LECTIN-BASED DEODORANT AND/OR ANTIPERSPIRANT COSMETIC

COMPOSITIONS

Field of the invention

The present invention relates to lectin-based deodorant and/or antiperspirant cosmetic compositions and the cosmetic use thereof in humans.

State of the art

Lectins are proteins or glycoproteins binding specifically with glycans.

Lectins are frequently used in research for detecting the presence of glycans in biological samples. For example, B. Mehul et al. (Experimental Dermatology, 2003, 12 537-545) used lectins to characterize glycans expressed in a reconstructed epidermis compared to a normal human epidermis.

Perspiration, also referred to as sweating, is a physiological mechanism for regulating body temperature. Perspiration corresponds to the excretion of water and solutes in the form of sweat via skin pores. Sweat is excreted by the sweat glands which include the eccrine glands and the apocrine glands. The eccrine glands, situated all over the surface of the body, secrete transparent, aqueous and odorless sweat. This eccrine sweat plays a role in hydrating the stratum corneum, microbiological protection, gripping and percutaneous absorption. For their part, the apocrine glands, the excretory part thereof is situated at the upper level of the hair follicles, are more particularly located in the armpits, ear canals, breast areolae, peri-anal regions and feet. These glands only become functional after puberty and secrete viscous sweat that is milky in appearance and low in odor. This apocrine sweat is rich in lipids and also contains proteins. Its secretion is regulated psychologically. In fact, sweat, whether of eccrine or apocrine origin, provided that it is not generated excessively, has little or no odor when secreted. It is the contact of sweat and particularly of apocrine sweat with bacteria which causes the formation of malodorous and volatile compounds, via the bacterial degradation of this sweat. The compounds obtained from the degradation of sweat and giving rise to unpleasant odors are particularly represented by C2-C5 short-chain volatile acids, malodorous steroids, such as the steroid 16-androstene, C6-C12 fatty acids, such as the trans (E) isomer of 3-methyl-2-hexenoic acid (3M2H), or sulfanylalkanols. In this way, the axillary region (armpit) is the area of the body where the most bacteria are found (approximately 10 ⁶ cells/cm2).

Eccrine sweat is made up of 99% water and salt (NaCI) and smaller quantities of potassium, calcium and magnesium ions. It may contain lactic acid in varying quantities in the case of physical exercise and may also comprise urea, antibodies, volatile fatty acids and cholesterol. Sweat contains 0.5% organic substances including proteins, the most plentiful being N-glycoproteins, such as apolipoprotein D (ApoD), prolactin-inducible protein (PIP) and Zinc alpha2-glycoprotein (ZAG).

Deodorants and antiperspirants represent the two categories of products used to reduce perspiration and the adverse effects thereof, in particular in respect of axillary perspiration.

The use of deodorants and antiperspirants is very common, equally well for the purposes of image, visual appeal and comfort, for example to "smell nice", remove damp sensations in the armpits and remove the rings and stains associated with perspiration on clothing.

The most effective antiperspirants at the present time are based on aluminum salts. Given the controversies associated with the use of aluminum salts in cosmetic products intended for human use, and the existence of cases of skin irritations associated with aluminum salts, there is a genuine need for alternative solutions suitable for totally or partially replacing aluminum salts, while enabling effective protection in terms of odor and/or perspiration per se.

Detailed description

The present invention is based, firstly, on the discovery by the inventors of a preferential precipitation of the N-glycoproteins contained in sweat by aluminum salts. This effect on N-glycoprotein precipitation suggests that the antiperspirant effect of aluminum salts could result from the production of plugs consisting of N-glycoproteins on the sweat pores, thus reducing perspiration in the axillary regions.

The inventors then detected specific lectins capable of recognizing and precipitating the main N-glycoproteins in human sweat and/or reducing the production of odorous molecules.

A first aim of the invention is that of providing a method for selecting a lectin-based deodorant and/or antiperspirant agent.

A second aim of the invention is that of providing cosmetic compositions comprising at least one deodorant and/or antiperspirant agent, for example as obtained using the selection method above. The cosmetic compositions according to the invention are preferably intended for topical use on the perspiration areas of the skin.

A third aim of the invention consists of a cosmetic treatment method comprising a step for applying a cosmetic composition as defined above onto the skin. A fourth aim of the invention is the use of one or a plurality of specific lectins as a deodorant agent and/or antiperspirant agent.

The deodorant and/or antiperspirant agents according to the invention thus make it possible to reduce, or even completely replace, conventionally used antiperspirant agents, such as aluminum salts.

The terms "sweat" and "perspiration" are synonymous herein.

Lectins

Lectins are proteins binding specifically with glycans.

A lectin may for example be specific for glucose, fructose, lactose, mannose, branched mannose, sialic acid (including N-acetylneuraminic acid and/or N- glycolylneuraminic acid), galactose, melibiose, fucose, galactoseamine, glucosamine, N- acetylmannosamine, mannosamine, alpha galactose N-acetylgalactosamine, beta galactose N-acetylgalactosamine, N-acetylglucosamine and/or N-acetylgalactosamine.

A lectin may be of animal, plant or microbial origin.

An example of lectin of animal origin is galectin or collectin.

An example of plant lectin is the lectin Con A, the lectin SNA 1 , the lectin MAA, the lectin AIA/Jacalin, the lectin ACA/ACL, the lectin IRA, the lectin LcH, the lectin HPA or the lectin Griffithsin.

An example of microbial lectin is cyanovirin or actinohivin.

Method for selecting a deodorant and/or antiperspirant agent

A first aim of the invention is that of providing novel lectin-based deodorant and/or antiperspirant agents.

The term "deodorant agent" denotes herein an agent suitable for reducing odorous molecule production.

The term "antiperspirant agent" denotes herein an agent suitable for limiting or suppressing perspiration per se. For this reason, the antiperspirant agent makes it possible to indirectly reduce odorous molecule production.

The present invention thus relates to a method for selecting a deodorant and/or antiperspirant agent, comprising the following steps:

a) providing a test compound comprising at least one lectin,

b) carrying out at least one of the following substeps:

b1 . placing said test compound in the presence of at least one sweat glycoprotein and measuring the precipitation of said glycoprotein by said test compound, and/or b2. placing said test compound in the presence of at least one sweat sample and measuring the odorous molecule production, and c) selecting, as a deodorant and/or antiperspirant agent, a compound capable of precipitating at least one sweat glycoprotein and/or capable of reducing odorous molecule production.

Step a) consists of providing a test compound comprising at least one lectin.

The test compound may for example be a purified lectin, a recombinant lectin, a mixture of at least two purified and/or recombinant lectins or a plant extract.

The lectin(s) to be tested are particularly as defined above.

Step b) consists of testing the antiperspirant agent properties in a step b1 . and/or testing the deodorant agent properties in a step b2.

In step b1 , the test compound is placed in the presence of at least one sweat glycoprotein.

The test compound may, for example, be added in an aqueous solution comprising a sweat glycoprotein or a mixture of sweat glycoproteins, preferably a sweat sample.

For this purpose, the sweat glycoprotein used may be chosen among the apolipoprotein D, the glycoprotein PIP, the glycoprotein ZAG or combinations thereof. The test compound is, preferably, placed in the presence of at least one sweat sample.

The sweat sample may be a natural sweat sample or a synthetic sweat sample. The sweat sample is, preferably, a natural sweat sample.

A natural sweat sample may be obtained from one or a plurality of subjects who may be males or females of any age.

The natural sweat sample is, preferably, obtained from the axillary area.

The natural sweat sample is advantageously obtained after the subject has perspired, for example after more or less intense physical exercise or after placing the subject for some time, for example 1 to 2 hours, in a warm environment, such as for example in a room at a temperature greater than 30°C, a sauna or a steam bath.

The natural sweat sample may also be a mixture of sweat samples from the same subject and/or from a plurality of different subjects, for example at least 10 subjects.

Before testing the ability to precipitate a sweat glycoprotein in a natural sweat sample, it is preferable to centrifuge the sweat sample, in order to remove the pellet comprising corneocytes, and to perform the glycoprotein precipitation test on the supernatant of the sweat sample thus obtained. In a further embodiment, the sweat sample is a synthetic sweat sample.

A synthetic sweat sample is a sweat sample obtained by mixing various constituents of natural sweat. A synthetic sweat sample has a similar or identical composition to that of a natural sweat sample.

A natural or synthetic sweat sample may, for example, comprise the following inorganic compounds: Na ⁺ , Cl ⁺ , K ⁺ , Ca ²⁺ , HC0 ₃ ^" , NH ₄ ⁺ , P0 ₄ ⁺ and/or S0 ₄ ²⁺ ; the following organic compounds: glucose, gentamicin sulfate, urea, lactic acid and/or glycerol; the following amino acids: serine, alanine, glycine, citrulline and/or histidine; and/or the following lipids: triglyceride (for example triolein), free fatty acids, mono and di-glycerides and/or surfactants (for example Tween 80).

A natural or synthetic sweat sample may, for example, comprise the following inorganic compounds: 1200 to 1700 mg/l of Na ⁺ , 3200 to 3400 mg/l of Cl ⁺ , 400 to 600 mg/l of K ⁺ , 10 to 60 mg/l of Ca ²⁺ , 100 to 150 mg/l of HC0 ₃ ^" , 100 to 300 mg/l of NH ₄ ⁺ , 3 to 8 mg/l of P0 ₄ ⁺ and/or 50 to 80 mg/l of S0 ₄ ²⁺ ; the following inorganic compounds: 150 to 200 mg/l of glucose, 0 to 200 mg/l of gentamicin sulfate, 250 to 350 mg/l of urea, 2100 to 2400 mg/l of lactic acid and/or 0 to 2 mg/l of glycerol; 1200 to 1500 mg/l of amino acids, including for example 20% to 40% of serine, 10% to 20% of alanine, 10% to 20% of glycine, 8% to 15% of citrulline and/or 5% to 15% of histidine; and/or the following lipids: 100 to 300 mg/l of triglyceride (for example triolein), 0 to 10 mg/l of free fatty acids, 30 to 100 mg/l of mono and di-glycerides and/or surfactants (for example Tween 80).

The precipitation of at least one sweat glycoprotein may be measured using any test suitable for measuring the precipitation of a protein, and in particular a glycoprotein.

The precipitation of at least one sweat glycoprotein may be measured, for example, by electrophoresis (in particular, including protein separation on a gel by electrophoresis followed by the detection thereof, for example using a protein marker, such as a SYPRO type luminescent marker), by Western Blot or by a turbidity measurement.

The result of the precipitation measurement may be compared to that obtained with at least one control, for example a positive control wherein the antiperspirant agent is an aluminum salt, for example ACH (aluminum hydroxy chloride), and/or a negative control comprising the same solution or the same sample, but in the absence of the test compound.

An increase in turbidity in the presence of the test compound with respect to the same sample in the absence of said test compound indicates that said compound is capable of precipitating the sweat glycoproteins. In step b2., the test compound is placed in the presence of at least one sweat sample.

The sweat sample is as defined above. Preferably, the sweat sample is used without prior processing, in particularly without a prior centrifugation step.

Step b2. consists of measuring odorous molecule production in a sweat sample placed in the presence of the test compound.

The odorous molecule production may be measured using direct or indirect methods.

An example of a direct method consists of measuring the emission of volatile odorous molecules, particularly alcohols and ketones, by a sweat sample placed in the presence of the test compound, using any suitable technique, such as gas chromatography, mass spectrometry, a Sniff test, the sniff test preferably being performed with a panel of noses and preferably at the end of a gas chromatography.

For this purpose, the sweat samples are placed in the presence of the test compound for at least 5hrs, preferably at least 10hrs, for example 24hrs at a temperature greater than or equal to 25°C, preferably greater than or equal to 30°C, preferentially greater than or equal to 35°C, for example at 37°C.

The emission of volatile odorous molecules by the sample in the presence of the test compound may be compared to that obtained in the absence of the test compound.

Since bacteria are responsible for odorous molecule production from the molecules present in sweat, an example of an indirect method consists of measuring the bacterial growth in a sweat sample placed in the presence of said test compound.

For this purpose, the bacterial growth is measured after incubating a sweat sample with the test compound for a time greater than or equal to 10hrs, preferably greater than or equal to 24hrs, more preferentially greater than or equal to 48hrs, for example 72hrs, and at a temperature greater than or equal to 25°C, preferably greater than or equal to 30°C, preferentially greater than or equal to 35°C, for example at 37°C.

The bacterial growth may be assessed using any suitable known method, such as measuring the quantity of live bacteria and that of dead bacteria, measuring their consumption of at least one metabolite or energy source, such as glucose, maltose and/or ATP, measuring the turbidity in the sweat sample containing said bacteria, measuring radial diffusion, measuring microplate growth.

The bacterial growth in a sweat sample in the presence of the test compound may be compared to the growth obtained in the absence of said compound. A decrease in live bacteria and/or an increase in dead bacteria, a decrease in the consumption of a metabolite or energy source and/or a decrease in turbidity, a decrease in radial diffusion, in a sweat sample placed in the presence of the test compound compared to a sample in the absence of said test compound, corresponds to an inhibition of bacterial growth.

Inhibition of bacterial growth in the sweat sample placed in the presence of the test compound conveys a decrease in odorous molecule production.

Examples of tests for measuring the precipitation of glycoproteins of sweat and/or odorous molecule production are described in more detail in the examples section.

Finally, step c) consisting of selecting a compound capable of precipitating at least one sweat glycoprotein and/or capable of reducing odorous molecule production.

Lectin-based deodorant and/or antiperspirant agent

A deodorant and/or antiperspirant agent is, preferably, a lectin obtained or likely to be obtained by a method for selecting a deodorant and/or antiperspirant agent as defined above.

A deodorant agent according to the invention has the property of reducing odorous molecule production. As described above and in the examples, this property may be detected via inhibition of bacterial growth in a sweat sample placed in the presence of said agent and/or via a reduction in the emission of volatile odorous molecules by a sweat sample placed in the presence of said agent.

An antiperspirant agent according to the invention has the property of precipitating at least one sweat glycoprotein. As described above and in the examples, this property may be detected via a direct measurement of the precipitation of the sweat glycoprotein(s), for example by electrophoresis, Western Blot, or by measuring the turbidity of a solution comprising at least one sweat glycoprotein and said agent.

A preferred deodorant and/or antiperspirant agent according to the invention is a lectin selected from the group consisting of a lectin Con A, a lectin SNA 1 , a lectin MAA, a lectin AIA/Jacalin, a lectin ACA/ACL, a lectin IRA, a lectin HPA and a lectin LcH.

Preferably, the deodorant agent is selected from the group consisting of the lectin Con A, the lectin SNA 1 , the lectin MAA and the lectin LcH.

Preferably, the antiperspirant agent is selected from the group consisting of the lectin Con A, the lectin SNA 1 , the lectin AIA/Jacalin, the lectin ACA/ACL, the lectin HPA and the lectin IRA. The lectin Con A is a lectin specific for mannose, branched mannose, glucose and N-acetylglucosamine, found for example in Canavalia ensiformis.

The lectin SNA 1 is a lectin specific for sialic acid, particularly N-acetylneuraminic acid found for example in Sambucus nigra.

The lectin MAA is a lectin specific for sialic acid, particularly N-acetylneuraminic acid found for example in Maackia nigra.

The lectin AIA / Jacalin is a lectin specific for galactose found for example in Artocarpus intergrifolia.

The lectin ACA / ACL is a lectin specific for N-acetylgalactosamine and beta galactose/ N-acetylgalactosamine found for example in Amaranthus caudatus.

The lectin LcH is a lectin specific for mannose and glucose found for example in Lens culinaris.

The lectin IRA is a lectin specific for alpha galactose / N-acetylgalactosamine, N- acetylgalactosamine and galactose (in decreasing order of specificity), found for example in Iris hybrid.

The lectin HPA is a lectin specific for N-acetylgalactosamine, found for example in Helix pomatia.

The deodorant and/or antiperspirant agents according to the invention have an effect locally at the area of the skin where they are applied.

The deodorant and/or antiperspirant agents according to the invention are particularly suitable for topical use at perspiration areas, such as the armpits and/or shiny areas of the skin.

The expressions "skin application" and "topical application" are synonymous herein.

Skin denotes, broadly, the skin, scalp and mucosa. In one preferred embodiment, the term "skin" denotes the skin and scalp, but does not include the mucosa.

A lectin may be provided in pure form, in plant extract form and/or in recombinant form.

The expressions "lectin in recombinant form" and "recombinant lectin" are synonymous herein and denote a lectin produced by a cell wherein the genetic material has been modified to produce said lectin. The cell used to produce the lectin may be an animal cell, a plant cell, a yeast cell or a bacterial cell.

The expressions "lectin in pure form", "lectin provided in pure form", "purified lectin", "lectin in purified form" are synonymous herein and indicate that the lectin is provided in a composition comprising at least 85% of said lectin, preferably at least 90% of said lectin, more preferentially at least 95% of said lectin, more preferentially at least 98% of said lectin, the percentages being expressed by weight in respect of the weight of the total composition.

A lectin in pure form may be a natural lectin, i.e. purified from a plant or a plant extract, a lectin obtained by chemical synthesis or a recombinant lectin.

A plant extract denotes herein an extract of the whole plant or of a part of the plant, such as the leaf, trunk, stem, seed, root, flower, nectar, fruit.

A preferred plant extract is a seed extract.

The plant extract may comprise one or a plurality of lectins of interest.

The plant extract may also be a mixture of at least two plant extracts, said plant extracts being obtained from the same plant and/or from a different plant.

The plant extract may have undergone one or a plurality of prior processing steps, such as extraction, precipitation, ultra-purification and/or purification by means of affinity chromatography.

A preferred plant extract according to the invention is a plant extract comprising a lectin Con A, a lectin SNA 1 , a lectin MAA, a lectin AIA/Jacalin, a lectin AC A/ACL, a lectin IRA, a lectin HPA, a lectin LcH, or combinations thereof.

In one advantageous embodiment, the lectin(s) may be in immobilized form, for example to stabilize the lectin(s) and/or enhance the antiperspirant and/or deodorant properties thereof.

A lectin in immobilized form is a lectin bound covalently or non-covalently with at least one substrate.

The substrate may be soluble or non-soluble.

The substrate used for immobilization may be an organic substrate or an inorganic substrate.

An organic substrate is for example, a polysaccharide or a polymer.

A polysaccharide type organic substrate is, for example, selected from the group consisting of cellulose acetate, cellulose nitrate, dextran, agarose, alginate, chitosan, starch and combinations thereof.

A polymer type organic substrate is, for example, selected from the group consisting of polyethylene, polyethylene glycol, polyalanine, polypropylene, polystyrene, polyethylene terephthalate, collagen, hyaluronic acid and combinations thereof.

An inorganic substrate is, for example, selected from the group consisting of porous glass, silica, metals, alumino-silicate, titanates, perlite, talc and combinations thereof. The lectin may be immobilized by physical retention or by physical interaction or chemical binding.

A lectin immobilized by physical retention is, preferably, immobilized by inclusion and/or by confinement.

Immobilization by inclusion may be obtained in a matrix consisting of the three- dimensional network of a polymer, for example hyaluronic acid.

Immobilization by confinement may be performed in a microcapsule, for example a liposome, or in a hollow fiber.

A lectin immobilized by physical interaction or chemical binding is, preferably, immobilization by fixing on a substrate, for example by adsorption or by covalent binding, or by cross-linking, for example by creating a network by means of intermolecular covalent bonds.

Cosmetic composition comprising a lectin-based deodorant and/or antiperspirant agent

The present invention also relates to a cosmetic composition comprising at least one lectin-based deodorant and/or antiperspirant agent, as defined above.

The cosmetic composition according to the invention is preferably a deodorant and/or antiperspirant composition.

The present invention thus relates to a cosmetic composition comprising, as deodorant and/or antiperspirant agent, at least one lectin obtained or likely to be obtained by the selection method defined above.

The present invention thus relates to a cosmetic composition comprising, as deodorant and/or antiperspirant agent, at least one lectin selected from the group consisting of a lectin Con A, a lectin SNA 1 , a lectin MAA, a lectin AIA/Jacalin, a lectin AC A/ACL, a lectin IRA, a lectin HPA and a lectin LcH.

In one preferred embodiment, the cosmetic composition does not comprise lectin other than a lectin Con A, a lectin SNA 1 , a lectin MAA, a lectin AIA/Jacalin, a lectin AC A ACL, a lectin IRA, a lectin HPA or a lectin LcH.

A preferred cosmetic composition according to the invention comprises at least one deodorant and/or antiperspirant agent selected from the group consisting of a lectin Con A, a lectin SNA 1 , a lectin MAA, a lectin AIA/Jacalin, a lectin ACA ACL, a lectin IRA, a lectin HPA and a lectin LcH.

The deodorant and/or antiperspirant agent is, preferably, selected from the group consisting of a lectin Con A, a lectin SNA 1 and a lectin MAA. One particularly preferred deodorant and/or antiperspirant agent according to the invention is a lectin SNA 1 . In one advantageous embodiment, the cosmetic composition according to the invention comprises at least two deodorant and/or antiperspirant agents according to the invention.

For example, the cosmetic composition according to the invention may comprise two or at least two deodorant and/or antiperspirant agents selected from the group consisting of a lectin Con A, a lectin SNA 1 , a lectin MAA, a lectin AIA/Jacalin, a lectin AC A/ACL, a lectin IRA, a lectin HPA and a lectin LcH.

One preferred cosmetic composition comprises the lectin Con A and/or the lectin

SNA 1 .

The cosmetic composition preferably comprises from 0.00001 % to 10% of said deodorant and/or antiperspirant agent or said deodorant and/or antiperspirant agents when the composition comprises at least two agents, preferentially from 0.0001 % to 10%, preferentially from 0.001 % to 10%, more preferentially from 0.01 % to 10%, more preferentially from 0.1 % to 10%, even more preferentially from 1 % to 10%, the percentage being given by weight with respect to the total weight of the composition.

As mentioned above, the deodorant and/or antiperspirant agent may be provided in a pure form, in a recombinant form and/or in the form of a plant extract.

In one particular embodiment, the cosmetic composition does not comprise plant extract.

In one advantageous embodiment, the composition comprises, as deodorant and/or antiperspirant agent, at least one lectin in immobilized form.

A lectin in immobilized form is particularly as defined above.

In one advantageous embodiment, the cosmetic composition comprises one or at least one co-factor of said deodorant and/or antiperspirant agent(s).

The term "co-factor" denotes herein a compound which is not a deodorant and/or antiperspirant agent per se and which is suitable for enhancing the deodorant and/or antiperspirant properties of a deodorant and/or antiperspirant agent according to the invention.

An example of a co-factor is a salt, for example a magnesium salt, a manganese salt, a calcium salt, a dextran, a PEG (polyethylene glycol) or combinations thereof.

The co-factor is thus not an aluminum salt.

The cosmetic composition may further comprise one further antiperspirant agent and/or one further deodorant agent. The term "further antiperspirant agent" or "further deodorant agent" denotes an agent which is not a lectin.

An example of a further antiperspirant agent is an aluminum salt, for example aluminum hydroxychloride (ACH), aluminum bromohydrate, aluminum zirconium octachlorohydrate, aluminum chloride, aluminum zirconium Octachlorohydrex Gly, aluminum chlorohydrate, aluminum zirconium pentachlorohydrate, aluminum chlorohydrex, aluminum zirconium pentachlorohydrex Gly, aluminum chlorohydrex Peg, aluminum zirconium tetrachlorohydrate, aluminum chlorohydrex Pg, aluminum Zirconium Tetrachlorohydrex Gly, aluminum citrate, aluminum zirconium trichlorohydrate, aluminum dichlorohydrate, aluminum zirconium trichlorohydrex Gly, aluminum dichlorohydrex Peg, aluminum dichlorohydrex Pg, aluminum sesquichlorohydrate, aluminum sesquichlorohydrex Peg, aluminum sesquichlorohydrex Pg, aluminum sulfate, ammonium alum, sodium alum and sodium aluminum chlorohydroxy lactate.

A preferred further antiperspirant agent according to the invention is an aluminum hydroxychloride (ACH).

An example of a further deodorant agent is an antimicrobial agent, for example triclosan, an amino acid derivative, for example capryloyi glycine, an odorizing agent, for example an essential oil or a mixture of essential oils, an astringent agent such as aluminum chlorohydrate or tannic acid, an adsorbent agent, such as talc. Said further deodorant agent is thus defined more broadly than the deodorant agent according to the invention.

A preferred composition according to the invention comprises a lectin chosen from the lectin Con A and the lectin SNA 1 and an aluminum salt, preferably an aluminum hydroxychloride (ACH). In this case, the aluminum salt concentration in the cosmetic composition is, preferably, less than the concentrations routinely used in antiperspirant compositions; in particular the aluminum salt concentration in the cosmetic composition is less than 15%, preferentially less than 7.5%, preferentially less than 5%, preferentially less than 1 %, more preferentially less than 0.6%, even more preferentially less than 0.3%, more preferentially less than 0.2%, even more preferentially less than 0.1 %, the percentage being expressed by weight with respect to the total weight of the composition.

The cosmetic composition is in any form suitable for topical use, particularly at the perspiration areas of the skin, for example the armpits, and/or shiny areas of the skin.

The cosmetic composition is for example in liquid, solid, gel, aerosol, semi-liquid, paste, cream, mask, powder or serum form. The cosmetic composition may thus be contained in a spray, stick, container or bottle.

A cosmetic composition according to the invention may also comprise at least one excipient selected from the group consisting of: mica titanium dioxide, perfume, dimethicone, ceteryl alcohol, ceteareth, EDTA, glycerin, propylene glycol, sodium stearate, steareth, behenic acid and sodium hydroxide.

In one particularly advantageous embodiment, the composition according to the invention does not comprise lectin from Moringa oleifera, more generally does not comprise Moringa oleifera seed extract, or more generally again does not comprise Moringa oleifera extract.

In one particularly advantageous embodiment, the composition according to the invention does not comprise lectin from Astragalus membranaceus, or more generally does not comprise Astragalus membranaceus root extract, or more generally again does not comprise Astragalus membranaceus extract.

In one preferred embodiment, the cosmetic composition according to the invention does not comprise lauric acid.

Cosmetic treatment method

The present invention also relates to a method of cosmetic treatment comprising a step of applying a cosmetic composition as defined above onto the skin.

The cosmetic composition is preferably applied on the perspiration area(s) of the skin, for example on the armpits or the shiny areas of the skin.

The cosmetic composition is preferably applied on clean skin.

The frequency of application of the composition is preferably once to twice daily.

The cosmetic treatment method according to the invention enables partial or total inhibition of perspiration and/or a reduction in odorous molecule production, in particular at the areas of the skin where the cosmetic composition is applied, particularly via the use of cosmetic compositions not comprising aluminum salts or a lower aluminum salt concentration.

Use of a cosmetic composition according to the invention

The present invention also relates to the topical use of a cosmetic composition as defined above for partially or totally inhibiting perspiration and/or for reducing odorous molecule production. The cosmetic composition is preferably applied on the perspiration area(s) of the skin, for example on the armpits or the shiny areas of the skin.

Partial or total inhibition of perspiration and/or reduction of odorous molecule production is obtained at the areas of the skin where the cosmetic composition according to the invention is applied.

Use of a lectin as deodorant and/or antiperspirant agent

The present invention also relates to the use of a lectin selected from the group consisting of a lectin Con A, a lectin SNA 1 , a lectin MAA, a lectin AIA/Jacalin, a lectin ACA/ACL, a lectin IRA, a lectin HPA, a lectin LcH and another lectin obtained by the selection method defined above, as a deodorant agent and/or antiperspirant agent.

The present invention also relates to the use as defined above of a lectin in association with a further antiperspirant agent, for example an aluminum salt such as an aluminum hydroxychloride. The association of said lectin with said further antiperspirant agent particularly makes it possible to obtain a synergistic effect on the reduction of perspiration, for example by increasing the precipitation of sweat glycoproteins.

The present invention also relates to the use as defined above, wherein the lectin is in immobilized form. A lectin in immobilized form is particularly as defined above. Further features and advantages of the invention will emerge more clearly from the following examples, given by way of illustration and not limitation.

Figures

Figure 1 : Western Blot analysis of the binding of the lectins Con A (2) and SNA 1 (3) with the three main N-glycoproteins of sweat: PIP, ZAG and ApoD. In 1 , the proteins stained using Sypro Ruby in human sweat are shown.

Figure 2: Effect of lectins on the precipitation of apolipoprotein D (ApoD) of sweat (using sweat sample supernatants). In A, the results are given as a percentage of precipitation with respect to the result obtained with aluminum salts. In B, the Western Blot results show the precipitation of the proteins of sweat with various lectins. A1 : Con A - 500 μg/ml, A2: Con A - 250 μg/ml, A3: Con A - 125 μg/ml, B1 : SNA 1 - 500 μg/ml, B2: SNA 1 - 250 μg/ml, C1 : HPA - 500 μg/ml, C2: HPA - 250 μg/ml, D2: MAA 250 μg/ml, E2: AAA 250 μg/ml, T: aluminum salts.

Figure 3: Percentage of turbidity activity at 600nm of various lectins at an equivalent quantity of molecules, with respect to the turbidity obtained with aluminum salts (ACH). The test is performed at a final concentration of 1 mM, after 60 minutes of incubation at 37°C with axillary sweat. The lectins tested are: Con A, SNA 1 , UEA 1 , MAA, AC A/ACL, CSA, IRA and AIA/Jacalin.

Figure 4: OD at 600 nm obtained with the lectin SNA 1 alone at a concentration of 0.025% (A), aluminum salts ACH alone at a concentration of 0.01 % (B) and with SNA 1 at a concentration of 0.025% in association with ACH at a concentration of 0.01 % (C). T represents the sweat control in the absence of lectin or ACH.

Figure 5: Effect of the lectins Con A, LcH, SNA 1 , MAA, EEA and AIA, and of a Moringa oleifera extract (Mol) on bacterial growth in sweat, after incubation, measured by means of the dead/live bacterial assay. T: "sweat" control with no lectin.

Example 1 : Deodorant and antiperspirant effect of lectins

Materials and Methods

(i) Sweat sample collection and preparation

Axillary sweat samples are obtained from volunteers (after sauna for 2hrs at 60- 80°C) in glass jars, as per the clinical protocol FtCL-COS-0619 {RAD Chevilly). The sweat samples are pooled and stored at -20°C until use. Before analysis, the sweat samples are centrifuged for 8 minutes at 8000g at 4°C to remove corneocytes. The resulting supernatant is collected in a microtube. (ii) SDS-PAGE

The protein precipitates and the supernatants from the sweat samples are (re)- solubilized in a Laemmli buffer, and subjected to denaturation (for 5 minutes at 95°C) and centrifugation at 10000g for 1 minute. They are deposited on polyacrylamide gel (Criterion TGX 10-20%). Migration is performed with the Protean III system (Biorad) at 200V for around 1 hr in SDS 1 X migration buffer. The gels are stained using SYPRO RUBY reagent (Invitrogen) as per the manufacturer's protocol. The fluorescent images of the gels are acquired with a ProExpress Imager (Perkin Elmer) using excitation (480 nm) / emission (620 nm) filters. (iii) Western blot

10 μg of the aliquots of soluble proteins (supernatants) or protein precipitates of purified sweat (i.e. centrifuged sweat as described above) are separated by SDS- PAGE as described in (ii). The proteins are transferred to PVDF membranes (Immobilon P, Millipore Molsheim la France) according to standard conditions. The membranes are then blocked with a 1 % milk (v/v) PBS 1 X 0.05% Tween 20 buffer for 1 hour at ambient temperature. Then the membranes are incubated in the presence of a primary antibody (NCL-ApoD Novacastra) in 1 % milk (v/v) PBS 1 X 0,05% Tween 20 buffer; then washed 3 times in PBS 1 X 0.05% Tween 20 buffer.

Finally, the membranes are incubated in the presence of a secondary antibody coupled with horseradish peroxidase. After the successive washes in PBS 1 X 0,05% Tween 20 buffer, the membranes are detected with ECL Plus® reagent (GE Healthcare Orsay France) and viewed with the Fluor-S Multimager system (Biorad, Marnes-la coquette France). The signal intensity is quantified using Quantity One® software (Biorad, Marnes-la coquette France).

The precipitation percentages are obtained based on the following formula: (Control ApoD band intensity - Test Apo D band intensity) / Control ApoD band intensity) X 100.

(iv) Lectin blot

10 μg of the aliquots of soluble proteins or protein precipitates of purified sweat (i.e. centrifuged sweat as described above) are separated by SDS-PAGE. The proteins are transferred to PVDF membranes (Immobilon P, Millipore Molsheim la France) according to standard conditions. The membranes are then blocked with a 1 % milk (v/v) PBS 1 X 0.05% Tween 20 buffer for 1 hour at ambient temperature. The membranes are then incubated in the presence of a 5 μg/ml biotinylated lectin.

Finally, the membrane is incubated in the presence of streptavidin conjugated with horseradish peroxidase diluted to 1 :20,000. After the successive washes with PBS 1 X 0.05% Tween 20 buffer, the membrane is detected with ECL Plus® reagent (GE Healthcare) and viewed with the Fluor-S Multimager system (Biorad). The signal intensity is quantified using Quantity One® software (Biorad, Marnes-la coquette France).

(v) Sweat turbidity test

The native sweat is centrifuged for 10 minutes at 10,000 g at 4°C. The supernatant retrieved is incubated for 1 hour in the presence of lectins at a suitable concentration, with stirring, in a 96-well plate at 37°C. The final volume in each well is ~\ 00μ\- (1 :1 volume centrifuged sweat: lectin). Finally, the plate is read with a spectrophotometer (SpectraMax5 ^e - Molecular Devices) at 600 nm. For the kinetic test, the incubation is performed directly on the instrument at 37°C, at 5 minute intervals.

(vi) Microflora viability test

The live and dead bacteria content in the sweat samples is evaluated using the Baclight Live/Dead bacterial viability kit (Molecular Probes-lnvitrogen). The supernatants of the centrifuged sweat samples (100 μΙ) are incubated in a 96-well plate for 72 hours at 37°C, with stirring. 100 μΙ_ of 2X dye solution containing SYTO (which inserts itself in the DNA of live bacteria) and propidium iodide (PI) (which only inserts itself in the DNA of bacteria having damaged membranes) are added to each well, mixed and incubated at ambient temperature in darkness for 15 min. The plate is read with a spectrophotometer (SpectraMax5 ^e - Molecular Devices) at 485 nm (excitation wavelength) and, respectively, at 530 nm and 630 nm for SYTO and PI (emission wavelength). The inhibition percentage is calculated on the basis of the fluorescence measured with the treated sweat sample with respect to the fluorescence measured with the untreated control sweat sample.

(vii) Gas Chromatography (GO

1 ml sweat samples are incubated in the presence or absence of 200 μΙ of 1 mg/ml lectins at 37°C, under stirring, for 24 hours. The samples are then analyzed for the presence of volatile odorous compounds (essentially alcohols and ketones), by Gas Chromatography (CP3800 Varian) and mass spectrometry (HS-SPME-GC/MS). Using a previously defined model, an efficacy score is assigned to each agent used (i.e. to each lectin used), based on the quantity placed in contact with 1 ml of sweat.

As the scores relating to the agents are dependent on the type of sweat, an assessment is given in respect of efficacy:

Agent score and Control sweat score 4°C > 1 .3 => Very effective agent

1 < Agent score and Control sweat score 4°C < 1 .3 => Effective agent

0.75 < Agent score and Control sweat score 4°C < 1 => Relatively ineffective agent

Agent score and Control sweat score 4°C < 0.75 => Ineffective agent.

Results

Over 30 lectins in pure form are evaluated for the ability thereof to bind and precipitate sweat glycoproteins.

The Western Blot analysis using the biotinylated lectins Con A (from Canavalia ensiformis) and SNA 1 (from Sambucus nigra) demonstrates that these lectins recognize the main N-glycoproteins of sweat, more particular ApoD (see Figure 1).

The lectins Con A and SNA 1 , which respectively bind with mannoses and sialic acids, are then analyzed for their potential to precipitate sweat proteins, with respect to aluminum salts (ACH). For this purpose, the lectins Con A and SNA 1 are added to the supernatant of a centrifuged sweat sample, at different concentrations. After incubating at 37°C for one hour and after centrifugation, the presence of the lipoprotein ApoD in the supernatant is evaluated. The results demonstrate that the lectins Con A and SNA 1 at the final concentration of 0.05% are capable of precipitating at least 80% and at least 55% of the lipoprotein ApoD of sweat, respectively, compared to the complete precipitation of the lipoprotein ApoD by aluminum salts at 0.01 % (see Figure 2).

Table 1 : Summary of the results obtained, at an equivalent quantity of molecules, when selecting lectins

Glc: glucose, Man: mannose, GlcNAc: N-acetylglucosamine, NeuAc: sialic acid (N- acetylneuraminic acid), Fuc: Fucose, Gal33GalNAca-0-R: beta galactose/ N- acetylgalactosamine; GalNAc: N-acetylgalactosamine, GalNAca3Ga: alpha galactose/ N- acetylgalactosamine.

A broader range of lectins was then evaluated in various turbidity tests. These results confirm Con A and SNA 1 as effective flocculation agents in human sweat with respect to aluminum salts (ACH). The lectins specific for galactose (AIA) and N-acetylgalactosamine (ACA, IRA) are also effective (see Figure 3, Table 1).

The effect of lectins in association with aluminum salts or further lectins on the precipitation of glycoproteins in sweat was also analyzed. The results demonstrate that the association of SNA 1 with the aluminum salts ACH gives the highest precipitation with respect to the lectin SNA 1 alone or the aluminum salts ACH alone (see Figure 4 and Table 2). The efficacy of SNA 1 in association with aluminum salts for precipitating sweat proteins is also greater than the effect obtained with other lectin/lectin or lectin/aluminum salt associations (see Table 2).

Table 2: Summary table of the results obtained with the lectins SNA 1 , Con A and ACA, aluminum salts and the association of said lectins with aluminum salts ACH at 0.5 mMol/L final

% of turbidity

Final Mean activity with

Lectins and

concentration actual OD at respect to ACH

range mMol/L 600nm final 0.5

mMol/L ACH

Aluminum salts ACH 0.5 0.036 100 alone

Lectins alone Con A 0.5 0.027 75

SNA 1 0.5 0.040 111

ACA 0.5 0.022 61

Lectin / lectin or Con A - ACH 0.5 0.043 119 lectin / aluminum SNA 1 - ACH 0.5 0.052 144 salt associations Con A - SNA 1 0.5 0.040 1 1 1

Con A - ACA 0.5 0.028 78

SNA 1 - ACA 0.5 0.042 1 17

ACH - ACA 0.5 0.037 103

SNA 1 - Con A 0.5 0.040 1 1 1 - ACA Lectins such as SNA 1 or plant extracts containing said lectins, which target sialic acids at the terminal part of mannose chains are the most effective in precipitating sweat proteins. The effect of lectins on odor production is also analyzed by measuring in a first stage the effect thereof on bacterial growth in human sweat. The live / dead bacterial assay results demonstrate that the presence of the lectins Con A, LcH, SNA 1 or MAA inhibits bacterial growth at a final concentration of 0.05% (see Figure 5). Lectins specific for mannose or sialic acid are particularly effective (81 to 90% inhibition) compared to untreated sweat (see figure 5).

The effect of some lectin families is analyzed in a second stage on odor emission in human sweat. The four lectins tested are as follows: Con A and LcH (specific for mannose) and SNA 1 and MAA (specific for sialic acid). Sweat samples treated or untreated with lectins are analyzed by gas chromatography (GC) for the presence of odorous volatile substances, i.e. essentially alcohols and ketones.

Table 3: Summary table of the results obtained with various pure lectins on odorous volatile molecule production

Samples tested Relative score Effect

(Test result / control

result 4°C)

Sweat control at 4°C 1

Sweat control at 0.1

37°C

Control buffer 0.41

(PBS/MgCI ₂ /CaCI ₂₎

Con A 0.55 No effect

LcH -0.3 No effect

SNA 1 1 .06 Effective

MAA 1 .15 Effective The result in respect of efficacy for each lectin tested is calculated with respect to the volatile substances present in the following two controls: one sweat sample stored at 4°C and another incubated at 37°C under the same experimental conditions. The results demonstrate that the lectins SNA 1 (from Sambucus nigra) and MAA (from Maackia amurensis) are effective for reducing the concentrations of volatile odorous substances in human sweat (see Table 3).

Conclusions

Aluminum salts (ACH) preferentially precipitate N-glycoproteins of human sweat, which could be the essential mechanism whereby aluminum salts produce sweat pore plugs reducing perspiration in axillary regions.

The lectins Con A and SNA 1 recognize and bind the N-glycoproteins of human sweat.

The lectins, Con A (specific for mannose) and SNA 1 (specific for sialic acid) and, to a lesser degree, the lectin AIA/Jacalin (specific for galactose) precipitate effectively bound N-glycoproteins of human sweat.

In the turbidity test, the lectins Con A, SNA 1 , AIA/Jacalin and the lectins targeting

N-acetyl-galactosamine (ACA and IRA) are effective.

The lectins Con A and LcH which are specific for mannose and the lectins SNA 1 and MAA which are specific for sialic acid inhibit bacterial growth in whole sweat samples in vitro.

The lectins SNA 1 (from Sambucus nigra) and MAA (from Maackia amurensis) are effective in reducing levels of odorous volatile substances in whole human sweat.

Example 2: Lectin-based deodorant and/or antiperspirant cosmetic compositions

Two examples of deodorant and/or antiperspirant cosmetic compositions are shown in tables 4 and 5.

Table 4: Roll-on deodorant or antiperspirant composition

Raw material Content

(% by weight with respect to total weight of composition)

Lectin 10.00

Mica Titanium dioxide 0.70

Fragrance 1 .00

Dimethicone 0.50

Ceteryl alcohol 2.50

Ceteareth-33 1 .25

Water 84.05

Table 5: Stick deodorant or antioersoirant composition

Raw material Content

(% by weight with respect to total weight of composition)

Lectin 10

Mica Titanium dioxide 0.5

EDTA 0.5

Glycerin 20

Propylene glycol 50

Sodium stearate 5

Steareth-100 1

Behenic acid 2

Perfume 1

Sodium hydroxide 1

Water 9 The lectin used in the compositions in tables 4 and 5 is, for example, the lectin Con A, the lectin SNA 1 , the lectin MAA, the lectin AIA/Jacalin, the lectin ACA/ACL, the lectin IRA, the lectin HPA, the lectin LcH, or a mixture of said lectins.