FR3139467A1 - Use of a Sichuan pepper extract for skin treatment and a composition adapted to this use. - Google Patents

Abstract

The present invention relates to a use of a Sichuan pepper extract and a composition comprising said extract, for a cosmetic and/or dermatological treatment, in particular a topical treatment. The extract according to the invention has moisturizing, soothing, anti-aging, anti-odor and care effects for oily skin including the scalp and acne-prone skin, in particular by acting on the production of different molecules such as hyaluronic acid, elastin, collagen, laminins, on the inhibition of oxidation, radicals, lipases, inflammation factors, all while preserving the skin microbiota.

Description

Use of Sichuan pepper extract for skin treatment and a composition suitable for this use.

The present invention relates to the use of an extract of Sichuan pepper, more particularly of berries, and a composition comprising said extract for a cosmetic or dermatological treatment, in particular a topical treatment.

By "cosmetic or dermatological treatment" is meant a treatment that treats healthy skin and/or appendages, said treatment being intended to improve or beautify their appearance and condition. Such treatment has no therapeutic purpose.

The present invention relates in particular to the cosmetics and dermopharmacy industries, which manufacture and/or use products intended for the treatment of the skin, including the scalp, mucous membranes and appendages (such as hair, eyelashes, eyebrows, nails, hair) of mammals, animals or humans, to improve their appearance and/or general condition.

These industries are in increasing demand for new products, in particular in demand for new active ingredients which are derived from plants because these make it possible to combine efficacy, limitation of the risks of irritation and allergy, reduction of side effects, biodegradability, with the possibilities of labelling/certifications and suitability with a logic of sustainable development and/or fair trade.

So-called "holistic" products are appearing on the market to improve the daily care routine. The objective is to act on the user's well-being, to treat their emotions globally. The product must provide the user with a good overall feeling, both in terms of the beautification of their skin but also in terms of the sensory and olfactory comfort provided. The beautification of the skin will typically result, for example, in smoother skin with less roughness, more elastic, less sagging, a better radiance of the complexion, more even with fewer imperfections, while sensory comfort will result, for example, in softness to the touch, soothing of the skin with less redness and tightness, and olfactory comfort by neutralizing or masking body odors.

Human odor is genetically controlled and systematically influenced by gender and ethnicity, as well as emotional, physiological and environmental factors (influence of sweat composition and quantity). Bad body odor affects social interactions by decreasing self-confidence.

Body odor comes from sweat secreted by sweat glands, which can be of two types:

- Eccrine sweat glands are present mainly in the palms of the hands and the soles of the feet. Eccrine sweat is responsible for regulating body temperature.

- Apocrine sweat glands are larger, limited to hairy areas, mainly the armpits, perineal region and scalp. Apocrine sweat is responsible for body odor.

Apocrine sweat contains large, odorless, non-volatile molecules. Odor is generated when these molecules are lysed into small, volatile, odorous entities. This lysis is carried out by enzymes including lipases, which are themselves produced by skin bacteria.

To combat odors, a large part of the population uses some form of antiperspirant or deodorant on a daily basis. These products are mostly antibacterial and/or contain aluminum salts. The latter have the ability to tighten the pores of the skin, and therefore reduce the flow of sweat, but have many disadvantages including an increased risk of breast cancer.

The present invention aims to meet all of these demands, namely to offer a complete cosmetic product and treatment, in particular “holistic” and more specifically to meet the demand for treatment of body odors.

By “body odor treatment” we mean an anti-odor treatment intended to reduce, mask, eliminate, absorb and/or limit the generation of bad odor resulting from the decomposition of sweat by bacteria.

For this purpose, it proposes the use of an extract of Sichuan berries of the species Zanthoxylum bungeanum or Zanthoxylum simulans for a non-therapeutic cosmetic treatment of the skin and its appendages. Preferably according to the invention, the treatment is topical.

According to the invention, by “topical treatment” or “topical use” is meant an application which is intended to act at the place where it is applied: skin, mucous membrane and/or appendages.

The Sichuan pepper tree is defined by one of two species of shrubs: Zanthoxylum bungeanum and Zanthoxylum simulans . It is native to the Sichuan region (China).

The Sichuan pepper plant used according to the invention is preferably Zanthoxylum bungeanum .

It is a thorny and robust shrub of the Rutaceae family, which is the citrus family. The main ethnobotanical properties attributed to this plant species are the relief of dental problems and gastrointestinal disorders, the effectiveness against rheumatism and its analgesic power. The husks of the fruits of the Sichuan pepper tree are widely used in Japan as a spice for their powerful, aromatic odors and their flavors of ripe citrus fruits.

Sichuan pepper berries come from the pericarp of the fruit of the Sichuan pepper tree.

In vitro test results are given further in the description demonstrating beneficial cosmetic activities for the skin and its appendages:

- On the olfactory level, a treatment to reduce body odors, thanks to the reduction of the lipase activity of bacteria, while advantageously preserving the skin microbiota, while the majority of products currently on the market are antibacterial. An in vivo olfactory test of sweat confirms these results.

- On the general beautification or improvement of the skin:

1) an anti-aging treatment, for denser, plumper, firmer, more supple and elastic skin with, as a result, a more homogeneous, smoother skin texture and a better complexion, thanks to the preservation and stimulation of the synthesis of different molecules of the dermal extracellular matrix (ECM), such as:

- elastin responsible for the resilience and elasticity of the skin, and

- collagen VII and laminins reinforcing the dermo-epidermal junction (DEJ) which ensures good anchoring of keratinocytes to the basal lamina and better communication between cells allowing polarization of keratinocytes, therefore a better skin barrier. It is therefore less fragile, which contributes to the suppleness of the epidermis.

- an increase in the anti-radical and anti-oxidant effect, particularly on the DPPH radical (2,2-diphenyl-1-picrylhydrazyl) and singlet oxygen, as well as an anti-glycation effect, helping to preserve the dermal ECM.

The increase in oxidative and radical forms is linked to age and/or repeated environmental stresses, for example UV. Radicals and reactive oxygen species attack in particular membrane lipids and/or functional proteins of the dermal ECM, such as collagens or elastin.

As for glycation, it also affects proteins and/or reducing sugars. These interactions with proteins and/or reducing sugars have the consequence of altering the mechanical and elastic properties of the ECM of the dermis, which becomes less flexible, more rigid, but also more flaccid and less reactive. This visually translates into the appearance of wrinkles and fine lines, dull, flaccid, lackluster skin, with a tired appearance, skin lacking tone and suppleness.

2) treatment of oily and acne-prone skin, thanks to the reduction of sebum production and/or astringent action, which together have the effect of reducing the size of pores and making them less visible, reducing the shiny appearance characteristic of oily skin and/or making the scalp healthier, with less dandruff and the itching associated with it.

3) an anti-dandruff scalp treatment, thanks to the reduction of sebum production and/or the lipase activity of bacteria, while advantageously preserving the skin microbiota.

Sebum causes excessive growth of dandruff-causing bacteria such as Malassezia fungi on the scalp. This fungus is responsible for the formation of dandruff and the itching associated with it.

On the scalp surface, Malassezia release lipases to support their growth, which hydrolyze triacylglycerols into monoacylglycerols and then into fatty acids and glycerol. These free fatty acids inhibit the growth of other microorganisms, promote the proliferation of Malassezia and penetrate the stratum corneum causing damage to the skin barrier.

4) a moisturizing treatment, thanks to an increase in the synthesis of hyaluronic acid and the differentiation of keratinocytes.

Hyaluronic acid is a major constituent of the epidermis, participating in the barrier function as well as maintaining satisfactory hydration of the skin. It is capable of capturing 1000 times its weight in water. It comes in the form of an aqueous and nourishing gel that fills the spaces between the keratinocytes. In this way, it prevents skin dryness, which is known to alter the texture of the skin, giving it a rough and rough feel.

Keratinocyte differentiation occurs from the basal layer of the epidermis. The epidermis is divided into several layers:

- the deepest is the basal layer, made up of keratinocytes, and

- the most superficial is the horny layer (or stratum corneum), consisting of several layers of keratinocytes at the terminal stage of their differentiation and called corneocytes. This highly water-insoluble horny envelope is composed of anucleated horny cells, without cytoplasmic organelles and totally keratinized.

Keratinocytes migrate from the deepest layer to the most superficial layer, differentiating into corneocytes.

The stratum corneum is a semi-permeable protective layer that prevents water loss and maintains skin hydration. It is composed of corneocytes as well as lipids, lamellar bodies, and natural moisturizing factors (NMFs) - substances capable of binding water in the stratum corneum - that maintain the skin's hydration level at an optimal level.

Transepidermal Water Loss (TEWL) characterizes the evaporation of water contained in the skin: the more the skin barrier is damaged, the more easily water evaporates.

Better differentiation of keratinocytes leads to a strengthening of the skin barrier and therefore contributes to better protection of the epidermis and better hydration.

- On the sensory level:

A soothing treatment for the skin, thanks to the reduction of inflammatory mediators responsible for the appearance of micro-inflammations in the skin causing feelings of tightness and discomfort, and also a moisturizing treatment as described above, allowing for softer, less rough skin.

These cosmetic effects can be envisaged according to the invention separately or in a combined manner, advantageously making it possible in particular to propose a “holistic” type use combining olfactory, beautifying and sensory effects.

Thus, the present invention preferentially proposes a use of the Sichuan berry extract of the species Zanthoxylum bungeanum or Zanthoxylum simulans , for at least one treatment chosen from:

- treatment of body odors; and/or

- treatment of oily skin including the scalp and acne-prone skin; and/or

- an anti-aging treatment; and/or

- a moisturizing treatment; and/or

- a soothing treatment.

According to other preferred characteristics, the Sichuan berry extract, of the species Zanthoxylum bungeanum or Zanthoxylum simulans , used according to the invention can be obtained by the usual solid/liquid extraction techniques including for example maceration, simple decoction, infusion, leaching, reflux extraction, extraction by subcritical or supercritical fluid, extraction by means of ultrasound or microwaves or any other physical and/or chemical method such as percolation, digestion, cryoextraction, enzymatic digestion, etc.

Preferably according to the invention, the extract is obtained by extraction under reflux, by maceration, by decoction, by infusion, by leaching, by subcritical or supercritical fluid, by means of ultrasound or microwaves, by percolation, by digestion, by cryoextraction or by enzymatic digestion.

By "maceration" we mean a process consisting of soaking a plant or part of a plant in an extraction solvent at room temperature.

By "infusion" we mean a process of bringing the extraction solvent to a boil before pouring it onto the plant or plant part.

By "decoction" we mean a process consisting of mixing the plant or part of the plant with the extraction solvent, then bringing the whole to the boil.

By "digestion" we mean a process of mixing the plant or plant part with the extraction solvent and then heating the mixture below boiling temperature.

More preferably according to the invention, the extract is obtained by digestion of Sichuan berries in at least one aqueous, alcoholic, hydroalcoholic, glycolic, lipidic solvent or mixture, such as water, a C1, C2, C3 or C4 alcohol, or a polyol chosen from pentane-diol, sorbitol, butylene glycol, pentylene glycol, propylene glycol, hexanol, caprylic/capric triglyceride (GTCC), a vegetable oil, or a mixture thereof.

Preferably, the extraction solvent is a glycolic solvent, more preferably propylene glycol.

The Sichuan berry extract, Zanthoxylum bungeanum or Zanthoxylum simulans , used according to the invention mainly contains xanthoxylin, polyphenols including flavonoids, carbohydrates and proteins.

This extract can be used as is or in a composition, diluted in a physiologically acceptable medium. The nature of the medium is defined according to the properties of the Sichuan berry extract and also according to the destination of the composition formed: a simple ingredient or a more sophisticated galenic form of a final composition for the consumer.

By “physiologically acceptable medium” is meant according to the present invention, without being limiting, an aqueous or hydroalcoholic solution, a water-in-oil emulsion, an oil-in-water emulsion, a microemulsion, an aqueous gel, an anhydrous gel, a serum, a dispersion of vesicles, a powder.

“Physiologically acceptable” means that the compositions are suitable for topical or transdermal use, in contact with the mucous membranes, nails, scalp, hair, fur and skin of mammals and more particularly humans, compositions that can be ingested or injected into the skin, without risk of toxicity, incompatibility, instability, allergic response, and others. This “physiologically acceptable medium” forms what is classically called the excipient of the composition.

Preferably according to the invention, the physiologically acceptable medium may be an aqueous, hydroglycolic or hydroalcoholic medium, or formed by a water-in-oil emulsion, an oil-in-water emulsion, or a microemulsion. More preferably, it is hydroglycolic. More preferably, the physiologically acceptable medium is a mixture of propylene glycol and water.

The present invention therefore covers a non-therapeutic, cosmetic or nutraceutical topical treatment method for beautifying or improving the appearance and general condition of the skin and/or appendages and treating imperfections, in a subject who needs an effective amount of at least one Sichuan berry extract according to the invention or a composition comprising it, in a physiologically acceptable excipient.

The "effective" amount of Sichuan berry extract according to the invention, i.e. its dosage, depends on the intended use of the composition. It depends on various factors, such as age, condition of the patient, severity of the disorder. An effective amount means a non-toxic amount sufficient to achieve the desired effect.

In a composition containing at least one Sichuan berry extract according to the invention, to be present in an effective amount, it is generally found in proportions of between 0.000001% and 15% relative to the total weight of the composition, more preferably between 0.00001% and 10%, depending on the intended use of the composition and the more or less pronounced desired effect. More preferably, the effective amount is between 0.0001% and 5% relative to the total weight of the composition.

All percentages and ratios used in this application are by weight of the total composition and all measurements are made at 25°C, unless otherwise specified.

According to other particularities, the cosmetic treatment method according to the invention can be associated with one or more other treatment methods targeting the skin, such as for example light therapy, heat or aromatherapy treatments.

According to the invention, it is possible to propose multi-compartment devices or kits intended for implementing the method described above, and which could comprise, by way of example, and without this being limiting, in a first compartment a composition containing an extract of Sichuan berries according to the invention in an excipient, and in a second compartment a second extract of Sichuan berries according to the invention in an excipient and/or additional active agent, the compositions contained in said first and second compartments being considered here as a combination composition for simultaneous, separate or spread out use over time, in particular in one of the treatments defined above.

For example, for a cosmetic facial treatment, the European Cosmetics Directive has set a standard application quantity of 2.72 mg/cm²/day/person for a cream and 0.5 mg/cm²/day/person for a body lotion.

According to other advantageous characteristics, the Sichuan berry extract according to the invention can be used in combination with one or more additional active ingredients at effective concentrations advantageously making it possible to offer a synergistic or reinforcing effect to achieve the desired effects described for the invention. The additional active ingredients may be chosen, for example, from lightening, anti-redness, anti-blemish, calming active ingredients for the treatment of sensitive, reactive skin, filtering radiation, in particular UVA, UVB, IR, from blue light, moisturizing, humectant, exfoliating, smoothing, toning, anti-aging, anti-wrinkle and fine lines active ingredients, improving mechanical and elastic properties, radiance of the complexion, detoxifying, anti-hair regrowth active ingredients, acting on the skin barrier, anti-acne, acting on sebum secretion, mattifying, unifying, anti-inflammatory, antioxidant, anti-radical, anti-glycant, eye contour (anti-dark circles and anti-puffiness), promoting blood circulation, peptides, vitamins, etc. These active ingredients may be obtained from plant materials, such as plant extracts or plant culture or fermentation products.

The "International cosmetic ingredient dictionary & handbook" published by "the Cosmetic, Toiletry, and Fragrance Association, Inc.", Washington, D.C. describes a wide variety, without limitation, of cosmetic and pharmaceutical ingredients commonly used in the skin care industry, which are suitable for use as additional ingredients in the compositions according to the present invention, as long as they are physically and chemically compatible with the other ingredients of the composition and especially with the actives of the present invention. Furthermore, the nature of these additional ingredients must not unacceptably alter the benefits of the actives of the invention. These additional ingredients may be synthetic or natural, such as plant extracts or come from a biofermentation process.

Other skin care active ingredients which are particularly useful in combination with the composition according to the invention can be found in the commercial documentation of Crodarom and Sederma, and on the website www.croda.fr.

More specifically, the Sichuan berry extract according to the invention can be combined with at least one of the compounds chosen from vitamin compounds, group B, C, E, F, D and A in particular, compounds such as niacinamide or tocopherol, retinoid compounds such as retinol, hyaluronic acid, hexamidine, α-lipoic acid, resveratrol, or DHEA, peptides, aluminum salts, phosphate salts, magnesium oxide, zirconium hydrochloride salts, which are conventional active ingredients used in cosmetic or dermo-pharmaceutical topical compositions.

The present invention also provides the use of an extract of Sichuan berries of the species Zanthoxylum bungeanum or Zanthoxylum simulans , as described above, for the manufacture of a composition for a cosmetic treatment, as also described above.

The composition for use according to the invention may be provided in any galenic form (examples are given further in the description) defined according to the destination of the composition and its site of application.

A composition according to the invention may be applied to the face, body, or décolleté, in any form or vehicle known to those skilled in the art, in particular in the form of a solution, dispersion, emulsion, paste or powder, individually or as a premix, or may be conveyed individually or as a premix by vectors such as macrocapsules, microcapsules or nanocapsules, macrospheres, microspheres, or nanospheres, liposomes, oleosomes or chylomicrons, macroparticles, microparticles or nanoparticles, macrosponges, microsponges or nanosponges, microemulsions or nanoemulsions, or adsorbed on powdery organic polymers, talcs, bentonites, spores or exines and other mineral or organic supports.

In cosmetics in particular, applications may be proposed, in particular in facial and/or body skin care ranges and makeup-care ranges, in particular eyelashes and eyebrows.

For example, the galenic form of the composition may be a lotion, a cream, a butter, a milk, a solid form, a foam, a gel, a deodorant, an antiperspirant, a shampoo, a conditioner, a hair mask, a face mask, a shower gel, etc.

For example, in the case of an anti-odor use, the galenic form of the composition may be a cream suitable for the face, feet, and/or the entire body, a spray, an aerosol, a roll-on or a deodorant stick. For a “holistic” product, the preferred galenic form may be a body lotion.

Galenic formulas may be included in ranges of personal care products and/or beauty products, in particular ranges of skin care, cleansing, makeup, makeup removal, anti-sun, artificial tanning, pre-shave, shaving, or after-shave, moisturizer, humectant, emollient, conditioner, exfoliant, astringent, depilatory, antiperspirant or antiperspirant, deodorant, air freshener, etc.

The composition may also be incorporated on a non-woven or woven material, made of natural or synthetic fibers, wool, or on any material intended to come into contact with the skin and which may be used in clothing, in particular tights and socks, shorts, day or night underwear, handkerchiefs, or fabrics, in order to exert its cosmetic effect via this skin/textile contact and allow continuous topical delivery (cosmetotextiles).

According to the invention, there is also provided a woven or non-woven material comprising at least one Sichuan berry extract, for use in a non-therapeutic cosmetic treatment.

DETAILED DESCRIPTION

1. Description des figures:

The present invention will be better understood in light of the detailed description of exemplary embodiments, studies and figures described below.

1. Description of figures:

represents a photograph to illustrate the anti-lipase test, showing an agar medium containing glyceryl tributyrate which has not been brought into contact with the extract according to the invention (control case).

1. Exemples d’obtention d’un extrait de baies de Sichuan selon l’invention et d’une composition en comprenant

represents a similar photograph, but having been placed in contact with the extract according to the invention.

1. Examples of obtaining a Sichuan berry extract according to the invention and a composition comprising it

Plant material: Sichuan berries of the species Zanthoxylum bungeanum.

Maceration solvent: Propylene glycol

Procedure: The berries are macerated in propylene glycol for 1 to 6 hours between 70 and 100 °C, then the macerate is filtered to remove solid debris from the plant. Several successive filtrations are carried out, with a size between 5 and 0.1 µm. The filtered macerate constitutes the berry extract according to the invention.

This procedure advantageously makes it possible to obtain a Sichuan berry extract rich in xanthoxyline, titrated between 10 and 20% by weight of xanthoxyline relative to the weight of the dry extract. The identification of this molecule is carried out by high-performance thin-layer chromatography (“HPTLC: High-performance thin-layer chromatography”).

1. Evaluations des différentes activités de l’extrait selon l’invention, par des tests in vitro et in vivo
   1. Odeurs corporelles
      1. Activité des lipases

To obtain a composition comprising the extract according to the invention, the filtered macerate obtained above is mixed with a physiologically acceptable matrix, consisting of a water-propylene glycol mixture. The water-propylene glycol ratio in the composition is between 0:100 and 30:70.

1. Evaluations of the different activities of the extract according to the invention, by in vitro and in vivo tests
   1. Body odors
      1. Lipase activity

Principle

Lipases are able to hydrolyze long-chain triglycerides of fatty acids into glycerol and the corresponding fatty acids, which are volatile and responsible for the bad odor.

Lipases are widely distributed in bacteria, yeasts, and filamentous fungi. They are produced by Gram-positive bacteria such as Corynebacterium, Propionibacterium, Cutibacterium, and Staphylococcus, as well as by Gram-negative bacteria such as Pseudomonas .

At the axillary level, the genus Corynebacterium is mainly responsible for the bad odor. Gram-negative bacteria hardly contribute to the axillary bad odor.

Protocol

The substrate glycerol tributyrate (fat-soluble) is hydrolyzed by lipases into glycerol and butyric acid (water-soluble). The fat-soluble substrate, mixed with the liquid medium or deposited on the agar, causes a cloudiness of the culture medium. When the substrate becomes water-soluble following the action of the lipase, this cloudiness of the culture medium disappears proportionally to the hydrolysis.

• une méthode en milieu liquide pour laquelle le trouble est mesuré à 620nm par spectrophotométrie.
• une méthode sur plaque de gélose, pour laquelle le trouble est évalué visuellement.

Two methods were used to demonstrate the anti-lipase activity of the extract according to the invention:

• a liquid medium method for which the turbidity is measured at 620nm by spectrophotometry.
• an agar plate method, for which the turbidity is assessed visually.

Results

1. Liquid medium analysis

Evolution of the lipase activity of Corynebacterium xerosis over time. Effect of a composition comprising 3% of the extract according to the invention compared to the control (n=3):

Control case 3% of the extract according to the invention Variation (%); significance 0 hours 0.00 ± 0.00 0.00 ± 0.00 0.0% ; dns* 48 hours 57.00 ± 0.06 43.70 ± 1.31 - 23.3%; p<0.0001 72 hours 77.40 ± 0.19 40.50 ± 0.20 - 47.7%; p<0.0001

*dns: non-significant data

1. Analyse en milieu gélosé

The results show that 3% of the extract according to the invention significantly reduces the lipase activity of Corynebacterium xerosis at 48 hours (-23.3%; p<0.0001) and at 72 hours (-47.7%; p<0.0001).

1. Analysis in agar medium

represents a photograph of the agar medium containing glycerol tributyrate which has not been brought into contact with the extract according to the invention (control case), after 48 hours at 37°C in anaerobic conditions.

represents a photograph of the agar medium containing glycerol tributyrate having been placed in contact with 3% of the extract according to the invention, after 48 hours at 37°C in anaerobic conditions.

By comparing these two figures, we observe that the cloudiness is less present in the agar medium containing 3% of the extract according to the invention compared to the control case, which means that the glyceryl tributyrate has been more hydrolyzed and therefore that the extract according to the invention reduces the lipase activity of Corynebacterium xerosis under the test conditions.

Conclusion

1. Protection de la flore bactérienne

Both methods, in liquid medium and in agar medium, demonstrate that the lipase activity of Corynebacterium xerosis is reduced by the presence of the extract according to the invention.

1. Protection of bacterial flora

Principle

Corynebacterium xerosis and Staphylococcus epidermis are important commensal strains of the axillary region. Staphylococcus epidermis is a beneficial probiotic strain of the skin and Corynebacterium xerosis is the main strain responsible for malodor.

The Minimum Inhibitory Concentration (MIC) is the lowest concentration of an antibacterial agent required to inhibit visible growth of the test strain. The higher this percentage, the less antibacterial activity the test product has.

Protocol

The culture medium is inoculated with the strains Corynebacterium xerosis and Staphylococcus epidermis, and different percentages of the extract according to the invention or of a market product known for its antibacterial action (Triclosan or Phenonip ^TM ). The test is then incubated for 48 hours at 32.5°C ± 2.5°C. After incubation, the presence or absence of a disorder is observed, the presence of a disorder revealing bacterial growth.

Results

Minimum inhibition concentration of the extract according to the invention, compared to two market products (n=3).

CMI Extract according to the invention Corynebacterium xerosis 14% S. epidermidis > 20% Triclosan Corynebacterium xerosis > 0.05% S. epidermidis 0.013% Phenonip ^TM (Phenoxyethanol (and) Methylparaben (and) Ethylparaben (and) Propylparaben (and) Butylparaben (and) Isobutylparaben) Corynebacterium xerosis 0.63% S. epidermidis 0.63%

The results show that Triclosan and Phenonip ^TM , two preservatives used in cosmetic products, have a very low MIC, respectively 0.013% and 0.63% on S.epidermidis while the extract according to the invention has a MIC greater than 20%.

Similarly, Phenonip ^TM shows a MIC of 0.63% on Corynebacterium xerosis while the extract according to the invention has a MIC of 14%.

Conclusion

1. Evaluation in vivo olfactive de la sueur

The extract according to the invention is therefore respectful of the skin microbiome, because a high concentration is required to obtain a result equivalent to 0.63% Phenonip ^TM or 0.013% Triclosan and alter the two most important microbial strains in the axillary region. The extract according to the invention therefore does not alter bacterial growth.

1. In vivo olfactory evaluation of sweat

Protocol

The study was conducted on 14 volunteers: 3 men and 11 women. Among the panelists of this trial, 5 experts participated in the study. They are trained and accustomed to the evaluation of odors and perfumes.

The intensity of underarm odor is assessed every hour, from 0 to 8 hours after the application of a composition comprising 3% of the extract according to the invention or a placebo. The test takes place over 3 days: the first day for odor removal, the second day with application of the placebo on one side and the composition comprising 3% of the extract according to the invention on the other side, and the reverse on the third day.

Volunteers were asked one day before the test and until the end of the third day not to eat spicy foods, onions or garlic and not to use perfumed products.

Once the product is applied, the odor is assessed every hour and a score is given for each armpit, ranging from 0 to 5: 0 (no pleasant odor), 1 (very slight pleasant odor), 2 (slight pleasant odor), 3 (normal odor), 4 (pleasant odor) and 5 (very pleasant odor).

Results

Variation of odor intensity over time. Effect of a composition containing 3% of the extract according to the invention compared to the same composition not containing the extract according to the invention (placebo):

Panel of volunteers (average) Expert panel (average) T= 0 hours T = 8 hours T= 0 hours T = 8 hours Placebo 4.7 ± 0.55 1.7 ± 0.71 4.5 ± 0.53 2.1 ± 0.53 3% of product according to the invention 4.9 ± 0.31 2.3 ± 0.94 5.0 ± 0.00 3.3 ± 0.00 Variation (%); significance - 4.6%; p <0.01 - 35.4%; p <0.01 - 11.1%; p <0.01 - 57.1%; p <0.01

The results show that for the panel of volunteers, the composition comprising 3% of the extract according to the invention significantly reduces bad axillary odor, 8 hours after application (-35.4%; p <0.01).

The results show that for the panel of experts, the composition comprising 3% of the extract according to the invention significantly reduces bad axillary odor, 8 hours after application (-57.1%; p <0.01).

Conclusion

1. Anti-vieillissement
   1. Synthèse d’élastine

These results show that the extract according to the invention reduces body odors at the axillary level.

1. Anti-aging
   1. Elastin synthesis

Principle

Elastin is a molecule of the extracellular matrix. The elasticity of the skin is modified during aging due to the decrease in the amount of elastin produced, due to an inappropriate assembly of elastin fibers.

Protocol

Once the cells reached confluence, they were placed or not (for control cases) in contact with the extract according to the invention in their culture medium, then incubated at 37°C and 5% _CO2 for 5 days. At the end of this contact, the cell mats were rinsed, fixed and labeled with an anti-elastin antibody. The labeling was revealed using a fluorescent secondary antibody and photos captured under a microscope. An image analysis of these photos made it possible to quantify the production of elastin. A counterlabeling of the nuclei was carried out using the fluorescent dye Hoesch, labeling DNA, to evaluate the cell population and thus weight the fluorescence data obtained.

Results

Variation in elastin production by fibroblasts. Effect of the extract according to the invention at 0.14% compared to the control (n=3):

Elastin
(Average area / number of cells) Variation (%); significance Control 103 ± 17 Reference 0.14% of the extract according to the invention 249 ± 13 + 141%; p<0.01

1. Renforcement de la jonction dermo-épidermique

The results show that 0.14% of the extract according to the invention significantly increases by 141% (p<0.01) the production of elastin by fibroblasts. The extract according to the invention can thus improve skin aging linked to a loss of skin elasticity.

1. Strengthening of the dermo-epidermal junction

Principle

The DEJ ensures cohesion between the epidermis and the dermis. During aging, there is a decrease in the synthesis of these components (notably collagen VII and laminins). The aging of the DEJ has repercussions on the resilience of the skin and the loss of its dynamism.

Protocol

Human keratinocytes (KH) were cultured to sub-confluence and then were placed or not (for control cases) in contact with the extract according to the invention. After this contact, the synthesis of collagen VII and laminins were evaluated using ELISA type kits. An estimation of the quantity of cells by Hoechst method made it possible to standardize the results.

Results

Variation in the production of collagen VII and laminins by keratinocytes. Effect of the extract according to the invention at 0.26% compared to the control (n=5):

Collagen VII
(µg/mL/10 ⁶ cell.) Variation (%); significance Control 5.8 ± 0.3 Reference 0.26% of the product according to the invention 8.5 ± 0.4 + 47%; p<0.01

The results show that 0.26% of the product according to the invention significantly increases by 47% (p<0.01) the production of collagen VII in keratinocytes.

Variation in the production of laminins by keratinocytes. Effect of the extract according to the invention at 0.26% compared to the control (n=5):

Laminin
(µg/mL/10 ⁶ cell.) Variation (%); significance Control 32 ± 2 Reference 0.26% of the product according to the invention 51 ± 3 + 58%; p<0.01

The results show that 0.26% of the product according to the invention significantly increases by 58% (p<0.01) the production of laminins in keratinocytes.

Conclusion

These results show that the extract according to the invention has a direct action of reinforcing the JDE by stimulating the synthesis of laminins and collagen VII.

1. Capacité antioxydante

The extract according to the invention can thus improve skin aging linked to a disorganization of the DEJ, by counteracting the loss of suppleness and elasticity that it causes.

1. Antioxidant capacity

Principle

Oxidative stress plays a central role in the skin's response to various stresses. Free radicals (H ₂ O ₂ , OH•, O ₂ -, O ₂ , ¹ O ₂ …) cause damage to proteins, lipids and DNA, leading to premature aging of the skin.

Protocol

For the DPPH test: DPPH (2,2-diphenyl-1-picrylhydrazyl) is a stable free radical that is widely used for the detection of free radical scavengers. This molecule, by losing its radical character, is converted into DPPH-H (2,2-diphenyl-1-picrylhydrazine). This conversion is accompanied by a discoloration by redox, from violet to yellow, which can be followed over time by spectrophotometry at 490nm.

For the singlet _O2 test: A singlet _O2 generating system receives product according to the invention. To measure the evolution, the degradation of uric acid is monitored at 292nm by spectrophotometry.

Results

Variation in DDPH production (n=6). Effect of the extract according to the invention at 2% and 3% compared to the control:

DDPH
(Average of optical density variation) Variation (%); significance Control 0.268 ± 0.005 Reference 2% of product according to the invention 0.101 ± 0.037 - 62%; p<0.01 3% of product according to the invention 0.082 ± 0.059 - 69%; p<0.01

The results show that the product according to the invention significantly reduces the presence of DPPH.

Variation in singlet _O2 production (n=2). Effect of the extract according to the invention at 2% and 3% compared to the control:

O ₂ singlet
(Average of optical density variation) Variation (%); significance Control 32.33 ± 0.81 Reference 2% of product according to the invention 20.02 ± 0.95 - 38%; p<0.01 3% of product according to the invention 14.87 ± 1.49 - 54%; p<0.01

The results show that the product according to the invention significantly reduces the presence of singlet _O2 .

Conclusion

1. Glycation non enzymatique de la BSA

The extract according to the invention has a strong antioxidant capacity, making it possible to effectively combat premature aging of the skin.

1. Non-enzymatic glycation of BSA

Principle

Glycation of proteins with reducing sugars in the skin is also responsible for skin aging.

Indeed, the formation of numerous glycated proteins whose functional, enzymatic and structural properties are altered, has consequences on the proper functioning of the cell or the organism.

This results in altering the mechanical and elastic properties of the extracellular matrix of the dermis, which becomes less flexible, more rigid, but also more flaccid and less reactive, also reflected by a dull complexion.

Protocol

The study of non-enzymatic glycation is carried out between a model protein, serum albumin, which serves as a target and an edible reducing sugar from fruits. The protein is progressively glycated (linked to the sugar) irreversibly, in the presence or absence of the product according to the invention. This modification is monitored by fluorescence.

Results

Variation of glycation (n=2). Effect of the extract according to the invention at 2% and 3% compared to the control:

Non-enzymatic glycation
(Average of the variation of the fluorescence unit) Variation (%); significance Control 84540 ± 1803 Reference 2% of product according to the invention 7926 ± 403 - 92%; p<0.01 3% of product according to the invention 5304 ± 667 - 95%; p<0.01

The results show the strong anti-glycating potential of the product according to the invention.

Conclusion

1. Peaux grasses et/ou à tendance acnéique
   1. Astringence

The extract according to the invention has an anti-glycating power which also helps combat skin aging.

1. Oily and/or acne-prone skin
   1. Astringency

Principle

• Resserrer les pores dilatés et réduire la sécrétion excessive d'huile et de sueur ;
• Enlever l'excès de sébum ;
• Précipite les acides aminés provoquant une mauvaise odeur corporelle. Ces acides aminés sont formés de cornéocytes, protéines secrétées ou d’acides aminés libres issus du catabolisme bactérien.

An astringent substance, capable of shrinking or tightening body tissues by aggregation, is perfectly suited for:

• Tighten enlarged pores and reduce excessive oil and sweat secretion;
• Remove excess sebum;
• Precipitates amino acids that cause bad body odor. These amino acids are formed from corneocytes, secreted proteins or free amino acids from bacterial catabolism.

The in tubo test to determine the astringency capacity of a product is based on the tannin-polymer interaction which results in the formation of insoluble tannin-polymer complexes which then precipitate. The difference in optical density (OD) before and after precipitation corresponds to the astringency capacity of the product.

Protocol

The extract according to the invention on the one hand, and red wine on the other hand, are incubated with and without methylcellulose and the optical density at 280nm is measured by spectrophotometer. The precipitated part of the extract corresponds to the astringent tannic fraction.

Results

Astringency of the extract according to the invention compared to the astringency of red wine (n=4 for the extract according to the invention and n=12 for red wine):

Medium Methylcellulose Precipitated Tannins (MCP)
(ppm epicatechin equivalent) % Variation Tannic red wine 1,334 ±222 Reference Extract according to the invention 5,276 ± 161 295%*

1. Anti-séborrhéique

Tannic red wine is considered as the positive control of astringency. Its astringency is measured at 1334 ppm. The product according to the invention shows a statistically different astringent capacity of 295% higher than that of tannic red wine.

1. Anti-seborrheic

Principle

The skin has many pores on its surface whose function is to evacuate excess sebum and impurities from the skin such as dead cells and sweat. Oily skin is associated with excessive sebum production by sebocytes. Too much sebum causes changes in the properties of the skin and scalp, for example by increasing the formation of pimples, blackheads and by clogging the pores which will then dilate, become more visible and make the skin texture irregular and/or lead to excessive growth of bacteria responsible for dandruff such as fungi of the genus Malassezia on the scalp.

An anti-seborrheic cosmetic active ingredient will counteract this development by reducing sebum production; which will have the effect of tightening the pores of the skin, smoothing it and reducing the oily/shiny appearance with an irregular grain, characteristic in particular of oily skin and/or making the scalp healthier, with less dandruff and itching associated with it.

Protocol

Sebocytes were seeded in their growth medium. At confluence, the cells are placed in contact or not (control case) with the product according to the invention for 48 hours. After removal of the media, the cell lawns are incubated with neutral red, a marker of intracellular lipids, which makes it possible to estimate the quantity of lipids present in the cells by measuring the fluorescence emitted. The viability estimate is carried out in parallel on the same lawns using a fluorescent dye.

Results

Variation of lipid synthesis by sebocytes. Effect of the extract according to the invention at 2% compared to the control (n=2):

(UFA/10 ⁶ cells) Variation (%); significance Control 160962 ± 6691 Reference 2% of product according to the invention 77026 ± 5163 - 52% ; p<0.01

The results show that exposing sebocytes to 2% of the extract according to the invention makes it possible to reduce the quantity of lipids in sebum-producing cells.

Conclusion

1. Hydratation de la peau
   1. Synthèse d’acide hyaluronique

The extract according to the invention can therefore be used to treat skin disorders linked to oily or oily skin such as shiny, glossy appearance, size and number of pores, to give the skin a smoother, more uniform, more harmonious appearance and/or make the scalp healthier, with less dandruff and itching linked to it.

1. Skin hydration
   1. Synthesis of hyaluronic acid

Protocol

Human keratinocytes were cultured to sub-confluence and then placed in contact with the ingredient according to the invention. Following this contact, the synthesis of hyaluronic acid was assessed using an ELISA-type assay. An estimation of the quantity of cells by the Hoechst method made it possible to homogenize the results.

Results

Variation in hyaluronic acid production by keratinocytes. Effect of the extract according to the invention at 0.26% compared to the control (n=5):

Hyaluronic acid
(µg/mL/10 ⁶ cell.) Variation (%); significance Control 340 ± 37 Reference 0.26% of the product according to the invention 1215 ± 106 + 257 ; p<0.01

1. Différentiation des kératinocytes

The results show that the product according to the invention significantly increases the synthesis of hyaluronic acid in keratinocytes.

1. Differentiation of keratinocytes

Protocol

Human keratinocytes were cultured to sub-confluence and then placed in contact or not with the product according to the invention. Differentiation is monitored visually by observing the phenotype of the keratinocytes over 4 days.

Results

Condition Pro-differentiating effect observed after 4 days Control case 0 3% of the product according to the invention ++

Visually, in the control case, the untreated cells show a carpet of joined keratinocytes with well-marked cellular contours whereas in the presence of 3% of the product according to the invention. The cells retract and the contacts between the cells decrease until there are empty spaces between the cells, an aspect characteristic of advanced differentiation of the keratinocytes.

Conclusion

1. Apaisement de la peau

The extract according to the invention increases the differentiation of keratinocytes participating in better protection of the epidermis and better hydration.

1. Skin soothing

Principle

Inflammatory mediators are very present in microinflammatory phenomena. Reducing the presence of these mediators results in reducing the sensations of discomfort in sensitive skin and redness.

Protocol

Normal human dermal fibroblasts (NHDF) were cultured until a confluent lawn was obtained. At this stage, they were placed in contact with the test products for 24 hours, then the lawns were irradiated with UVB and placed in contact with the test products again for 24 hours. The amounts of PGE ₂ and IL-6 synthesized were measured in the culture supernatants by ELISA assay. The number of cells was evaluated in order to be able to normalize the data. A variance study and a Student t test for paired series were performed in order to judge the significance of the results.

Results

Variation in IL-6 and PGE ₂ production in fibroblasts. Effect of the extract according to the invention at 3% compared to the control (n=3):

IL-6 (pg/10 ⁶ cell) PGE ₂ (pg/10 ⁶ cell) Not irradiated Control 1244 ± 32 1613 ± 49 3% of product according to the invention 627 ± 54 334 ± 35 Variation (%); significance -50%; p<0.01 -79%; p<0.01 Irradiated by UVB Control 9238 ± 323 8539 ± 683 3% of product according to the invention 3399 ± 181 1053± 208 Variation (%); significance -63%; p<0.01 - 88%; p<0.01

Thus, advantageously the extract according to the invention strongly and significantly reduces the 2 pro-inflammatory messengers.

Conclusion

1. Exemple de formulation galénique

These results show that the extract according to the invention reduces micro-inflammations of the skin. The extract according to the invention can therefore be used to soothe skin discomforts of sensitive skin such as redness, tightness, etc.

1. Example of galenic formulation

Various cosmetic formulations are described below, containing the extract according to the invention, in particular as formulated in the active ingredient described in point B. above.

Additional active ingredients, where appropriate supporting and/or complementing the activity of the extract according to the invention, may be added in the appropriate phase according to their hydrophobic, hydrophilic or amphiphilic character. These ingredients may be of any category according to their function(s), the place of application (body, face, neck, bust, hands, hair, eyelashes, eyebrows, hair, etc.), the desired final effect and the targeted consumer, for example anti-aging, anti-wrinkle, moisturizing, anti-dark circles, firming, anti-glycation, slimming, soothing, muscle relaxant, anti-redness, sensitive skin, anti-stretch marks, mattifying, etc.

Active ingredient according to the invention: Sichuan berry extract according to the invention in a water-propylene glycol matrix.

This ingredient is recommended in a composition at a rate of 1 to 5%, preferably a rate of 3%. It goes without saying that these rates could vary without departing from the scope of the present invention, depending on the more or less pronounced effects sought.

Examples of formulations are described below. Additional active ingredients may support and/or complement the activity of the active ingredient according to the invention. These ingredients may be of any category depending on their function(s), the place of application (body, face, neck, bust, hands, etc.), the desired final effect and the targeted consumer.

Example 1: Roll-on Deodorant

Raw materials % INCI name Phase A SP CITHROL GMS 40 MBAL 3.9 Glyceryl stearate SP CRODACOL C90 MBAL 1.9 Cetyl Alcohol SP BRIJ CS12 MBAL 1.2 Ceteareth-12 SP BRIJ CS20 MBAL 1.2 Ceteareth-20 Cetiol V 3.9 Decyl Oleate Phase B Aqua 63.6 Water Vegetable Glycerin PH 4.9 Glycerin Kathon CG 0.1 Methylchloroisothiazolinone Phase C REACH 103 19.4 Aluminum chlorohydrate Phase D Extract according to the invention 3.0

Operating mode

Weigh phase A and heat. Weigh phase B, homogenize and then heat. Pour phase B into phase A, while stirring. Weigh phase C. Add phase C to phase B+A, while stirring. Leave to cool. Weigh phase D and add it to the cooled phase C+B+A, while stirring.

Example 2: Deodorant spray

Raw materials % INCI name Phase A Ethanol 96% 60.4 Alcohol Phase B SP Crodamol IPM MBAL 1.5 Isopropyl Myristate Phase C D-Panthenol 0.1 Panthenol Phase D Extract according to the invention 3.0 Phase E Propellent gas 35

Operating mode:

Weigh phase A, and add phase B, while stirring. Add phase C to phase B+A, while stirring. Add phase D to phase C+B+A, while stirring. In a container, add phase E to phase D+C+B+A.

Example 3: Body and face balm

Raw materials % INCI name Phase A Aqua QSP Water Potassium sorbate 0.10 Potassium sorbate Phase B Incroquat™ Behenyl TMS-50 4.00 Behentrimonium Methosulfate (and) Cetyl Alcohol (and) Butylene Glycol Crodacol™ CS90 3.00 Cetearyl Alcohol Crodamol™ CP 6.00 Cetyl Palmitate Crodamol™GTIS 4.00 Triisostearin NG Shea Unsaponifiable™ 0.50 Butyrospermum Parkii (Shea) Butter (and) Butyrospermum Parkii (Shea) Butter Unsaponifiables Dl Alpha Tocopherol 0.20 Tocopherol Phase C Glycerin 5.00 Glycerin Phenoxyethanol 0.80 Phenoxyethanol Phase D Extract according to the invention 2.00 Phase E Aqua 1.00 Water Sodium Hydroxide 30% 0.10 Sodium Hydroxide

Operating mode:

Weigh phase A, and heat to 85°C in a water bath. Weigh phase B, and heat to 85°C in a water bath. Add phase B to phase A, while stirring vigorously. Weigh phase C, and add it to phase A+B while stirring. Add phase D to phase A+B+C, at approximately 45°C, and homogenize. Weigh phase E and adjust the pH with it.

Examples of other ingredients that can be added to these formulations:

Crodarom® Nordic Cotton: active ingredient marketed by Crodarom, to help strengthen the skin's barrier function, soothe the most sensitive skin, moisturize and protect hair.

Phytessence ^TM Blue Daisy: Ingredient marketed by Crodarom, soothes sensitive skin and reduces feelings of discomfort by acting on the synthesis of inflammation mediators.

Crodarom® Bamboo Tea EC: ingredient marketed by Crodarom, stimulates cellular metabolism while maintaining the skin's hydration level, protects the skin from external aggressions.

Claims (8)

Use of an extract of Sichuan berries of the species Zanthoxylum bungeanum or Zanthoxylum simulans for non-therapeutic cosmetic treatment of the skin and its appendages. Use according to claim 1, characterized in that the treatment is topical. Use according to claim 1 or 2, characterized in that the treatment is chosen from:
- treatment of body odors; and/or
- treatment of oily skin including the scalp; and/or
- an anti-aging treatment; and/or
- a moisturizing treatment. Use according to one of the preceding claims, characterized in that the treatment allows is an anti-aging treatment to prevent and/or treat the loss of elasticity and suppleness of the skin. Sichuan berry extract of the species Zanthoxylum bungeanum or Zanthoxylum simulans for dermatological treatment of the skin and its appendages. Sichuan berry extract according to claim 5, for treatment of acne-prone skin. Sichuan berry extract according to claim 5, for anti-dandruff treatment. Sichuan berry extract according to claim 5, for a soothing treatment.