Cleome Droserifolia Extract and its Therapeutic or Cosmetic Use

Abstract

The invention relates to an extract of Cleome Droserifolia intended for topical application to the skin and / or mucous membranes. The invention also relates to the use of such an extract as well as a method of obtaining it and the compositions including it.

Description

Title of the invention: Extract of Cleome Droserifolia and its therapeutic or cosmetic use

[0001] The invention relates to an extract of Cleome droserifolia intended for topical application to the skin and / or mucous membranes having a therapeutic or cosmetic effect. The invention also relates to the use of such an extract as well as a method of obtaining it and the compositions including it.

[0002] The skin constitutes the envelope of the human body, in continuity with the mucous membranes which cover the cavities of the organism.

[0003] The skin and mucous membranes form a protective barrier against the external environment and perform many vital functions. They are regularly attacked and can be subject to various pathologies.

[0004] To deal with skin disorders, there are specific active ingredients and compositions aimed at treating a given skin pathology or a particular unsightly manifestation. Regarding therapeutic or dermatological products for topical application, a majority of them are synthetic products and are not of natural origin. As for cosmetic products, there are products of natural origin but these are not always effective.

[0005] Also, an objective of the invention is to propose a solution of natural origin which can be used effectively in topical application for the treatment of numerous pathological disorders on diseased skin and / or mucous membranes but also for cosmetic effects on healthy skin.

[0006] To answer this, the invention proposes to use an extract of Cleome droserifolia.

[0007] Cleome droserifolia is a natural plant present in large quantities in several countries of the Near and Middle East.

[0008] However, surprisingly: - extracts obtained with a solvent comprising at least water, namely: * aqueous extracts (solvent consisting exclusively of water) or * obtained using water-based solvent(s) (hydroalcoholic solvent or solvent consisting of a mixture of water and organic solvent), or - extracts obtained by supercritical CO2 extraction, have powerful effects on many skin or mucous membrane pathologies but also cosmetic effects when applied to healthy skin, in particular an antioxidant effect which can act as an anti-aging agent.

[0009] The invention therefore specifically relates to an extract of Cleome droserifolia intended for topical application (for therapeutic or cosmetic purposes) on the skin and / or mucous membranes, said extract being an aqueous, hydroalcoholic extract or obtained using a solvent comprising water and an organic solvent.

[0010] Advantageously, the extract according to the invention comes from a little-used plant, available at low cost over a large area extending over several countries, without requiring fertilizers, pesticides or any other chemical product. In addition, water is the main solvent or even the only solvent used to obtain the extracts according to the invention, so that the extract according to the invention is used in green chemistry products. This is also the case in the case of supercritical CO2 extraction. Furthermore, Cleome droserifolia is not toxic.

[0011] Another advantage of the active extract according to the invention is that the product is highly soluble in water so that it can be formulated in various formulations for topical application.

[0012] The invention also relates to this extract for its use in topical application in the treatment of the skin and / or mucous membranes, in particular for an anti-coagulant and / or healing and / or anti-oxidant and / or anti-inflammatory and / or anti-itching and / or anti-bacterial and / or anti-histamine effect.

[0013] The invention also relates to the cosmetic use of this extract on healthy skin for cosmetic applications, in particular anti-aging and / or restorative effect of healthy skin after exposure to UV.

[0014] Another subject of the invention consists of compositions comprising at least 0.1% by weight of such an active extract, as well as the therapeutic (dermatological in particular) or cosmetic uses of these compositions.

[0015] Finally, the invention relates to a specific process for obtaining an extract according to the invention, comprising at least the implementation of the following steps: - drying then grinding of aerial parts of Cleome droserifolia, - extraction: aqueous extraction (with a solvent consisting exclusively of water) or using a hydroalcoholic solvent or using a solvent consisting of a mixture of water and organic solvent or supercritical CO2 extraction, with a proportion of Cleome droserifolia of between 5 and 20% and a proportion of solvent of between 80 and 95%, the percentages being given by weight relative to the total weight of the mixture of dried and ground aerial parts of Cleome droserifolia and solvent, - filtration to remove suspended particles, - obtaining a liquid extract.

[0016] The invention is now described in detail with regard in particular to non-limiting examples, test results and the following appended figures:

[0017] [fig.la] which is a photograph of a severed finger, taken at D0 at 1:36 a.m. on which a composition comprising an extract according to the invention has just been applied.

[0018] [fig.lb] which is a photograph of a view of the same finger as Figure 1a taken at 0100 at 11:50.

[0019] [fig.lc] which is a photograph of another view of the same finger as Figures 1a and 1b, taken at 00 at 11:50.

[0020] [fig.ld] which is a photograph of a view of the same finger as Figures 1a, 1b and 1c, taken at D0 at 8:05 p.m.

[0021] [fig.1e] which is a photograph of a view of the same finger as Figures 1a, 1b, 1c and 1d, taken on D+1 at 8:44 a.m.

[0022] [fig.If] which is a photograph of a view of the same finger as Figures 1a, 1b, 1c, 1d and 1e, also taken at D+l at 8:44 a.m. from a different angle than Figure 1d.

[0023] [fig.2a] which is a photograph of a forearm presenting urticaria, taken on D0 at 3:44 p.m. on which a composition comprising an extract according to the invention has just been applied.

[0024] [fig.2b] which is a photograph of the same forearm as that in Figure 2a, taken at D0 at 3:56 p.m.

[0025] [fig.2c] which is a photograph of the same forearm as that of Figures 2a and 2b, taken at D0 at 4 p.m.

[0026] [fig.2d] which is a photograph of the same forearm as that of Figures 2a, 2b and 2c, taken at D0 at 5:22 p.m.

[0027] [fig.2e] which is a photograph of the same forearm as that of Figures 2a, 2b, 2c and 2d, taken at D0 at 19hl3.

[0028] [fig.3] which is a photograph of a lyophilisate obtained by lyophilization of a liquid extract according to the invention.

[0029] Definitions

[0030] By “extract” of Cleome droserifolia within the meaning of the invention, we mean at least one molecule, preferably a set of molecules obtained from Cleome droserifolia.

[0031] By "therapeutic effect" or "therapeutic use" within the meaning of the invention is meant a curative or preventive effect or a use in treatment or prevention, to treat a pathology, a pathological state or a symptom of a pathological state or a symptom of a pathology. It may be a dermatological effect or use in the case of application to diseased skin.

[0032] For the purposes of the invention, the term “aqueous extract” means an extract obtained from a solvent consisting exclusively of water.

[0033] For the purposes of the invention, the term “hydroalcoholic extract” means an extract obtained from a solvent consisting of water and at least one alcohol.

[0034] By "healthy skin" within the meaning of the invention, we mean skin which is not diseased or which does not have pathological disorders, but which may possibly contain non-pathological unsightly manifestations such as wrinkles, fine lines, dryness, pigmentation, redness, etc. Detailed description of the invention

[0035] According to a first object, the invention relates to an extract of Cleome droserifolia, intended for topical application to the skin and / or mucous membranes, said extract being an extract obtained with a solvent comprising at least water and / or obtained by supercritical CO2 extraction. Preferably, it is an aqueous, hydroalcoholic extract or obtained using a solvent comprising water and an organic solvent.

[0036] The extract according to the invention is therefore obtained by an extraction carried out with one or more water-based solvent(s) consisting of: - water alone (aqueous extraction), or - a mixture of water and alcohol (hydroalcoholic extraction), preferably a mixture consisting of 60 to 80% water and 20 to 40% alcohol by volume; the alcohol is preferably chosen from ethanol, methanol and isopropanol, preferably ethanol, or - or water + organic solvent mixture, preferably a mixture consisting of 60 to 80% water and 20 to 40% organic solvent by volume; the organic solvent is preferably chosen from triols such as glycerol, or esters such as ethyl acetate.

[0037] Advantageously, the use of such solvents has no negative impact on the environment and human beings, and makes it possible to obtain extracts exhibiting very high efficacy in the curative or preventive treatment of healthy or diseased skin and / or mucous membranes.

[0038] The extract according to the invention can also be obtained by supercritical CO2 extraction. Supercritical fluid extraction is a process for extracting a solute from a substance using a supercritical fluid as an extraction solvent. This process can be compared to a liquid extraction process. Supercritical carbon dioxide (CO2) is the most widely used fluid. Its advantages are mainly its non-toxicity and chemical inertness. During extraction, the conditions for using supercritical carbon dioxide are above its critical points (critical temperature 31°C and critical pressure 73 atm). The carbon dioxide passes into an intermediate state between the liquid state and the gaseous state, called the "supercritical" state. In this state, the CO2 effectively solubilizes certain categories of active ingredients. By restoring the initial pressure, the CO2 returns to the gaseous state and releases the desired ingredients.Low operating temperatures preserve all active molecules. Supercritical CO2 extraction can be used alone or combined with hydro-ethanolic extraction, accompanied by co-solvents such as ethanol or methanol. The action is complementary if certain desired active ingredients have a high polarity. This efficient extraction technique is environmentally friendly because the low operating temperatures preserve all of the active molecules.

[0039] The extract according to the invention can be presented in any form: - in liquid form, - in the form of a water-soluble resinous gum, or - in the form of a lyophilisate (freeze-dried powder), soluble, which can be solubilized in water

[0040] These different forms allow different uses, in different types of compositions, depending on the intended uses. In addition, they allow optimal effectiveness of the active ingredient according to the invention and to preserve its initial properties.

[0041] Preferably, the Cleome droserifolia extract according to the invention is an extract obtained from aerial parts of Cleome droserifolia, and in particular from leaves and / or bulbs containing the seeds.

[0042] The extract according to the invention can be obtained by any suitable process using a solvent comprising water and / or by supercritical CO2 extraction. According to a particularly suitable embodiment, the extract according to the invention is obtained by a process comprising at least the implementation of the following steps: - drying then grinding of aerial parts of Cleome droserifolia; - aqueous extraction or using a hydroalcoholic solvent or using a solvent consisting of a mixture of water and organic solvent, with a proportion of Cleome droserifolia of between 5 and 20% and a proportion of solvent (water or mixture of water and alcohol or mixture of water and organic solvent) of between 80 and 95%, the percentages being given by weight relative to the total weight of the mixture of dried and ground aerial parts of Cleome droserifolia and solvent, - filtration to remove suspended particles, - obtaining a liquid extract.

[0043] The drying and grinding step can be carried out by any suitable method known to those skilled in the art. Preferably, the leaves, if possible separated from the bronchioles, are subjected to fine grinding.

[0044] Drying can be carried out according to conventional methods for drying plant raw materials.

[0045] The grinding is preferably carried out to a particle size preferably less than 2 mm.

[0046] If the extraction is an aqueous extraction, it can be carried out: - by decoction at room temperature for 24 to 48 hours, or - by infusion in water at a temperature between 40 and 60°C, then infusion and decantation for at least 24 hours at a temperature between 2 and 8°C, or - by infusion in water at a temperature of at least 100°C for 4 to 10 minutes, then infusion and decantation, preferably for at least 24 hours at a temperature between 2 and 8°C; this last extraction method makes it possible to reduce the level of microorganisms present in the extract from the raw material.

[0047] The filtration step to remove suspended particles is preferably carried out with a filtration of between 1 and 1000 μm, in particular between 100 and 200 μm. It can be carried out on standard filter paper. The filter or filter mesh (cloth adapted to the size of the filter) can be made in particular of Polyethylene (PE), Polypropylene (PP), Polyamide / nylon (PA), Polyester (PES), Fluorinated polymer type PTFE (Teflon), PVDF (Polyvinylidene fluoride), E-CTFE or PEEK (Poly EtherEtherKetone).

[0048] Industrial filtration fabrics and filter mesh can be made from several types of materials, synthetic materials as above but also stainless steel.

[0049] After the filtration step, the method may also comprise the following steps: - possibly a storage step either between 2 and 8°C, or at a temperature below -20°C until use, - a sterilizing filtration step before use, for example sterilizing filtration under vacuum using a 0.2 pm filter. This may include ultrafiltration and / or nanofiltration to possibly allow the retention of viruses.

[0050] The extract obtained is a liquid extract. The method may comprise additional steps for obtaining an extract in the form of a resin or an extract in the form of a lyophilisate, from this liquid extract.

[0051] Thus, if the extract is in the form of a resinous gum, the method preferably also comprises a step of drying the liquid extract. This drying step can be carried out at a temperature between room temperature and 80°C. Preferably, the liquid extract is subjected to hot air drying at a temperature between 45 and 80°C. The drying can be carried out, for example, in a closed enclosure equipped with a hot air evacuation system. After evaporation of the liquids contained in the extract, a blackish resinous gum is obtained. This resin or resinous gum is soluble in water.

[0052] If the extract is in the form of a lyophilisate, the process preferably comprises also includes a freeze-drying step of the liquid extract. Freeze-drying can be carried out by implementing the following steps: - freezing the liquid extract at a temperature between -20°C and -40°C for 24 to 72 hours, - freeze-drying for 100 to 150 hours at a temperature between -20°C and -80°C and a pressure between 0.10 mbar - 0.40 mbar. The freeze-drying product is preferably in the form of a yellow powder cake in the form of needle-like crystals. The lyophilized form of the active ingredient according to the invention is soluble in water. The apparent solubility is preferably greater than 43 mg / g.

[0053] Thus the extract according to the invention is preferably presented either in liquid form (before or after filtration), or in the form of a dry extract (resin or lyophilisate).

[0054] The extract according to the invention can be used in a composition but it can also be used as such, either in the form of a liquid extract, or in the form of a lyophilized powder or resin.

[0055] The invention therefore also relates to a composition comprising an extract of Cleome droserifolia, in particular a composition comprising at least 0.1% by weight of an extract according to the invention, preferably comprising between 0.1% and 5%.

[0056] The compositions according to the invention are suitable for topical application. They are either a medicament, a dermatological composition, or a cosmetic composition.

[0057] The compositions according to the invention comprise, in addition to the active ingredient obtained from Cleome droserifolia, a medium suitable for use on the skin and / or mucous membranes.

[0058] Advantageously, the high solubility in water of the Cleome droserifolia extracts according to the invention makes it possible to produce galenic formulations to combat infections of the mucous membranes such as the oral mucosa, the vaginal mucosa, the anal mucosa and the skin.

[0059] The compositions according to the invention may be presented in different galenic forms suitable for topical application to the skin and / or mucous membranes. They may in particular be a cream, an ointment, a gel, a paste, a lotion, an ointment, a toothpaste, a soap, a mouthwash, a shampoo, a shaving foam, a patch or dressing.

[0060] The compositions according to the invention, in addition to the active ingredient, also comprise a physiologically acceptable medium.

[0061] The extract according to the invention and the compositions including it have remarkable properties, on the one hand, on healthy skin and / or mucous membranes to improve the appearance aesthetics of the skin and, on the other hand, on skin which presents a pathological disorder (burn, wound, itching, canker sores etc.).

[0062] In particular, the extract according to the invention presents: - an antioxidant activity, - plasma coagulant activity, - anti-bleeding activity, - a healing activity, - antibacterial activity, in particular inhibitory activity against fast-growing mycoplasma, - antihistamine activity - anti-itch activity, - anti-inflammatory activity.

[0063] Thus, the invention also relates to an extract comprising an extract of Cleome droserifolia according to the invention or a composition including it for its use in topical application for the treatment of the skin and / or mucous membranes, and in particular for an anti-coagulant and / or healing and / or anti-oxidant and / or anti-inflammatory and / or anti-itching and / or anti-bacterial and / or anti-histamine effect.

[0064] An extract according to the invention or a composition according to the invention can in particular be used: - as an active ingredient or anti-inflammatory composition for the skin and mucous membranes, and / or - as an active ingredient or composition for soothing and / or anti-itching the skin and mucous membranes, and / or - as an active ingredient or healing and / or coagulant composition for cutaneous, oral, vaginal or anal use, and / or - as an active ingredient or composition for skin hygiene and / or intimate hygiene (oral, vaginal, anal), in particular to prevent infections such as mycoplasmas, and / or - as an active ingredient or anti-allergic composition.

[0065] The extract according to the invention or the compositions including it can therefore be used for a therapeutic effect in the treatment of various pathological skin disorders and in particular in the treatment of urticaria, infections of the skin and / or mucous membranes, mycoses of the skin and / or mucous membranes, atopic dermatitis, psoriasis, acne, rosacea, herpes, mouth ulcers, burns, insect bites, sunburn burns, itching and / or wounds.

[0066] Furthermore, on healthy skin, the extract according to the invention has an effect which improves the aesthetic appearance of the skin. In particular, its antioxidant effect allows it to be used for numerous cosmetic applications and in particular for anti-aging effects, i.e. to prevent or combat the appearance of unsightly signs due to skin aging, such as wrinkles, fine lines, dry skin, etc.

[0067] An object of the invention is therefore the cosmetic use of an extract of Cleome dro-serifolia according to the invention or of a composition containing it, on healthy skin, in topical application, in particular for an anti-aging cosmetic effect and / or to reduce redness (anti-redness effect) of the skin caused by exposure to the sun or UV rays.

[0068] In order to illustrate the cosmetic effects on the skin and mucous membranes of active ingredients comprising extracts obtained from Cleome droserifolia, examples and test results are presented below. Examples

[0069] Example 1: example of active ingredient according to the invention in liquid form (aqueous extract)

[0070] The aerial part of Cleome droserifolia is collected and dried. The leaves are separated from the bronchioles to be subjected to fine grinding.

[0071] An aqueous extract is produced. The extraction is carried out for 36 hours in water at room temperature.

[0072] The proportion of dried and ground raw materials is 10% for 90% water mass / mass. We obtain a brown-colored infusion with aromatic flavors.

[0073] The decoction is then subjected to filtration on a standard paper filter in order to eliminate suspended particles.

[0074] The aqueous extract is stored either in a refrigerator between 2 and 8°C or in a freezer at -20°C until use. Before use, the aqueous extract is subjected to sterilizing filtration under vacuum using a 0.2 pm Nalgen type filter.

[0075] The liquid extract obtained is soluble in water.

[0076] Example 2: example of active ingredient according to the invention in liquid extract form aqueous)

[0077] The aerial part of Cleome droserifolia is collected and dried then finely ground.

[0078] An aqueous extract is produced. The extraction is carried out by infusion in hot water brought from 40°C to 60°C, then infusion and decantation for 24 hours in the refrigerator at 4°C.

[0079] The proportion of dried and ground raw materials is 12% for 88% water mass / mass. We obtain a brown-colored infusion with aromatic flavors.

[0080] The decoction is then subjected to filtration on a standard paper filter in order to eliminate suspended particles.

[0081] The aqueous extract is stored either in a refrigerator between 2 and 8 °C or in a freezer at -20 °C until use. Before use, the aqueous extract is subjected to sterilizing vacuum filtration using a 0.2 μm filter.

[0082] The liquid extract obtained is soluble in water.

[0083] Example 3: example of active ingredient according to the invention in liquid form (aqueous extract)

[0084] The aerial part of Cleome droserifolia is collected and dried then finely ground. An aqueous extract is made. The extraction is carried out as follows: the dried and crushed Cleome Dorserifolia leaves are added to boiling water (100°C) and boiled for 5 minutes to extract the water-soluble compounds. It is then left to infuse and decant in the refrigerator for 24 hours between 2 and 8°C.

[0085] The proportion of dried and ground raw materials is 9% for 91% water mass / mass. A brown-colored infusion with aromatic flavors is obtained.

[0086] The decoction is then subjected to filtration on a standard paper filter in order to eliminate suspended particles.

[0087] The aqueous extract is stored either in a refrigerator between 2 and 8 °C or in a freezer at -20 °C until use. Before use, the aqueous extract is subjected to sterilizing filtration under vacuum using a 0.2 pm Nalgen type filter.

[0088] The liquid extract obtained is soluble in water.

[0089] Example 4: example of active ingredient according to the invention in liquid form (hydroalcoholic extract)

[0090] The aerial part of Cleome droserifolia is collected and dried then finely ground. A hydroalcoholic extract is made. The extraction is carried out as follows: the dried and crushed Cleome Dorserifolia leaves are added to a boiling hydroalcoholic mixture (70% water - 30% ethanol) and allowed to boil for 5 minutes to extract the water-soluble compounds. The mixture is then left to infuse and decant in the refrigerator for 24 hours between 2 and 8°C.

[0091] The proportion of dried and ground raw materials is 10% for 90% of mass / mass hydroalcoholic mixture. We obtain a brown-colored infusion with aromatic flavors.

[0092] The decoction is then subjected to filtration on a standard paper filter in order to eliminate suspended particles.

[0093] The hydroalcoholic extract is stored either in the refrigerator between 2 and 8 °C, or in the freezer at -20 °C until use. Before use, the extract is subjected to sterilizing filtration under vacuum using a 0.2 pm Nalgen type filter.

[0094] The liquid extract obtained is soluble in water.

[0095] The liquid extract obtained is soluble in water or in hydroalcoholic solution.

[0096] Example 5: example of active ingredient according to the invention in resin form

[0097] The liquid extract of examples 1 to 3 is subjected to drying with hot air at 60°C in a closed enclosure equipped with a system for evacuating hot air pulsed by dryers.

[0098] After evaporation, a blackish resinous gum is obtained.

[0099] The resinous gum obtained is soluble in water.

[0100] Example 6: example of active ingredient according to the invention in the form of lyophilisate

[0101] A LIOBRAS brand model L1O1 laboratory freeze dryer was used with a capacity of 3.0 liters of ice per 24-hour cycle and a total capacity of 5.0 liters at a working temperature of -55°C.

[0102] The liquid extract of Examples 1 to 3 was frozen at -30°C for 48 hours. It was then lyophilized for 120 hours at an internal temperature and pressure of -45°C and 200 pmHg, respectively.

[0103] At the end of the freeze-drying cycle, a cake of yellowish powder in the form of needle crystals is obtained as shown in Figure 3.

[0104] The lyophilized form is soluble in water.

[0105] Example 7: example of composition in the form of a gel

[0106] An example of a composition in gel form is presented below.

[0107] In this example the composition is made up of: Hydroxyethylcellulose 1.8% Propylene glycol 5.0% Imidazolidinyl urea 5 drops (Sol 5% in propylene glycol) Cleome droserifolia (example 6) between 0.1 and 5% preferably between 0.12 and 2% Distilled water qsp 100.0%

[0108] This composition can be obtained according to the procedure described below: - dissolve the Cleome Droserifolia extract in distilled water by heating to 40°C while stirring - add the required amount of hydroxyethylcellulose and continue stirring until a gel is obtained - let cool while maintaining stirring, then add the propylene glycol and imidazolidinyl urea.

[0109] Example 8: Example of composition

[0110] An example of a composition in the form of a non-ionic cream is presented below.

[0111] In this example the composition is made up of: Polawax™ NF 9.0% Cetyl alcohol 5.0% Cetiol® V 4.0% Propylene glycol 4.0% Imidazolidinyl urea 3 drops (5% sol. in propylene glycol) Cleome droserifolia (example 6) between 0.1 and 5% preferably between 0.12 and 2% Distilled water qsp 100.0%

[0112] This composition can be obtained according to the procedure described below: - in a bowl, add the polawax, cetyl alcohol and cetiol V and heat on a hot plate to a temperature of 75°C - in a beaker, add distilled water and propylene glycol at a temperature of 70°C, - then slowly pour the aqueous phase into the oily phase while stirring -remove from the heating system and stir slowly until cool. - at 50°C, add the imidazolidinyl urea solution then add the Cleome Droserifolia extract previously dissolved in water.

[0113] Example 9: example of composition in the form of a soap

[0114] An example of a composition in the form of a soap is presented below.

[0115] In this example the composition is made up of (for a mixture of 625g): Olive oil: 330.0 g (52.80%) Bayberry Oil: 84.0 g (13.44%) H2O: 134.4 g (21.51%) KOH or NaOH: 51.6 g (8.26%) Cleome droserifolia (example 6) 25.0 g (between 0.5 and 4%)

[0116] The liquid extract of one of examples 1 to 4 can also be used. In this case the dry extract must represent 25% by mass of the extract.

[0117] The equipment used to produce this composition is as follows: - Balance - Benchtop thermometer with probe - 1 liter stainless steel container - 500 ml stainless steel container - Porcelain weighing dish - Propeller agitator

[0118] The operating protocol is described as follows: - Weigh the oils in the 1 liter stainless steel container - Weigh the water in the 500 ml stainless steel container - Weigh the freeze-dried or resin-form Cleome Droserifolia extract into a cup - Weigh the NaOH or KOH in a cup - Dissolve the KOH or NaOH in the water and wait until the temperature drops to 60°-80°C - Add the Cleome Droserifolia extract and shake until the compounds are completely dissolved. - Pour the solution thus prepared into the 1 litre stainless steel container containing the previously homogenised oils. - Stir with the propeller stirrer until the trace is obtained - Pour into the molds and let stand for 24 to 48 hours - Unmold and let the soap mature for 4 weeks to 9 months before use

[0119] Variants can be prepared by varying the total oil content as well as the nature of the oils. The total oil value can vary between 70 and 80%.

[0120] Quantities of aromatic essential oils may be added such as rosemary, cypress, peppermint, ylang-ylang, patchouli etc.

[0121] The Cleome Droserifolia extract content for this example can vary between 0.1% and 5%

[0122] Evaluation of the effect of the extract according to the invention

[0123] Evaluation of the effect on activity on blood coagulation and healing:

[0124] The aim of this test is to evaluate the activity of the Cleome droserifolia extract according to the invention (extract of the example solubilized in water at a concentration of 2 mg / ml) on coagulation and to observe the effect compared to the reference standard of unfractionated heparin, using the in vitro model based on the determination of the response by measurement of the coagulation time.

[0125] Two in vitro and human methods were used:

[0126] In vitro coagulant activity:

[0127] The material used is described as follows: - The 6th International Standard for Unfractionated Heparin (WHO 07; 328) - Dade® Actin® Activated Cephaloplastin Reagent - Siemens - Calcium chloride - Siemens®. - Thermo Scientific Coagulometer - 8.7% Citrated Sheep Plasma. - Aqueous extract of lyophilized Cleome Droserifolia

[0128] The operating protocol is described as follows: The lyophilized extract was dissolved in ultrapure water to obtain a stock concentration of 2 mg / ml. Dilution was carried out to obtain a concentration of 0.6 mg / ml.

[0129] In a first experiment 100 μl of each of the following solutions was added to the coagulometer cuvettes (n=2): * Concentrated lyophilisate solution at 0.6 mg / ml, * Heparin solution at a concentration of 0.6 IU / ml * Physiological solution for the control group, - Then, 100 μl of sheep plasma was added to all wells. - After 2 minutes, 100 µl of dade® Actin cephalosplatin-activated reagent was added. - After 5 minutes of incubation, 100 µl of calcium chloride solution was added and the clotting times were recorded. The invention (Cleome droserifolia lyophilisate (example 7) diluted to 0.6 mg / ml) gave mean clotting times of 30.3 seconds, while the control group showed a clotting time of 32.4 seconds.

[0130] In a second experiment, the Cleome droserifolia solution at a concentration of 2 mg / ml reduced clotting times to 14.6 seconds, while the control had a clotting time of 32.2 seconds. The tests were compared to heparin, which showed clotting times of 54.3 seconds at a concentration of 0.6 IU / ml.

[0131] A summary of the results obtained is presented in Table 1.

[0132] [Tables 1] Coagulant activity Concentration Coagulation time Physiological solution 0.9% NaCl 32.4 Invention (example 6) 2.0mg / ml 14.6 Heparin 54.3U / ml 54.3

[0133] The data obtained therefore demonstrate the pro-coagulant activity of an extract of Cleome dorserifolia according to the invention, by the significant reduction in the coagulation time of the ovine plasma used as a substrate compared with the control.

[0134] Coagulant and healing activity in humans:

[0135] The coagulating and healing effect described below has been confirmed in a human experiment.

[0136] To carry out this study, a small quantity of dry extract of Cleome Droserifolia in lyophilized form was deposited on wounds.

[0137] An example is given below and shown in Figures 1a, 1b, 1c, 1d, 1c and 1f, for an incision of a person's left index finger with drip bleeding, caused by mishandling of a kitchen knife.

[0138] Fourteen minutes after depositing the extract on the wound, it was rinsed using gauze soaked in mineral water. The evolution of the wound was monitored in time-dependent, with taking photographs.

[0139] It is noted that after depositing the lyophilisate, the bleeding stops immediately (Figure 1a).

[0140] After rinsing at 14 minutes post application, the wound has a very clean, clean, bleeding-free skin incision and skin suggesting the beginning of healing. (Figures 1b and 1c).

[0141] The finger was re-examined for 9.5 hours after application of the product. The wound was well healed with a crust of coagulated blood. (Figure 1d).

[0142] The same observations were made the next day, i.e. after 21 hours and 8 minutes post application (Figures 1e and 1f).

[0143] No other product was used in parallel.

[0144] This result confirms the coagulating and healing effect of an extract of Cleome Dro-serifolia according to the invention demonstrated in vitro.

[0145] Evaluation of the antioxidant effect:

[0146] The objective of this study is to determine the antioxidant activity of an extract of Cleome Dorserifolia according to the invention (extract of the example solubilized in water at a concentration of 2 mg / ml) using the cell line NCTC clone 929, after oxidative stress induced by hydrogen peroxide to determine cell viability.

[0147] The material used is described as follows: - Hydrogen peroxide - Ascorbic acid as a positive antioxidant control - NCTC 929 clone cell line (ATCC number CCL-1) - MEM culture medium, from Sigma-Aldrich® - MTT coloring - PBS solution, Sigma-Aldrisch® - 96-well plates - Water Jacket CO2 Incubator, Thermo Fisher Scientific® - Varioskan Flash plate reader, Thermo Fisher Scientific® - IKA® MS 1 Shaker plate stirrer - Concentrated stock solution at 2 mg / ml of a lyophilisate of 10% aqueous extract of Cleome Dorserifolia.

[0148] The operating protocol is described as follows: - In a 96-well plate, place the cells at a concentration of 3.5 x 105 cells / ml. - Incubate for 24 hours in an incubator set to 5% CO2 and 37°C. - Remove the supernatant after 24 hours of incubation - In each well, place 200 µl of the lyophilisate stock solution respectively diluted in MEM medium at 1 / 3 and 1 / 6 (i.e. respectively 0.67 mg / ml and 0.33 mg / ml of lyophilized extract), 200 ml of ascorbic acid at 0.56 mg / ml in MEM medium as a positive control and 200 μl of MEM culture medium as a negative control. - Incubate again for 24 hours, then add 100 µl of 25 µM hydrogen peroxide diluted in sterile, apyrogenic water. - Incubate for a further 3 hours, then wash with PBS and add 20 µl of a MTT dye solution at a concentration of 5 mg / ml in MEM medium - Incubate for a further 2 hours then add 100 µl of SDS at a concentration of 24% to each well and shake the plate using the plate shaker for 10 minutes - Perform the optical density reading on the plate reader at the wavelength of 570 nm.

[0149] The method for calculating antioxidant activity (AO) is calculated as follows: AO = Average absorbance obtained by the sample x 100 / Average absorbance obtained by the negative control with MEM medium.

[0150] The cell viability results presented in Table 2 show a powerful capacity of ascorbic acid at 0.56 mg / ml and of the extract according to the invention at concentrations of 0.67 mg / ml and 0.33 mg / ml to prevent the oxidative effect induced by hydrogen peroxide at 25 pM on cells of the NCTC line. The Cleome Dorserifolia extract according to the invention demonstrates an anti-oxidative capacity comparable to that of ascorbic acid.

[0151] [Tables 2] Evaluation of the antioxidant activity of aqueous extract of lyophilized Cleome droserifolia on NCTC cell line (3.5 x 105 cells / ml) Without H 2 O 2 With H 2 O 2 (25 pM) Negative control MEM medium Lyophili sat 0.67 mg / ml Lyophili sat 0.33 mg / ml Positive control ascorbic acid 0.56 mg / ml H2O2 25p M Lyophili sat 0.67 mg / ml Lyophili sat 0.33 mg / ml Positive control ascorbic acid 0.56 mg / ml H2O2 25p M Lyophili sat 0.67 mg / ml Lyophili sat 0.33 mg / ml Positive control ascorbic acid 0.56 mg / ml H2O2 25p M ascorbic 0.56 mg / ml Absorbance 0.4824 0.3208 0.4271 0.401 0.042 0.2774 0.4242 0.3713 0.4702 0.3123 0.398 0.3561 0.040 7 0.2701 0.3849 0.3502 0.4848 0.2816 0.4622 0.3585 0.045 9 0.3024 0.3709 0.3477 Average 0.4791 0.3049 0.4291 0.3719 0.042 9 0.2833 0.3933 0.3564 Standard deviation 0.0078 0.0206 0.0321 0.0253 0.002 7 0.0169 0.0276 0.0130 CV (%) 1.6340 6.7632 7.4917 6.7924 6.313 0 5.9792 7.0252 3.6375 Cell viability (%) 63.64 89.56 77.61 8.95 59.13 82.09 74.38

[0152] These results show that Cleome Dorserifolia extract exhibits antioxidant and protective activity related to the oxidative effects induced by hydrogen peroxide. The 1 / 6 dilution exhibits similar results to those observed with ascorbic acid as a positive control.

[0153] Evaluation of the anti-inflammatory / anti-itching effect

[0154] This study was carried out in the clinical context described below. A hives-like rash was visible on the wrist of a subject with symptoms characteristic of urticaria with itching, inflammation of the skin with localized blisters.

[0155] The operating protocol of the study is described as follows: After visualization, a liquid extract of Cleome Droserifolia according to the invention (extract of the example solubilized in water at a concentration of 2 mg / ml) was applied to the inflammatory skin area using a cotton pad. Observations were followed as a function of time post application. The application of the aqueous extract was carried out 10 minutes after observation. Then observations were carried out as a function of time with taking of photographs of the inflammatory area and questioning of the subject.

[0156] The results are shown in Figures 2a, 2b, 2c, 2d and 2e. Almost instantly after application of the Cleome Droserifolia solution the subject no longer had any skin itching. Signs of reduced inflammation (redness and vesicles) were visible to the naked eye within 3 minutes of application. The anti-inflammatory phenomenon continued over time as shown in the photos in the figures.

[0157] These results show that an extract according to the invention significantly and very quickly reduces itching and inflammatory skin symptoms.

[0158] Evaluation of the effect on fast-growing mycobacterial strains

[0159] The objective of the invention is to evaluate the in vitro activity of an extract of Cleome Dro serifolia according to the invention extracted from the example solubilized in water at a concentration of 2 mg / ml) on the growth of fast-growing mycobacteria (MCR) belonging to the group of non-tuberculous mycobacteria (NTM) by the micro-dilution technique.

[0160] The material used is described as follows: - Standard strains of MCR: M. fortuitum (ATCC 6841), M. massiliense (ATCC 48898) and M. abscessus (ATCC 19977) - medium Mueller Hinton (Himedia®) Agar Lowenstein-Jensen (HiMedia Laboratories Pvt. Ltd, India) - Dimethylsulfoxide (VETEC®) - TTC revealing colorant - Midfielder Mueller Hinton ((Himedia®) - Precision Ependorf pipettes and tips - 96-well culture plate with flat bottom and lid.

[0161] The operating protocol is described as follows: The susceptibility test was performed using the broth microdilution method and according to CLSI M07-A08 (CLSI, 2009). The lyophilized extract was solubilized in dimethyl sulfoxide for the stock solution, for the preparation of working solutions which were diluted in Mueller Hinton Broth. In 96-well plates, different concentrations of the extract were used from serial dilutions ranging from 1775 pg / mL to 27.73 pg / mL. Then, the bacterial suspension of each of the studied bacteria was prepared from colonies maintained on Lowenstein-Jensen agar. A final inoculum of 5 x 105 CFU / mL was obtained from a standard inoculum at the McFarland scale of 0.5. The inoculum was transferred into sterile microplates already added to the extract. The final volume of each well was 200 pL (100 pL of inoculum + 100 pL of extract) Plate reading for determination of the minimum inhibitory concentration (MIC) was performed after incubation at 37°C for 72 hours. To reveal the growth or not of colonies of microorganisms the test (2,3,5-triphenyltetrazolium chloride) at 1% was used.

[0162] The results obtained are presented in Table 3 below.

[0163] [Tables3] MCR (fast growing micro-bacteria) Minimum Inhibitory Concentration (MIC in pg / ml) M. Abscessus 443.75 M. Massiliense 443.75 M. Fortiutum 443.75

[0164] It is found that the Cleome droserifolia extract according to the invention is capable of inhibiting the growth of fast-growing mycobacteria at inhibitory concentrations equal to 443.75 pg / ml.

Claims

Claims

1. Extract of aerial parts of Cleome droserifolia intended for application to the skin and / or mucous membranes, characterized in that it is an extract obtained using a solvent comprising at least water and / or obtained by supercritical CO2 extraction, said extract obtained using a solvent comprising at least water being an aqueous extract or a hydroalcoholic extract or an extract obtained using a solvent comprising water and an organic solvent, for use in topical application for the treatment of the skin and / or mucous membranes.

2. Extract for use according to claim 1, characterized in that it is in liquid form.

3. Extract for use according to claim 1, characterized in that it is in the form of a water-soluble resinous gum.

4. Extract for use according to claim 1, characterized in that it is in the form of a water-soluble lyophilisate.

5. Extract for its use according to one of claims 1 to 4, in topical application on the skin and / or mucous membranes for an anticoagulant and / or healing and / or antioxidant and / or anti-inflammatory and / or anti-itching and / or anti-bacterial and / or anti-histamine effect.

6. Extract for use according to one of claims 1 to 5, in the treatment of urticaria, infections of the skin and / or mucous membranes, mycoses of the skin and / or mucous membranes, atopic dermatitis, psoriasis, acne, rosacea, herpes, mouth ulcers, burns, insect bites, sunburn burns, itching and / or wounds.

7. Cosmetic use of an extract of aerial parts of Cleome droserifolia, or of a composition containing it, on healthy skin, in topical application, characterized in that the extract of Cleome droserifolia is an extract obtained using a solvent comprising at least water and / or obtained by supercritical CO2 extraction, said extract obtained using a solvent comprising at least water being an aqueous extract or a hydroalcoholic extract or an extract obtained using a solvent comprising water and an organic solvent.

8. Cosmetic use according to claim 7 of an extract of Cleome droserifolia or of a composition containing it, on healthy skin, in topical application for an anti-aging and / or anti-cosmetic effect redness.

9. Process for obtaining an extract according to one of claims 1 to 4, characterized in that it comprises at least the implementation of the following steps: - drying then grinding of aerial parts of Cleome droserifolia, - aqueous extraction using a solvent consisting solely of water or using a hydroalcoholic solvent or using a solvent consisting of a mixture of water and organic solvent, with a proportion of Cleome droserifolia of between 5 and 20% and a proportion of solvent of between 80 and 95%, the percentages being given by weight relative to the total weight of the mixture of dried and ground aerial parts of Cleome droserifolia and solvent, - filtration to remove suspended particles, - obtaining a liquid extract.

10. Method according to claim 9, characterized in that the extraction is an aqueous extraction carried out: - by decoction at room temperature for 24 to 48 hours, or - by infusion in water at a temperature between 40 and 60°C, then infusion and decantation for at least 24 hours at a temperature between 2 and 8°C, or - by infusion in water at a temperature of at least 100°C for 4 to 10 minutes, then infusion and decantation for at least 24 hours at a temperature between 2 and 8°C.

11. Method according to one of claims 9 or 10, characterized in that after the filtration step the method comprises: - optionally a storage step either between 2 and 8°C, or at a temperature below -20°C, - a sterilizing filtration step before use.

12. Method according to one of claims 9 to 11, characterized in that the extract is in the form of a resinous gum and in that the method also comprises a step of drying the liquid extract.

13. Method according to one of claims 9 to 12, characterized in that the extract is in the form of a lyophilisate and in that the method also comprises a step of lyophilization of the liquid extract.

14. Composition comprising at least 0.1% by weight of an extract of aerial parts of Cleome droserifolia, characterized in that the extract of Cleome droserifolia is an extract obtained using a solvent comprising at least water and / or obtained by extraction with super-CO2

15.

16.

17.

18. critical, said extract obtained using a solvent comprising at least water being an aqueous extract or a hydroalcoholic extract or an extract obtained using a solvent comprising water and an organic solvent. Composition according to the preceding claim, characterized in that it is in the form of a cream, an ointment, a gel, a paste, a lotion, an ointment, a toothpaste, a soap, a mouthwash, a shampoo, a shaving foam. Composition according to one of claims 14 to 15, for its use in the treatment of the skin and / or mucous membranes. Composition according to one of claims 14 to 15, for its use in topical application to the skin and / or mucous membranes for an anti-coagulant and / or healing and / or anti-oxidant and / or anti-inflammatory and / or anti-itching effect. Composition for its use according to one of claims 16 or 17, in the treatment of urticaria, infections of the skin and / or mucous membranes, mycoses of the skin and / or mucous membranes, atopic dermatitis, psoriasis, acne, rosacea, herpes, mouth ulcers, burns, insect bites, sunburn burns, itching and / or wounds.

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