lotion, a serum, a paste, a foam or of a solid. 24. Composition according to any one of claims 7 to 23, characterized in that the composition has a pH of between 3 and 8, preferably between 5 and 7. 25. Use of a cell extract as defined in any one of claims 1 to 3, in or for the preparation of an anti-polluting or depolluting composition. 26. Use of a cosmetic composition as defined in any one of claims 7 to 24, for the protection of keratin materials, the integuments and the skin, from the harmful effects of heavy metals. 27. Use of a cosmetic composition as defined in any one of claims 7 to 24, for improving cellular respiration and / or reducing flaking and / or avoiding tarnishing or dirtying keratin materials, dander and the skin. 28. Cosmetic treatment method intended to obtain protection of the organism against the effects of pollution, characterized in that it consists in applying to the keratin materials, the integuments or the skin, a cosmetically effective amount of at least a cell extract as defined in any one of claims 1 to 3. 29. Cosmetic treatment process intended to obtain protection of the body against the effects of pollution, characterized in that it consists in applying to keratin materials, dander or the skin, a composition as defined in any one of claims 7 to 24.